no one will bury himno one will mourn himno one will utter a cry
O: The difference between how Kreon speaks of both of Oidipous' sons is shown by how he honors Eteokles by speaking with a very elevated form of language while he uses harsh words in repetition while speaking of Polyneikes.
repetition
many things strang
... to than Man." O: The Chorus lists humanity's achievements and contradictions, praising human intelligence and creativity while also warning of arrogance. This contrasts with Antigone's opening emphasis on divine law and familial duty.
you are someone in love with the impossible
O: Is Ismene saying Antigone is unrealistic, or is she blaming her of engaging in a hopeless rebellion?
if a man puts familyor friends ahead of fatherland1 count him absolutely goodfor nothin
O: Kreon here emphasizes the supremacy of law and the state, placing loyalty to the state above family and personal emotions. He believes that anyone who prioritizes family or friends over the interests of the state is useless, which reflects the conflict between power and personal morality. At the same time, this also indicates that Antigone and Kreon will have conflicts due to different values.
suicidal
O: Antigone's future act of disobeying this law to bury her brother could be considered a suicidal act when we know that Antigone is willing to quite literally die in honor of her family.
the man who honours the laws of the landthe man who keeps the justice of godswalkshigh in his high citycityless
O: The chorus seems to suggest that the ideal person obeys both human law and divine justice. But in the play, Antigone and Kreon stand for opposite extremes one follows divine law, the other human law. This contrast shows the central conflict—no one can fully satisfy both sets of rules.
he shattered his own eyeballs so as not to see it
O: Referencing the past play Oedipus Rex.
Antigone says it's not Kreon's business
O: Whatever Antigone says goes!!!
In all seriousness, I love this particular translation because it shows Antigone's unshakable nature and her willingness to be blunt about what she believes in.
your heart's so hot O sisterANTIGONE: the dead are coldthey'll welcome me
O: There's a pattern of antigone and death. Obviously that's relevant when her brothers died, but she keeps talking about how the dead will treat her well ('they'll welcome me' and 'they'll forgive me.' This could be foreshadowing or become relevant later on.
girls cannot force their way against men
O: There's a clear contrast between Antigone's moral duty to the family and Ismene’s obedience to the law and social rules. Thus, becomes representation of the play’s central conflict between divine law and human authority.
for death is longdeath is very long
O: Antigone repeats the word “death” twice, which creates a heavier tone. This shows she is determined to bury her brother, even if it means losing her own life. These two lines give me the feeling that she has already decided and nothing can change her mind.
sevengated
O: OBSERVATION
The word sevenagted or just seven in general tends to come up a lot here, "seven gates and in each gate a man and in each man a death"
dear sister, my dead are mineand yo= as well as minedo what you like butI will not betray him
O: here reflects the conflict between two kindnesses - ethics and civic obligations. Antigone choose to take family responsibility which was against the edict. It shows Antigone‘s fearless of power and heroism.
Antigone says it's not Kreon's busines
O: adding on to previous observation - further proof for Antigone's boldness (audience can see how it might get her into trouble as Kreon seems to be of consoldated power)
you have your thunder look
O: Could be a insight into the kind of person Antigone is (thunder is loud and sharp and bold)
We included data from the following species for analysis: grizzly bears, black bears (Ursus americanus), wolves (Canis lupus), coyotes (Canis latrans), cougar (Puma concolor), lynx (Lynx canadensis), moose (Alces alces), elk (Cervus canadensis), white-tailed deer (Odocoileus virginianus), mule deer (O. hemionus), unspecified deer, and bighorn sheep (Ovis canadensis) (Table 1).
These can be discussed in summary as the three broad category grouping
O, she knew well Thy love did read by rote and could not spell. 1150But come, young waverer, come, go with me, In one respect I'll thy assistant be; For this alliance may so happy prove, To turn your households' rancour to pure love.
The Friar suggests rosaline recognized Romeo’s love was mechanical (“read by rote and could not spell” means lacking depth). However he agrees to help because it maybe good for future family feuds.
Hist! Romeo, hist! O, for a falconer's voice, To lure this tassel-gentle back again! 1020Bondage is hoarse, and may not speak aloud; Else would I tear the cave where Echo lies, And make her airy tong
Juliet compares Romeo to a falcon that is well trained, she wishes for some to call Romeo for him
área ocupada
Se o Município tiver mais de 50% da sua área ocupada por UC's de domínio público ou por terra indígena homologada, poderá haver redução do percentual de RL. Com isso: - Amazônia legal: 40% do imóvel situado em floresta; - Cerrado: 17,5% do imóvel situado em cerrado; - Campos Gerais: 10% do imóvel situado em campos gerais.
If there is no mysterious, magical spring water hidden beneath the surface of the work, the most elegantly crafted, clever dialogue will not make the work live.
o: the way the author worded the sentence is very particular
because we are mesmerized by a writer’s unique voice.
O: this is a great way of explaining why we like reading novels
allows us entry into the mystery to be explored.
O: This form of literature and art allows for lots of interpretation. The lack of information and concrete facts allow for the viewers to form there own connections and visualize/place themselves into the character. In plays, less is more.
Envision a play dramatically by seeing the characters enter this emptiness, as they would a stage in a theater; as much “background” is provided as the playwright has given you in his notes and asides.
O: Here it emphasizes the virtuality of plays and the participation of reader's imagination. Readers need to visualize this drama in their minds according to the author's notes or "instructions".
But isn’t that everything? you may ask. Why would a writer give up so much, in exchange for the unknown? Precisely because the theater is an adventure, a challenge, a risk; because it is not only always present tense in the theater, but because the theater itself has no memory.
O: These sentences really capture what theater represents. Theater has represented adventure, challenge, and risks. And clearly shows how theater works when they write: “Present tense”.
Familiar names are juxtaposed with the less familiar and the newly emergent; realistic, accessible works are juxtaposed with the bizarre and perhaps unstageable
O: I notice that this collection of plays puts really different kinds of theater side by side. Some are realistic and easy to imagine on stage, while others are described as "bizarre" or "unstageable." I think the editor wants to show how theater can be both normal and really strange at the same time. This contrast makes me believe that theater should bring us surprises, rather than merely telling simple stories.
Have we not all been instructed in the sacrosanct virtues of beginning, middle, end? Complication, climax, resolution? A respect for Aristotelian unities? For the clarity of chronology? For clarity itself? As if the limitations of realism were not in fact limitations.
O: This quote is very interesting to me because it stresses the structure of plays, books, and literature that we believe is important to follow, like we have been told. Also, we are told that literature is limitless and can express anything, however this structure we have been taught to follow in most english classes for years, does in fact limit our writing.
Where in prose fiction the writer is accustomed to shaping subtleties of meaning by way of carefully composed language, in which the very punctuation and the spacing of paragraphs may be a part of the meaning, in drama the writer must provide for his characters ‘living” roles.
O: I noticed that the author compares the role of language in prose and drama. In prose, the author talks to us directly, and we can understand their thoughts easily. However, in drama, the author's voice and thoughts are shown through the characters instead of being told to us directly. Because of this, I feel that drama is more like an open space. It needs actors to perform and the audience to understand it on their own. This difference makes me think that drama relies more on performance than just the words themselves.
So we discover, to our astonishment, that theater is not “about” dialogue after all, but what might be called fields of dramatic tension; force-fields of human relationships beneath the level of language, and perhaps even of consciousness, at which dialogue hints in the way that a divining rod hints at a subterranean spring.
O: On stage, dialogue is merely a probe, pointing to the flow of emotions and underlying meanings, but not the flow itself. When writers choose to enter the realm of theater, they challenge and compromise, choosing to strip away elaborate strategies in language to emphasize responses to psychological and social dynamics that lie deep below consciousness. The core of theater is an invisible energy space – the tug, conflict, or attraction/repulsion between people. It creates drama, often beyond language. It is also a way to open up new ways of feeling for the audience, not only feelings on the page, but also intuitions when watching human contact.
Theater is present tense, the printed word is past tense.
O: Prose depends on the author's voice, but theatre depends on actors, directors, sets and lighting, and the audience's collective interpretation. This sentence summarizes the contrast by showing that theatre is a living thing that cannot be fixed in the same way prose can. I feel this difference is described quite precisely, and I never realized it before reading this.
. What shimmers with life on the page may die within minutes in the theater precisely because prose is a language to be spoken to an individual, recreated in an individual reader’s consciousness, usually in solitude, while dramatic dialogue is a special language spoken by living actors to one another, a collective audience overhearing.
O: Oates seems to be trying to enforce the idea that prose fiction is a space for readers to take in the 'art' of the novel/story/etc., and grow an interpretation. I think the earlier statement, "Where in prose fiction the writer is accustomed to shaping subtleties of meaning by way of carefully composed language," explains this best. While the play on the other hand is given directly to the viewer, with little space to fill in imaginative blanks (although the depth behind acting should not be disregarded.) I find this comparison impactful because it demonstrates the depth of prose fiction that may not be initially realized. A play becomes a predominately collective experience and interpretation, while prose fiction becomes a living document. I can think back to times when I was so immersed in a piece of prose fiction that I began to imagine it in my free time, the characters coming alive in my head, my imagination becoming incredibly involved in the story.
Entity resolution (ER) aims at matching records that refer to the same real-world entity. Although widely studied for the last 50 years,ER still represents a challenging data management problem, and several recent works have started to investigate the opportunityof applying deep learning (DL) techniques to solve this problem. In this paper, we study the fundamental problem of explainabilityof the DL solution for ER. Understanding the matching predictions of an ER solution is indeed crucial to assess the trustworthinessof the DL model and to discover its biases. We treat the DL model as a black box classifier and – while previous approaches toprovide explanations for DL predictions are agnostic to the classification task – we propose the certa approach that is aware ofthe semantics of the ER problem. Our approach produces both saliency explanations, which associate each attribute with a saliencyscore, and counterfactual explanations, which provide examples of values that can flip the prediction. certa builds on a probabilisticframework that aims at computing the explanations evaluating the outcomes produced by using perturbed copies of the input records.We experimentally evaluate certa’s explanations of state-of-the-art ER solutions based on DL models using publicly available datasets,and demonstrate the effectiveness of certa over recently proposed methods for this problem
A resolução de entidades (RE) visa corresponder registros que se referem à mesma entidade do mundo real. Embora amplamente estudada nos últimos 50 anos, a RE ainda representa um problema desafiador de gerenciamento de dados, e diversos trabalhos recentes começaram a investigar a oportunidade de aplicar técnicas de aprendizado profundo (ADL) para resolver esse problema. Neste artigo, estudamos o problema fundamental da explicabilidade da solução de ADL para RE. Compreender as previsões correspondentes de uma solução de RE é de fato crucial para avaliar a confiabilidade do modelo de ADL e descobrir seus vieses. Tratamos o modelo de ADL como um classificador de caixa-preta e – embora abordagens anteriores para fornecer explicações para previsões de ADL sejam agnósticas à tarefa de classificação – propomos a abordagem certa, que considera a semântica do problema de RE. Nossa abordagem produz explicações de saliência, que associam cada atributo a uma pontuação de saliência, e explicações contrafactuais, que fornecem exemplos de valores que podem inverter a previsão. A abordagem certa baseia-se em uma estrutura probabilística que visa computar as explicações avaliando os resultados produzidos usando cópias perturbadas dos registros de entrada. Avaliamos experimentalmente certas explicações sobre soluções de ER de última geração com base em modelos DL usando conjuntos de dados disponíveis publicamente e demonstramos a eficácia de certos métodos propostos recentemente para esse problema.
Kirk o tym, jak to przyjęcie „Civil Rights Act” (prawa, które zniosło segregację rasową w USA) to był „wielki błąd”: https://www.snopes.com/fact-check/kirk-civil-rights-act-mistake/
Owszem, Kirk mocno krytykował Civil Rights Act, w szczególności sposób, w jaki został wdrożony. Jednak WIELOKROTNIE chwalił fakt zniesienia segregacji rasowej oraz ubolewał, że dzisiejsza lewicowa polityka de facto ją przywraca, w szczególności na uczelniach przez affirmative-action i DEI.
Kirk mówi, że biblijna kara śmierci dla gejów przez kamieniowanie jest właściwa: https://x.com/danielsogay/status/1965887308454101427
To nieprawda. Za to samo twierdzenie dostał wiele krytyki - i przeprosił - autor Stephen King. Jest to wycinek wypowiedzi Kirka o "selektywnym cytowaniu Biblii".
Podane źródło też już nie istnieje.
Podejście Kirka do osób homoseksualnych na wiecach politycznych jest bardzo normalne. Jedyne co, to mówi im, że "jakby spytali się go o perspektywę chrześcijańską, to nie zgadza się z ich stylem życia, ale nie jest to jego spraw
Mike Lee przy wsparciu Donalda Trumpa Juniora
Trump Jr podał pewne nieprawdziwe informacje w trakcie wywiadu NewsNation:
Te same informacje podawał Mike Lee.
Tych rzeczy jednak w art OKOPress nie ma.
Są sugestie jakoby informacja o "zleceniodawcu Walzie" była wymyślona przez Republikan, a była to informacja z oficjalnego śledztwa - tak napisał w liście sam zamachowca.
Sam Trump nie postował żadnych nieprawdziwych informacji na ten temat, ale artykuł OKOPress tego slusznie - nie sugeruje.
Niesmaczny mem o "koszmarze z Walz Street" zapostował Mike Lee.
teorie spiskowe, jakoby zleceniodawcą morderstwa był gubernator Minnesoty i kandydat na wiceprezydenta Tim Walz
To słowa zamachowcy, które z pewnością są kłamstwem które ma wprowadzić zamęt, ale jakoś artykuł nic o tym nie mówi.
https://www.washingtonpost.com/nation/2025/07/15/vance-boelter-melissa-hortman-murder-charges/
nie możemy pozwolić się emocjonalnie terroryzować rodzinom ofiar
Charlie nie powiedział "nie możemy pozwolić się emocjonalnie terroryzować rodzinom ofiar".
Co powiedział naprawde:
Nie możemy dać porwać się emocjom pod wpływem takich tragedii.
"We need to be very careful not to let our emotions get the best of us in the wake of these tragedies."
"I respect their pain, but policy shouldn't be based on emotion alone."
Po tych tragediach emocje są wysokie, co jest zrozumiałe, ale nie możemy pozwolić, by emocje dyktowały politykę. Potrzebujemy spokojnej, racjonalnej dyskusji na temat tego, jak rzeczywiście zmniejszyć przemoc, nie naruszając naszych wolności.
"After these tragedies, emotions run high, and that's understandable, but we can't let emotions dictate policy. We need calm, rational discussion about how to actually reduce violence without infringing on our freedoms."
Ciezko sie odniesc do konkretnego wydarzenia, bo artykul OKO Press mowi tylko o "którejś z kolei strzelaninie w szkole w USA", jednak słowa podane w artykule nigdy nie padły - przynajmniej w takiej formie.
Charlie Kirk ogłosił wszem wobec, że... jakiś prawdziwy patriota powinien wpłacić kaucję za napastnika
To prawda powiedział to - w kontekście.
Potępił też atak, a o patriocie mówił w kontekście nierównego stosowania prawa, i zwalniania z aresztu niebezpiecznych recydywistów. (Można nawiązać tu do wyżej wspomnianego zabójcy Iryny)
I’m not qualifying it. I think it’s awful. It’s not right... But why is it that in Chicago you’re able to commit murder and be out the next day? Why is it that you’re able to trespass, second-degree murder, arson, threaten a public official, cashless bail. This happens all over San Francisco. But if you go after the Pelosis, oh, you’re [not] let out immediately. Got it.
Oczywiście, jak na prawicowego Amerykanina przystało, był onucą (powtarzał wszystkie standardowe teksty o nazistach w Ukrainie, Zełenskim jako marionetce USA, żądał anulowania pomocy dla Ukrainy i oddania Krymu Rosji),
Tak, Kirk ma podobne stanowisko co konserwatywna czesc amerykanskiej polityki, w tym Donald Trump. Z naszego punktu widzenia nazwac to mozna "onucyzm" Powiedzial tez ze nie lubi ani Rosji ani Putina, ktorego nazwal dyktatorem, zbrodniarzem i osilkiem, i potepil atak na Ukraine.
“does not like the Russian Federation or Russian dictator Vladimir Putin.” https://www.kyivpost.com/post/59880
Said that the invasion of Ukraine was "wrong" and called Putin a gangster and a thug https://kyivindependent.com/american-right-wing-activist-charlie-kirk-killed-in-utah-media-reports/
Yes, Putin is a thug and his aggression in Ukraine is a giant geopolitical problem, but we rely on our presidents to solve difficult problems. Instead we’re hurtling toward another endless, costly quagmire like Afghanistan, Iraq, and Vietnam. Have we learned nothing? https://x.com/charliekirk11/status/1628040413738307586
czarnoskórzy (ostatnie dni przed śmiercią spędził, organizując rasistowskie szczucie na czarnoskórych Amerykanów poprzez przypisanie im zbiorowej odpowiedzialności za zamordowanie uchodźczyni z Ukrainy, którą zabił czarnoskóry przestępca-recydywista)...
Główną krytyką Kirka w tej sprawie był fakt, że ten recydywista został wypuszczony na wolność, mimo że oczywistym było, że jest niebezpieczny i nawet jego własna matka błagała władze, by nie był na wolności. Krytykował przede wszystkim wymiar sprawiedliwości, który tutaj zawiódł.
A "uchodźczyni z Ukrainy" to Iryna Zarutska - lewica bardzo nie lubi wypowiadać jej imienia, ani w ogóle o niej wspominać, i media w stanach milczały na temat tego zabójstwa dopóki nie zostały zmuszone do przerwania milczenia przez nacisk ze strony konserwatywnej - m.in. Kirka.
Milczały bo nie wpisywało sie to w ich narracje.
单片机内部有 PLL（锁相环倍频电路） 和 分频器。
单片机芯片内部不是只有两个存储器,I/O接口,和CPU吗?怎么又来了个PLL和分屏器?
O
I really like how the pictures can tell the story in ways words can't and how the words add to the story that pictures cannot
La pasiva morfológica, que funcionaba tan bien en latín y que se usa a diestro y siniestro en inglés o en alemán, nunca cuajó en las lenguas románticas. Sólo la influencia humanística y culta la estabilidad (Alcina y Blecua, 1989), contra la tendencia natural del habla, que la utiliza muy raramente. El
Siento que en español se entiende mejor cuando decimos las cosas de forma directa y clara, como hablamos normalmente.
Para terminar, las ramas del final son más fáciles de leer y de escribir, porque reproduce el pensamiento natural de las personas —según afirma la misma autora con un ejemplo parecido al anterior—. Primero leemos la frase principal, que contiene la información más importante, y después los datos secundarios. El pel
Considero que sí, es bueno siempre seguir esa estructura para leer. Porque es mas fácil comprender el mensaje o la lectura.
Una última cuestión más filosófica sobre los incisos se refiere a las causas que los provocativo. Cada persona elige su estilo de escritura y, por lo tanto, parece lícito que alguien se inclina por el inciso abundante y el estilo rebuscado. Pero s
Me parece perfecto que cada persona tiene su estilo de escribir, pero en lo personal pienso que es mejor amontonadora o incisos, cuando el texto es claro y directo, se conecta mejor con quien lo lee, y lo comprende mejor.
El inciso puede estorbar o incluso confundir la lectura, si se inserta torpemente entre dos palabras que deben aparecer juntas. Este
Puede interrumpir lo natural de la oración, y generar confusión. Debemos tener cuidado al momento de escribir.
La retahila de frases de la derecha ayuda poco o nada a comprender el significado del fragmento, porque no relaciona las ideas entre sí como hacen las otras dos versiones.
Creo que cuando las ideas no están bien organizadas o conectadas, el mensaje se vuelve confuso de comprender.
* get attached = encariñarse.
* Each and every one = todos y cada uno.
* 📌 complemento proposicional "to" = Indica dirección o acción como reacción del sujeto de la oración hacia el objeto de esta. Usualmente se usa para complementar a algunos verbos intransitivos ya que estos carecen de OD y por consecuencia de OI eso es porque estos verbos no indican una modificación ni beneficio al objeto de la oración. Solo indican un cambio de estado del sujeto en reacción a objeto.
👉 Identifican exactamente de qué persona o cosa hablamos. 👉 Son necesarias para entender la oración. 👉 No llevan comas.
Ejemplos:
The man who lives next door is very friendly. → El hombre que vive al lado es muy amable.
This is the book that I bought yesterday. → Este es el libro que compré ayer.
Students who study hard usually get good grades. → Los estudiantes que estudian mucho suelen sacar buenas notas.
Visit O*Net (http://online.onetcenter.org/find) and look up one of the careers you may be considering. What kinds of things does O*Net tell you about a career? ________________________________________________________________ /*<![CDATA[*/#mt-toc-container {display: none !important;}/*]]>*//*<![CDATA[*/ $(function() { if(!window['autoDefinitionList']){ window['autoDefinitionList'] = true; $('dl').find('dt').on('click', function() { $(this).next().toggle('350'); }); } });/*]]>*/ /*<![CDATA[*/window.addEventListener('DOMContentLoaded', function () { $('iframe').on('load', function () { $(this).iFrameResize({ warningTimeout: 0, scrolling: 'omit' }); }) })/*]]>*/ /*<![CDATA[*/ var front = "auto"; if(front=="auto"){ front = "12.2: Career Exploration"; if(front.includes(":")){ front = front.split(":")[0]; if(front.includes(".")){ front = front.split("."); front = front.map((int)=>int.includes("0")?parseInt(int,10):int).join("."); } front+="."; } else { front = ""; } } front = front.replace(/_/g," "); MathJaxConfig = { TeX: { equationNumbers: { autoNumber: "all", formatNumber: function (n) { if(false){ return front + (Number(n)+false); } else{return front + n; } } }, macros: { PageIndex: ["{"+front+" #1}",1], test: ["{"+front+" #1}",1] }, Macros: { PageIndex: ["{"+front+" #1}",1], test: ["{"+front+" #1}",1] }, SVG: { linebreaks: { automatic: true } } } }; MathJax.Hub.Config(MathJaxConfig); MathJax.Hub.Register.StartupHook("End", ()=>{if(activateBeeLine)activateBeeLine()}); /*]]>*/
O*Net explains the training I have to do to become a hairsylist
O
O*Net explains income with jobs and duties.
Visit O*Net (http://online.onetcenter.org/find) and look up one of the careers you may be considering. What kinds of things does O*Net tell you about a career?
O*Net explains the duties that are given when you become a medical assistant
Un verbo ditransitivo es aquel que requiere dos objetos para completar su significado:
Objeto Indirecto (OI): Indica para quién o a quién se realiza la acción. Responde a las preguntas "to/for whom?" o "to/for what?".
Objeto Directo (OD): Indica qué recibe directamente la acción. Responde a la pregunta what?.
La Prueba Definitiva: La Estructura de la Oración
La forma más infalible de identificarlo es verificar si el verbo puede aparecer en dos estructuras gramaticales sin cambiar su significado básico.
Estructura 1: Verbo + Objeto Indirecto + Objeto Directo Sujeto + Verbo + [¿Para quién?] + [¿Qué?]
Ejemplo:
"She gave her friend (OI) a book (OD)."
Estructura 2: Verbo + Objeto Directo + to/for + Objeto Indirecto Sujeto + Verbo + [¿Qué?] + [to/for] + [¿Para quién?]
Ejemplo:
"She gave a book (OD) to her friend (OI)."
Si un verbo puede usar AMBAS estructuras, es ditransitivo.
Algunos verbos solo pueden tener un objeto directo (transitivos) o no pueden cambiar de estructura.
Ejemplo 1: Explain (explicar)
❌ "She explained me the problem." (Incorrecto - suena mal en inglés).
✅ "She explained the problem (OD) to me." (Correcto).
Conclusión: Explain no es ditransitivo; es monotransitivo y necesita la preposición to.
Ejemplo 2: Say (decir)
❌ "She said me hello." (Incorrecto).
✅ "She said hello (OD) to me." (Correcto).
Conclusión: Say no es ditransitivo.
* Such as = tal como.
* one = alguien.
* Harbor = alberga.
* towards = hacia.
* 📌 Diferencia principal
to = indica dirección literal (movimiento físico o transferencia).
I go to school.
Give the book to her.
toward = indica orientación, actitud, relación o dirección general, no un destino físico puntual.
She is walking toward the park. (dirección aproximada)
He has a positive attitude toward his job. (sentimientos / actitud)
En tu ejemplo
harbor a feeling toward cats = “albergar un sentimiento hacia los gatos” → habla de una actitud emocional dirigida a un grupo, no de entregar algo físico.
* Shitshow = desastre o inconveniente.
* sorta (contracción) de sort of.
* Sort of = tipo de.
* no funciona para indicar imposibilidad de.
* there's no working (falso sujeto) = es imposible de trabajar no hay forma de trabajar.
* lying (verbo irregular) de mintiendo
* pair subject up (modismo) = forma una pareja, es una matiz de completitud que le de el up, sino diría emparejar o unir.
* trying to get = intentando que llegue.
* To quit = a renunciar.
* don't fuck with me = no te metas conmigo.
* slander = calumnia, difamación.
* Something or other (modismo) sirve para expresar algo parecido al termino que no sabes o no te acuerdas nombrar pero quieres dar a entender eso.
* character something or other = esa cosa del carácter o como se llame, esa cosa de carácter o como se diga. Queriendo decir, character defamation.
* Dirty = sucio.
* damn = de mierda. Sirve como intensificador y aclaración de insulto.
* asshole = Cabrón, pendejo.
caso 25.06.2.00786.C Pontua o PP4? A medica assistente suspeitou de Coffin Lowrys @renatabtenorio
La formación que reciben los estudiantes de administración en México no va acorde conlas tendencias y las exigencias que demandan los nuevos entornos internacionales. Eneste artículo se hace un análisis de los programas de licenciatura en Administración quese imparten en veintidós universidades mexicanas, reconocidas por la agencia oficialcertificadora en esa disciplina (CACECA) para comprobar la hipótesis de que los actualescurrículos están estructurados con base en el proceso administrativo tradicional y susáreas funcionales, mientras que las materias relacionadas con las habilidades que unprofesional de la administración debería tener en este mundo globalizado (como elliderazgo, la toma de decisiones, el conocimiento de otras culturas y la creatividad, porcitar algunas de las más señaladas por los expertos en la materia) sólo figuran de maneramarginal en los programas de estudio de las carreras certificadas en el nivel nacional.
La hipotesis del autor es que la mayoría de los planes de estudio en administración en México siguen enfocados en lo clásico: el proceso administrativo y las áreas como finanzas, recursos humanos y mercadotecnia. Pero casi no incluyen materias que hoy en día son súper importantes para trabajar en un mundo global, como liderazgo, tomar decisiones, ser creativo o entender otras culturas. Tambien en México, la administración tiene poca tradición en investigación y está enfocada más en la docencia que en generar conocimiento nuevo. Se enseñan modelos idealizados (como la optimización o la excelencia) que son fáciles de transmitir pero difíciles de aplicar en la realidad. Además, la administración no puede estudiarse en abstracto, porque depende de las condiciones concretas de cada organización. El autor dice que el liderazgo es una de las habilidades más importantes que deberían enseñarse en las carreras de Administración, sobre todo por el entorno internacional actual.
la hipótesis central del presente trabajo es que los actuales planes de estudio de las licenciaturas en Administración en México están articulados en función de los procesos administrativos tradicionales y el estudio de cuatro o cinco de las áreas denominadas "funcionales" de la administración. Esta estructura curricular se complementa con una serie de materias básicas de carácter cuasipropedéutico y otro conjunto de materias especializadas, ya sean optativas o pertenecientes a un núcleo introductorio, que dependen más del capricho de las autoridades universitarias en turno o de las modas administrativas, que de un análisis sustentado en las necesidades de los mercados de trabajo o las realidades locales, regionales o nacionales. Pág. 4 Pdf y Pág. 125
$$Opinión Personal: $$El planteamiento de este articulo es acertado, la administración, al estar tan vinculada con la práctica profesional, debe actualizarse constantemente según las demandas del entorno, social, económico y profesional. Mas sin embargo, los programas universitarios tienden a ser sistemas cerrados ya que repiten esquemas que ya no corresponden a la realidad. En mi opinión, el artículo señala un problema que trasciende incluso hoy en día muchas universidades aún forman administradores con un perfil demasiado teórico y poco eficaz, dejando de lado habilidades requeridas en este campo, pensamiento crítico, visión global y responsabilidad social. Para responder a la globalización, se necesita un modelo más flexible, práctico e interdisciplinario.
La Administración es una disciplina relativamente reciente en México, con poca tradición en investigación y orientada principalmente hacia las áreas docentes. Se enseñan generalmente modelos idealizados desde la optimización hasta la excelencia ; los cuales representan la parte "dura", "científica" relativamente fácil de transferir, pero difícil de operar. La Administración está asentada en organizaciones concretas, alejadas de las condiciones de idealidad de dichos modelos. No se puede estudiar la Administración en abstracto, ésta tiene que ser referida siempre a las condiciones particulares de las organizaciones que aplican esos dispositivos. Pág. 7 Pdf y Pág. 128
$$$$
¿Qué opina el autor acerca del liderazgo?
El autor opina que el liderazgo es una habilidad esencial para los administradores en el nuevo entorno laboral, ya que con este es de las partes clave que debería formar parte central de los programas de Administración, junto con la toma de decisiones, creatividad y solución de problemas.
hipótesis es que los actuales planes de estudio de las licenciaturas de administración en México están articulados en funciones de los procesos administrativos tradicionales y el estudio de cuatro o cinco de las áreas denominadas "funcionales" de la administración y esas están en estructura curricular es una serie de materias básicas y un conjunto de materias especializadas ya sea en operativas o pertenecientes a un núcleo introductorio. Mi opinión es de que ese plan de estudio no esta alineado con las necesidades del mercado laboral actual lo que le causaría problema a los recién egresados.
Luis Montaño que tiene poca tradición en investigación y se basan en procesos administrativos y tradicionales si base solida en investigación y análisis critico, la orientación va enfocada solo a la docencia.
¿Qué opina el autor acerca del liderazgo? La capacidad de liderar y gestionar equipos de manera efectiva en entornos complejos y globales, se trata de habilidades que permiten a los profesionales de la administración .
la hipótesis del autor es que las carreras de Administración en México todavía se basan mucho en lo tradicional, o sea en el proceso administrativo clásico (planeación, organización, dirección y control), y casi no incluyen materias más actuales como liderazgo, creatividad, toma de decisiones o trabajo en equipo. Lo que él dice es que por eso los estudiantes no salen tan preparados para el mundo globalizado en el que vivimos. Yo pienso que tiene razón, porque si solo nos enseñan lo viejo y no se actualiza el plan de estudios, al salir puede que no tengamos las herramientas necesarias para enfrentar lo que de verdad pasa en las empresas.
Sobre Luis Montaño, él afirma que la Administración en México es una disciplina relativamente nueva, con poca tradición en investigación, y que está más enfocada en la parte de dar clases que en generar conocimiento propio. También critica que se usan muchos modelos “ideales” que vienen de otros países, y que no siempre sirven para la realidad mexicana. Eso me parece cierto, porque muchas veces lo que vemos en libros extranjeros no encaja con lo que pasa en nuestras empresas o nuestra cultura.
Finalmente, acerca del liderazgo, el autor dice que en la mayoría de los programas de estudio casi no se enseña, aparece solo como optativa o muy de lado. Sin embargo, él lo considera súper importante, porque el administrador no solo debe saber de números o procesos, también necesita dirigir personas, motivar y trabajar con valores. Yo estoy de acuerdo, porque sin liderazgo un administrador difícilmente va a poder guiar a un equipo o generar un buen ambiente de trabajo.
¿Qué opina el autor acerca del liderazgo? El autor considera que el liderazgo, tal como se enseña tradicionalmente en la administración, suele ser un concepto idealizado y alejado de la realidad. Para él, el liderazgo debe entenderse como una práctica contextualizada, que requiere habilidades concretas y adaptabilidad para enfrentar situaciones complejas y específicas del entorno, en lugar de seguir un modelo rígido o universal.
Derivado de lo anterior, la hipótesis central del presente trabajo es que los actualesplanes de estudio de las licenciaturas en Administración en México están articuladosen función de los procesos administrativos tradicionales y el estudio de cuatro o cincode las áreas denominadas "funcionales" de la administración. Esta estructura curricularse complementa con una serie de materias básicas de carácter cuasipropedéutico yotro conjunto de materias especializadas, ya sean optativas o pertenecientes a un nú-cleo introductorio, que dependen más del capricho de las autoridades universitarias enturno o de las modas administrativas, que de un análisis sustentado en las necesidadesde los mercados de trabajo o las realidades locales, regionales o nacionales.
El autor critica que los planes de estudio de la carrera de Administración en México están estancados. Básicamente, se enfocan en lo mismo de siempre: procesos administrativos clásicos y unas ciertas áreas como finanzas, mercadotecnia o recursos humanos. Además, muchas materias que se agregan parecen elegirse por moda o por decisión de los que están a cargo, más que por un análisis real de lo que el país o el mercado laboral necesitan. Considero que su critica es válida, ya que la formación profesional debe responder a las transformaciones sociales, económicas y organizacionales actuales. De lo contrario, se corre el riesgo de formar profesionales poco preparados para enfrentar los retos del entorno real.
los actuales planes de estudio de las licenciaturas en Administración en México están articulados en función de los procesos administrativos tradicionales y el estudio de cuatro o cinco de las áreas denominadas "funcionales" de la administración. Esta estructura curricular se complementa con una serie de materias básicas de carácter cuasipropedéutico y otro conjunto de materias especializadas, ya sean optativas o pertenecientes a un núcleo introductorio... - página 4 del .pdf y 125 del artículo
En paráfrasis la hipótesis que expone el autor es que el sistema educativo ha tomado un enfoque más centrado en formas obsoletas de enseñar la administración, esto en vez de adaptarse a los contextos temporales, económicos y políticos que le concierne a nuestra sociedad.
Se enseñan generalmente modelos idealizados desde la optimización hasta la excelencia ; los cuales representan la parte "dura", "científica" relativamente fácil de transferir, pero difícil de operar. La Administración está asentada en organizaciones concretas, alejadas de las condiciones de idealidad de dichos modelos. No se puede estudiar la Administración en abstracto, ésta tiene que ser referida siempre a las condiciones particulares de las organizaciones que aplican esos dispositivos. - página 7 del .pdf y 128 del artículo
Este fragmento indica que a la hora de realizarse el proceso de enseñanza los subtemas de administración es en base a presunciones de utopías organizacionales, mientras que la materia debería de estar aterrizada en las aplicaciones reales en rubros laborales.
La dirección por valores (DpV) es una herramienta de liderazgo que va más allá de los conceptos de dirección por instrucciones y dirección por objetivos que la precedieron, y pretende darle un sentido más ético y emocional a la construcción de empresas. - página 10 del .pdf y 131 del artículo
Aquí el autor plantea un enfoque innovador acerca del liderazgo, consiste de un acercamiento al empleado en base a los axiomas de moral social, con reciprocidad, como se sobreentiende, en lugar de dar el enfoque anticuado de presentar indicaciones y objetivos. Sin embargo quiero recalcar que no se debe de confundir un enfoque humanístico de una empresa con ciertas practicas de manipulación emocional que toman determinadas compañías para atar a sus empleados, esta propuesta del autor busca un compromiso real para generar una relación con limites establecidos entre empleador y empleado donde se tenga siempre presente el contrato entre ambas partes (obligaciones y derechos laborales) y asegurarse de que se cumplan mediante el uso de esta estrategia novedosa.
los actuales planes de estudio de las licenciaturas en Administración en México están articulados en función de los procesos administrativos tradicionales y el estudio de cuatro o cinco de las áreas denominadas "funcionales" de la administración. Esta estructura curricular se complementa con una serie de materias básicas de carácter cuasipropedéutico y otro conjunto de materias especializadas, ya sean optativas o pertenecientes a un núcleo introductorio, que dependen más del capricho de las autoridades universitarias en turno o de las modas administrativas, que de un análisis sustentado en las necesidades de los mercados de trabajo o las realidades locales, regionales o nacionales.
Justificación:
El autor analizó 22 programas de licenciatura en Administración acreditados por CACECA.
Encontró que la mayoría de las materias se centran en:
Proceso administrativo (planeación, organización, dirección, control).
Áreas funcionales (mercadotecnia, finanzas, producción, recursos humanos).
Las materias relacionadas con habilidades blandas (liderazgo, toma de decisiones, ética, etc.) aparecen de forma marginal o optativa.
Critica la dependencia de bibliografía extranjera (principalmente estadounidense), que no siempre se adapta a la realidad mexicana.
Señala que los egresados no están preparados para enfrentar problemas complejos, multiculturales y no cuantificables.
Opinión: La hipótesis es sólida y está bien sustentada. El autor evidencia una desconexión entre la formación académica y las necesidades reales del mercado global. Es crucial que los programas de administración integren más contenidos sobre habilidades transversales, contexto local e internacional, y ética profesional.
La administración en México tiene poca tradición investigadora y está orientada principalmente a la docencia.
Se enseñan modelos idealizados (optimización, excelencia) que son difíciles de aplicar en la realidad organizacional mexicana.
Existe una falta de contextualización: los modelos extranjeros no siempre se adaptan a las condiciones sociales, económicas y culturales de México.
Hay una escasa producción académica nacional en administración, lo que obliga a depender de literatura traducida, often poco adecuada.
Reflejo en el texto: Pariente respalda esta visión al señalar la ausencia de investigaciones y publicaciones de autores mexicanos, así como la falta de materias que aborden la realidad nacional y regional.
El liderazgo es una materia poco frecuente en los planes de estudio analizados (solo aparece en el 20% de los programas).
Cuando se incluye, often forma parte de bloques optativos o complementarios, no del núcleo central.
Propone que el liderazgo debe integrarse junto con otras habilidades como la toma de decisiones, la creatividad y el trabajo en redes.
Además, el autor valora el enfoque de dirección por valores (Blanchard y O’Connor) como una herramienta clave para un liderazgo ético y efectivo.
Hipótesis del autor y opinión personal
-los actuales planes de estudio de las licenciaturas en Administración en México están articulados en función de los procesos administrativos tradicionales y el estudio de cuatro o cinco de las áreas denominadas "funcionales" de la administración…- Página 4.
Los autores Pariente Fragoso y José Luis proponen como su hipótesis que, las licenciaturas de administración de México no se encuentran actualizadas antes las nuevas situaciones y factores que pasa el país y el mundo, manteniendo una enseñanza obsoleta. Hipótesis la cual yo me encuentro de acuerdo, debido a que, es muy usual que varias licenciaturas se vean en la misma posición de solo centrarse en la enseñanza teórica e introductoria (como ejemplos de licenciaturas podemos usar, pedagogía, filosofía, historia, etc) afectando no solo al aprendizaje de los estudiantes y egresados al momento de tener que enfrentarse al campo laboral real, no solo eso, si no que también afectando al mismo sistema (ya sea económico, politico y/o el social) que nos rige debido a la ineficiencia que ellos presentan, creando un ciclo de ineptitud de progresión en nuestro país.
¿Por qué Luis Montaño afirma que la Administración es una disciplina relativamente reciente en México, con poca tradición en investigación y orientada principalmente hacia las áreas docentes?
-con poca tradición en investigación y orientada principalmente hacia las áreas docentes. Se enseñan generalmente modelos idealizados…- Página 7.
Porque Luis Montaño explica que la forma la forma en la que se lleva a cabo la enseñanza de la administración es debido a solamente se enseña de forma aislada de situaciones reales (las cuales cuentan con cambios graduales o repentinos), solo ofreciendo la vista de un modelo idealizado y aislado. Es decir, que solo se enseña lo que es la administración como un concepto, forma que solo funciona cuando se trata de enseñar para la formación básica o futura con un enfoque de docente, y que se encuentra errónea cuando se supone que el objetivo es la enseñanza de este para su aplicación general y no exclusivamente enfocada en una area de desempeño laboral; teniendo que incluir de forma eficiente otras areas para ejercer, sus condiciones y cambios que se puedan presentar en el mundo laboral.
¿Qué opina el autor acerca del liderazgo?
-las materias de vanguardia que deberían integrar la currícula de los estudios administrativos como liderazgo, solución de problemas y toma de decisiones, creatividad, trabajo en redes, conocimiento del entorno e incluso calidad total sólo se encuentran de manera aislada y marginal en la mayor parte de todos los programas estudiados…- Página 15.
Los autores opinan que el liderazgo se trata de una competencia que deben de contar los egresados de cualquier licenciatura de administración, la cual Mexico cuenta pero no formando de forma clave en las aptitudes de los egresados, al tratarse de una materia opcional que se puede o no cursar. Contradiciendo no solo la demanda si no que también, las mismas habilidades que debe de poseer un administrador.
La Administración es una disciplina relativamente reciente en México, con pocatradición en investigación y orientada principalmente hacia las áreas docentes. Seenseñan generalmente modelos idealizados desde la optimización hasta laexcelencia ; los cuales representan la parte "dura", "científica" relativamentefácil de transferir, pero difícil de operar. La Administración está asentada enorganizaciones concretas, alejadas de las condiciones de idealidad de dichosmodelos. No se puede estudiar la Administración en abstracto, ésta tiene que serreferida siempre a las condiciones particulares de las organizaciones que aplicanesos dispositivos.
El autor plantea que las universidades se enfocan demasiado en lo técnico y lo tradicional, descuidando formar líderes. Esto es crucial, porque aunque se domine la teoría, sin capacidad de dirigir equipos o decidir bajo presión, se corre el riesgo de quedar rezagado frente a otros.
¿Qué opina el autor acerca del liderazgo? Ve el liderazgo como una habilidad clave para el administrador moderno, aunque casi no se enseña en los programas. Además, menciona nuevas corrientes como la dirección por valores, que plantean un liderazgo más humano y ético, que no solo mande o fije metas, sino que inspire y dé sentido a las organizaciones.
La Administración es una disciplina relativamente reciente en México, con pocatradición en investigación y orientada principalmente hacia las áreas docentes. Seenseñan generalmente modelos idealizados desde la optimización hasta laexcelencia ; los cuales representan la parte "dura", "científica" relativamentefácil de transferir, pero difícil de operar. La Administración está asentada enorganizaciones concretas, alejadas de las condiciones de idealidad de dichosmodelos. No se puede estudiar la Administración en abstracto, ésta tiene que serreferida siempre a las condiciones particulares de las organizaciones que aplicanesos dispositivos
Hay una escasez de investigaciones originales y publicaciones académicas nacionales.
Predomina el uso de traducciones de textos extranjeros, sobre todo de Estados Unidos, esto dificulta la generación de conocimiento adaptado a la realidad mexicana.
No hay suficientes revistas, artículos o libros hechos por investigadores nacionales; eso limita el debate teórico.
Se produce muy poco conocimiento propio, lo cual limita que la administración se consolide como disciplina académica.
Derivado de lo anterior, la hipótesis central del presente trabajo es que los actualesplanes de estudio de las licenciaturas en Administración en México están articuladosen función de los procesos administrativos tradicionales y el estudio de cuatro o cincode las áreas denominadas "funcionales" de la administración. Esta estructura curricularse complementa con una serie de materias básicas de carácter cuasipropedéutico yotro conjunto de materias especializadas, ya sean optativas o pertenecientes a un nú-cleo introductorio, que dependen más del capricho de las autoridades universitarias enturno o de las modas administrativas, que de un análisis sustentado en las necesidadesde los mercados de trabajo o las realidades locales, regionales o nacionales.
La hipótesis se justifica porque muchos planes de estudio de administración en México siguen un esquema tradicional, centrado en procesos clásicos y unas cuantas áreas funcionales, complementadas con materias básicas y optativas que a menudo responden más a decisiones del momento o modas que a un análisis serio de las necesidades del mercado laboral. Provoca que los egresados tengan una formación general pero poco adaptada a los retos del momento. En mi opinión, esta crítica es válida, mientras la administración evoluciona con temas como digitalización o sostenibilidad, limitan la preparación real de los estudiantes.
La hipótesis se enfoca principalmente en como el estudio de la Administración en México es tradicional y como se complementa con materias básicas y personalmente creo que el seguir toda esta base o estructura es bastante importante ya que seguimos una estructura ya previamente analizada y trabajada
Luis Montaño nos dice que relativamente es nueva la administración y que tiene poca tradición en investigación y la orientación dirigida principalmente a docentes y que es fácil de transferir pero difícil de operar
¿Qué opina el autor acerca del liderazgo? es la toma de decisiones y el conocimiento hacia los objetivos
Derivado de lo anterior, la hipótesis central del presente trabajo es que los actualesplanes de estudio de las licenciaturas en Administración en México están articuladosen función de los procesos administrativos tradicionales y el estudio de cuatro o cincode las áreas denominadas "funcionales" de la administración. Esta estructura curricularse complementa con una serie de materias básicas de carácter cuasipropedéutico yotro conjunto de materias especializadas, ya sean optativas o pertenecientes a un nú-cleo introductorio, que dependen más del capricho de las autoridades universitarias enturno o de las modas administrativas, que de un análisis sustentado en las necesidadesde los mercados de trabajo o las realidades locales, regionales o nacionales.
Estoy de acuerdo en la parte de innovar la educación con el tiempo ya que muchas veces se mantienen planes de estudio que cuando el alumno egresa son obsoletos en el mercado, solo que creo que se deberían de basar mejor en lo actual y preservar en el plan nuevo solo lo que se considere necesario del plan anterior, no que se enseñe a las nuevas generaciones lo que aprendieron generaciones de 20 años atrás, porque el mercado de trabajo no es el mismo.
Derivado de lo anterior, la hipótesis central del presente trabajo es que los actualesplanes de estudio de las licenciaturas en Administración en México están articuladosen función de los procesos administrativos tradicionales y el estudio de cuatro o cincode las áreas denominadas "funcionales" de la administración. Esta estructura curricularse complementa con una serie de materias básicas de carácter cuasipropedéutico yotro conjunto de materias especializadas, ya sean optativas o pertenecientes a un nú-cleo introductorio, que dependen más del capricho de las autoridades universitarias enturno o de las modas administrativas, que de un análisis sustentado en las necesidadesde los mercados de trabajo o las realidades locales, regionales o nacionales
El autor hace una critica a los planes de estudios y el sistema educatico obsoleto ya que llevan mas de 50 años con la misma estrategia, tambièn menciona sobre los docentes que tienen poca experiencia practica lo cual hace que los egresados esten poco preparados para problemas reales y complejos. A mi parecer tiene razon en lo que dice, creo que cosas tan importantes como lo es la educacion deben evolucionar junto con las nuevas tecnologias, creando nuevas estrategias para asi poder adaptarnos al entorno real
La Administración es una disciplina relativamente reciente en México, con pocatradición en investigación y orientada principalmente hacia las áreas docentes. Seenseñan generalmente modelos idealizados desde la optimización hasta laexcelencia ; los cuales representan la parte "dura", "científica" relativamentefácil de transferir, pero difícil de operar. La Administración está asentada enorganizaciones concretas, alejadas de las condiciones de idealidad de dichosmodelos
El autor dice básicamente que las escuelas siguen demasiado enfocadas en lo técnico y lo tradicional, pero se olvidan de enseñar a los futuros administradores a ser líderes. Lo cual es importante ya que puedes saber mucha teoría, pero si no sabes guiar un equipo o tomar decisiones bajo presión, te vas a quedar atrás en comparación con algunas personas
a hipótesis central del presente trabajo es que los actualesplanes de estudio de las licenciaturas en Administración en México están articuladosen función de los procesos administrativos tradicionales y el estudio de cuatro o cincode las áreas denominadas "funcionales" de la administración.
En mi opinión, pienso que el autor tiene razón ya que suena lógico que si se sigue enseñando como hace muchos años los egresados puedan salir no tan preparados. Ya que el mundo cambio bastante desde ese entonces ahora las empresas piden mas requisitos y muchos programas siguen con lo mismo de antes pienso que deberían de implementar nuevas materias que ayuden al egresado a prepararse mejor como lo pueden ser materia de liderazgo o de innovación
Derivado de lo anterior, la hipótesis central del presente trabajo es que los actualesplanes de estudio de las licenciaturas en Administración en México están articuladosen función de los procesos administrativos tradicionales y el estudio de cuatro o cincode las áreas denominadas "funcionales" de la administración. Esta estructura curricularse complementa con una serie de materias básicas de carácter cuasipropedéutico yotro conjunto de materias especializadas, ya sean optativas o pertenecientes a un nú-cleo introductorio, que dependen más del capricho de las autoridades universitarias enturno o de las modas administrativas, que de un análisis sustentado en las necesidadesde los mercados de trabajo o las realidades locales, regionales o nacionales
Aquí esta la hipótesis. Y estoy en un punto medio de estar de acuerdo con el porque siento que esta bien que en los planes de estudio se vean materias que se relacionen con los procesos administrativos tradicionales ya que esto seria como la base que deberían de ver. Pero al igual que el estoy de acuerdo con que también deberían de dar enseñanza a las necesidades mas actuales al mercado y a lo que es el mundo real y no solo lo que seria en teoria.
se comprueba la hipótesisplanteada de que la estructura de los programas de estudio de las licenciaturas enAdministración en México, representadas por aquéllas certificadas por CACECA,tienen como eje central de su diseño curricular las funciones y el proceso adminis-trativo tradicional. Prácticamente todas ellas incluyen un núcleo de materias bási-cas en las áreas disciplinares de las matemáticas, la informática, la contabilidad,la economía y el derecho, así como una serie de materias complementarias de lamás diversa índole a veces agrupadas en un núcleo inicial formativo o como op-ciones optativas de la más variopinta conformación.— —
Esto refuerza la crítica del autor. Las universidades mexicanas no están formando a los administradores con las competencias que demanda la realidad laboral. En mi opinión, actualizar el currículo es urgente para no rezagarse frente a los estándares internacionales.
la hipótesis central del presente trabajo es que los actualesplanes de estudio de las licenciaturas en Administración en México están articuladosen función de los procesos administrativos tradicionales y el estudio de cuatro o cincode las áreas denominadas "funcionales" de la administración.
El autor critica que los planes de estudio estén centrados en un modelo obsoleto. Coincido en que esto limita a los futuros administradores frente a un mundo globalizado, pues hoy se requieren competencias como liderazgo, creatividad y manejo intercultural que no aparecen de forma sólida en los programas.
¡Hola a tod@s!
Con el gusto de saludarles, esperando se encuentren muy bien, les pido que por favor instalen la extensión hypothes.is en su navegador (ver tablón de Classroom).
Posteriormente, en el espacio designado en , identifiquen la hipótesis del autor y la explicación o justificación que ofrece para sustentarla. Coméntenla y emitan su opinión al respecto.
También, encuentren por qué Luis Montaño afirma que la Administración es una disciplina relativamente reciente en México, con poca tradición en investigación y orientada principalmente hacia las áreas docentes.
Finalmente, respondan la siguiente pregunta: ¿Qué opina el autor acerca del liderazgo?
¡Comenzamos!
A Teoria da Base Exportadora, formulada por Douglass North na década de 1950, propõe que o crescimento econômico de uma região está diretamente relacionado à sua capacidade de exportar bens e serviços para outras
Interessante
It took years to acquire a Model O at a price I could afford. It's my dream machine. The other 8 or 9 machines are now being donated to Goodwill, where most came from. I only need one machine and this is it.
quote of u/RickBuxton at https://www.reddit.com/r/typewriters/comments/1nfg9tt/im_new_but_obssessed/
Example of someone who likely should have gone to a typewriter shop and bought a well-adjusted and clean typewriter from the start and it would have saved them time, effort, and money.
8 machines * $30 per machine = $240 plus time, energy, travel, shipping, etc.
Отбрось светильники, отбрось дефиниции.Говори, как видишь, чтó есть в темноте.То есть то. Это есть это.Отринь, отвергни прогнившие имена.Как можно пространствомШествовать, не внимаяЛитые фантасмагорииЕго созидательного шутовства?Вон светильники! НичтоМежду тобой и формами, ты вольешься в,Вставать не должно, когдаХитин закоснелогоС хрустью расколется, лопнет.Быть собою?! Еси.Есмь.
Threads & Concurrency
Threads are independent paths of execution within a program that allow multiple tasks to run concurrently. Concurrency refers to the ability of a system to make progress on multiple tasks during the same period, often by interleaving execution or using multiple cores. This improves performance, especially in I/O-bound or parallelizable tasks. However, when threads share resources, issues like race conditions can occur, requiring synchronization mechanisms such as locks or semaphores.
Es cierto que nosotros como seres humanos nos vemos inmersos en diferentes áreas donde podemos aprender diferentes tipos de información, hay que tener en cuenta que la investigación científica o el conocimiento científico no es el único que se puede utilizar.
En el subtema “El diseño de la investigación”, el autor presenta una breve definición del modelo no lineal, aludiendo al proceso investigativo como un espiral (se puede observar en la figura 1.). Esta metáfora permite observar que cada "reinicio" es retomado con conocimiento previo, también desglosa el proceso, teniendo como puntos clave el reflexionar, cuestionar, analizar, interpretar, etc.
En la semana hemos hablado sobre la formulación de las preguntas que añadiremos en nuestra investigación, si lo relacionamos con el artículo, podemos incluirlo en el subtema "Cómo se debe preguntar", ya que nuestras preguntas son muy "generales y abstractas" y las necesitamos aclarar y delimitar para hacerlas específicas y pertinentes. El autor menciona que es preciso jerarquizar las preguntas reelaboradas, esto dependiendo de la pregunta central formulada: 1. Peguntas fundamentales, 2. preguntas accesorias y 3. preguntas accidentales.
Menciona también que la justificación se debe explicar en forma precisa y clara y por qué la investigación es necesaria y/o conveniente, para esto, la justificación debe responder las siguientes preguntas :¿Por qué es objeto de preocupación este problema específico?, ¿Es un problema importante? y, ¿Qué beneficios aporta?
es muy importante resaltar que el autor menciona que una de las principales dificultades al iniciar una investigación o una tesis es saber escoger el nombre del título y concuerdo con el ya que como hemos mencionado en clase guía a nosotros como investigadores a relacionarlo con todo el proyecto.
COMENTARIO 1: El texto explica muy bien el proceso de investigación, pero resulta demasiado extenso y cargado de información esto puede hacer que uno como lector se abrume o nos perdamos un poco y no nos quede muy claro
Responder a las preguntas sobre la importancia, relevancia y beneficios del problema permite no solo darle sentido al trabajo, sino también mostrar el impacto que puede tener en la sociedad, en un área del conocimiento o en la práctica profesional. En mi opinión, una buena justificación le da credibilidad al estudio y motiva tanto al investigador como a quienes consulten sus resultados.
Me parece fundamental lo que señala el texto, porque en toda investigación la calidad de la pregunta inicial marca el rumbo de los resultados. Una pregunta mal planteada lleva a respuestas confusas o poco útiles, mientras que una bien formulada abre la puerta a un proceso con más claridad yorden
Coincido con Ander-Egg respecto a que la investigación realmente nace de la necesidad de resolver dudas o problemáticas que enfrentamos en nuestro día a día. El hecho de que todo proceso investigativo parta de una situación-problema nos recuerda que la investigación no es algo lejano ni exclusivo de los académicos, sino que está presente en cualquier persona que se cuestiona el porqué de las cosas y busca respuestas.
Coincido con este punto ya que una vez definido el tema se tiene que profundizar buscando información en diferentes sitios o diferentes fuentes esto ayuda a delimitar y sustentar el problema, es de gran ayuda ya que podemos evitar la duplicación de trabajos.
Aquí se presentan dos aspectos sumamente importantes el objetivo y la justificación, se sabe que los objetivos son nuestros guías para saber hacia dónde llevar nuestra investigación, por otra parte, la justificación responde el porqué del trabajo o investigación, en general estos dos apartandos garantizan que nuestra investigacion tenga un proposito.
Primero se tiene que tener un interés del investigador ya que esto sirve de motivación para saber hacia donde va su trabajo o hacia donde lo quiere llevar, este interés despertara curiosidad por aprender más le invertirá tiempo y esfuerzo para profundizar y alcanzar su objetivo.
El texto describe como la investigación no es un proceso lineal, sino una dinámica más compleja y avanzada y se va corrigiendo según lo encontrado se decide si se corrige o si se sigue avanzando, en este aspecto se resalta la importancia de la reflexión contante y la adaptación a la investigación.
por lo que entendi del tema de La situación y el campo problemático son la base para poner en contexto o en situacion una investigación, ya que permiten entender de dónde surge el problema y cuáles son los factores que lo rodean
El proceso de investigación científica es fundamental porque permite transformar dudas o problemas en conocimiento válido y confiable
El artículo explica que no se avanza de forma lineal, sino que se retrocede constantemente para replantear preguntas, ajustar objetivos o redefinir enfoques. Esto refleja que la investigación no es estática, sino dinámica, y permite ver los retrocesos como parte del aprendizaje y no como errores definitivos.
Estas preguntas servirán de guía para estructurar un buen anteproyecto o protocolo de investigación.
This map is a vast simplification of how capitalism actually works.
I think it's amazing how something this complex can even be made so digestible in the first place.
Step 1: Investment (I) Before anything else happens in capitalism, the capitalistmust decide to make an initial investment: establishing their company and startingproduction. This requires an initial expenditure on fixed capital (includingthe workplace itself, and all the machinery and tools inside). The capitalist mustalso provide some working capital, to pay the initial wages of the company’semployees, buy raw materials and supplies, and meet other day-to-day expenses;hence the capitalist needs a certain revolving fund of cash to get production started.(After a full cycle of production has occurred, the company can use some of itsrevenues to pay for those expenses in the next production cycle.)
I just recently had to experience this exact step actually, and I can say that when you look at it like this it really doesn't seem that bad. It can get a lot more frighting when you have to start adding up all the money that it'll cost to do this initial investment. I think depending on the type of business you end up running some steps might not (Step 2 for instance), but over all I defiantly think that this first step will always apply.
Worker households This is where wage-labourers live and reproducethemselves. They raise and educate children; feed, clothe, and care for eachother (including sick and elderly family members); and spend essentiallyall of their wages and salaries to buy the consumer goods and services thatthey need to survive and enjoy life. A great deal of unpaid, unmarketed workoccurs inside the household, most of it performed by women.
I just really love the way that this is worded, it describes humans like some sort of insect that gathers in hives to survive. Thought it does make me wonder if this is how people who are higher in the social hierarchy see those of use that are lower
Economists actually try to use the production function toexplain productivity differences between entire countries orto explain the historical path of productivity within a country.
I think that's really interesting how they can use the past of a country and maybe even see where it'll go in the future simply due to their production history.
Sociologists use a framework that emphasizes group
The idea of this using group framework reminds me of the other reading specifically at the top of chapter 10 where it talks about the different groups that function in the economy.
Is economics a science? Some people, mostly economists,believe that it is. On the other hand, other people, mostlynoneconomists, are skeptical or even scornful of what econ-omists teach.
I think that in some ways it is and isn't, but more so in the way of like how you'd apply different math equations to problems you encounter in the real world. Overall I personally think that economics is more of a psychological than anything, even so, I don't really see any reason it can't be a mixture of both, or is there some reason that I'm just not understanding?
which says that demand goes down as the price goes up.10 Has thelaw of demand been found to be false?Neither I nor any other economist would be willing to con-cede that the law of demand fails to hold. Instead, we wouldlook for factors that might account for the Ursinus Collegeapplication anomaly
I can actually say that this has happened to me in real life where this anomaly occurred. This video game called "R.U.S.E." was removed from Steam awhile ago and me as well as some of my friends really enjoyed it, so when we got computers we decided to go looking for digital keys for it. When we found them before a few years back they were like $60 and now they are $200 - $250 even though the demand for the game has not grown since it is so old. I just wanted to show that this anomaly is more common than one might think.
Explicación o justificación: para poder consolidar un verdadero y sustantivo cambio en la formación de nuestros futuros administradores es imprescindible partir de un nuevo paradigma que deseche radicalmente los obsoletos enfoques de funciones y proceso administrativo además deberá articularse con una visión realista de los entornos multiculturales y globaliza dotes de la sociedad en la que se desenvolverán los futuros profesionales de la administración en México y en el mundo.
HIPÓTESIS: la hipótesis central del presente trabajo es que los actuales planes de estudio de las licenciaturas en Administración en México están articulados en función de los procesos administrativos tradicionales y el estudio de cuatro o cinco de las áreas denominadas "funcionales" de la administración. Esta estructura curricular se complementa con una serie de materias básicas de carácter casi propedéutico y otro conjunto de materias especializadas, ya sean optativas o pertenecientes a un núcleo introductorio, que dependen más del capricho de las autoridades universitarias en turno o de las modas administrativas, que de un análisis sustentado en las necesidades de los mercados de trabajo o las realidades locales, regionales o nacionales.
Mi opinión: de acuerdo a lo ya leído es que se necesitan nuevos enfoques de enseñanza que cada cierto tiempo vayan actualizándose y cambiando de acuerdo a como cambian los tiempos y las personas ya que así se tendría una mejor enseñanza y una más clara definición de lo que es o será ser un administrador ya sea en las diferentes áreas en las que se puede administrar o que tiene relación con la administración.
documentary of its making, Memory: The Origins of Alien;
Tengo que verlo. Necesito verlo.
Modelos cognitivo-behavioristas são mais claramente teorias de ensino e modelossocioconstrutivistas são mais claramente teorias de aprendizagem, mas ambos aindase traduzem bem em métodos e processos para ensino. Os modelos conectivistas sãomais distintamente teorias do conhecimento, o que torna difícil traduzi-los em maneirasde aprender – e ainda mais difícil traduzi-los em maneiras de ensinar. (ANDERSON;DRON, 2011, p. 89-90).
Síntese muito boa. Onde tudo á disponível, o que devemos escolher para/como aprender e ensinar.
As teorias de aprendizagem tradicionais,utilizadas como suporte à educação presencial, nãoforam produzidas tendo em mente ambientes virtuais.
Aqui, talvez fosse importante estudar quais seriam as novas teorias desejáveis para o e-learning.
In my beginner’s guide to Roam, I completely left out the Daily Notes section to keep things simple. Let’s now have a look together. This is what a daily note with interstitial journaling looks like. <img fetchpriority="high" decoding="async" width="674" height="717" src="https://viahtml.hypothes.is/proxy/im_/https://nesslabs.com/wp-content/uploads/2020/04/interstitial-journaling-example.png" alt="" class="wp-image-8473" srcset="https://viahtml.hypothes.is/proxy/https://nesslabs.com/wp-content/uploads/2020/04/interstitial-journaling-example.png 674w, https://viahtml.hypothes.is/proxy/https://nesslabs.com/wp-content/uploads/2020/04/interstitial-journaling-example-282x300.png 282w" sizes="(max-width: 674px) 100vw, 674px"/> Track time. Type /time to insert the current time, then type whatever you are thinking about.Track tasks. Type /todo to create to-do items. Check off these to-do items when done.Track content. When you stumble upon something interesting that would disturb your workflow, add it to master lists such as [[To read]]. You can see I have done it in this screenshot with an interesting-looking article that had nothing to do with the essay I was trying to write.Track ideas. Similarly, if you think of something else you’d like to do today, just add it as a to-do where and when you think about it. For people using the [[Today]], [[Tomorrow]], [[Someday]] system, you can also add that to the to-do items, or add a specific date, as I have done with “call Morgane.”Track well-being. I like to start my work day with a quick note checking in on how I feel, anything that’s been sometimes literally keeping me up at night, any major roadblock I’m anticipating for the day. It’s rarely longer than one bullet point, but it’s a great way to take care of my general well-being. I also finish the work day with a similar quick closing note.
Me parece curiosa esta parte y es que muestra cómo el diario intersticial convierte hasta lo más simple como es la hora, una tarea o una idea en algo organizado. Sin imaginar, terminas con un registro completo de tu día sin tener que hacer demasiado y eso es lo que buscamos la mayoría algo practico.
write a few lines every time you take a break, and to track the exact time
Esta práctica me parece cercana a los diarios de investigación o a las bitácoras de campo. Permite capturar micro-evidencias de lo que ocurre en los intersticios del trabajo académico, fortaleciendo la trazabilidad del proceso de aprendizaje.
Proactive breaks: reflect on your previous task, plan for the next one, take your own mental pulse, jot down anything else that comes to mind so as to reduce your cognitive load.Procrastination breaks: become aware of these breaks and how long they actually take. When you create the habit of writing down all your breaks, it becomes easier to not open a new tab to “quickly” check Twitter. You don’t want to have to admit that failure to yourself.
Escribir tus descansos cambia un momento perdido en un recordatorio para seguir enfocado en tus proyectos o metas.
Notice the mix of goals (“finish the first draft”), self-awareness (“fell into a Twitter blackhole”, “feeling anxious”), self-review (“good progress”), and actionable items (“call Anna”)? I love interstitial journaling because it’s a great way to make your breaks more mindful.
Lo que me gusta de la idea del diario es que mezcla varias cosas a la vez como por ejemplo lo que planeas hacer, como te sientes, si has avanzado o aun no, y lo mas importante lo que queda pendiente. Me parece una forma sencilla de volver más conscientes los descansos.
Doe s no t pass i nt o [a view of] existence
I think I do not understand the wording. Is this perhaps referring to his definition of emptiness of 'emptiness'? That emptiness by itself is not a universal, or independent truth/essence/existence?
T h e m e nt al a nd physical aggregates ariseFr om t he c once pti on o f I whi ch is false i n fact.bH o w c ould wh at is grownFr om a false seed be true?
Nagarjuna, belonging to the Madhyamika school of thought, argued for emptiness (even of emptiness). In doing so, he contradicted the brahmanical schools of thought, which believed in the existence of atman and brahman, and a 'permanence'. Nagarjuna on the contrary, being a mahayana buddhist, believed in 'impermanence'. This impermanence extended to that of 'being' as well, with all existence being 'momentary', and a superimposition of our cognition, of sorts.
O n e who sees how cause a nd effectAre produce d a nd destroyedDoe s no t r egard t he wor ldAs really existent or really non-exist ent
My takeaway from this explanation is that the crux of exiting the cycle of birth and rebirth is in a complete and total belief that it and everything it comprises is nonexistent. Upon arriving at the conclusion that is it nonexistent, there is no way to truly verify if it exists or not. It's a situation where you have decided that it is nonexistent and that action is what removes you from it altogether. So it makes sense that the person who understands all this ultimately becomes indifferent to whether it is "really existant or really nonexistant" which in action is as good as saying you believe it does not exist.
But like t he i mage o f one ’s faceT h e I does no t a t all really exist.*
This is interesting, but what if we switched the analogy from the face/I being a reflection dependent on a mirror, to the face/I being perceived by another person. Would this logic still hold steady? How do we reconcile this with one human perceiving another? Would we then say that the I is nonexistant but the other is?
mérito
Ramo do Direito DIREITO PROCESSUAL CIVIL
TemaPaz, Justiça e Instituições Eficazes <br /> Embargos de divergência. Incidência da Súmula n. 7 do STJ no exame do recurso especial. Ausência de apreciação do mérito do recurso. Não cabimento.
Destaque - Não é cabível a interposição de embargos de divergência quando o mérito do recurso especial não foi apreciado devido à incidência da Súmula n. 7 do STJ.
Informações do Inteiro Teor - A questão em discussão consiste em saber se é cabível a interposição de embargos de divergência quando o mérito do recurso especial não foi apreciado devido à incidência da Súmula n. 7 do STJ.
No caso, entendeu-se pela aplicação da Súmula n. 7 do STJ porque o conjunto probatório, notadamente o relato da vítima, confirmado pelos depoimentos das testemunhas, não deixou dúvidas quanto à autoria dos delitos sob apuração.
Foi registrado que não havia como se acolher a pretensão de absolvição se o Tribunal a quo, ao concluir pela autoria do recorrente no cometimento do delito em questão, sopesou as provas colhidas e os depoimentos obtidos em juízo. Desfazer tal conclusão implicaria revolver conteúdo fático-probatório dos autos, vedado pelo óbice sumular.
Vale anotar que, na hipótese, "as peculiaridades do caso concreto ensejaram a incidência da Súmula 7 do STJ, circunstância que inviabiliza o cabimento dos embargos de divergência, ante a impossibilidade de harmonizar o juízo de conhecimento realizado no acórdão embargado, com o do paradigma quanto à aplicação de regra técnica de conhecimento do recurso especial." (AgInt nos EREsp n. 1.998.469/PE, rel. Ministro Gurgel de Faria, Primeira Seção, julgado em 14/11/2023, DJe de 23/11/2023).
Sendo assim, não há divergência jurisprudencial a ser reconhecida, pois, consoante a jurisprudência do STJ, "os embargos de divergência não se constituem de instrumento processual adequado para discussão sobre técnica de admissibilidade do recurso especial" (STJ, AgInt nos EAREsp 893.726/SP, Rel. Ministro Mauto Campbell Marques, Corte Especial, DJe de 7/11/2018).
Author response:
Reviewer #1, Comment (1): Terminology
We fully acknowledge the importance of terminological consistency and will align our usage with established literature. Specifically, we will revise as follows,
(1) Replace “sinusoidal analysis” with either “sinusoidal modulation” (Doeller et al., 2010; Bao et al., 2019; Raithel et al., 2023) or “GLM with sinusoidal (cos/sin) regressors” (Constantinescu et al., 2016).
(2) Replace “1D directional domain” with either “angular domain of movement directions (0–360°)” or “directional modulation analysis”.
Reviewer #1, Comment (2): Spectral analysis and 3-fold periodicity
We agree that the presentation of our spectral analysis and the theoretical motivation underlying our expectation of a three-fold periodicity within hippocampal data requires further clarification.
In our revised manuscript, we will:<br /> (1) Clearly articulate the theoretical motivation for anticipating a three-fold signal, explicitly linking it to the known hexagonal grid structure encoded by the entorhinal cortex.
(2) Clarify our methodological rationale for using Fourier analysis (FFT).
a) FFT allows unbiased exploration of multiple candidate periodicities (e.g., 3–7-fold) without predefined assumptions.
b) FFT results cross-validate our sinusoidal modulation results, providing complementary evidence supporting the 6-fold periodicity in EC and 3-fold periodicity in HPC.
c) FFT uniquely facilitates analysis of periodicities in behavioral performance data, which is not feasible via standard sinusoidal GLM approaches. This consistency allows us to directly compare periodicities across neural and behavioral data.
(3) Further, we will expand our discussion to provide:
a) A deeper interpretation of potential biological bases for the observed hippocampal three-fold periodicity.
b) A careful examination of alternative explanations within existing hippocampal modeling frameworks.
Reference:
Doeller, C. F., Barry, C., & Burgess, N. (2010). Evidence for grid cells in a human memory network. Nature, 463(7281), 657-661.
Constantinescu, A. O., O'Reilly, J. X., & Behrens, T. E. J. (2016). Organizing conceptual knowledge in humans with a gridlike code. Science, 352(6292), 1464-1468.
Bao, X., Gjorgieva, E., Shanahan, L. K., Howard, J. D., Kahnt, T., & Gottfried, J. A. (2019). Grid-like neural representations support olfactory navigation of a two-dimensional odor space. Neuron, 102(5), 1066-1075.
Raithel, C. U., Miller, A. J., Epstein, R. A., Kahnt, T., & Gottfried, J. A. (2023). Recruitment of grid-like responses in human entorhinal and piriform cortices by odor landmark-based navigation. Current Biology, 33(17), 3561-3570
Author response:
The following is the authors’ response to the original reviews.
Reviewer #1 (Public Review):
Summary:
This paper sets out to achieve a deeper understanding of the effects of hydrogen sulfide on C. elegans behavior and physiology, with a focus on behavior, detection mechanism(s), physiological responses, and detoxification mechanisms.
Strengths:
The paper takes full advantage of the experimental tractability of C. elegans, with thorough, welldesigned genetic analyses.
Some evidence suggests that H<SUB>2</SUB>S may be directly detected by the ASJ sensory neurons. The paper provides interesting and convincing evidence for complex interactions between responses to different gaseous stimuli, particularly an antagonistic role between H<SUB>2</SUB>S and O2 detection/response. Intriguing roles for mitochondria and iron homeostasis are identified, opening the door to future studies to better understand the roles of these components and processes.
We thank the reviewer for the supportive comments.
Weaknesses:
The claim that worms' behavioral responses to H<SUB>2</SUB>S are mediated by direct detection is incompletely supported. While a role for the chemosensory neuron ASJ is implicated, it remains unclear whether this reflects direct detection. Other possibilities, including indirect effects of ASJ and the guanylyl cyclase daf-11 on O2 responses, are also consistent with the authors' data.
We thank the reviewer for the insightful comment and agree that the role of ASJ neurons in H<SUB>2</SUB>S detection was not clear. We included new experiments and revised our text to make it clearer.
Since our initial analyses suggest a role of ASJ neurons in H<SUB>2</SUB>S-evoked locomotory responses (Figure 2F and G), We thought that this would offer us a starting point to dissect the neuronal circuit involved in H<SUB>2</SUB>S responses. Expression of the tetanus toxin catalytic domain in ASJ, which blocks neurosecretion, inhibited H<SUB>2</SUB>S-evoked locomotory speed responses (Figure 2H), suggesting that neurosecretion from ASJ promotes H<SUB>2</SUB>S-evocked response (Lines 162–165). We then performed calcium imaging of ASJ neurons in response to H<SUB>2</SUB>S exposure. However, while we observed CO<SUB>2</SUB>-evoked calcium transients in ASJ using GCaMP6s, we did not detect any calcium response to H<SUB>2</SUB>S, under several conditions, including animals on food, off food, and with different H<SUB>2</SUB>S concentrations and exposure times (Figure2—Figure supplement 2E and F) (Lines 166–170). Since signaling from ASJ neurons regulates developmental programs that modify sensory functions in C. elegans (Murakami et al., 2001), the involvement of ASJ neurons is not specific to H<SUB>2</SUB>S and ASJ neurons are unlikely to serve as the primary H<SUB>2</SUB>S sensor (Discussed in Line 449–458). Therefore, the exact sensory neuron, circuit and molecular triggers mediating acute H<SUB>2</SUB>S avoidance remain to be elucidated.
Our subsequent investigation on mitochondrial components suggests that a burst of mitochondrial ROS production may be the trigger for H<SUB>2</SUB>S avoidance, as transient exposure to rotenone substantially increases baseline locomotory speed (Figure 7E) (Line 391–396). However, to initiate avoidance behavior to H<SUB>2</SUB>S, mitochondrial ROS could potentially target multiple neurons and cellular machineries, making it challenging to pinpoint specific sites of action. Nevertheless, we agree that further dissection of the neural circuits and mitochondrial signaling in H<SUB>2</SUB>S avoidance will be important and should be explored in future studies.
The role of H<SUB>2</SUB>S-mediated damage in behavioral responses, particularly when detoxification pathways are disrupted, remains unclear.
We thank the reviewer for the insightful comment and fully agree with the concern raised. The same issue was also noted by the other reviewers. We agree that decreased locomotory responses in H<SUB>2</SUB>S-sensitized animals can arise from distinct causes, either systemic toxicity or behavioral adaptation, and distinguishing between these is critical. We have included new experiments and revised the text to clarify this issue.
Our data suggest that increased initial omega turns and a rapid loss of locomotion in hif-1 and detoxification-defective mutants including sqrd-1 and ethe-1 likely reflect an enhanced sensitivity to H<SUB>2</SUB>S toxicity due to their failure to induce appropriate adaptative responses (Figure 5D–F, Figure 5J–L, Figure 5—Figure supplement 1F–P). Supporting this, hif-1 mutants become less responsive to unrelated stimuli (near-UV light) after 30 minutes of H<SUB>2</SUB>S exposure (Figure 5I).
In contrast, egl-9 and SOD-deficient animals show reduced initial omega-turn and reduced speed responses (Figure 5B, Figure 7G, Figure 5—Figure supplement 1A and B, and Figure 7—Figure supplement 1F and G), although both egl-9 and sod mutants respond normally to the other stimuli prior or after H<SUB>2</SUB>S exposure (Figure 5I, Figure 5—Figure supplement 1C, and Figure 7—Figure supplement 1H). Since disrupting egl-9 stabilizes HIF-1 and upregulates the expression of numerous genes involved in cellular defense against H<SUB>2</SUB>S toxicity, the enhanced detoxification capacity in egl-9 mutants likely increases animals’ tolerance to H<SUB>2</SUB>S, thereby reducing avoidance to otherwise toxic H<SUB>2</SUB>S levels. Similarly, persistently high ROS in SOD deficient animals activates a variety of stress-responsive signaling pathways, including HIF-1, NRF2/SKN-1 and DAF-16/ FOXO signaling (Lennicke & Cocheme, 2021; Patten et al., 2010), facilitating cellular adaptation to redox stress and reducing animals’ responsiveness to toxic H<SUB>2</SUB>S levels. Taken together, these findings support the view that reduced locomotory speed during H<SUB>2</SUB>S exposure can arise from distinct mechanisms: early systemic toxicity in hif-1 and detoxificationdefective mutants, versus enhanced cellular adaptation in egl-9 and SOD mutants. We have integrated the relevant information across the result section and discussed this in Lines 485-536.
The findings of the paper are somewhat disjointed, such that a clear picture of the relationships between H<SUB>2</SUB>S detection, detoxification mechanisms, mitochondria, and iron does not emerge from these studies. Most importantly, the relative roles of H<SUB>2</SUB>S detection and integration, vs. general and acute mitochondrial crisis, in generating behavioral responses are not convincingly resolved.
We thank the reviewer for this comment and agree that our presentation did not fully connect different findings into a cohesive picture. To address this, we have acquired new data, and revised the abstract, results and discussion sections to clarify two phases of H<SUB>2</SUB>S-evoked responses: an initial avoidance behavior upon H<SUB>2</SUB>S exposure, followed by a later phase of adaption and detoxification when the escape is not successful.
In brief, we began with the basic characterization of H<SUB>2</SUB>S-induced locomotory speed response, followed by a candidate gene screen to identify key molecules and pathways involved in initial speed response to H<SUB>2</SUB>S. Subsequently, we focused on three major intersecting pathways that contributed to the acute behavioral response to H<SUB>2</SUB>S. These include cGMP signaling, which led to the identification of ASJ neurons; nutrient-sensitive pathways that modulate behavioral responses to both H<SUB>2</SUB>S and CO2; and O2sensing signaling, whose activation inhibits responses to H<SUB>2</SUB>S. However, the molecules and neurons in these pathways, including ASJ, likely play modulatory roles and are unlikely to serve as the primary H<SUB>2</SUB>S sensors. Our subsequent analysis, however, suggests that mitochondria play a critical role in triggering avoidance behavior upon H<SUB>2</SUB>S exposure. Brief treatment with rotenone, a potent inducer of ROS, led to marked increase in locomotory speed (Figure 7E). This suggests the possibility that a burst of ROS production triggered toxic levels of H<SUB>2</SUB>S (Jia et al., 2020) may initiate the avoidance behavior.
When the initial avoidance fails, H<SUB>2</SUB>S detoxification programs are induced as a long-term survival strategy. The induction of detoxification programs appears to enhance tolerance to H<SUB>2</SUB>S exposure and contributes to the gradual decrease of locomotory speed in H<SUB>2</SUB>S. We now provide a clearer image of how different pathways modulate H<SUB>2</SUB>S detoxification and adaptation (see our responses to other comments). Briefly, mutants defective in detoxification, such as hif-1 and other detoxification-defective mutants, showed stronger initial omega-turn response and a rapid loss of locomotion. This loss of locomotion is likely caused by early cellular toxicity as the mutants failed to respond to other unrelated stimuli (nearUV light) after 30 minutes of H<SUB>2</SUB>S exposure (Figure 5I). Likewise, smf-3 mutants and BP-treated animals were hypersensitive to H<SUB>2</SUB>S (Figure 6D and E, and Figure 6—Figure supplement 1G and I), likely due to impaired H<SUB>2</SUB>S detoxification under low iron conditions, as iron is a co-factor required for the activity of the H<SUB>2</SUB>S detoxification enzyme ETHE-1 (Figure 5K and Figure 5—Figure supplement 1E).
In contrast, reduced locomotion and response in other contexts such as egl-9 mutants and SODdeficient animals reflect H<SUB>2</SUB>S-induced adaptive mechanism rather than toxicity as they remain responsive to the other stimuli after H<SUB>2</SUB>S exposure. Since disrupting egl-9 stabilizes HIF-1 and upregulates the expression of numerous genes involved in cellular defense against H<SUB>2</SUB>S toxicity, the enhanced detoxification capacity in egl-9 mutants likely increases animals’ tolerance to H<SUB>2</SUB>S, thereby reducing avoidance to otherwise toxic H<SUB>2</SUB>S levels. Similarly, persistently high ROS in SOD deficient animals activates a variety of stress-responsive signaling pathways, including HIF-1, NRF2/SKN-1 and DAF-16/ FOXO signaling (Lennicke & Cocheme, 2021; Patten et al., 2010), facilitating cellular adaptation to redox stress and reducing animals’ responsiveness to toxic H<SUB>2</SUB>S levels. Therefore, different animals decline their locomotory speed to the effects of H<SUB>2</SUB>S through distinct mechanisms. We have integrated the relevant information across the result section and discussed this in Lines 485-536.
Reviewer #2 (Public Review):
Summary:
H<SUB>2</SUB>S is a gas that is toxic to many animals and causes avoidance in animals such as C. elegans. The authors show that H<SUB>2</SUB>S increases the frequency of turning and the speed of locomotion. The response was shown to be modulated by a number of neurons and signaling pathways as well as by ambient oxygen concentrations. The long-term adaptation involved gene expression changes that may be related to iron homeostasis as well as the homeostasis of mitochondria.
Strengths:
Overall, the authors provide many pieces that will be important for solving how H<SUB>2</SUB>S signals through neuronal circuits to change gene expression and physiological programs. The experiments rely mostly on a behavioral assay that measures the increase of locomotion speed upon exposure to H<SUB>2</SUB>S. This assay is then combined with manipulations of environmental factors, different wild-type strains, and mutants. The mutants analyzed were obtained as candidates from the literature and from transcriptional profiling that the authors carried out in worms that were exposed to H<SUB>2</SUB>S. These studies imply several genetic signaling pathways, some neurons, and metabolism-related factors in the response to H<SUB>2</SUB>S. Hence the data provided should be useful for the field.
We thank the reviewer for the supportive comments.
Weaknesses:
On the other hand, many important aspects of the underlying mechanisms remain unsolved and the reader is left with many loose ends. For example, it is not clear how H<SUB>2</SUB>S is actually sensed, how sensory neurons are activated and signal to downstream circuits, and what the role of ciliated and RMG neurons is in this circuit. It remains unclear how signals lead to gene expression and physiological changes such as metabolic rewiring. Solving all this would clearly be beyond the scope of a single manuscript. Yet, the manuscript also does not focus on understanding one of these central aspects and rather is all over the place, which makes it harder to understand for readouts that are not in this core field. Multiple additional methods and approaches exist to dig deeper into these mechanisms in the future, such as neuronal calcium imaging, optogenetics, and metabolic analysis. To generate a story that will be interesting to a broad readership substantial additional experimentation would be required. Further, in the current manuscript, it is often difficult to understand the rationales of the experiments, why they were carried out, and how to place them into a context. This could be improved in terms of documentation, narration/explanation, and visualization.
We thank the reviewer for the comment, which has also been raised by the other reviewers. We agree that our initial submission was poorly presented. We also acknowledge the fact that some aspects, such as detailed neural circuit and sensory transduction, still remain unresolved. We have now included additional experiments and revised the manuscript to clarify the logic of our experiments, provided better context for our findings, and improved both the narrative flow and data visualization to make the manuscript more accessible to readers. We now provide a clearer image of how different pathways interact to modulate the initial avoidance response, and the H<SUB>2</SUB>S detoxification and behavioral habituation during prolonged H<SUB>2</SUB>S exposure. The following response is similar to the one for reviewer #1.
In brief, we began with the basic characterization of H<SUB>2</SUB>S-induced locomotory speed response, followed by a candidate gene screen to identify key molecules and pathways involved in initial speed response to H<SUB>2</SUB>S. Subsequently, we focused on three major intersecting pathways that contributed to the acute behavioral response to H<SUB>2</SUB>S. These include cGMP signaling, which led to the identification of ASJ neurons; nutrient-sensitive pathways that modulate behavioral responses to both H<SUB>2</SUB>S and CO2; and O2sensing signaling, whose activation inhibits responses to H<SUB>2</SUB>S. However, the molecules and neurons in these pathways, including ASJ, likely play modulatory roles and are unlikely to serve as the primary H<SUB>2</SUB>S sensors. Our subsequent analysis, however, suggests that mitochondria play a critical role in triggering avoidance behavior upon H<SUB>2</SUB>S exposure. Brief treatment with rotenone, a potent inducer of ROS, led to marked increase in locomotory speed (Figure 7E). This suggests the possibility that a burst of ROS production triggered toxic levels of H<SUB>2</SUB>S (Jia et al., 2020) may initiate the avoidance behavior.
When the initial avoidance fails, H<SUB>2</SUB>S detoxification programs are induced as a long-term survival strategy. The induction of detoxification programs appears to enhance tolerance to H<SUB>2</SUB>S exposure and contributes to the gradual decrease of locomotory speed in H<SUB>2</SUB>S. We now provide a clearer image of how different pathways modulate H<SUB>2</SUB>S detoxification and adaptation (see our responses to other comments). Briefly, mutants defective in detoxification, such as hif-1 and other detoxification-defective mutants, showed stronger initial omega-turn response and a rapid loss of locomotion. This loss of locomotion is likely caused by early cellular toxicity as the mutants failed to respond to other unrelated stimuli (nearUV light) after 30 minutes of H<SUB>2</SUB>S exposure (Figure 5I). Likewise, smf-3 mutants and BP-treated animals were hypersensitive to H<SUB>2</SUB>S (Figure 6D and E, and Figure 6—Figure supplement 1G and I), likely due to impaired H<SUB>2</SUB>S detoxification under low iron conditions, as iron is a co-factor required for the activity of the H<SUB>2</SUB>S detoxification enzyme ETHE-1 (Figure 5K and Figure 5—Figure supplement 1E).
In contrast, reduced locomotion and response in other contexts such as egl-9 mutants and SODdeficient animals reflect H<SUB>2</SUB>S-induced adaptive mechanism rather than toxicity as they remain responsive to the other stimuli after H<SUB>2</SUB>S exposure. Since disrupting egl-9 stabilizes HIF-1 and upregulates the expression of numerous genes involved in cellular defense against H<SUB>2</SUB>S toxicity, the enhanced detoxification capacity in egl-9 mutants likely increases animals’ tolerance to H<SUB>2</SUB>S, thereby reducing avoidance to otherwise toxic H<SUB>2</SUB>S levels. Similarly, persistently high ROS in SOD deficient animals activates a variety of stress-responsive signaling pathways, including HIF-1, NRF2/SKN-1 and DAF-16/ FOXO signaling (Lennicke & Cocheme, 2021; Patten et al., 2010), facilitating cellular adaptation to redox stress and reducing animals’ responsiveness to toxic H<SUB>2</SUB>S levels. Therefore, different animals decline their locomotory speed to the effects of H<SUB>2</SUB>S through distinct mechanisms. We have integrated the relevant information across the result section and discussed this in Lines 485-536.
Reviewer #3 (Public Review):
Summary:
The manuscript explores the behavioral responses of C. elegans to hydrogen sulfide, which is known to exert remarkable effects on animal physiology in a range of contexts. The possibility of genetic and precise neuronal dissection of responses to H<SUB>2</SUB>S motivates the study of responses in C. elegans. The manuscript is well-written in communicating the complex physiology around C. elegans behavioral responses to H<SUB>2</SUB>S and in appropriately citing prior and related relevant work.
There are three parts to the manuscript.
In the first, an immediate behavioral response-increased locomotory rate-upon exposure to H<SUB>2</SUB>S is characterized. The experimental conditions are critical, and data are obtained from exposure of animals to 150ppm H<SUB>2</SUB>S at 7% O2. The authors provide evidence that this is a chemosensory response to H<SUB>2</SUB>S, showing a requirement for genes encoding components of the cilia apparatus and implicating a role for tax-4 and daf-11. Neuron-specific rescue in the ASJ neurons suggests the ASJ neurons contribute to the response to H<SUB>2</SUB>S. One caveat is that previous work has shown that the dauer-constitutive phenotype of daf-11 mutants can be suppressed by ASJ ablation, suggesting that there may be pervasive changes in animal nervous system signaling that are ASJ-dependent in daf-11 mutants, which may indirectly alter chemosensory responses to H<SUB>2</SUB>S. More direct methods to assess whether ASJ senses H<SUB>2</SUB>S, e.g. using calcium imaging, would better assess a direct role for the ASJ neurons in a behavioral response to H<SUB>2</SUB>S. The authors also point out interesting parallels between the response to H<SUB>2</SUB>S and CO2 though provide some genetic data separating the two responses. Importantly, the authors note that when aerotaxis (O2sensing and movement) in the presence of bacterial food is intact, as in npr-1 215F animals, the response to H<SUB>2</SUB>S is abrogated. Mutation in gcy-35 in the npr-1 215F background restores the H<SUB>2</SUB>S chemosensory response.
There is a second part of the paper that conducts transcriptional profiling of the response to H<SUB>2</SUB>S that corroborates and extends prior work in this area.
The final part of the paper is the most intriguing, but for me, also the most problematic. The authors examine how H<SUB>2</SUB>S-evoked locomotory behavioral responses are affected in mutants defective in the stress and detoxification response to H<SUB>2</SUB>S, most notably hif-1. Prior genetic studies have established the pathways leading to HIF-1 activation/stabilization, as well as potential downstream mechanisms. The authors conduct logical genetic analysis to complement studies of the hif-1 mutant and in part motivated by their transcriptional profiling studies, examine the role of iron sequestration/free iron in the locomotory response to H<SUB>2</SUB>S, and further speculate on how the behavior of mutants defective in mitochondrial function might be affected by exposure to H<SUB>2</SUB>S.
In some regard, this part of the manuscript is interesting because the analysis begins to connect how the behavior of an animal to a toxic compound is affected by mutations that affect sensitivity to the toxic compound. However, what is unclear is what is being studied at this point. In the context, of noting that H<SUB>2</SUB>S at 150ppm is known to be lethal, its addition to mutants clearly sensitized to its effects would be anticipated to have pervasive effects on animal physiology and nervous system function. The authors note that the continued increased locomotion of wild-type animals upon H<SUB>2</SUB>S exposure might be due to the byproducts of detoxification or the detrimental effects of H<SUB>2</SUB>S. The latter explanation seems much more likely, in which case what one may be observing is the effects of general animal sickness, or even a bit more specifically, neuronal dysfunction in the presence of a toxic compound, on locomotion. As such, what is unclear is what conclusions can be taken away from this part of the work.
Strengths:
(1) Characterization of a motor behavior response to H<SUB>2</SUB>S
(2) Transcriptional profiling of the response to H<SUB>2</SUB>S corroborating prior work.
We thank the reviewer for the supportive comments.
Weaknesses:
Unclear significance and experimental challenges regarding the study of locomotory responses to animals sensitized to the toxic effects of H<SUB>2</SUB>S under exposure to H<SUB>2</SUB>S.
We thank the reviewer for the comment, which has also been raised by the other reviewers. We agree that our initial submission left several important questions open, and we acknowledge the fact that some aspects, such as detailed neural circuit and sensory transduction, still remain unresolved. Nevertheless, we acquired new data and revised our text, aiming to clarify the distinct mechanisms underlying the reduced locomotion in different mutants during prolonged H<SUB>2</SUB>S exposure.
Our data suggest that increased initial omega turns and a rapid loss of locomotion in hif-1 and detoxification-defective mutants including sqrd-1 and ethe-1 likely reflect an enhanced sensitivity to H<SUB>2</SUB>S toxicity due to their failure to induce appropriate adaptative responses (Figure 5D–F, Figure 5J–L, Figure 5—Figure supplement 1F–P). Supporting this, hif-1 mutants become less responsive to unrelated stimuli (near-UV light) after 30 minutes of H<SUB>2</SUB>S exposure (Figure 5I).
In contrast, egl-9 and SOD-deficient animals show reduced initial reorientation and reduced speed responses (Figure 5B, Figure 7G, Figure 5—Figure supplement 1A and B, and Figure 7—Figure supplement 1F and G), although both egl-9 and sod mutants respond normally to the other stimuli prior or after H<SUB>2</SUB>S exposure (Figure 5I, Figure 5—Figure supplement 1C, and Figure 7—Figure supplement 1H). Since disrupting egl-9 stabilizes HIF-1 and upregulates the expression of numerous genes involved in cellular defense against H<SUB>2</SUB>S toxicity, the enhanced detoxification capacity in egl-9 mutants likely increases animals’ tolerance to H<SUB>2</SUB>S, thereby reducing avoidance to otherwise toxic H<SUB>2</SUB>S levels. Similarly, constant high ROS in SOD deficient animals activates a variety of stress-responsive signaling pathways, including HIF-1, NRF2/SKN-1 and DAF-16/ FOXO signaling (Lennicke & Cocheme, 2021; Patten et al., 2010), facilitating cellular adaptation to redox stress and reducing animals’ responsiveness to toxic H<SUB>2</SUB>S levels. Taken together, these findings support the view that reduced locomotory speed during H<SUB>2</SUB>S exposure can arise from distinct mechanisms: early systemic toxicity in hif-1 and detoxification-defective mutants, versus enhanced cellular adaptation in egl-9 and SOD mutants. We have integrated the relevant information across the result section and discussed this in Lines 485-536.
Reviewer #1 (Recommendations For The Authors):
To better substantiate a role for H<SUB>2</SUB>S detection, it would be useful for the authors to image Ca responses to H<SUB>2</SUB>S in ASJ in WT and unc-13, and to rule out the possibility that the requirement for daf-11 in ASJ reflects a role in O2 rather than H<SUB>2</SUB>S detection.
We thank the reviewer for this comment. As suggested, we performed calcium imaging of ASJ neurons using GCaMP6s. As previously described, 3% CO<SUB>2</SUB> evoked a calcium transient in ASJ (Figure 2—figure supplement 2F). To investigate whether H<SUB>2</SUB>S evoked a calcium transient in ASJ neurons, we tested several conditions, including animals on food or off food, with different H<SUB>2</SUB>S concentrations (~75 or ~150ppm), and different exposure time (4 or 8 mins). However, we did not detect a calcium response to H<SUB>2</SUB>S in ASJ under any of the conditions tested (Figure2—figure supplement 2E) (Lines 166–168). Given that neuronspecific rescue of daf-11 or tax-4 mutants pointed to a role of ASJ neurons in promoting H<SUB>2</SUB>S responses, we sought to determine how ASJ neurons were involved. Expression of the tetanus toxin catalytic domain in ASJ neurons, which blocks neurosecretion, inhibited H<SUB>2</SUB>S-evoked locomotory speed responses (Figure 2H), similar to the phenotypes observed in daf-11 and daf-7 mutants (Figure 2C and D) (Lines 162–165). These results confirm that ASJ activity and neurosecretion contribute to the H<SUB>2</SUB>S responses, although ASJ is unlikely to serve as the primary H<SUB>2</SUB>S sensor. One potential explanation is that DAF-7 released by ASJ controls the starvation program, which in turn modulates the animal’s response to H<SUB>2</SUB>S. We also discussed this in Lines 449–458.
The paper would be significantly strengthened by testing the possibility (as the authors acknowledge in lines 348-52) that disruption of detoxification mechanisms reduces sustained behavioral responses to H<SUB>2</SUB>S because of physiological damage. Authors use acute exposure to high O2 for this purpose earlier in the paper, but not to probe the consequences of loss of hif-1 and detoxification factors.
We thank the reviewer for the valuable suggestion. As the reviewer highlighted, we attributed the brief locomotory speed responses to H<SUB>2</SUB>S observed in hif-1 mutants to the lack of detoxification response, leading to the rapid intoxication of the animals. Several lines of evidence support this conclusion. First, we observed that hif-1 and the detoxification mutants displayed a stronger initial reorientation response (omega turns) and a more rapid decline in speed and reversals compared to wild type (Figure 5 D–F). Second, to test if hif-1 mutants were indeed more susceptible to H<SUB>2</SUB>S toxicity, we exposed WT and hif-1 animals to H<SUB>2</SUB>S for 30 mins and subsequently tested their ability to respond to near-UV light. Unlike WT animals, the speed response to near-UV light was inhibited in hif-1 mutants (Figure 5I), suggesting that exposure to H<SUB>2</SUB>S for 30 min causes a stronger toxicity in animals deficient of HIF-1 signaling. Third, hif-1 and detoxification mutants displayed a sustained high speed in response to 1% O<SUB>2</SUB> , suggesting the specific impairment of H<SUB>2</SUB>S response. The data were presented in Lines 318–347, and were further discussed this in Lines 485–508.
To better understand whether mitochondrial damage has a role in H<SUB>2</SUB>S-evoked behavior, it might be useful for the authors to determine whether general ROS response pathways are important for H<SUB>2</SUB>S behavioral responses.
We thank the reviewer for this insightful comment. As suggested, we investigated whether ROS detoxification pathways contribute to H<SUB>2</SUB>S-evoked locomotory speed responses by analyzing mutants in the superoxide dismutase (SOD) family. These experiments, together with other observations, suggest that mitochondrial ROS play a dual role in H<SUB>2</SUB>S-evoked locomotion. The relevant results were presented in Lines 401–425, and were further discussed in Lines 509–536.
First, we found that increased mitochondrial ROS formation, either induced pharmacologically by rotenone or genetically in mitochondrial electron transport chain (ETC) mutants (Ishii et al., 2013; Ochi et al., 2016; Ramsay & Singer, 1992; Yang & Hekimi, 2010; Zorov, Juhaszova, & Sollott, 2014), suppressed the behavioral response to toxic H<SUB>2</SUB>S (Figure 7A–E). This indicates that mitochondrial ROS plays a significant role in H<SUB>2</SUB>S-evoked responses. One likely explanation is that high ROS formation may dampen the H<SUB>2</SUB>S-triggered ROS spike, or may impair other H<SUB>2</SUB>S signaling processes required to initiate avoidance. Second, consistent with previous reports (Onukwufor et al., 2022), we observed that shortterm rotenone exposure (<1 hour) significantly increased baseline locomotory speed. Given that toxic H<SUB>2</SUB>S levels promote ROS formation (Jia et al., 2020), our findings suggest that acute mitochondrial ROS production by toxic levels of H<SUB>2</SUB>S exposure may serve as a trigger for the avoidance response.
In contrast, animals with sustained mitochondrial ROS production do not have an increased baseline locomotory speed. This effect was observed after 2 hours of rotenone exposure, in mitochondrial ETC mutants, and in animals lacking all SOD enzymes (Figures 7A–K). A likely explanation for the reduced basal locomotory speed during sustained mitochondrial ROS production is the activation of ROSresponsive signaling pathways including HIF-1, NRF2/SKN-1, and DAF-16/FOXO (Lennicke & Cocheme, 2021; Patten, Germain, Kelly, & Slack, 2010), which may promote adaptation to prolonged oxidative stress (Figure 7H). Notably, unlike hif-1 mutants, SOD-deficient animals remained as responsive as WT to other stimuli after 30 minutes of H<SUB>2</SUB>S exposure (Figure 7—figure supplement 1H), indicating that elevated ROS levels do not compromise overall viability or the ability to detoxify H<SUB>2</SUB>S.
Taken together, these results support a model in which mitochondrial ROS exerts a biphasic effect on H<SUB>2</SUB>S-induced avoidance. It enhances detection and avoidance under acute stress but contributes to locomotory suppression when ROS levels remain elevated chronically.
Reviewer #2 (Recommendations For The Authors):
The way the manuscript is presented could be improved without much effort by rewriting/editing. For the reader, it is hard at present to understand the rationales of the experiments, why they were carried out, and how to place them into a context. This could be improved on three levels:
(1) Documentation
(2) Narration/Explanation
(3) Visualization
(1) Documentation
Not all of the results in the text are well documented. The results should be described with more details in the written text and improved documentation and quantification of the results. Example:
Turning behavior is mentioned as an important aspect of the response to H<SUB>2</SUB>S. There is no citation given but this effect is not well documented. The authors image the animals and could provide video footage of the effect, could quantify eg turning/pirouettes, and provide the data. At the moment the manuscript largely relies on measuring the increase in speed, but the reader is left wondering what other behavioral effects occur and how this is altered in all of the mutant and other conditions tested. Just quantifying speed reduces the readout and seems like an oversimplification to characterize the behavioral response.
We are grateful for this comment. We now provide a video footage of the H<SUB>2</SUB>S effects (Figure 1—Video 1). As suggested, we analyzed the recordings to extract reorientation (omega-turns) and reversals. These analyses are now included in the Supplemental file 1 with representative panels displayed in Figure 5 and supplements to Figures 2, 3, 5, 6 and 7. Even though the mutant effects on omega-turns were often subtle, and reversal responses showed considerable variability (likely due to differences in population density, food availability, or animals’ physiological state prior to the assay), this analysis has proven valuable for distinguishing mutants that exhibit adaptation from those that display hypersensitivity to H<SUB>2</SUB>S toxicity. For instance, although both SOD-deficient and BP-treated animals failed to increase their locomotory speed in H<SUB>2</SUB>S (Figure 6E and Figure 7G), they exhibited distinct omega-turn responses (Figure 6—figure supplement 1I and Figure 7—figure supplement 1F), suggesting that different mechanisms likely underlie the locomotory defects of these two animals. We have integrated the omega-turn and reversal data into the text and discussed under relevant contexts.
(2) Narration/Description.
Generally, the description of the results part is very brief and it is often not clear why a certain experiment was carried out and how. Surely it is possible to check the methods but this interrupts the flow of reading and it would be easier for the reader to be guided through the results with more information what the initial motivation for an experiment is, what the general experimental outline is, and what specific experiments are carried out.
We apologize for the lack of clarity and logical structure in the initial submission. In the revised manuscript, we have thoroughly revised the text to improve its organization and readability.
Examples:
Line 97ff: The authors performed a candidate screen yet it is not described why which genes were chosen. Are there also pathways that were tested that turned out to not be involved?
We thank the reviewer for the suggestion. To address this, we have added a new section, explaining the rationale for selecting genes and pathways in our candidate screen. Briefly, we focused on genes known or predicted to be involved in sensory responses to gaseous stimuli in C. elegans and mammals, including globins and guanylate cyclases (21% O<SUB>2</SUB> sensing), potassium channels (acute hypoxia), and nutrientsensitive pathways (CO<SUB>2</SUB> responses). We also included mutants defective in sensory signal transduction and neurotransmission. In addition, mitochondrial mutants were analyzed because mitochondria play a central role in H<SUB>2</SUB>S detoxification. The pathways that contributed to the acute H<SUB>2</SUB>S response included cGMP, insulin, and TGF-β signaling, as well as mitochondrial components. In contrast, globins, potassium channels, and biogenic amine signaling did not appear to play significant roles under our assay conditions. The results of the candidate screen are described in Lines 106–138 and summarized in Supplementary File 1.
line 262ff: the paragraph starts with explaining ferritin genes that are important for iron control but the reader does not yet know why. Then it is explained that a ferritin gene is DE in the H<SUB>2</SUB>S transcriptomes. then a motivation to look into the labile iron pool is described. Why not first explain what genes are strongly regulated and why they are selected based on their DE? Then explain what is known about these genes and pathways, and then motivate a set of experiments.
We agree with the reviewer that our initial description could have been more logically organized. We reframed this section to first present the RNA-seq data, followed by an explanation of their known biological functions and the motivation for the subsequent experiments (Lines 350–357).
nhr-49 appears suddenly in the results part and it is not clear why it was tested and how the result links. Is nhr-49 a key transcription factor that is activated by H<SUB>2</SUB>S sensory or physiological response, and does it control the signaling or protective changes induced by H<SUB>2</SUB>S?
We thank the reviewer for the comment. As suggested, we revised the text to present the information more clearly. In our candidate gene screen, a set of mutants exhibiting reduced speed responses to H<SUB>2</SUB>S has previously been shown to be defective in response to CO<SUB>2</SUB> stimulation (Hallem & Sternberg, 2008). These included animals deficient in nutrient-sensitive pathways, including insulin, TGF-beta, and NHR49, which were reported by Sternberg’s lab to exhibit dampened responses to CO<SUB>2</SUB> (Hallem & Sternberg, 2008) (Lines 173–179). We also included a simply cartoon to further illustrate this (Figure 3C).
The nuclear hormone receptor NHR-49 has been implicated in a variety of stress responses, including starvation (Van Gilst, Hadjivassiliou, & Yamamoto, 2005), bacterial pathogen (Naim et al., 2021; Wani et al., 2021), and hypoxia (Doering et al., 2022). The nhr-49 mutants exhibited a rapid decline in locomotory speed during H<SUB>2</SUB>S exposure, implicating a role in sustaining high speed in the presence of H<SUB>2</SUB>S. Furthermore, we observed that fmo-2, a well-characterized target gene of NHR-49, was significantly upregulated after 1 hour of exposure to 50 and 150 ppm H<SUB>2</SUB>S (Supplementary file 2), suggesting that NHR-49 signaling is rapidly activated by H<SUB>2</SUB>S exposure. Exactly how NHR-49 contributes to H<SUB>2</SUB>S response requires further investigation.
(3) Visualization
Adding a model/cartoon summary that describes the pathways tested and their interaction would be helpful in some of the figures for the reader to keep an overview of the pathways that were tested. Also, a final summary cartoon that integrates all the puzzle pieces into one larger picture would be helpful. Such a final cartoon overview could also point to the key open questions of the underlying mechanisms.
We thank the reviewer for this suggestion. We have added a series of models/cartoons to illustrate the different pathways and their interactions. These include starvation regulatory mechanisms (Figure 3C), 21% O<SUB>2</SUB> sensing mechanisms (Figure 3G), HIF-1 signaling and detoxification (Figure 5—figure supplement 1E), HIF-1 signaling and the regulation of labile iron (Figure 6H), as well as ROS signaling and regulation (Figure 7L). To help interpretation and to elaborate on these models, we have also included explanatory sentences in the corresponding figure legends.
Other comments:
Introduction and line 93: The authors mention that 50 ppm H<SUB>2</SUB>S has beneficial effects on lifespan yet does not have a detectable phenotype." Are there any concentrations of H<SUB>2</SUB>S that cause attraction of C. elegans and what is the preferred range if it exists? Could this be measured in an H<SUB>2</SUB>S gradient?
We thank the reviewer for the insightful comment. We performed an H<SUB>2</SUB>S gradient assay, which suggests that wild type animals are attracted toward low concentrations of H<SUB>2</SUB>S around 40 ppm (Figure 1G and H) (Lines 95–104). These results suggest that H<SUB>2</SUB>S acts as a strong repellent for C. elegans at high concentrations but as an attractant at low levels. This dual role may be ecologically relevant, as wild C. elegans lives in complex and dynamic environments where H<SUB>2</SUB>S levels likely fluctuate over short distances (Adams, Farwell, Pack, & Bamesberger, 1979; Budde & Roth, 2011; Morra & Dick, 1991; Patange, Breen, Arsuffi, & Ruvkun, 2025; Rodriguez-Kabana, Jordan, & Hollis, 1965; Romanelli-Cedrez, Vairoletti, & Salinas, 2024).
Line 146: "Local H<SUB>2</SUB>S concentrations could also be significantly higher in decomposing substances where wild C. elegans thrives" please provide a citation.
As suggested, we included a set of references that have described the H<SUB>2</SUB>S enrichment in the natural environment in early field studies (Adams et al., 1979; Morra & Dick, 1991; Rodriguez-Kabana et al., 1965), as well as references that have discussed and implied this in C. elegans studies (Budde & Roth, 2011; Patange et al., 2025; Romanelli-Cedrez et al., 2024). They can be found in the introduction (Lines 59–62) and in the result (Lines 197–199).
Line 311 "Wild C. elegans isolates thrive in the decomposing matters, where the local concentrations of O2 are low while the levels of CO2 and H<SUB>2</SUB>S could be high. These animals have adapted their behavior in such an environment, displaying increased sensitivity to high O2 exposure but dampened responses to CO2." Please provide citations for these statements.
As suggested, we cited the relevant articles or books describing the variation of O<SUB>2</SUB> and CO<SUB>2</SUB> levels in the decomposing matters including several C. elegans papers that mentioned this in Lines 197–199 (Bretscher, Busch, & de Bono, 2008; Gea, Barrena, Artola, & Sanchez, 2004; Hallem & Sternberg, 2008; Oshins, Michel, Louis, Richard, & Rynk, 2022), and the above-mentioned articles for H<SUB>2</SUB>S (Adams et al., 1979; Budde & Roth, 2011; Morra & Dick, 1991; Patange et al., 2025; Rodriguez-Kabana et al., 1965; Romanelli-Cedrez et al., 2024).
For C. elegans’ sensitivity to O2 and CO2, these articles were cited in Lines 201–203 (Beets et al., 2020; Bretscher et al., 2008; Carrillo, Guillermin, Rengarajan, Okubo, & Hallem, 2013; Hallem & Sternberg, 2008; Kodama-Namba et al., 2013; McGrath et al., 2009).
Reviewer #3 (Recommendations For The Authors):
More work could be conducted establishing the neuronal circuitry involved in the initial, tractable response to H<SUB>2</SUB>S.
We thank the reviewer for the insightful comment. Since our initial analyses suggest a role of ASJ neurons in H<SUB>2</SUB>S-evoked locomotory responses (Figure 2F and G), We thought that this would offer us an entry point to dissect the neuronal circuit involved in H<SUB>2</SUB>S responses. Expression of the tetanus toxin catalytic domain in ASJ, which blocks neurosecretion, inhibited H<SUB>2</SUB>S evoked locomotory responses (Figure 2H), suggesting that neurosecretion from ASJ promotes the speed response to H<SUB>2</SUB>S (Lines 162– 165). We then performed calcium imaging of ASJ neurons in response to H<SUB>2</SUB>S exposure. However, while we observed CO<SUB>2</SUB> -evoked calcium transients in ASJ using GCaMP6s, we did not detect any calcium response to H<SUB>2</SUB>S, under several conditions, including animals on food, off food, and with different H<SUB>2</SUB>S concentrations and exposure times (Figure2—Figure supplement 2E and 2F) (Lines 166–168). Since signaling from ASJ neurons regulates developmental programs that modify sensory functions in C. elegans, including CO<SUB>2</SUB> and O<SUB>2</SUB> responses (Murakami, Koga, & Ohshima, 2001), the involvement of ASJ neurons is not specific to H<SUB>2</SUB>S responses and ASJ neurons are unlikely to serve as a primary H<SUB>2</SUB>S sensor (Discussed in Line 449–458). Therefore, the exact sensory neuron, circuit and molecular triggers mediating acute H<SUB>2</SUB>S avoidance behavior remain to be elucidated.
Our subsequent investigation on mitochondrial components suggests that a burst of mitochondrial ROS production may be the trigger for H<SUB>2</SUB>S avoidance, as transient exposure to rotenone substantially increases baseline locomotory activity (Figure 7E) (Line 391–396). However, mitochondrial ROS could potentially target multiple neurons and cellular machineries to initiate avoidance behavior to H<SUB>2</SUB>S, making it challenging to pinpoint specific sites of action. Nevertheless, we agree that further dissection of the neural circuits and mitochondrial signaling in H<SUB>2</SUB>S avoidance will be important and should be explored in future studies. We discussed this in Lines 509–536.
I am not sure how to overcome the challenges involved in reaching conclusions from the decreased locomotory responses of animals that are sensitized to the effects of H<SUB>2</SUB>S. Perhaps this conundrum could be discussed in more detail in the text.
We thank the reviewer for this important comment. We agree that decreased locomotory speed during H<SUB>2</SUB>S exposure can arise from distinct causes, either systemic toxicity or adaptation, and distinguishing between these is critical. We have included new experiments and revised the text to clarify this issue.
Our data suggest that increased initial omega turns and a rapid loss of locomotion in hif-1 and detoxification-defective mutants including sqrd-1 and ethe-1 likely reflect an enhanced sensitivity to H<SUB>2</SUB>S toxicity due to their failure to induce appropriate adaptative responses (Figure 5D–F, Figure 5J–L, Figure 5—Figure supplement 1F–P). Supporting this, hif-1 mutants become less responsive to unrelated stimuli (near-UV light) after 30 minutes of H<SUB>2</SUB>S exposure (Figure 5I).
In contrast, egl-9 and SOD-deficient animals show reduced initial reorientation and reduced speed responses (Figure 5B, Figure 7G, Figure 5—Figure supplement 1A and B, and Figure 7—Figure supplement 1F and G), although both egl-9 and sod mutants respond normally to the other stimuli prior or after H<SUB>2</SUB>S exposure (Figure 5I, Figure 5—Figure supplement 1C, and Figure 7—Figure supplement 1H). Since disrupting egl-9 stabilizes HIF-1 and upregulates the expression of numerous genes involved in cellular defense against H<SUB>2</SUB>S toxicity, the enhanced detoxification capacity in egl-9 mutants likely increases animals’ tolerance to H<SUB>2</SUB>S, thereby reducing avoidance to otherwise toxic H<SUB>2</SUB>S levels. Similarly, persistently high ROS in SOD deficient animals activates a variety of stress-responsive signaling pathways, including HIF-1, NRF2/SKN-1 and DAF-16/ FOXO signaling (Lennicke & Cocheme, 2021; Patten et al., 2010), facilitating cellular adaptation to redox stress and reducing animals’ responsiveness to toxic H<SUB>2</SUB>S levels. Taken together, these findings support the view that reduced locomotory speed during H<SUB>2</SUB>S exposure can arise from distinct mechanisms: early systemic toxicity in hif-1 and detoxificationdefective mutants, versus enhanced cellular adaptation in egl-9 and SOD mutants. We have integrated the relevant information across the result section and discussed this in Lines 485–536.
References
Adams, D. F., Farwell, S. O., Pack, M. R., & Bamesberger, W. L. (1979). Preliminary Measurements of Biogenic Sulfur-Containing Gas Emissions from Soils. Journal of the Air Pollution Control Association, 29(4), 380-383. doi:Doi 10.1080/00022470.1979.10470805
Beets, I., Zhang, G., Fenk, L. A., Chen, C., Nelson, G. M., Felix, M. A., & de Bono, M. (2020). NaturaL Variation in a Dendritic Scaffold Protein Remodels Experience-Dependent Plasticity by Altering Neuropeptide Expression. Neuron, 105(1), 106-121 e110. doi:10.1016/j.neuron.2019.10.001
Bretscher, A. J., Busch, K. E., & de Bono, M. (2008). A carbon dioxide avoidance behavior is integrated with responses to ambient oxygen and food in Caenorhabditis elegans. Proc Natl Acad Sci U S A, 105(23), 8044-8049. doi:10.1073/pnas.0707607105
Budde, M. W., & Roth, M. B. (2011). The response of Caenorhabditis elegans to hydrogen sulfide and hydrogen cyanide. Genetics, 189(2), 521-532. doi:10.1534/genetics.111.129841
Carrillo, M. A., Guillermin, M. L., Rengarajan, S., Okubo, R. P., & Hallem, E. A. (2013). O-2-Sensing Neurons Control CO2 Response in C. elegans. Journal of Neuroscience, 33(23), 9675-9683. doi:10.1523/Jneurosci.4541-12.2013
Doering, K. R. S., Cheng, X., Milburn, L., Ratnappan, R., Ghazi, A., Miller, D. L., & Taubert, S. (2022). Nuclear hormone receptor NHR-49 acts in parallel with HIF-1 to promote hypoxia adaptation in Caenorhabditis elegans. Elife, 11. doi:10.7554/eLife.67911
Gea, T., Barrena, R., Artola, A., & Sanchez, A. (2004). Monitoring the biological activity of the composting process: Oxygen uptake rate (OUR), respirometric index (RI), and respiratory quotient (RQ). Biotechnol Bioeng, 88(4), 520-527. doi:10.1002/bit.20281
Hallem, E. A., & Sternberg, P. W. (2008). Acute carbon dioxide avoidance in Caenorhabditis elegans. Proc Natl Acad Sci U S A, 105(23), 8038-8043. doi:10.1073/pnas.0707469105
Ishii, T., Miyazawa, M., Onouchi, H., Yasuda, K., Hartman, P. S., & Ishii, N. (2013). Model animals for the study of oxidative stress from complex II. Biochim Biophys Acta, 1827(5), 588-597. doi:10.1016/j.bbabio.2012.10.016
Jia, J., Wang, Z., Zhang, M., Huang, C., Song, Y., Xu, F., . . . Cheng, J. (2020). SQR mediates therapeutic effects of H(2)S by targeting mitochondrial electron transport to induce mitochondrial uncoupling. Sci Adv, 6(35), eaaz5752. doi:10.1126/sciadv.aaz5752
Kodama-Namba, E., Fenk, L. A., Bretscher, A. J., Gross, E., Busch, K. E., & de Bono, M. (2013). Crossmodulation of homeostatic responses to temperature, oxygen and carbon dioxide in C. elegans. PLoS Genet, 9(12), e1004011. doi:10.1371/journal.pgen.1004011
Lennicke, C., & Cocheme, H. M. (2021). Redox metabolism: ROS as specific molecular regulators of cell signaling and function. Mol Cell, 81(18), 3691-3707. doi:10.1016/j.molcel.2021.08.018
McGrath, P. T., Rockman, M. V., Zimmer, M., Jang, H., Macosko, E. Z., Kruglyak, L., & Bargmann, C. I. (2009). Quantitative mapping of a digenic behavioral trait implicates globin variation in C. elegans sensory behaviors. Neuron, 61(5), 692-699. doi:10.1016/j.neuron.2009.02.012
Morra, M. J., & Dick, W. A. (1991). Mechanisms of h(2)s production from cysteine and cystine by microorganisms isolated from soil by selective enrichment. Appl Environ Microbiol, 57(5), 14131417. doi:10.1128/aem.57.5.1413-1417.1991
Murakami, M., Koga, M., & Ohshima, Y. (2001). DAF-7/TGF-beta expression required for the normal larval development in C-elegans is controlled by a presumed guanylyl cyclase DAF-11. Mechanisms of Development, 109(1), 27-35. doi:Doi 10.1016/S0925-4773(01)00507-X
Naim, N., Amrit, F. R. G., Ratnappan, R., DelBuono, N., Loose, J. A., & Ghazi, A. (2021). Cell nonautonomous roles of NHR-49 in promoting longevity and innate immunity. Aging Cell, 20(7). doi:ARTN e13413 10.1111/acel.13413
Ochi, R., Dhagia, V., Lakhkar, A., Patel, D., Wolin, M. S., & Gupte, S. A. (2016). Rotenone-stimulated superoxide release from mitochondrial complex I acutely augments L-type Ca2+ current in A7r5 aortic smooth muscle cells. Am J Physiol Heart Circ Physiol, 310(9), H1118-1128. doi:10.1152/ajpheart.00889.2015
Onukwufor, J. O., Farooqi, M. A., Vodickova, A., Koren, S. A., Baldzizhar, A., Berry, B. J., . . . Wojtovich, A. P. (2022). A reversible mitochondrial complex I thiol switch mediates hypoxic avoidance behavior in C. elegans. Nat Commun, 13(1), 2403. doi:10.1038/s41467-022-30169-y
Oshins, C., Michel, F., Louis, P., Richard, T. L., & Rynk, R. (2022). Chapter 3 - The composting process. In R. Rynk (Ed.), The Composting Handbook (pp. 51-101): Academic Press.
Patange, O., Breen, P., Arsuffi, G., & Ruvkun, G. (2025). Hydrogen sulfide mediates the interaction between C. elegans and Actinobacteria from its natural microbial environment. Cell Reports, 44(1), 115170. doi:10.1016/j.celrep.2024.115170
Patten, D. A., Germain, M., Kelly, M. A., & Slack, R. S. (2010). Reactive oxygen species: stuck in the middle of neurodegeneration. J Alzheimers Dis, 20 Suppl 2, S357-367. doi:10.3233/JAD-2010100498
Ramsay, R. R., & Singer, T. P. (1992). Relation of superoxide generation and lipid peroxidation to the inhibition of NADH-Q oxidoreductase by rotenone, piericidin A, and MPP+. Biochem Biophys Res Commun, 189(1), 47-52. doi:10.1016/0006-291x(92)91523-s
Rodriguez-Kabana, R., Jordan, J. W., & Hollis, J. P. (1965). Nematodes: Biological Control in Rice Fields: Role of Hydrogen Sulfide. Science, 148(3669), 524-526. doi:10.1126/science.148.3669.524
Romanelli-Cedrez, L., Vairoletti, F., & Salinas, G. (2024). Rhodoquinone-dependent electron transport chain is essential for Caenorhabditis elegans survival in hydrogen sulfide environments. J Biol Chem, 300(9), 107708. doi:10.1016/j.jbc.2024.107708
Van Gilst, M. R., Hadjivassiliou, H., & Yamamoto, K. R. (2005). A Caenorhabditis elegans nutrient response system partially dependent on nuclear receptor NHR-49. Proc Natl Acad Sci U S A, 102(38), 13496-13501. doi:10.1073/pnas.0506234102
Wani, K. A., Goswamy, D., Taubert, S., Ratnappan, R., Ghazi, A., & Irazoqui, J. E. (2021). NHR- 49/PPAR-α and HLH-30/TFEB cooperate for host defense via a flavin-containing monooxygenase. Elife, 10. doi:ARTN e62775 10.7554/eLife.62775
Yang, W., & Hekimi, S. (2010). A mitochondrial superoxide signal triggers increased longevity in Caenorhabditis elegans. PLoS Biol, 8(12), e1000556. doi:10.1371/journal.pbio.1000556
Zorov, D. B., Juhaszova, M., & Sollott, S. J. (2014). Mitochondrial reactive oxygen species (ROS) and ROS-induced ROS release. Physiol Rev, 94(3), 909-950. doi:10.1152/physrev.00026.2013
Reviewer #2 (Public review):
Summary:
The authors aimed to investigate whether advanced microstructural diffusion MRI modeling using the SANDI framework could reveal clinically relevant tissue alterations in the subcortical structures of individuals with Huntington's disease (HD). Specifically, they sought to determine if SANDI-derived parameters-such as soma density, soma size, and extracellular diffusivity-could detect abnormalities in both manifest and premanifest HD stages, complement standard MRI biomarkers (e.g., volume, MD), and correlate with disease burden and motor impairment. Through this, they hoped to demonstrate the feasibility and added biological specificity of SANDI for early detection and characterization of HD pathology.
Strengths:
(1) Novelty and relevance:
This is, to the best of my knowledge, the first clinical deployment of SANDI in HD, offering more biophysically interpretable and specific imaging biomarkers than standard DTI or volumetric features.
(2) More specific microstructural insight: Traditional approaches have used volumetric features (e.g., striatal volume loss) or DTI metrics (like FA and MD), which are indirect and non-specific markers. They can indicate something is "wrong" but not what is wrong.
(3) SANDI parameters permit establishing clearer links with microstructure:
o Apparent soma density (fis): proxy for neuronal/glial cell body density.
o Apparent soma size (rs): reflects possible gliagl hypertrophy or neuronal shrinkage.
o Neurite density (fin): linked to dendritic/axonal integrity.
o Extracellular fraction and diffusivity: sensitive to edema, gliosis, and tissue loss.
In this way, a decrease in soma density can be related to neural loss (e.g., medium spiny neurons), and an increase in soma size and extracellular fraction could be related to glial reactivity (astrocytes, microglia). This enables differentiating between atrophy due to neuron loss vs reactive gliosis, which volumetrics or DTI cannot do.
(4) Integration of modalities: The inclusion of motor impairment (Q-Motor), HD-ISS staging, and multi-compartment diffusion modeling is a methodological strength.
(5) Early detection potential: SANDI metrics showed abnormalities in premanifest HD, sometimes even when volume loss was mild or absent. This suggests the potential for earlier, more sensitive biomarkers of disease progression.
(6) Predictive power: Regression models showed that SANDI metrics explained up to 63% of the variance in striatal volumes in HD. And this correlated strongly with motor impairment and disease burden (CAP100). This shows they are not just redundant with volume or DTI, but they are complementary and potentially more mechanistically meaningful.
Weaknesses:
Certain aspects of the study would benefit from clarification:
(1) Scanner and acquisition consistency: While HD data are from the WAND study, it is not clear whether controls were scanned on the same scanner or protocol. Given the use of model-derived metrics (especially SANDI), differences in scanner or acquisition could introduce confounds. Also, although it offers novel and biologically informative markers, widespread clinical translation still faces hurdles. For instance, the study used a 3T Connectom scanner (300mT/m gradients), which is not widely available. Reproduction of these results in standard 3T clinical scanners would be a great addition, in scenarios with lower resolution, less precise parameter recovery, and longer scans if SNR needs to be maintained.
(2) HD-ISS staging and group comparisons:<br /> a) Only 26-27 out of 56 gene-positive participants could be assigned HD-ISS stages, and none were classified into stages 0 or 4.
b) Visual overlap between stages 1 and 2 in behavioral and imaging features suggests that staging-based group separation may not be robust.
c) The above may lead to claims based on progression across HD-ISS stages to be overinterpreted or underpowered
(3) Regression modeling choices:<br /> a) SANDI metrics included in the models differ between HC and HD groups, reducing comparability.
b) The potential impact of multicollinearity (e.g., between fis and rs) is not discussed.
c) Beta coefficients could reflect model instability or parameter degeneracy rather than true biological effects.
These issues do not undermine the study's main conclusions, which effectively demonstrate the feasibility and initial clinical relevance of applying SANDI to HD. Nonetheless, addressing them more thoroughly would enhance the clarity and interpretability of the manuscript.
RRID:BDSC_9782
DOI: 10.1186/s12860-019-0198-z
Resource: RRID:BDSC_9782
Curator: @scibot
SciCrunch record: RRID:BDSC_9782
¿es la extensión el atributo más importante en la valoración de la calidad de un título?
Un título bien formulado actúa como una síntesis estratégica del planteamiento, orientando la lectura y evitando interpretaciones erróneas sobre el alcance o propósito del trabajo Esta coherencia asegura que el lector comprenda desde el inicio qué se investiga, por qué y en qué contexto.
los títulos de los artículos científicos, proyectos de investigación o informes finales de tesis.
No sabía que un título de algún documento científico, podría llegar a ser tan extenso teniendo entre 15 a 20 palabras aunque hay que tomar en cuenta que son necesarias a veces un poco más, por ejemplo: “Evolución de indicadores de salud seleccionados en la provincia de Cienfuegos durante el decenio 1991-2000”<br /> indica tiempo y espacio, mientras que: “Evolución de indicadores de salud seleccionados” tiene información incompleta que se podría prestar a otros tiempos y otro espacio o podría ser inútil ya que no te serviría como referencia.
Adicionalmente, y también derivado del problema y el objetivo, el título contextualiza en tiempo y espacio ese objeto de estudio
El titulo es de suma importancia definirlo de una manera clara ya que este nos ayuda a entender el porque o el objetivo de la investigación, ya que todos están conectados para que los lectores comprenda la idea de la investigación
Aportación 1: Considero que es un tema del que se habla poco y por lo tanto se me hace relevante ya que habla de la calidad del titulo. Generalmente se da mayor énfasis a la extensión, cuando en realidad lo central es la precisión, pues esta permite reflejar de forma fiel el objeto de estudio y orientar al lector sobre la pertinencia del trabajo.
Aportación 2: El texto se apoya en ejemplos concretos como el de la investigación sobre leucemias agudas o el estudio del impacto de la COVID-19 para demostrar cómo la reducción de palabras en un título puede sacrificar información clave. Estos fortalecen el argumento y evidencian que la correspondencia entre problema, objetivo y título debe prevalecer sobre una regla rígida de extensión.
Aportación 3:La conclusión del artículo es práctica y aplicable: un título no debe juzgarse por ser corto o largo, sino por su capacidad de expresar con exactitud el objeto de estudio. Este enfoque resulta útil para investigadores, tesistas y editores, ya que orienta hacia una valoración más justa y científica de los títulos.
1-Se enfatiza que un buen título no puede formularse correctamente si no se ha definido antes el problema y el objetivo de la investigación. El título debe surgir como una consecuencia lógica de estos elementos, asegurando que exista una correspondencia clara entre ellos. 2- El texto sostiene que, al evaluar la calidad de un título en una investigación, la prioridad debe ser la precisión, es decir, qué tan bien refleja el contenido del estudio, y no su brevedad o número de palabras. Un título largo puede ser perfectamente válido si expresa claramente el objeto del estudio. 3-Aunque es deseable que los títulos no sean innecesariamente extensos, no se debe recortar información.
No es imprescindible que un título contenga referencia directa y explícita al contexto de tiempo y espacio en que se desarrolla la investigación. La referencia “al cuándo” y “al dónde” se justifica solo cuando contribuye a la necesaria precisión del objeto de estudio, por lo que no debe ser una exigencia rutinaria.
Esto es muy rescatable y de mucha ayuda, ya que podemos ver que existe una flexibilidad en la construcción de los títulos y que esta abre la posibilidad de enfocarnos en lo importante, y que no forzar la inclusión del tiempo o espacio nos da la oportunidad que el título sea más universal y que el lector se centre en el problema investigado, sin ponerle un límite con un contexto de título no necesario. APORTACIÓN 2
Aunque los que se inician en el campo de la investigación suelen intentar comenzar sus proyectos precisamente con el título, en realidad la elaboración de este transita por una adecuada formulación del problema de investigación (con la complejidad que ello implica) y, consecutivamente, por la correcta enunciación del o los objetivos de investigación.
Esto es interesante ya que menciona la idea de que primero hay que entender bien el problema y los objetivos de una investigación, y ya después crear el título. Esto nos ayuda a que el trabajo que realizamos sea más coherente, organizado y sólido. APORTACIÓN 1
La incorporación esquemática y no justificada del contexto de espacio y tiempo del estudio, puede contribuir a la excesiva extensión del título de la investigación o de un artículo científico.
Consejo valido, sintetizar el titulo y explicar más en el resumen. Me gustaría que indagara más que clase de cosas puede contribuir el reasumen para también evitarlo en el titulo.
El ejemplo de la investigación sobre leucemias ilustra bien cómo un título puede derivarse con precisión del problema y del objetivo. Esto resalta que la coherencia entre estos tres elementos es clave para evitar títulos vagos o poco informativos.
1.- El título no debe ser un punto de partida improvisado, sino el resultado de un proceso reflexivo.
2.- Un título extenso no es un defecto en sí, si aporta claridad y especificidad. Sin embargo, la extensión debe ser funcional, evitando redundancias o detalles irrelevantes.
3.- Diferenciación entre objeto de estudio y variables. Esto ayuda a evitar títulos que mencionan solo el fenómeno general sin especificar qué se está midiendo o analizando.
USKORENIE, GLASNOST' AND PERESTROIKA: THE PATTERN O
Group 1 has no summary posted
Group 2: Gorbachev’s Non-Violent Revolution. Key points: Perestroika was about empowering the people and breaking away from totalitarian power. Gorbachev had a very humanistic approach that was against violence His attitude towards women was of true respect There was a lack of a clear plan from Gorbachev in the beginning
Conclusions: How and why was he able to gain power under communism? Gorbachev’s goal was to reform the system in which the people had continuously been oppressed by, and his reform was called Perestroika. His upbringing was full of hardships and famine, making him more understanding towards the people in his policies and shaping the way he took on his non-violent revolution and New Thinking.
Group 4 has no summary posted
Group 5: Linking Gorbachev’s Domestic and Foreign Policies Key points: The Soviet Union has a neutral position regarding the conflict between ex-Union countries. Gorbachev’s policies are seen as “ambitious” by the author of the article There was an anti-alcohol campaign that failed Gorbachev’s international goals are focused on improving relations with countries like the US, UK and Israel, which was unusual in Russian policies.
Conclusions: Gorbachev’s domestic reforms are linked to his foreign policy actions during this crucial time in Soviet history. The ongoing tension between Armenia and Azerbaijan, both of which used to be part of the Union, bring the Soviet Union at a neutral state, and shifting its attention towards foreign relations, driven by the realization that China was no longer a dependable ally, so there was a need to form new allays and have stability.
But if you go into a classroom, you may notice a teacher calling on a shy student and not calling on another student who usually dominates the discussion. Is this equal
Although I had always experienced this situation, I had never thought about this question and if we look closely, in reality my teacher wasn't equal to all of us because when she called us in-front of the class to answer the question we no longer had the option if whether we wanted to o not.
A pesar de ser zambo y de llamarse López, quería parecerse cada vez menos a un zaguero de Alianza Lima y cada vez más a un rubio de Filadelfia. La vida se encargó de enseñarle que si quería triunfar en una ciudad colonial más valía saltar las etapas intermediarias y ser antes que un blanquito de acá un gringo de allá. Toda su tarea en los años que lo conocí consistió en deslopizarse y deszambarse lo más pronto posible y en americanizarse antes de que le cayera el huaico y lo convirtiera para siempre, digamos, en un portero de banco o en un chofer de colectivo. Tuvo que empezar por matar al peruano que había en él y por coger algo de cada gringo que conoció. Con el botín se compuso una nueva persona, un ser hecho de retazos, que no era ni zambo ni gringo, el resultado de un cruce contranatura, algo que su vehemencia hizo derivar, para su desgracia, de sueño rosado a pesadilla infernal.
En esta sección...
o build a te reo speech recognition model, it needed anabundance of transcribed audio. To transcribe the audio, it needed the advanced speakers whose smallnumbers it was trying to compensate for in the first place. There were, however, plenty of beginningand intermediate speakers who could read te reo words aloud better than they could recognize them ina recording.
Truly a Catch-22.
El volumen sanguíneo total constituye de 7% a 8% del peso corporal de un adulto, lo que indica que un individuo de 70 kilos tiene aproximadamente 5.5 L de sangre. Este volumen está compuesto tanto por el líquido extracelular ―que es el plasma― como por los elementos formes de la sangre ―que también contienen líquido en su interior―, sobre todo los eritrocitos, que contienen hemoglobina disuelta en agua. Teniendo en cuenta que el hematócrito representa 45% del volumen sanguíneo total, un adulto tiene aproximadamente 2000 a 2400 mL de eritrocitos, que representan de 3.2% a 3.5% del peso corporal y de 3000 a 3500 mL de plasma, que representa 5% del peso total. En los varones, el volumen de sangre es de 61 a 66 mL/kg, de los cuales, 24 a 30 mL/kg son eritrocitos, y 33 a 35 mL/kg son plasma. En las mujeres, estos valores son de 1 a 2 mL menos. En relación con la superficie corporal, el volumen sanguíneo de un adulto es de 2.89 L/m2 en el hombre y 2.44 L/m2 en la mujer. En los jóvenes y en los niños, el volumen es relativamente mayor. Al nacimiento, el volumen sanguíneo es de casi 300 mL, y se duplica durante el primer año de vida. En los lactantes, el volumen sanguíneo es hasta de 83 mL/kg. Más de la mitad del volumen total de sangre, esto es, 64%, se encuentra en las venas; 13% en las arterias, 9% en la circulación pulmonar, 7% en los capilares y arteriolas, y el otro 7% en las cavidades cardiacas. En condiciones normales, la frecuencia cardiaca es de 75 latidos por minuto, con una entrega de sangre a la aorta de 70 mL por latido, que da un total de aproximadamente 5.25 litros/minuto. La sangre se distribuye de la siguiente manera en los principales órganos: cerebro: 14%; circulación coronaria: 4%; riñones: 22%; hígado: 27%; músculos en reposo: 15%; huesos: 5%; piel: 6%; bronquios: 2%. ++ Durante el ejercicio, la frecuencia cardiaca se puede incrementar a 150 latidos por minuto, con lo que el gasto cardiaco aumenta a 20 litros por minuto y el volumen sanguíneo sufre variaciones. El volumen sanguíneo disminuye debido a la pérdida de líquidos y al paso de éstos hacia el espacio intersticial, con lo que el hematócrito sufre un incremento relativo. Vuelve a las condiciones normales durante los 30 minutos siguientes. Otros factores que influyen en el volumen son la temperatura corporal, la altitud sobre el nivel del mar, la cantidad de líquidos ingeridos y el grado de ejercicio o de reposo. Durante el embarazo existe un estado de hemodilución relativa que incrementa el volumen sanguíneo hasta en más de un litro.
encontre esta informacion, favor de checar si es de utilidad
petição
Súmula nº 416/TST - MANDADO DE SEGURANÇA. EXECUÇÃO. LEI Nº 8.432/1992. ART. 897, § 1º, DA CLT. CABIMENTO - Devendo o agravo de petição delimitar justificadamente a matéria e os valores objeto de discordância, não fere direito líquido e certo o prosseguimento da execução quanto aos tópicos e valores não especificados no agravo.
Obs.: Considerando a obrigação legal do agravante em delimitar as matérias e valores impugnados, sob pena de imediata execução da parte não impugnada, não há que se falar em violação de direito líquido e certo se o agravante não especifica adequadamente suas impugnações.
pagará em dobro
A não fruição das férias no período correto gera o dever de pagar o dobro da remuneração.
Obs.: Estava vigente a Súmula nº 450/TST, que previa o pagamento em dobro no caso de inobservância do prazo para pagamento das verbas relativas às férias, para além da hipótese legal do pagamento dobrado para fruição de férias fora do período adequado. Porém, foi julgada inconstitucional pela ADPF 501.
Ementa: ARGUIÇÃO DE DESCUMPRIMENTO DE PRECEITO FUNDAMENTAL. CONSTITUCIONAL E TRABALHISTA. SÚMULA 450 DO TRIBUNAL SUPERIOR DO TRABALHO. PAGAMENTO DA REMUNERAÇÃO DE FÉRIAS EM DOBRO QUANDO ULTRAPASSADO O PRAZO DO ART. 145 DA CLT. IMPOSSIBILIDADE DE O PODER JUDICIÁRIO ATUAR COMO LEGISLADOR POSITIVO. AUSÊNCIA DE LACUNA. INTERPRETAÇÃO RESTRITIVA DE NORMA SANCIONADORA. OFENSA À SEPARAÇÃO DE PODERES E AO PRINCÍPIO DA LEGALIDADE. PROCEDÊNCIA. - 1. Os poderes de Estado devem atuar de maneira harmônica, privilegiando a cooperação e a lealdade institucional e afastando as práticas de guerrilhas institucionais, que acabam minando a coesão governamental e a confiança popular na condução dos negócios públicos pelos agentes públicos. Precedentes. - 2. Impossibilidade de atuação do Poder Judiciário como legislador positivo, de modo a ampliar o âmbito de incidência de sanção prevista no art. 137 da CLT para alcançar situação diversa, já sancionada por outra norma. - 3. Ausência de lacuna justificadora da construção jurisprudencial analógica. Necessidade de interpretação restritiva de normas sancionadoras. Proibição da criação de obrigações não previstas em lei por súmulas e outros enunciados jurisprudenciais editados pelo Tribunal Superior do Trabalho e pelos Tribunais Regionais do Trabalho (CLT, art. 8º, § 2º). - 4. Arguição julgada procedente.
(ADPF 501, Relator(a): ALEXANDRE DE MORAES, Tribunal Pleno, julgado em 08-08-2022, PROCESSO ELETRÔNICO DJe-163 DIVULG 17-08-2022 PUBLIC 18-08-2022)
30 (trinta) dias
A empresa devera participar, avisar, comunicar, o empregado a respeito da concessão de férias com antecedência <u>mínima</u> de 30 dias.
Deverá haver recibo desse aviso de concessão de férias.
responderá solidariamente
O sindicato é solidariamente responsável com o empregado que não tenha recebido o benefício de Justiça gratuita para o pagamento das custas processuais, desde que o Sindicato tenha intervindo
depósito recursal
Súmula nº128/TST - DEPÓSITO RECURSAL (incorporadas as Orientações Jurisprudenciais nºs 139, 189 e 190 da SBDI- - I - É ônus da parte recorrente efetuar o depósito legal, integralmente, <u>em relação a cada novo recurso interposto</u>, sob pena de deserção. Atingido o valor da condenação, nenhum depósito mais é exigido para qualquer recurso. (ex-Súmula nº 128 - alterada pela Res. 121/2003, DJ 21.11.03, que incorporou a OJ nº 139 da SBDI-I - inserida em 27.11.1998)
Obs.: Essa súmula define que, na hipótese de condenação solidária, o depósito recursal efetuado por apenas uma devedora aproveita às demais acaso a empresa que efetuou o depósito não demande sua exclusão da lide. Isso porque o depósito recursal tem natureza de garantia do juízo, razão pela qual a empresa que demanda sua exclusão da lide, acaso se reconheça a sua ilegitimidade, o valor garantido seria perdido acaso as demais devedoras não tivesse realizado o depósito recursal.
No mais, a súmula determina o depósito recursal para cada novo recurso, sob pena de deserção, até o limite do valor da condenação.
Afirma-se que, na fase executória, é descabida a exigência de depósito recursal se o juízo já tiver sido garantido, não obstando, lado outro, a complementação da garantia na hipótese de alteração do valor da condenação.
Note que a súmula, sobre o depósito recursal em condenação de duas ou mais empresas, fala em aproveitamento de depósito recursal na hipótese de condenação solidária. Esse mesmo entendimento deve ser estendido à hipótese de condenação subsidiária, desde que a depositante não pretenda à sua exclusão da lide. Vide:
Tema 146/TST: - O depósito recursal efetuado pelo devedor principal, desde que não tenha requerido sua exclusão da lide, aproveita ao responsável <u>subsidiário</u>.
Alguns outros temas importantes aprovados pelo TST em relação ao depósito recursal:
Tema 158: - O comprovante de agendamento bancário não é suficiente para demonstrar o recolhimento das custas processuais e do depósito recursal e não cabe a concessão de prazo para regularização. Obs.: É considerado defeito insanável, insuscetível de concessão de prazo para regularização, a juntada de comprovante de agendamento de depósito para fins de pagamento de custas e depósito recursal, sendo insuficiente para se provar o efeito pagamento.
Via de regra, a jurisprudência consolidada do TST somente admite a concessão de prazo de regularização acaso haja algum valor pago, concedendo-se prazo para a complementação. No entanto, é tido por insanável a ausência de qualquer pagamento de custa ou depósito recursal, razão pela qual, nessas hipóteses, o recurso será tido por deserto. (Tema 271)
Tema 173: - A substituição do depósito recursal por seguro-garantia, nos termos do art. 899, § 11, da CLT, sem a inclusão do acréscimo de 30% exigido pelo art. 3º, II, do Ato Conjunto TST/CSJT/CGJT nº 1/2019, impõe a intimação do recorrente para complementação da garantia, sob pena de deserção, conforme dispõe o art. 1.007, § 2º, do CPC/2015.
Obs.: No processo do trabalho, é admissível a substituição do depósito recursal por seguro-garantia, desde que o valor do seguro abranja o principal + 30%, na mesma sistemática do CPC. O seguro que assegure apenas o valor nominal será considerado insuficiente, razão pela qual deve haver prazo para complementação, sob pena de deserção.
Tema 271: - É incabível a concessão de prazo para regularização do preparo nos casos de <u>total ausência</u> de comprovação do recolhimento das custas ou do depósito recursal no prazo do recurso, não se aplicando o disposto no art. 1.007, §§ 2º, 4º e 7º, do CPC.
embargos de declaração
RESOLUÇÃO Nº 203, DE 15 DE MARÇO DE 2016. - Edita a Instrução Normativa n° 39, que dispõe sobre as normas do Código de Processo Civil de 2015 aplicáveis e inaplicáveis ao Processo do Trabalho, de forma não exaustiva
Art. 9º O cabimento dos embargos de declaração no Processo do Trabalho, para impugnar qualquer decisão judicial, rege-se pelo art. 897-A da CLT e, supletivamente, pelo Código de Processo Civil (arts. 1022 a 1025; §§ 2º, 3º e 4º do art. 1026), excetuada a garantia de prazo em dobro para litisconsortes (§ 1º do art. 1023). - Parágrafo único. A omissão para fins do prequestionamento ficto a que alude o art. 1025 do CPC dá-se no caso de o Tribunal Regional do Trabalho, mesmo instado mediante embargos de declaração, recusar-se a emitir tese sobre questão jurídica pertinente, na forma da Súmula nº 297, item III, do Tribunal Superior do Trabalho.
Súmula nº 297/TST PREQUESTIONAMENTO. OPORTUNIDADE. CONFIGURAÇÃO
I. Diz-se prequestionada a matéria ou questão quando na decisão impugnada haja sido adotada, explicitamente, tese a respeito.
II. Incumbe à parte interessada, desde que a matéria haja sido invocada no recurso principal, opor embargos declaratórios objetivando o pronunciamento sobre o tema, sob pena de preclusão.
III. Considera-se prequestionada a questão jurídica invocada no recurso principal sobre a qual se omite o Tribunal de pronunciar tese, não obstante opostos embargos de declaração.
Súmula nº184/TST EMBARGOS DECLARATÓRIOS. OMISSÃO EM RECURSO DE REVISTA. PRECLUSÃO - Ocorre preclusão se não forem opostos embargos declaratórios para suprir omissão apontada em recurso de revista ou de embargos.
Súmula nº 278/TST EMBARGOS DE DECLARAÇÃO. OMISSÃO NO JULGADO - A natureza da omissão suprida pelo julgamento de embargos declaratórios pode ocasionar efeito modificativo no julgado.
Súmula nº421/TST EMBARGOS DE DECLARAÇÃO. CABIMENTO. DECISÃO MONOCRÁTICA DO RELATOR CALCADA NO ART. 932 do CPC DE 2015. ART. 557 DO CPC de 1973. (atualizada em decorrência do CPC de 2015) - I – Cabem embargos de declaração da decisão monocrática do relator prevista no art. 932 do CPC de 2015 (art. 557 do CPC de 1973), se a parte pretende tão somente juízo integrativo retificador da decisão e, não, modificação do julgado. - II – Se a parte postular a revisão no mérito da decisão monocrática, cumpre ao relator converter os embargos de declaração em agravo, em face dos princípios da fungibilidade e celeridade processual, submetendo-o ao pronunciamento do Colegiado, após a intimação do recorrente para, no prazo de 5 (cinco) dias, complementar as razões recursais, de modo a ajustá-las às exigências do art. 1.021, § 1º, do CPC
OJ-SDI1-142 EMBARGOS DE DECLARAÇÃO. EFEITO MODI-FICATIVO. VISTA PRÉVIA À PARTE CONTRÁRIA (cancelado o item II em decorrência do CPC de 2015) - - É passível de nulidade decisão que acolhe embargos de declaração com efeito modificativo sem que seja concedida oportunidade de manifestação prévia à parte contrária.
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Su proyecto personalizado y su plano en 3D ofrecidos por su diseñador.Disfrute de un acompañamiento a medida para crear la cocina de sus sueños. Un asesor dedicado le explicará cada etapa para realizar su cocina, le ayudará a elegir los muebles, la configuración ideal para su espacio, los accesorios y los electrodomésticos según sus usos y necesidades… todo ello adaptado a SU presupuesto.El asesor le ayudará a tener una visión más clara, especialmente gracias a un diseño en 3D de su cocina que le permitirá proyectarse mejor. Podrá tomarse el tiempo que necesite: su asesor se adapta a USTED. Además, también podrá acompañarle en la búsqueda de soluciones de financiación a corto, medio o largo plazo.
Un diseñador realiza tu proyecto a medida y el plano en 3D Disfruta de un asesoramiento personal para crear la cocina de tus sueños. Un asesor te acompañará en casa paso, te ayudará a elegir los muebles, la configuración ideal, los accesorios y los electrodomésticos según tus usos y necesidades… todo ello adaptado a TU presupuesto. El asesor te ayudará a tener una visión más clara, especialmente gracias a un diseño en 3D de tu cocina que te permitirá proyectarla mejor. Podrás tomarte el tiempo que necesites: tu asesor se adapta a TI. Además, también podrá ayudarte a encontrar opciones de financiación a corto, medio o largo plazo.
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Reviewer #1 (Evidence, reproducibility and clarity):
This manuscript described the translational responses to single and combined BCAA shortages in mouse cell lines. Using Ribo-seq and RNA-seq analysis, the authors found selective ribosome pausing at codons that encode the depleted amino acids, where the pausing at valine codons was prominent at both a single and triple starvations whereas isoleucine codons showed pausing only under a single depletion. They analyzed the mechanisms of the unexpected selective pausing and proposed that the positional codon usage bias could shape the ribosome stalling and tRNA charging patterns across different amino acids. They also examined the stress responses and the changes in the protein expression levels under BCAA starvation.
The manuscript was well-written, and the findings are interesting, especially their model that positional codon usage bias could be a regulator of ribosome pausing and tRNA charging levels. Although different translational responses to distinct amino acid starvation have been widely documented, the positional codon usage bias is an interesting aspect. The manuscript's central message could have been made clearer. The authors may consider emphasizing this point more explicitly in the abstract. The rich multi-omics dataset in this work provides valuable resources for the translation field.
We thank the reviewer for the thoughtful and positive evaluation of our work.
Major comments
- The abstract may need to be revised since it is hard to immediately catch the authors' main point. If the authors regard this work as a resource paper, the current version is fine. But it could be better to point out the positional codon usages the authors found, which is a strong point of the current manuscript.
Response: We thank the reviewer for highlighting the importance of positional codon usage, which indeed represents a key finding of our study. We revised the abstract, and we now emphasize this aspect more clearly. However, in response to review #2, we have framed the observed positional effects and the idea of an elongation bottleneck as one possible contributing mechanism among others and relate it specifically to the attenuation of isoleucine-specific stalling under triple starvation.
- Page 18 "Beyond these tRNA dynamics, our data also highlight the importance of the codon positional context within mRNAs, indicating that where a codon is located within the CDS can influence both the extent of ribosomal stalling and overall translation efficiency during nutrient stress." This idea is interesting. To what extent the authors think this could be generalized? The authors may discuss whether they think their proposed model is specific to the different ribosome stalling patterns between valine and isoleucine codons or generalized to other codon combinations. For example, the positional codon usage bias will be different among different organisms, and are there any previous reports on ribosome behaviors that align with their model?
Response: We thank the reviewer for raising these important points. While our study primarily focuses on the differential stalling patterns of valine and isoleucine codons, we believe the underlying principle, that the position of codons within the CDS can modulate the extent of ribosome stalling, may under very specific circumstances extend beyond this amino acid pair. We expect this positional effect to be potentially relevant for combinations in which one amino acid has considerable enrichment near the 5′ end of coding sequences, coupled with starvation-sensitive tRNA isoacceptors, while the other does not. In our case, valine meets these criteria (see Fig. S11A and Fig. 6). In contrast, isoleucine and leucine codons, although also relatively frequent, show more variable positional distributions and are both decoded by isoacceptors that appear more resistant to starvation, as illustrated in Fig. 6 and reported for mammals and bacteria in Saikia et al. 2016; Darnell, Subramaniam, and O’Shea 2018; Elf et al. 2003; Dittmar et al. 2005. To explore the generalizability of this model, we have now included a transcriptome-wide analysis of codon position biases in mouse for all codons in the revised manuscript (Supplementary Figures 10 and 11). This analysis may serve as a basis to identify additional candidate codons for future studies. Furthermore, we now mention in the Discussion that amino acids with similar properties to valine regarding their positional distribution and tRNA isoacceptors, such as phenylalanine, and glutamine, whose tRNA isoacceptors are predicted to be fully deacylated under their respective starvation in bacteria (Elf et al. 2003), could be promising candidates for testing this model, in combination with amino acids, whose tRNAs are expected to remain partially charged under starvation or to be depleted at the start of the CDS such as i.e. His (Supplementary Fig.11C).
Even if the authors think this model can be applied to BCAA starvation, would it be possible to explain the different isoleucine codon responses between single and double starvation? The authors may discuss why the ribosome stalling at isoleucine AUU and AUC codons was slightly attenuated under double starvation. And how about the different leucine codon responses among single, double, and triple starvations, although the pausing is not as strong as isoleucine and valine codons?
Response: Regarding the attenuated isoleucine stalling under double starvation, we believe this is primarily due to stronger inhibition of the mTORC1 pathway when leucine is co-depleted (i.e., in the double starvation condition; Fig. 2D–F). This results in a more substantial suppression of global translation, reducing overall tRNA demand and thereby mitigating stalling (Darnell, 2018). A similar effect may explain the only mild leucine codon stalling observed under single leucine starvation, which also triggers strong mTORC1 inhibition and reduced initiation. In contrast, triple starvation does not suppress mTORC1 to the same extent, and thus reduced initiation alone cannot explain the absence of leucine codon stalling. Instead, we propose that additional features, such as the relative sensitivity of tRNA isoacceptors to starvation and their aminoacylation dynamics, must be considered. Valine tRNAs, for example, are known to be highly sensitive and become strongly deacylated under starvation in bacteria (Elf et al. 2003), a pattern that we also find in our own data (Fig. 6). Leucine tRNAs, by contrast, appear more resistant, possibly due to better amino acid recycling or isoacceptor-specific differences in charging kinetics, though further validation would be needed. However, combined with the strong stalling at 5′-enriched valine codons, this could reduce downstream ribosome traffic and limit exposure of leucine codons, thus preventing stalling. However, our new analysis of the positional relationship between valine and leucine codons within individual transcripts (now shown in Supplementary Figure 11B) did not reveal as strong a pattern as we observed for valine and isoleucine codons. We now discuss these points and their implications in the revised Discussion.
Experimental validation using artificial reporters carrying biased sequences may also be considered.
Response: We appreciate the reviewer’s suggestion. In fact, we explored this experimentally using a dual-fluorescent reporter system (GFP–RFP) (Juszkiewicz and Hegde 2017) containing consecutive Val or Ile codons. However, the constructs yielded variable and non-reproducible results under starvation conditions. In addition, testing the role of codon position would require placing the same codons at multiple defined positions within a single transcript and performing ribosome profiling directly on the reporter. This type of targeted experimental validation is technically challenging and falls beyond the scope of the current study. We now mention this explicitly in the revised Discussion as an interesting direction for future work.
- Page 13 "Moreover, we noticed that DT changes extend beyond the ribosomal A-site, including the P-site, E-site, and even further positions (Supplementary Fig. 2A), consistent with other studies on single amino acid starvation 39 (Supplementary Fig. 2B-C)." Could the widespread DT changes be due to Ribo-DT pipeline they used or difficulties in offset determination? Indeed the authors showed that this feature was found in other datasets, but it seems that the datasets were processed and analyzed in the same way as their data. The original Ribo-DT paper (Gobet and Naef, 2022, Methods) also showed some widespread DT changes even from RNA-seq. Another analysis method like the codon subsequence abundant shift as a part of diricore analysis (Loayza-Puch et al., 2016, Nature) did not show that broad changed regions. The authors are encouraged to re-analyze the data sets using different methods.
Response: We agree with the reviewer that the fact that DT changes beyond the ribosomal A-site is puzzling, but this has already been seen in other papers using other approaches (Darnell, Subramaniam, and O’Shea 2018). To validate that this shift is not due to our A-site assignment, enrichment analysis, or DT method, we applied the Diricore pipeline to our Ribo-Seq data. The output of the pipeline provides either 5’-end ribosome density or “subsequence” analysis using an A-site offset for each read size based on the metagene profile at the start codon. Both analyses show the same enriched codons across the different conditions as in our analyses, and the broad shift is similar, with the maximum signal at E, -1 position (Fig. R1).
- Page 13 "Intriguingly, only two of the three isoleucine codons (AUU and AUC) showed increased DTs upon Ile starvation (p < 0.01), while just one leucine codon (CUU) exhibited a modest but significant DT increase (p < 0.01) under Leu starvation (Figure 1A-B, Supplementary Figure 2A)." How can the authors explain the different strengths of ribosome pausing at Ile codons under Ile and double starvation? The AUA codon did not show any pausing under either of the starvation conditions. Throughout the manuscript, the authors mainly describe the difference between amino acids but it is desirable to discuss the codon-level difference as well.
Response: Thank you for raising this point. The observed differences in stalling between the isoleucine codons can likely be explained by differences in tRNA isoacceptor charging and positional bias within transcripts. The AUA codon is decoded by a distinct tRNAIle isoacceptor (tRNAIleUAU), which, according to our tRNA charging data (Fig. 6), remains largely charged during Ile starvation. This observation aligns with previous reports suggesting that this isoacceptor is more resistant to starvation-induced deacylation in mammalian cells and bacteria (Saikia et al. 2016; Elf et al. 2003). In contrast, the AUU and AUC codons are primarily decoded by the tRNAIleAAU isoacceptor, which we find to be strongly deacylated under Ile starvation, likely contributing to the observed codon-specific ribosome pausing. Additionally, we found that the AUA codons are relatively rare in general and particularly underrepresented near the 5′ ends of coding sequences. Our new spatial analysis (now included in Supplementary Figure 11B) confirms that AUA codons tend to occur downstream of AUU and AUC codons within transcripts. This potentially further reduces stalling on these codons and further diminishes their apparent DT increase under starvation. In order to better explain these important points, we have now expanded the codon-level discussion of these differences in the revised manuscript.
- Page 13 "We examined the effects of single amino acid starvations (-Leu, -Ile and -Val), as well as combinations, including a double starvation of leucine and isoleucine (hereafter referred to as "double") and a starvation of leucine, isoleucine, and valine ("triple"), allowing us to identify potential non-additive effects." The different double starvations, isoleucine and valine, and leucine and valine, will further support their hypothesis on the effects of the positional codon usage bias on ribosome pausing and tRNA charging patterns. Although this could be beyond the scope of the current manuscript, the authors are encouraged to provide a rationale for the chosen combination.
Response: Our experimental design evolved stepwise: we initially focused on leucine and isoleucine depletion as we found that despite their structure similarity these had respectively short and long dwell times in our previous work in the mouse liver (Gobet et al. 2020). Valine was included at a later stage to cover all the BCAAs. At the time, we did not anticipate valine to yield particularly striking effects in cells, and therefore we did not include systematic pairwise depletions involving valine. However, the strong and unexpected stalling observed at valine codons, especially under triple starvation, became a central aspect of the study. Thus, we agree that additional combinations, such as Leu/Val or Val/Ile, could be informative and now mention this in the Discussion as a potential direction for future studies.
Minor comments
Page 16 "these results imply that BCAA deprivation lowers protein output through multiple pathways: a combination of reduced initiation, direct elongation blocks (stalling), and possibly an increased proteolysis" This conclusion is totally right but may be too general. Could the authors summarize BCAA-specific features of the events including reduced initiation, stalling, and proteolysis that all contribute to protein outputs? This is not well discussed in the latter sections including Discussion.
Response: We thank the reviewer for this helpful suggestion. We agree that the original statement was too general and have revised the relevant section to more clearly delineate the distinct responses observed under each BCAA starvation condition. Specifically, we now summarize that valine starvation is characterized by strong, positionally biased ribosome stalling; leucine starvation primarily impacts translation initiation, likely via mTORC1 repression; and isoleucine starvation shows a mixed phenotype, with features of both impaired initiation and codon-specific elongation delays. We also clarify that while protein stability or degradation may contribute to the observed changes in protein output, our current data do not allow for quantitative assessment of proteolytic effects (e.g., changes in protein half-life). Therefore, we refrain from making direct quantitative conclusions about the differential modulations of proteolysis and instead focus our discussion on the translational mechanisms supported by our data.
Reviewer #1 (Significance):
The manuscript was well-written, and the findings are interesting, especially their model that positional codon usage bias could be a regulator of ribosome pausing and tRNA charging levels. Although different translational responses to distinct amino acid starvation have been widely documented, the positional codon usage bias is an interesting aspect. The manuscript's central message could have been made clearer. The authors may consider emphasizing this point more explicitly in the abstract. The rich multi-omics dataset in this work provides valuable resources for the translation field.
We thank the reviewer for the encouraging comments and share the view that positional codon-usage bias is an important result; accordingly, we now underscore this point explicitly in the revised Abstract. We also emphasise that our other observations are, to our knowledge, novel: only a handful of multi-omics studies have combined ribosome-pausing profiles with direct tRNA-aminoacylation measurements, and none has systematically examined multiple amino-acid-deprivation conditions as presented here.
Reviewer #2 (Evidence, reproducibility and clarity):
This study examines the consequences of starvation for the BRCAAs, either singly, for Leu & Ile, or for all three simultaneously in HeLa cells on overall translation rates, decoding rates at each codon, and on ribosome density, protein expression, and distribution of ribosome stalling events across the CDS for each expressed gene. The single amino acid starvation regimes specifically reduce the cognate intracellular amino acid pool and lead to deacylation of at least a subset of the cognate tRNAs in a manner dependent on continuing protein synthesis. They also induce the ISR equally and decrease bulk protein synthesis equally in a manner that appears to occur largely at the initiation level for -Leu and -Val, judging by the decreased polysome:monsome ratio, but at both the initiation and elongation levels for -Ile-a distinction that remains unexplained. Only -Leu appears to down-regulate mTORC1 and TOP mRNA translation.There is a significant down-regulation of protein levels for 50-200 genes, which tend to be unstable in nutrient-replete cells, only a fraction of which are associated with reduced ribosome occupancies (RPFs measured by Ribo-Seq) on the corresponding mRNAs in the manner expected for reduced initiation, suggesting that delayed elongation is responsible for reduced protein levels for the remaining fraction of genes. All three single starvations lead to increased decoding times for a subset of the cognate "hungry" codons: CUU for -Leu, AUU and AUC for -Ile, and all of the Val codons, in a manner that is said to correspond largely to the particular tRNA isoacceptors that become deacylated, although this correspondence was not explained explicitly and might not be as simple as claimed. All three single starvations also evoke skewing of RPFs towards the 5' ends of many CDSs in a manner correlated with an enrichment within the early regions of the CDSs for one or more of the cognate codons that showed increased decoding times for -Ile (AUC codon) and -Val (GUU, GUC, and GUG), but not for -Leu-of which the latter was not accounted for. These last findings suggest that, at least for -Val and -Ile, delays in decoding N-terminal cognate codons cause elongating ribosomes to build-up early in the CDS. They go on to employ a peak calling algorithm to identify stalling sites in an unbiased way within the CDS, which are greatest in number for -Val, and find that Val codons are enriched in the A-sites (slightly) and adjacent 5' nucleotides (to a greater extent) for -Val starvation; and similarly for Ile codons in -Ile conditions, but not for -Leu starvation-again for unknown reasons. It's unclear why their called stalling sites have various other non-hungry codons present in the A sites with the cognate hungry codons being enriched further upstream, given that stalling should occur with the "hungry" cognate codon in the A site. The proteins showing down-regulation are enriched for stalling sites only in the case of the -Val starvation in the manner expected if stalling is contributing to reduced translation of the corresponding mRNA. It's unclear why this enrichment apparently does not extend to -Ile starvation which shows comparable skewing of RPFs towards the 5'ends, and this fact diminishes the claim that pausing generally contributes to reduced translation for genes with abundant hungry codons. All of the same analyses were carried out for the Double -Ile/-Leu and Triple starvations and yield unexpected results, particularly for the triple starvation wherein decoding times are increased only at Val codons, skewing of RPFs towards the 5' ends of CDSs is correlated only with an enrichment for Val codons within the early regions of the CDSs, and stall sites are enriched only for Val codons at nearly upstream sites, all consistent with the finding that only Val tRNAs become deacylated in the Triple regime. To explain why only Val tRNA charging is reduced despite the observed effective starvation for all three amino acids, they note first that stalling at Val codons is skewed towards the 5'ends of CDS for both -Val and triple starvations more so than observed for Ile or -Leu starvation, which they attribute to a greater frequency of Val codons vs Ile codons in the 5' ends of CDSs. As such, charged Val tRNAs are said to be consumed in translating the 5'ends of CDSs and the resulting stalling prevents ribosomes from reaching downstream Ile and Leu codons at the same frequencies and thus prevents deacylation of the cognate Ile and Leu tRNAs. It's unclear whether this explanation is adequate to explain the complete lack of Ile or Leu tRNA deacylation observed even when amino acid recycling by the proteasome is inhibited-a treatment shown to exacerbate deacylation of cognate tRNAs in the single amino acid starvations and of Val tRNA in the triple starvation. As such, the statement in the Abstract "Notably, we could show that isoleucine starvation-specific stalling largely diminished under triple starvation, likely due to early elongation bottlenecks at valine codons" might be too strong and the word "possibly" would be preferred over "likely". It's also unclear why the proteins that are down-regulated in the triple starvation are not significantly enriched for stalling sites (Fig. 5B) given that the degree of skewing is comparable or greater than for -Val. This last point seems to undermine their conclusion in the Abstract that "that many proteins downregulated under BCAA deprivation harbor stalling sites, suggesting that compromised elongation contributes to decreased protein output." In the case of the double -Ile/-Leu starvation, a related phenomenon occurs wherein decoding rates are decreased for only the AUU Ile codon and only the AAU Ile tRNA becomes deacylated; although in this case increased RPFs in the 5' ends are not correlated with enrichment for Ile or Leu codons and, although not presented, apparently stall sites are not associated with the Ile codon in the double starvation. In addition, stalling sites are not enriched in the proteins down-regulated by the double starvation. Moreover, because Ile codons are not enriched in the 5'ends of CDS, it doesn't seem possible to explain the selective deacylation of the single Ile tRNA observed in the double starvation by the same "bottleneck" mechanism proposed to explain selective deacylation of only Val tRNAs during the triple starvation. This is another reason for questioning their "bottleneck" mechanism.
We thank the reviewer for their deep assessment, exhaustive reading, and constructive feedback, which have greatly contributed to improving the clarity and contextualization of our manuscript. We would first like to clarify that all experiments in this study were conducted in NIH3T3 mouse fibroblasts, not HeLa cells; we assume this was a misunderstanding and have verified that the correct cell line is consistently indicated throughout the manuscript. We also clarify that our data show that -Leu, double starvation, and to a lesser extent -Ile, downregulate mTORC1 signaling and TOP mRNA translation, whereas valine -Val and triple starvation had minimal effects on these pathways. We agree that some of our conclusions and observed phenomena were not explained in sufficient detail in the original version. To address this, we have significantly reworked the discussion, added complementary figures and clarified key points throughout the text, to better convey the underlying rationale and biological interpretation of our findings. We address each of the reviewer’s points in detail in the point-by-point responses below.
Specific comments (some of which were mentioned above):
-The authors have treated cells with CHX in the Ribo-Seq experiments, which has been shown to cause artifacts in determining the locations of ribosome stalling in vivo owing to continued elongation in the presence of CHX (https://doi.org/10.1371/journal.pgen.1005732 ). The authors should comment on whether this artifact could be influencing some of their findings, particular the results in Fig. 5C where the hungry codons are often present upstream of the A sites of called stalling sites in the manner expected if elongation continued slowly following stalling in the presence of CHX.
Response: We thank the reviewer for raising this important concern. We would like to clarify that our ribosome profiling protocol did not include CHX pretreatment of live cells. CHX was added only during the brief PBS washes immediately before lysis and in the lysis buffer itself. This approach aligns with best practices aimed at minimizing post-lysis ribosome run-off, and is intended to prevent the downstream ribosome displacement artifacts described by Hussmann et al. 2015, which result from pre-incubation of live cells with CHX for several minutes before harvesting. Furthermore, recent studies have demonstrated that CHX-induced biases are species-specific. For instance, Sharma et al. 2021 found that human (and mice) ribosomes are not susceptible to conformational restrictions by CHX, nor does CHX distort gene-level measurements of ribosome occupancy. This suggests that the use of CHX in the lysis buffer, as performed in our protocol, is unlikely to introduce significant artifacts in our ribosome profiling data. To further support this, we reanalyzed data from Darnell, Subramaniam, and O’Shea 2018, where the ribosome profiling samples were prepared without any CHX pretreatment or CHX in the wash buffer, and still observed similar upstream enrichments in their stalling profiles (see Supplementary Figure 2B-C in our manuscript). Additionally, in our previous work (Gobet et al. 2020), we compared ribosome dwell times with and without CHX in the lysis buffer and found no significant differences, reinforcing the notion that CHX use during lysis does not substantially affect the measurement of ribosome stalling. Given these considerations, we believe that CHX-related artifacts, such as downstream ribosome movement, are unlikely to explain the enrichment of hungry codons upstream of identified stalling sites in our data. We have now adjusted the Methods section to clarify this point.
-p. 12: "These starvation-specific DT and ribosome density modulations were also evident at the individual transcript level, as exemplified by Col1a1, Col1a2, Aars, and Mki67 which showed persistent Val-codon-specific ribosome density increases but lost Ile-codon-specific increases under triple starvation (Supplementary Figure 3A-D). " This conclusion is hard to visualize for any but Val codons. It would help to annotate the relevant peaks of interest for -Ile starvation with arrows.
Response: We agree and thank the reviewer for this observation. We have now annotated exemplary peaks in Supplementary Figure 3A–D to highlight ribosome pileups over Ile codons. However, we agree that it is still hard to visualize in the given Figure. Therefore, we added scatter plots for each of the transcripts that show the RPM of each position in the Ctrl vs starvation to allow for a better illustration of the milder effects upon Ile starvation (Supplementary Figure 4).
-To better make the point that codon-specific stalling under BCAA starvation appears to be not driven by codon usage, rather than the analysis in Fig. 1H, wouldn't it be better to examine the correlation between increases in DT under the single amino acid starvation conditions and the codon frequencies across all codons?
Response: We appreciate the suggestion. We have now added an additional analysis correlating the change in DT with codon usage frequency for each starvation condition. This is included in Supplementary Figure 5A-D and supports our interpretation that codon frequency alone does not explain the observed stalling behavior.
-p. 13, entire paragraph beginning with "Our RNA-seq and Ribo-seq revealed a general activation of stress response pathways across all starvations..." It is difficult to glean any important conclusions from this lengthy analysis, and the results do not appear to be connected to the overall topic of the study. If there are important conclusions here that relate to the major findings then these connections should be made or noted later in the Discussion. If not, perhaps the analysis should be largely relegated to the Supplemental material.
Response: We thank the reviewer for this comment. The paragraph in question is intended to provide a global overview of transcriptional and translational responses across the starvation conditions. It serves both as a quality control (e.g., PCA clustering and global shifts in RPF/RNA-seq profiles), and to confirm that expected starvation-induced responses are among the strongest detectable signals separating the starved samples from the control. Indeed, these observations establish that the perturbations are effective and that hallmark nutrient stress responses are globally engaged across conditions. Importantly, very few studies to date have examined transcriptional and translational responses under single or combined branched-chain amino acid (BCAA) starvation conditions. It therefore remains unclear to what extent BCAA depletion broadly remodels gene expression and translation. Our analysis contributes to addressing this gap, revealing that while certain stress pathways are commonly induced, others show condition-specific patterns such as we observed for -Ile starvation. To maintain focus, we have kept the detailed pathway analyses and transcript-level enrichments in the Supplement and rewritten the corresponding text in a more compact manner, reducing it by more than one third.
-p. 15: "Together, these findings highlight that BCAA starvation triggers a combination of effects on initiation and elongation, with varying dynamics by amino acid starvation." I take issue with this statement as it appears that translation is reduced primarily at the initiation step for all conditions except -Ile. As noted above, these data are never menitioned in the DISCUSSION as to why only -Ile would show a marked elongation component to the inhibition whereas -Val gives the greatest amount of ribosome stalling.
Response: We acknowledge the reviewer’s point. While the polysome profiles (Figure 3F-H) directly indicate that most conditions repress initiation, codon- and condition-specific elongation defects can still contribute to reduced protein output, even if they are not always detectable as global polysome shifts. Polysome profiles reflect the combined outcome of reduced initiation (which decreases polysome numbers) and ribosome stalling (which can, but does not always have to, increase ribosome density on individual transcripts, potentially counteracting the effects of reduced initiation). For valine starvation strong stalling occurs very early in the CDS (Figure 5F). This bottleneck restricts overall ribosome movement to downstream regions. Thus, while elongation is profoundly impaired, the total number of ribosomes per transcript (which polysome signals largely reflect) may appear low due to reduced overall ribosome traffic. In contrast, isoleucine codon stalling tends to occur also further downstream on the transcript (Figure 5F), allowing ribosomes to accumulate in larger numbers on the mRNA, leading to a clearer "elongation signature" in polysome profiles (Figure 3F, H). Additionally, we observed slightly higher inter-replicate variance for isoleucine starvation (Supplementary Figure 6B), which may have reduced the number of statistically significant stalling sites extracted compared to valine. We have revised the main text and discussion to clarify these points.
-I cannot decipher Fig. 4D and more detail is required to indicate the identity of each column of data.
Response: We thank the reviewer for pointing this out. Figure 4D (now Figure 4E) presents an UpSet plot, which is a scalable alternative to Venn diagrams commonly used to visualize intersections across multiple sets. Briefly, each bar in the upper plot represents the number of transcripts with increased 5′ ribosome coverage (Δpi < -0.15; p < 0.05) shared across the conditions indicated in the dot matrix below. Each column in the dot matrix highlights the specific combination of conditions contributing to a given intersection (e.g., dots under “Val” and “Triple” show the overlap between these two). To improve clarity, we have expanded the figure legend accordingly and now refer to the UpSetR methodology in the main text.
-In Fig. 4E, one cannot determine what the P values actually are, which should be provided in the legend to confirm statistical significance.
Response: Thank you for pointing that out. The legend in Figure 4E (now Figure 4F) for the p-values was accidentally removed during figure editing. We have added the legend back, so that the statistical significance is clear.
-It's difficult to understand how the -Leu condition and the Double starvation can produce polarized RPFs (Fig. 4A) without evidence of stalling at the cognate hungry codons (Fig. 4E), despite showing later in Fig. 5A that the numbers of stall sites are comparable in those cases to that found for -Ile.
Response: We appreciate this comment, which points to an important property of RPF profiles under nutrient stress. As shown in Figure 4A, all starvation conditions induce a degree of 5′ ribosome footprint polarization, a pattern that can be observed under various stress conditions and perturbations (Allen et al. 2021; Hwang and Buskirk 2017; Li et al. 2023). This general 5′ bias likely reflects a combination of slowed elongation and altered ribosome dynamics and is not necessarily linked to codon-specific stalling. However, Val and Triple starvation show a much stronger and more asymmetric polarization, characterized by pronounced 5′ accumulation and 3′ depletion of ribosome density. To better illustrate this, we have updated the visualization of polarity scores and added a new bar chart summarizing the number of transcripts showing strong 5′ polarization under each condition. This quantification highlights that the effect is markedly more prevalent under Val and Triple conditions than under Leu or Double starvation. In addition, Figure 4F demonstrates that this polarity is codon-specific under Val and Triple starvation. We clarify that this analysis tests for enrichment of specific codons near the start codon among the polarized transcripts and does not directly assess stalling. The observed enrichment of Val codons in the 5′ regions of polarized transcripts supports the interpretation that early elongation delays contribute to the RPF shift. In contrast, no such enrichment is observed for Leu starvation, reinforcing that Leu-induced polarity is not driven by stalling at Leu codons. While Figure 5 shows a similar number of peak-called stalling sites in -Leu, -Ile, and Double starvation, we note that Ribo-seq signal variability under Ile starvation was higher, which may have limited statistical power for detecting stalling sites, even though clear dwell time increases were observed at specific codons. Additionally, we have improved the metagene plots depicting total ribosome footprint density in Figure 4A. The previous version incorrectly showed sharp drops at CDS boundaries due to binning artifacts. The updated version more accurately reflects the density distribution and further highlights the stronger polarization in Val and Triple conditions. Together, these clarifications and improvements within the main text now more clearly distinguish between general polarity effects and codon-specific stalling.
-Fig. 5B: the P values should be given for all five columns, and it should be explained here or in the Discussion why the authors conclude that stalling is an important determinant for reduced translation when a significant correlation seems to exist only for the -Val condition and not even for the Triple condition.
Response: We thank the reviewer for this important observation. In response, we have revised both the text and the figures to provide a clearer and biologically more meaningful representation of the relationship between ribosome stalling and reduced protein output. Specifically, we have replaced the previous Figure 5B with a new analysis that stratifies transcripts based on the number of identified stalling sites. This updated analysis, now shown in Figure 5B, reveals that under Val and Triple starvation conditions, proteins that are downregulated tend to originate from transcripts with multiple stalling sites. Importantly, the corresponding p-values for all five conditions are now explicitly shown in the figure (as red lines). As the reviewer correctly notes, only the Val condition shows a statistically significant enrichment when considering overall overlap. Triple starvation shows a similarly high proportion of overlap (72.3%) but does not reach statistical significance, likely due to the more complex background composition under combined starvation, which increases the expected overlap and reduces statistical power. By stratifying transcripts by the number of stalling sites, we uncover that transcripts with ≥2 stalling sites are enriched among downregulated proteins specifically under Val and Triple conditions, providing a more robust indication of the link between stalling and translation repression under Valine deprivations. We believe this refined approach, prompted by the reviewer’s comment, offers a clearer and biologically more relevant perspective on the role of ribosome stalling. The original analysis previously shown in Figure 5B is now provided as Supplemental Figure 10C for transparency and comparison. We have clarified this in the revised text and now interpret the relationship more cautiously.
-p. 17: "Of note, in cases where valine or isoleucine codons were present just upstream (rather than at) the stalling position, we noted a strong bias for GAG (E), GAA (E), GAU (D), GAC (D), AAG (K), CAG (Q), GUG (V) and GGA (G) (Val starvation) and AAC (N), GAC (D), CUG (L), GAG (E), GCC (A), CAG (Q), GAA (E) and AAG (K) (Ile starvation) at the stalling site (Supplementary Figure 7B)." The authors fail to explain why these codons would be present in the A sites at stalling sites rather than the hungry codons themselves, especially since it is the decoding times of the hungry codons that are increased according to Fig. 1A-E. As suggested above, is this a CHX artifact?
Response: We agree that the observation that the listed codons are enriched at identified stalling positions (now Supplementary Figure 10C), while the depleted amino acid codon is located upstream, is a finding that needs more detailed explanation. Importantly, this phenomenon is not attributable to CHX artifacts, as our Ribo-seq protocol employs CHX solely during brief washes and lysis to prevent post-lysis ribosome run-off, rather than live-cell pre-treatment. Instead, we propose two hypotheses to explain this pattern: Firstly, many of these enriched codons are already inherently slow-decoded with longer DTs even under control conditions (Supplementary Figure 5H, newly added). Together with the upstream hungry codons they might form a challenging consecutive decoding environment, which results in an attenuated ribosome slowdown downstream after the hungry codon. Second, ribosome queuing may further explain this pattern. When a ribosome encounters a critically hungry codon and stalls, subsequent ribosomes can form a queue. The codon within the A-site of the queued ribosome would be (more or less) independent of the identity of the hungry codon itself that caused the initial stall. Since the listed codons have a high frequency within the transcriptome (Supp. Fig 5B), they therefore have an increased likelihood of appearing at this “stalling site”. Importantly, both of these phenomena are not necessarily represented by a general increase of DT on all of the listed codons and would therefore only be captured by the direct extraction of stalling sites but might be averaged out in the global dwell time analysis. We mention this phenomenon now in the Discussion.
-Fig. 5D: P values for the significance, or lack thereof, of the different overlaps should be provided.
Response: Thanks for pointing out this omission. We have now computed hypergeometric p-values for comparisons shown in Figure 5D and Figure 5E, and report them directly in the main text. As described, the overlap in stalling sites between Val and triple starvation is highly significant (2522 positions, p < 2.2×10⁻¹⁶), while overlaps involving Ile-specific stalling positions are smaller but still statistically robust (e.g., 149 positions for Ile – Triple, p = 1.77×10⁻⁵²). Notably, we also calculated p-values at the transcript level and found that a large fraction of transcripts with Ile-specific stalling under single starvation also stall under triple starvation, though often at different positions (1806 transcripts, p = 1.78×10⁻⁵⁸). These values are now included in the revised results section to support the interpretation of these overlaps.
-p. 17: "Nonetheless, when we examined entire transcripts rather than single positions, many transcripts that exhibited isoleucine-related stalling under Ile starvation also stalled under triple starvation, but at different sites along the CDS (Figure 5E). This finding is particularly intriguing, as it suggests that while Ile-starvation-specific stalling sites may shift under triple starvation, the overall tendency of these transcripts to stall remains." The authors never come back to account for this unexpected result.
Response: Thank you for highlighting this point. We've incorporated this finding as part of the proposed "bottleneck" scenario. While the isoleucine-specific stalling sites identified under Ile starvation do shift or disappear under triple starvation, we've observed that the same transcripts still tend to exhibit stalling. However, this now primarily occurs at upstream valine codons. We interpret this as a consequence of early elongation stalling caused by strong pausing at Val codons. This restriction on ribosome progression effectively prevents ribosomes from reaching the original Ile stalling sites. Therefore, the stalling sites identified under triple starvation are largely explained by the Val codons, reflecting a redistribution of stalling rather than its loss. To further clarify this crucial point, we've now explicitly mentioned Figure 5D-E again in the subsequent paragraph, which introduces the bottleneck theory.
-It seems very difficult to reconcile the results in Fig. 5F with those in Fig. 4A, where similar polarities in RPFs are observed for -Ile and -Val in Fig, 4A but dramatically different distributions of stalling sites in Fig. 5F. More discussion of these discrepancies is required.
Response: Thank you for pointing this out. The apparent discrepancy between the RPF profiles shown in Figure 4A and the stalling site distributions in Figure 5F likely reflects the fact that RPF polarization includes both general (unspecific) and codon-specific components. Figure 4A displays total ribosome footprint density, capturing both broad stress-induced effects and codon-specific contributions, whereas Figure 5F focuses specifically on peak-called stalling sites, representing localized and statistically significant pauses. Importantly, we would like to emphasise that Fig 4 shows that -Val and -Ile starvation exhibit different responses and not the same patterns. To make these differences even clearer, we have now updated the visualizations in Figure 4, including improved polarity plots and a new bar chart summarizing the number of transcripts with strong 5′ polarization. These additions highlight that the RPF profiles under -Val starvation are more pronounced and asymmetric, particularly due to 3′ depletion, while the polarity under -Ile is milder and a distinct, much smaller subset of transcripts appears to show polarity score shifts. We believe the updated figures and accompanying explanations now make these distinctions clearer.
- p. 18: " These isoacceptor-specific patterns correlate largely with the particular subsets of leucine and isoleucine codons that stalled (Figure 1A)." This correlation needs to be addressed for each codon-anticodon pair for all of the codons showing stalling in Fig. 1A.
Response: We thank the reviewer for this important comment. In the revised manuscript, we have expanded the relevant sections to address codon–anticodon relationships more thoroughly. We now explicitly match codons that exhibited increased dwell times under starvation to the corresponding tRNA isoacceptors whose charging was affected, and we provide a clearer discussion of the caveats involved. As noted by the reviewer, this correlation is not straightforward, as it is complicated by wobble base pairing, anticodon modifications, and the fact that multiple codons can be decoded by more than one isoacceptor, and vice versa. Moreover, in our qPCR-based tRNA charging assay, certain isoacceptors cannot be distinguished due to highly similar sequences (e.g., LeuAAG and LeuUAG, and LeuCAA and LeuCAG), which limits resolution for exact pairing. In addition, we did not assess absolute tRNA abundance, which may further influence decoding capacity. Nevertheless, where resolution is possible, the patterns align well: All tRNAVal isoacceptors became uncharged under Val and triple starvation, matching the consistent dwell time increases across all Val codons. Only tRNAIleAAU (decoding AUU and AUC) was deacylated, matching to these codons showing increased dwell times, while AUA (decoded by still-charged tRNAIleUAU) did not. Only CUU (decoded by uncharged tRNALeuGAA) showed increased dwell time. A mild deacylation of the other Leu isoacceptors was observed, but isoacceptor-level resolution is limited by assay constraints. However, these rather minimal tRNA and DT changes were consistent with more dominant initiation repression rather than elongation stalls. To support this analysis, we included an illustrative figure (now in Supplementary Figure 12F) summarizing the codon–anticodon matches.
-p. 19: "For instance, in our double starvation condition, unchanged tRNA charging levels (Figure 6E) may result from a pronounced downregulation of global translation initiation, likely driven by the activation of stress responses (Figure 2), subsequently lowering the demand for charged tRNAs as it has been observed previously for Leu starvation 39.” This seems at odds with the comparable down-regulation of protein synthesis for the Double starvation and -Leu and -Ile single starvations shown in Fig. 3C. Also, in the current study, Leu starvation does lower charging of certain Leu tRNAs.
Response: We thank the reviewer for raising this important point. In the revised manuscript, we have clarified this section and now offer a more refined interpretation of the tRNA charging patterns observed under double starvation. While Figure 3C shows a comparable reduction in global protein synthesis across the -Leu, -Ile, and double starvation conditions, it needs to be considered that the OPP assay has limited sensitivity. It operates in a relatively low fluorescence intensity range and is subject to background signal, which may obscure subtle differences between conditions. Moreover, other factors such as changes in protein stability or turnover could also contribute to the observed differences. Therefore, inter-condition differences in translation repression should be interpreted with caution. However, based on our stress response analysis (Figure 2), mTORC1 inactivation appears strongest under double starvation, likely leading to more profound suppression of translation initiation. This would reduce the overall demand for charged tRNAs and could explain why no detectable tRNA deacylation was observed under double starvation, even though mild uncharging of Leu isoacceptors occurred under -Leu, which exhibited a milder stress response. This distinction is consistent with the observed mild dwell time increases for one Leu codon under -Leu, but not in the double condition. Similarly, the absence of Ile codon stalling and tRNA deacylation under double starvation may be attributed to stress-driven reductions in elongation demand, preventing the tRNA depletion and codon-specific delays observed under single Ile starvation. A more direct clarification is now included in the revised manuscript.
Reviewer #2 (Significance):
The results here are significant in showing that starvation for a single amino acid does not lead to deacylation of all isoacceptors for that amino acid and in revealing that starvation for one amino acid can prevent deacylation of tRNAs for other amino acids, as shown most dramatically for the selective deacylation of only Val tRNAs in the triple BRCAA starvation condition. For the various reasons indicated above, however, I'm not convinced that their "bottleneck" mechanism is adequate to explain this phenomenon, especially in the case of the selective deacylation of Ile vs Leu tRNA in the Double starvation regime. It's also significant that deacylation leads to ribosome build-up near the 5'ends of CDS, which seems to be associated with an enrichment for the hungry codons in the case of Val and Ile starvation, but inexplicably, not for Leu or the Double starvations. This last discrepancy makes it hard to understand how the -Leu and Double starvations produce RPF buildups near the 5 ends of CDSs. In addition, the claim in the Discussion that "our data also highlight the importance of the codon positional context within mRNAs, indicating that where a codon is located within the CDS can influence both the extent of ribosomal stalling and overall translation efficiency during nutrient stress" overstates the strength of evidence that the stalling events lead to substantial decreases in translational efficiencies for the affected mRNAs, as the stalling frequency and decreased protein output are significantly correlated only for the -Val starvation, and the data in Fig. 3 D-H suggest that the reductions in protein synthesis generally occur at the level of initiation, even for -Val starvation, with a contribution from slow elongation only for -Ile-which is in itself difficult to understand considering that stalling frequencies are highest in -Val. Thus, while many of the results are very intriguing and will be of considerable interest to the translation field, it is my opinion that a number of results have been overinterpreted and that important inconsistencies and complexities have been overlooked in concluding that a significant component of the translational inhibition arises from the increased decoding times at hungry codons during elongation and that the selective deacylation of Val tRNAs in the Triple starvation can be explained by the "bottleneck" mechanism. The complexities and limitations of the data and their intepretations should be discussed much more thoroughly in the Discussion, which currently is devoted mostly to other phenomena often of tangential importance to the current findings. A suitably revised manuscript would clearly state the limitations and caveats of the proposed mechanisms and consider other possible explanations as well.
Again, we thank the reviewer for the valuable insights and constructive critiques. We believe that the concerns regarding potential overinterpretation and inconsistencies have now been addressed through clearer explanations and more cautious interpretation throughout the revised manuscript. We also agree that the original Discussion included aspects that, while interesting, were of secondary importance. In light of the reviewer’s suggestions, we have restructured and rebalanced the Discussion to focus more directly on the key findings and their implications. Importantly, we wish to clarify that we do not propose the elongation bottleneck model as a general mechanism across all conditions. In particular, for double (Leu/Ile) starvation, we attribute the observed effects primarily to stress response–mediated translational repression, and not to codon-specific stalling or tRNA depletion. We believe that this distinction is now more clearly conveyed in the revised manuscript.
Reviewer #3 (Evidence, reproducibility and clarity):
Summary
Worpenberg and colleagues investigated the translational consequences of branched-chain amino acid (BCAA) starvation in mouse cells. Limitation of individual BCAAs has been reported to cause codon-specific and global translational repression. In this paper, the authors use RNA-seq, ribosome profiling (Ribo-seq), proteomics, and tRNA charging assays to characterize the impacts of individual and combined depletion of leucine, isoleucine, and valine on translation. They find that BCAA starvation increases codon-specific ribosome dwell times, activates global translational stress responses and reduces global protein synthesis. They infer that this effect is due to decreased translation initiation and codon-specific translational stalling. They find that the effects of simultaneous depletion are non-additive. In valine and triple (valine, leucine, and isoleucine) depletion, they show that affected transcripts have a high density of valine codons early in their coding sequences, creating an "elongation bottleneck" that obscures the impact of starvation of other amino acids. Finally, they identify isoacceptor-specific differences in tRNA charging that help explain the codon-specific effects that they observe.
We find the major findings convincing and clear. We find that some results are incompletely explained. We suggest an additional experiment and also have some minor comments that we hope will improve clarity and rigor.
We thank the reviewer for the thorough and constructive feedback. We appreciate the recognition of our main findings and the helpful suggestions for improving the manuscript. Below we address each point in detail.
Major comments
Figure 3O: In this figure and the associated text, the authors try to determine whether differences in protein degradation can explain why some proteins have higher ribosome density but lower proteomic expression. However, since this analysis relies on published protein half-lives from non-starvation conditions and on the assumption that protein synthesis has entirely stopped, we are not convinced it is informative for this experimental context. It does not distinguish between a model in which protein synthesis has been reduced by stalling and a model in which both protein synthesis and degradation rate have increased, which are both consistent with their Ribo-seq and proteomic data. To address this issue, the authors should either perform protein half-life measurements under their starvation conditions, or more clearly explain these two models in the text and acknowledge that they cannot distinguish between them.
Response: We agree with the reviewer that our current analysis, which is based on protein half-lives obtained under non-starvation conditions, can not definitively separate the effects of reduced translation from those of increased protein degradation. We have revised the relevant section in the manuscript to more clearly state that this analysis is correlative in nature and serves only to explore one possible explanation for the observed disconnect between ribosome density and protein levels. We now also explicitly acknowledge that our dataset does not allow us to distinguish between a model in which protein output is reduced due to stalling and one in which both translation and degradation rates are altered. However, the observed log2FC in the proteomics data are often milder than expected based on complete-medium condition half-life alone, which would be difficult to reconcile with a dominant contribution from global protein destabilization. That said, we also acknowledge that protein degradation is highly context- and protein-specific, and that proteolytic regulation might still play a role. Performing a direct protein half-life measurement under our starvation conditions would indeed be required to rigorously test this, but such an experiment is outside the scope of this study. We now highlight this as a limitation and a valuable direction for future work, and we have softened any interpretations in the main text to reflect the uncertainty regarding the contribution of protein stability changes.
Minor comments
Figure 1G: Why does intracellular valine seem to be less depleted under starvation conditions than intracellular leucine or isoleucine? Are the limits of detections different for different amino acids? The authors should acknowledge this discrepancy and comment on whether it has any implications for interpretation of their results.
Response: We thank the reviewer for this important point. While valine appears slightly less depleted than leucine or isoleucine in Figure 1G, the fold changes and absolute reductions are strong for all three BCAAs, including valine. To further illustrate this, we have added a supplementary bar chart showing the measured intracellular concentrations in µmol/L, including mean and variance across five biological replicates (Supplementary Figure 5A). We believe that the variation may reflect technical factors, such as differences in detection sensitivity or ionization efficiency between amino acids in the targeted metabolomics assay and, therefore, that the observed difference does not have a meaningful impact on the interpretation of our results. We now directly acknowledge these differences in the main text.
Figure 1H: These data do not appear to meet the assumptions for linear regression. We suggest either reporting a Spearman R correlation (as the data appears linear in rank but not absolute value), or remove it entirely - we think the plot without statistics is sufficient.
Response: We thank the reviewer for the suggestion. In the revised manuscript, we removed the statistical annotation and retained only the trend line to illustrate the general pattern. We agree that this visualization alone is sufficient to support the qualitative point we aimed to convey.
Figure 2B: The in-text description of this figure states that "most" ISR genes show a "robust induction," but only three genes are shown in the figure, two of which are upregulated. The authors should instead specify that 2 out of the 3 genes profiled were robustly induced.
Response: We have rephrased the sentence to say “two of the three genes profiled…” for precision and consistency with the data shown.
Figure 2D: Please include the full, uncropped blots in the supplementary materials.
Response: We have now added the full, uncropped western blots to the supplementary material (Supplementary Figure 8).
Figure 2E: Swap the positions of the RPS6 and 4E-BP1 plots so they line up with their respective blots to make these figures easier to interpret. Authors should consider doing a one-way ANOVA and post-hoc analysis, if we correctly understand that they are making a conclusion about the difference between multiple groups in aggregate.
Response: We thank the reviewer for the suggestion. The alignment of the RPS6 and 4E-BP1 plots with their respective blots has been corrected. As this panel focuses on comparisons to the control condition only, we have retained the original presentation.
Figure 4B: Panel A in this figure is very convincing, and these plots don't add additional information. The authors could consider removing them. If this panel stays in, we suggest removing the "mid index" plot, since it is never referenced in the text and doesn't seem relevant to the message of the figure.
Response: We appreciate the feedback. While we considered removing panel B as suggested, we decided to retain it because it provides a useful summary of panel A. To improve clarity and visual interpretation, we replaced the original boxplot with a bar plot displaying mean values and SEM error bars. We believe the bar plot now nicely illustrates that Val and Triple starvation lead to stronger effects, especially in the reduction of the 3′ index. The “mid index” plot, which was not referenced in the text and did not contribute to the central message, has been removed as suggested.
Figure 4E: Why is there a reduction in frequency of a Leu and a Val codon under Ile starvation?
Response: Thank you for highlighting this observation. The reduction in the frequency of a specific Leu and Val codon under Ile starvation in Figure 4F (former Figure 4E) is indeed intriguing. This figure reflects codon usage in the first 20% of the CDSs among the subset of transcripts that exhibit a footprint polarization under each starvation condition. As such, the observed depletion likely arises from the specific transcript composition of the polarized subset under -Ile, which differs from that under -Val or other conditions. Importantly, this pattern is not consistently observed when analyzing the full transcripts (another Leu codon is affected), indicating that it is not a systematic depletion of these codons. One possibility is that an increased frequency of Ile codons (AUC) within the constrained region may lead to a relative underrepresentation of other codons, such as Leu and Val. Alternatively, this may reflect non-random codon co-occurrence patterns within specific transcripts. While our current data do not allow us to investigate this further, we acknowledge these as speculative explanations and now mention this point in the Discussion as a potential avenue for future study.
Figure 5G: There appears to be one Val codon early in the Hint1 transcript without much stalling under triple or valine starvation conditions. The authors should acknowledge this and comment on why this may be.
Response: We thank the reviewer for pointing this out. While the Hint1 transcript indeed contains a valine codon early in its CDS, no clear stalling peak was observed at that position under valine or triple starvation. Several factors may contribute to this: local sequence context can influence ribosome pausing, and not all cognate codons necessarily lead to detectable stalling even under amino acid starvation. Additionally, coverage at the 5′ end of Hint1 is relatively sparse in our dataset, and potential mappability limitations, such as regions with low complexity or repetitive elements, may further reduce resolution at specific sites. We now briefly mention this in the manuscript to clarify the possible causes.
Figure 5B: In the text referencing this figure, the authors state that "a high number of downregulated proteins with associated ribosome stalling sites did not show an overall decreased mean RPF count...as it would be expected from translation initiation defects, linking these stalling sites directly to proteomic changes." However, RPF is affected both by stalling (increases RPF) and initiation defects (decreases RPF). A gene with both stalling and decreased initiation may appear to have no RPF change. The data does suggest a contribution from stalling, but the authors should also acknowledge that reduced initiation may also be playing a role.
Response: We agree with the reviewer comment. Our cited statement should indeed be more nuanced. The reviewer correctly points out that RPFs are influenced by both increased ribosome density due to stalling and decreased ribosome density due to reduced initiation. Therefore, a gene experiencing both stalling and reduced initiation might appear to have no net change in RPF, or even a slight increase if stalling is dominant. Thus, while the presence of stalling sites strongly suggests a contribution from compromised elongation to reduced protein output, we cannot definitively rule out a concurrent role for reduced initiation, even in cases where RPF counts are not globally decreased. We revised this section in the manuscript to acknowledge this interplay.
Figure 5E: the black text on dark brown in the center of the Venn diagram is difficult to read. The diagram should either have a different color scheme, or the text in the center should be white instead of black for higher contrast.
Response: We have adjusted the text color for better contrast and improved readability.
Supplementary Figure 1C: The ribosome dwell time data in this study is described as "highly correlated" with another published dwell time dataset, but the P and E site data do not seem strongly correlated. The authors should remove the word "highly."
Response: We have removed the word “highly” to have a more cautious interpretation in the text.
Supplementary Figure 3E: Not all of the highlighted codons in this figure are ones with prolonged dwell times. To clarify the point that dwell time change is not related to codon frequency, this figure should only highlight codons that have a significantly prolonged dwell time in at least one starvation condition.
Response: We thank the reviewer for pointing this out. To improve clarity, we have revised the figure and now specifically highlight codons with significantly prolonged dwell times with stars.
Supplementary Figure 5C: The gene Chop is mentioned in the main text when referencing this figure, but is absent from the heatmap.
Response: We thank the reviewer for noting this. The gene Chop is annotated under its alternative name Ddit3 in the current version of the heatmap and is indeed present. To avoid confusion, we have now updated the label in the figure to display Chop (Ddit3) directly.
Supplementary Figure 7A: The authors could clarify this figure by adding additional language to either the figure panel or the figure legend specifying that the RPM metric being used comes from Ribo-seq.
Response: We have updated the legend to explicitly state that the RPM values shown are derived from Ribo-seq data.
Supplementary Figure 7D: The metric used to describe the spatial relationship between the first valine and isoleucine codons in transcripts in this figure seems to be describing something conceptually similar to the stalling sites in Figure 5G, but uses a different metric. These figures would be easier to interpret if these spatial relationships were presented in a consistent way throughout the manuscript.
Response: We thank the reviewer for this helpful observation. Supplementary Figure 7D (now Supplementary Figure 11B) originally used a gene-length-normalized metric to describe codon spacing, whereas Figure 5G depicted absolute nucleotide distances to stalling sites. To ensure consistency across the manuscript, we have now updated Supplementary Figure 11B to also use absolute distances. We believe this adjustment improves clarity and allows for a more direct comparison between spatial codon patterns and stalling events.
Discussion:
Reader understanding would be improved if the relevance of paragraphs were established in the first sentence. For instance, in the paragraphs about adaptive misacylation and posttranscriptional modifications, it is unclear until the end of the paragraph how these topics are relevant. Introducing the relevant aspects of the study (the fact that some starvation conditions have less severe effects and the observation about m6A-related mRNAs) at the beginning of these paragraphs would improve clarity.
Response: We thank the reviewer for this helpful comment. We agree that the flow and clarity of the Discussion can be improved by making the relevance of each paragraph clearer from the outset. In the revised manuscript, we have restructured these sections to better highlight the connection between each topic and our main findings. These changes also align with suggestions from Reviewer 2, and we believe they help to focus the Discussion more tightly around the core insights of our study.
The authors should provide more information and speculation about possible physiological relevance of their findings, particularly about the way that the effects of triple starvation are highly valine-dependent. Are there physiological conditions under which starvation of all three BCAAs is more likely than starvation of one or two of them? If so, are there any reasons why a valine-based bottleneck might be advantageous?
Response: We appreciate the reviewer's insightful question regarding the physiological relevance of our findings, particularly the valine-dependent bottleneck observed under triple BCAA starvation. This prompts a crucial discussion on the broader biological context of our work.
While complete starvation of all three BCAAs might be less frequent than individual deficiencies, such conditions are physiologically relevant in several contexts. In prolonged fasting, starvation, or severe cachectic states associated with chronic diseases (e.g., advanced cancer, critical illness), systemic amino acid pools, including BCAAs, can become significantly depleted due to increased catabolism and insufficient intake (Yu et al. 2021). Moreover, certain specialized diets or therapeutic strategies aim to modulate BCAA levels. For instance, in some Maple Syrup Urine Disease (MSUD) management protocols, BCAA intake is severely restricted to prevent the accumulation of toxic BCAA metabolites (Mann et al. 2021). Similarly, emerging cancer therapies sometimes explore nutrient deprivation strategies to selectively target tumor cells, which could involve broad BCAA reduction (e.g. Sheen et al. 2011; Xiao et al. 2016).
In these contexts, a valine-based bottleneck, as we describe, could indeed represent an adaptive strategy. If valine-tRNAs are particularly susceptible to deacylation and valine codons are strategically enriched at the 5' end of transcripts, stalling at these early positions could serve as a rapid "gatekeeper" for global translation. This early-stage inhibition would conserve cellular energy and available amino acids by quickly reducing the overall demand for charged tRNAs. Such a mechanism could potentially prioritize the translation of a subset of proteins that might have different codon usage biases or are translated via alternative, less valine-dependent mechanisms. This aligns with the concept of a multi-layered translational control where global initiation repression (as reflected in mTORC1 inhibition and polysome profiles) is complemented by specific elongation checkpoints, allowing for a more nuanced and adaptive response to severe nutrient stress.
Reviewer #3 (Significance):
Nature and significance of the advance
The main contribution of this work is to demonstrate that depletion of multiple amino acids simultaneously impacts translation elongation in ways that are not necessarily additive. These impacts can depend on the distribution of codons in a transcript. It adds to a growing body of work showing that essential amino acid starvation can cause codon-specific ribosome stalling. The authors suggest that the position-dependent stalling they observe could be a novel regulatory mechanism to alleviate the effects of multi-amino acid starvation. However, it is not fully clear from the paper what the significance of a valine-based regulatory adaptation to BCAA starvation is, or whether simultaneous starvation of all three BCAAs is of particular physiological relevance. The paper's primary contribution is mainly focused on the similarity between valine and triple BCAA starvation, and it provides limited insight into the effects of combined depletion of two BCAAs.
Context of existing literature
Although ribosome profiling does not distinguish between actively-elongating and stalled ribosomes, sites with higher read coverage, and thereby higher inferred dwell time, can be used to infer ribosome stalling (Ingolia 2011). Various downstream effects of essential amino acid depletion have been documented, such as leucine deficiency being sensed by mTORC1 via leucyl-tRNA synthetase (Dittmar 2005, Han 2012), and shared transcriptional responses among many amino acid depletion conditions (Tang 2015). These authors have previously measured the translational effects of nutrient stress using ribosome profiling (e.g., Gobet 2020), as have others (Darnell 2018, Kochavi et al. 2024). The present work represents the first study (to our knowledge) combining BCAA depletions, representing an incremental and useful contribution to our understanding of translational responses to stress conditions.
Audience
This work is of interest to investigators studying the response of human cells in stress conditions, such as in human disease, as well as investigators studying the basic biology of eukaryotic translational control.
Reviewer expertise: mRNA decay and translation regulation in bacteria.
We hope the authors have found our comments thoughtful and useful. We welcome further discussion or clarification via email: Juliana Stanley (julianst@mit.edu) and Hannah LeBlanc (leblanch@mit.edu).
We sincerely thank the reviewers for their thoughtful and constructive feedback, as well as for their careful and thorough reading of our manuscript. We also gratefully acknowledge the invitation for further discussion and would be happy to engage in future correspondence.
References
“¿Educamos para pensar o para obedecer?” En muchas aulas, el silencio es confundido con disciplina, y la repetición con aprendizaje. Se espera que los estudiantes memoricen, repitan y aprueben, pero no siempre que cuestionen. ¿Qué ocurre cuando un estudiante levanta la mano para disentir? ¿Lo celebramos como pensamiento crítico o lo corregimos como falta de respeto? Educar no debería ser domesticar. El pensamiento crítico nace cuando se permite la duda, cuando se enseña a leer entre líneas, a identificar intenciones, a reconocer que todo texto tiene un contexto y todo autor una postura. Si queremos formar ciudadanos libres, debemos enseñarles a pensar, no solo a responder.
* Give up on (modismo) = rendirme con.
* El with en el ingles es mas limitado que en el español. Solo se usa para indicar compañía como un adicional a la oración que complementa sin cambiar el significado de esta. Ejemplo: I wrote a thesis with my best friend.
ooperatin Teac het CI for SI rR are committed educators work- g s ( 8) ; i i ining the next ing i Francisco classrooms and are interested in a g eee ae tion of 5 have successfully established produc generation of teachers. ©P iti ossess strong classroom manage- 7 ‘ ommunities, p . : and caring learning ¢ ntiated student-centered instruction o i i differe t skills, and deliver ney ander ns Francisco neighborhoods that have been traditi . ve - . n srve hemselves as teacher educators, are i , served. CTs see t ar ed to aeep- . . e comfortable making their practice public, and ar ening their own skills as teachers and mentors.
For mentors: learn new technologies, fresh ideas, and strengthen reflective practice.
For schools: improved teaching quality, collaborative culture, and equity in underserved communities.
ons Tap to enable a layout that focuses on the article. Focus mode setTimeout(()=>{try{if(-1===document.cookie.indexOf("c_mId="))return;const e=window.localStorage.getItem("FocusMode");if(!e)return;if(!JSON.parse(e).enabled)return;const o=document.querySelector(".focus-toggle"),t=o?o.querySelector(".toggle-switch-button"):void 0;if(!o||!t)return;document.documentElement.classList.add("focus","focus-enabled"),o.classList.remove("hidden"),t.classList.add("is-checked")}catch(e){console.warn("Error retrieving data for Focus Mode",e)}},0) Subscribe or Log In Profile Sign Out Show Search Search Query Submit Search Advertisement California The 9 LGBTQ+ children’s books targeted in high court ruling upending education policy A selection of books featuring LGBTQ+ characters that are part of a Supreme Court case are pictured April 15 in Washington. (Pablo Martinez Monsivais / Associated Press) By Jenny GoldStaff Writer Follow June 27, 2025 8:01 PM PT 8 Share via Close extra sharing options Email Facebook X LinkedIn Threads Reddit WhatsApp Copy Link URL Copied! Print Picture books are not usually the stuff of Supreme Court rulings. But on Friday, a majority of justices ruled that parents have a right to opt their children out of lessons that offend their religious beliefs — bringing the colorful pages of books like “Uncle Bobby’s Wedding” and “Pride Puppy” into the staid public record of the nation’s highest court.The ruling resulted from a lawsuit brought by parents in Montgomery County, Md., who sued for the right to remove their children from lessons where LGBTQ+ storybooks would be read aloud in elementary school classes from kindergarten through 5th grade. The books were part of an effort in the district to represent LGBTQ+ families in the English language arts curriculum.In a 6-3 decision, the Supreme Court ruled that schools must “notify them in advance” when one of the disputed storybooks would be used in their child’s class, so that they could have their children temporarily removed. The court’s three liberals dissented. Advertisement Politics Parents may pull their children from classes that offend their religion, Supreme Court rules Supreme Court hands down a major victory for parents’ rights June 27, 2025 As part of the the decisions, briefings and petitions in the case, the justices and lawyers for the parents described in detail the story lines of nine picture books that were part of Montgomery County’s new curriculum. In her dissent, Justice Sonia Sotomayor even reproduced one, “Uncle Bobby’s Wedding,” in its entirety. Here are the nine books that were the subject of the case:Pride PuppyAuthor: Robin Stevenson Illustrator: Julie McLaughlin Book “Pride Puppy” published by Orca Book Publishers. (Orca Book Publishers) “Pride Puppy,” a rhyming alphabet book for very young children, depicts a little girl who loses her dog during a joyful visit to a Pride parade. The story, which is available as a board book, invites readers to spot items starting with each of the letters of the alphabet, including apple, baseball and clouds — as well as items more specific to a Pride parade.Lawyers representing the parents said in their brief that the “invites students barely old enough to tie their own shoes to search for images of ‘underwear,’ ‘leather,’ ‘lip ring,’ ‘[drag] king’ and ‘[drag] queen,’ and ‘Marsha P. Johnson,’ a controversial LGBTQ activist and sex worker.”The “leather” in question refers to a mother’s jacket, and the “underwear” to a pair of green briefs worn over tights by an older child as part of a colorful outfit. Advertisement The Montgomery County Public Schools stopped teaching “Pride Puppy” in the midst of the legal battle. California As children’s book bans soar, sales are down and librarians are afraid. Even in California Book bans are tanking sales of children’s books. Schools and libraries aren’t buying books about LGBTQ+ issues and race as they brace for culture war pushback. Dec. 12, 2024 Love, VioletAuthor: Charlotte Sullivan WildIllustrator: Charlene Chua Book “Love Violet” published by macmillan publishers. (macmillan) The story describes a little girl named Violet with a crush on another girl in her class named Mira, who “had a leaping laugh” and “made Violet’s heart skip.” But every time Mira tries to talk to her, Violet gets shy and quiet.On Valentine’s Day, Violet makes Mira a special valentine. As Violet gathers the courage to give it to her, the valentine ends up trampled in the snow. But Mira loves it anyway and also has a special gift for Violet — a locket with a violet inside. At the end of the book, the two girls go on an adventure together.Lawyers for the parents describe “Love, Violet” as a book about “two young girls and their same-sex playground romance.” They wrote in that “teachers are encouraged to have a ‘think aloud’ moment to ask students how it feels when they don’t just ‘like’ but ‘like like’ someone.” Advertisement Born Ready: The True Story of a Boy Named PenelopeAuthor: Jodie Patterson Illustrator: Charnelle Pinkney Barlow Book “Born Ready” published by Random House. (Random House) In “Born Ready,” 5-year-old Penelope was born a girl but is certain they are a boy. “I love you, Mama, but I don’t want to be you. I want to be Papa. I don’t want tomorrow to come because tomorrow I’ll look like you. Please help me, Mama. Help me be a boy,” Penelope tells their mom. “We will make a plan to tell everyone we know,” Penelope’s mom tells them, and they throw a big party to celebrate.In her dissent, Sotomayor notes, “When Penelope’s brother expresses skepticism, his mother says, ‘Not everything needs to make sense. This is about love.’ ” In their opening brief, lawyers for the families said that “teachers are told to instruct students that, at birth, people ‘guess about our gender,’ but ‘we know ourselves best.’ ”Prince and Knight Author: Daniel Haack Illustrator: Stevie Lewis “Prince and Knight” is a story about a prince whose parents want him to find a bride, but instead he falls in love with a knight. Together, they fight off a dragon. When the prince falls from a great height, his knight rescues him on horseback. When the king and queen find out of their love, they “were overwhelmed with joy. ‘We have finally found someone who is perfect for our boy!’ ” A great wedding is held, and “the prince and his shining knight would live happily ever after.”“The book Prince & Knight clearly conveys the message that same-sex marriage should be accepted by all as a cause for celebration,” said Justice Samuel Alito, who wrote the majority opinion, a concerning message for Americans whose religion tells them that same-sex marriage is wrong.
This is just about acceptance and not really conforming into certain views
Street
Creo que esta foto es diferente porque esta muestra amistad. Algo que no está en las fotos de las mujeres. Ellas están solas con sus hijos, sin amigas o hombres. En pequeño espacios no en la comunidad.
Library
Las mamás parecen triste o enojada y las caras de los niños son son felices realmente tampoco. Esta chica tiene un poco de sonrisa en la cara, pero tal vez alguien esta hablando con ella porque todavía hay un poco de tristeza a tras de los ojos.
workers
Siento que en cada foto las mujeres parecen más mayor. Las caras tienen arrugas profundas. ¿Pensé es que la vida es duro o estas mamás tuvieron hijo mas tarde en sus vidas?
o launch a little ship on love’s storm-tossed seas, then only ten more left like rows of beans.
Rhyming couplet
Note: This response was posted by the corresponding author to Review Commons. The content has not been altered except for formatting.
Learn more at Review Commons
Manuscript number: RC-2025-03111
Corresponding author(s): Qingyin Qian and Ryusuke Niwa
We would like to thank reviewers for their feedback on our initial submission. Changes in figures were noted in the point-to-point reply. For submission of our current revised manuscript, we provide two Word files, which are the “clean” and “Track-and-Change” files. Page and line numbers described below correspond to those of the “clean” file. The “Track-and-Change” file might be helpful for Reviewers to find what we have changed for the current revision.
In the revised manuscript, major changes in the text were tracked, while minor edits in figure numbers and legends were not tracked. In the Discussion, the section “Xrp1-mediated EE plasticity…” was moved before “Xrp1, a transcription factor …”, to follow the order of the Results, and was split into two: “EE plasticity …” and “Xrp1-mediated EE plasticity …”.
- The authors should investigate the regenerative growth of the adult midgut after irradiation. Is there an impact on ISCs proliferation or cell turn over. Is Xrp1 in EEs required in this adaptive response. It would be elegant to use the recently generated tracing method by Tobias Reiff lab to observe overall impact on tissue renewal (rapport-tracing esglexReDDM esg-lexA, 13xLexAop2-CD8::GFP, 13xLexAop2-H2B::mCherry::HA, tub-Gal80ts on the second chromosome. It can be combined with any EEs Gal4-driver (see Nat Commun 2025, https://doi.org/10.1038/s41467-024-55664-2, the stock is already existing, see table1). This reviewer thinks that it is a key experiment to support the proposed model.
2.1. Author response:
We will conduct the following experiments to answer these criticisms.
(1) We will investigate the ISC behavior, proliferation and differentiation, after 100 Gy of radiation by examining changes in the number of progenitor cells and their progenies, using esgtsF/O (esg-Gal4, UAS-GFP, tub-Gal80ts; Act>Cd2>Gal4, UAS-Flp) generated in the study (Jiang et al. Cell 2009 DOI: 10.1016/j.cell.2009.05.014) or esgReDDM (esg-Gal4, UAS-CD8::GFP; UAS-H2B::RFP, tubGal80ts) generated in the study (Antonello et al. EMBO J. 2015 DOI: 10.15252/embj.201591517). Flies will have progenitor cell lineages traced for 7 days, irradiated on day 6, and examined at different time points after radiation, following the design shown in Fig. 2A. Based on the previous findings (Sharma et al. Sci. Rep. 2020 DOI: 10.1038/s41598-020-75867-z; Pyo et al. Radiat. Res. 2014 DOI: 10.1667/RR13545.1), we anticipate that radiation compromises ISCs’ proliferation and differentiation. Should this be the case, our results can be interpreted in relation to those earlier studies.
(2) In parallel, we will examine whether Xrp1 expression in EEs affects radiation-induced ISC behaviors. As suggested, we will use “EE Rapport” (esg-lexA, 13xLexAop2-CD8::GFP, 13xLexAop2-H2B::mCherry::HA, tub-Gal80ts; Rab3-Gal4) generated in the study (Zipper et al. Nat. Commun. 2025 DOI: 10.1038/s41467-024-55664-2) and compare control flies to flies with Xrp1 knocked down in EEs to assess the impact on ISC behaviors.
- Is p53 required for Xrp1 induction in the gut after irradiation?
2.2. Author response:
To answer this point, we will perform immunostaining of anti-Xrp1 antibody to examine whether p53 is required for Xrp1 induction in irradiated flies with p53 knocked down in EEs.
- Xrp1 over expression has been shown to induce upd3 ligand and nutrient-driven dedifferentiation of enteroendocrine cells is occuring by activation of the JAK-STAT pathway (DOI: 10.1016/j.devcel.2023.08.022). Could the authors test the function of this signaling pathway during irradiation (upd3-lacZ and Stat-GFP can be used in parallel of upd3 RNAi and UAS Dome-DN.
2.3. Author response:
We will conduct the following experiments to answer these points.
(1) We will examine the cell type in which upd3 ligand induction occurs after radiation by using the upd3.1-LacZ reporter generated in the study (Jiang et al. Cell Stem Cell 2011 DOI: doi.org/10.1016/j.stem.2010.11.026).
(2) One possibility is that upd3.1-LacZ is detected in EEs. In this case, we will examine the requirement of upd3 in EEs for radiation-induced EE plasticity by knocking down upd3. Another possibility is that upd3.1-LacZ is detected in non-EE cells. If so, we will examine the requirement of the JAK-STAT pathway in EEs by overexpressing dome[△cyt] generated in the study (Brown et al. Curr. Biol. 2001 DOI: 10.1016/s0960-9822(01)00524-3) or knocking down Stat92E in EEs. Because these conditions are not mutually exclusive, both approaches may be pursued, with the latter relating our results to nutrient-driven EE dedifferentiation.
- Xrp1 is known for its role in cell competition and elimination of looser cells by induction of apoptosis. It would be interesting to check for induction of cell death and/or caspase activation in the fly gut after irradiation and verify a non apoptotic role of DRONC activation in this context using a Dronc RNAi (as proposed by Bergmann lab (https://doi.org/10.1038/s41598-021-81261-0) or Baena-Lopez lab (DOI: 10.15252/embr.201948892)). Overexpression of Xrp1 could be combined with UAS-p35.
2.4. Author response:
To address these points, we will investigate apoptosis induction following radiation with anti-cleaved Dcp-1 immunostaining. Based on the previous finding (Sharma et al. Sci. Rep. 2020 DOI: 10.1038/s41598-020-75867-z), we anticipate seeing increased cleaved Dcp-1 signals in all cell types after radiation. We intend to clarify whether radiation increases the ratio of apoptotic EEs among EEs; however, we cannot yet be certain whether it will be feasible.
Regarding Dronc activation, we previously requested the antibody used in the study (Wilson et al. Nat. Cell Biol. 2002 DOI: 10.1038/ncb799; Lindblad et al. Sci. Rep. 2021 DOI: 10.1038/s41598-021-81261-0) and tested it in our context, after radiation and by Xrp1-S O/E in EEs. We present our data below. In the anterior midgut, anti-Dronc signals were not observed under both control conditions. After radiation and by Xrp1-S O/E in EEs, anti-Dronc signals were seen in part of past EEs (#2 past) and progenitor cells (#3 prgn), implying their EB identity. However, anti-Dronc signals were never observed in current EEs (#1 current), suggesting Dronc does not act directly downstream to Xrp1.
We will address UAS-p35 in 3.3. Author response and Dronc-RNAi in 4.2. Author response.
- The authors do not justify or explain why they used 100 Gy of radiation. This is higher than doses used in comparable regeneration studies in adult Drosophila (e.g., PMID25959206, PMID: 28925355). The authors should clarify why this dose was chosen.
2.5. Author response:
Our initial rationale was based on the paper (Sharma et al. Sci. Rep. 2020 DOI: 10.1038/s41598-020-75867-z), where the authors claimed that ISC proliferation was inhibited and the ISC number was decreased by 100 Gy of radiation.
Nevertheless, we understand the reviewer’s concern and will examine 50 Gy of radiation as used in the papers the reviewer listed. We will examine radiation-induced changes in EE lineages and ISC behaviors. Depending on the results, we will evaluate whether and how they should be incorporated into the manuscript.
- Fig. 2C, the number of past EE’s increased transiently so that baseline number is restored at 18 hr after IR. The authors conclude that fate plasticity is a transient event. Can they rule out loss due to cell death?
2.6. Author response:
In our system, past EEs were detected transiently but did not persist. We agree that we cannot distinguish whether the transient appearance of past EEs reflects transient adoption of another identity that ends in cell death or reversible plasticity.
To partially address this criticism, as noted in 2.4. Author response, we will examine the apoptosis marker cleaved Dcp-1, which also tests whether cleaved Dcp-1-positive cells can be past EEs. However, regardless of detecting apoptosis markers in past EEs, we have changed “transient” into “temporary” to describe a short-lived cell state (see Page 8, Line 178; Page 15, Line 338).
- They authors interpret fate-conversion as beneficial for tissue repair but never test whether blocking this process impairs recovery or organismal survival or whether promoting it improves outcomes.
2.7. Author response:
We have removed this potentially misleading interpretation (see Page 4, removed the last part of the previous introduction, “and propose the possibility that such plasticity contributes to tissue repair”). We present below the data showing a severe reduction of the ISC number in 7-day post-radiation guts, suggesting the inability of tissue repair. We will add this to the manuscript together with results from the following experiments.
(1) We will examine if the blockage of radiation-induced EE plasticity, via knocking down Xrp1 in EEs, alters the epithelial cell number and cell junction protein localization.
(2) To complement the result of plasticity inhibition, we attempt to promote plasticity by overexpressing Xrp1 in EEs, to test whether this rescues ISC loss or restores junctions.
Should knockdown worsen ISC loss and junction integrity, or overexpression rescue them, we will describe EE plasticity as beneficial; otherwise, we will present it as a radiation-induced response without inferring benefits, while noting our limitations.
We will address organismal survival in 4.3. Author response.
- Related to the above, it would be helpful to know if fate-converted cells function as true ISCs or ECs (e.g., through proliferation or absorption assays).
2.8. Author response:
To partially answer this criticism, we will examine whether EE-derived ISCs are proliferative by examining whether they can be positive for the mitotic marker phospho-histone 3.
We will address absorption assays in 4.4. Author response.
- It is surprising to observe EEs dedifferentiation at a steady state during homeostasis, a condition in which Xrp1 is not detected in the gut. Can the authors comment this point in the discussion?
3.1. Author response:
We have added our thoughts in terms of Xrp1 being not detectable in homeostatic EE lineages (see Page 15, Line 350 - 356). We have also added our thoughts regarding observation of EE plasticity in homeostatic guts (see Page 14, Line 322 - 332).
- Xrp1 is existing as a short of long isoforms. The short form has been recently proposed to be required for cell competition (https://doi.org/10.1101/2025.06.15.659587) whereas Xrp1 long isoform may be responsible for reduced cell growth. Could the authors test which isoform is induced in the gut after irradiation? Is the overexpression of Xrp1 long isoform having the same effect that the short isoform used by the authors.
3.2. Author response:
We have added data on the effect of Xrp1 long isoform overexpression on EE plasticity (see Fig. 5A - 5B, Page 12, Line 276 - 278), showing that overexpression of the Xrp1 long isoform caused a similar increase in past EEs. In addition, we have changed Xrp1 O/E to Xrp1-S O/E in the contents related to Figs 4, 5, S4, and S5.
We will address radiation-induced Xrp1 isoforms in 4.1. Author response.
- Xrp1 is known for its role in cell competition and elimination of looser cells by induction of apoptosis. It would be interesting to check for induction of cell death and/or caspase activation in the fly gut after irradiation and verify a non apoptotic role of DRONC activation in this context using a Dronc RNAi (as proposed by Bergmann lab (https://doi.org/10.1038/s41598-021-81261-0) or Baena-Lopez lab (DOI: 10.15252/embr.201948892)). Overexpression of Xrp1 could be combined with UAS-p35.
3.3. Author response:
We have added data regarding p35 O/E combined with Xrp1 O/E, showing that p35 O/E did not further increase the number of past EEs, thereby suggesting that Xrp1-driven EE plasticity has a non-apoptotic nature (see Fig. 5C - 5D, Page 13, Line 293 - 297).
- Line 221: fig S3E should be S3F
- Line 230: fig S3F-G should be S3G-H
3.4. Author response:
We have fixed this error.
- The posterior gut region R4 is more proliferative than the anterior part and is usually used for testing regenerative growth. What is happening there after irradiation?
3.5. Author response:
We present below radiation-induced changes in EE lineages and ISC number in the R4bc gut region. Radiation did not alter the proportion of past EEs among EE lineages but reduced the ISC number. We acknowledge differences between anterior and posterior gut regions, but we do not plan to further analyze regional differences or underlying mechanisms.
- The authors’ explanation for cells with weak GFP in Figure 1 is not convincing. Induction of GFP is an all or nothing event as it results from Pros-driven FLPase and a recombination that removes the transcription stop signals to express GFP from a Ubi promotor. Once that happens, it should not matter how strong or weak Pros is, GFP should be the same. So, another explanation is needed. Nuclear staining of cell #2 in Fig 1B resembles a metaphase chromosome arrangement. Nuclear GFP may appear ‘weak’ in mitosis as the nuclear envelope breaks down. It is positive for the purple Pros/Dl stain, which makes it hard to tell if it is Pros+ or Pros- even though the authors state that cells with weak GFP are Pros- in line 104 (see the point above regarding confusing same-color stain for ISC and EE markers). Could cell #2 be a pre-EE that is undergoing mitosis since the lineage tracer marks both EE and pre-EE cells (line 119)? Or do the authors mean recombination on one or both homologs? This should not be possible since the cells are heterozygotes for the Ubi-GFP locus.
3.6. Author response:
For cell #5, RFP- GFPweak may result from the leakiness of the G-TRACE system. We have added our observations of the G-TRACE strains and changed our previous explanation (see Fig. S1B - S1C, Page 5, Line 94 - 97, 103 - 106).
For cell #2, we agree that RFP+ GFPweak cells may either be a cell turning on pros expression just before sample preparation or a pre-EE undergoing mitosis. Nevertheless, it is not a past EE that has lost the EE marker Pros, so it is considered a current EE. We have removed our previous interpretation of cell #2 (see Page 5, removed “which likely had not yet fully activated recombination”), and changed the image to avoid confusion (see Fig. 1C).
- Fig. 2C, if past-EE’s increased in number while current EE’s stayed the same, where are new past-EE’s coming from? There cannot be compensatory proliferations since EE’s are post-mitotic. For fate conversion, one would expect the generation of each past-EE to accompany loss of one current EE.
3.7. Author response:
We agree that the generation of one past EE should be accompanied by the loss of one current EE. We do not have a clear answer to this question. Our data showed cell numbers per ROI rather than the total cell number across the whole gut. To address this, we have changed the number to the proportion, calculated from [past EE] / ([past EE] + [current EE]), in experiments examining damage-induced EE plasticity, which provides a more informative measure for EE fate conversion (see Fig. 2C, also Fig. S2B and 3E).
- Fig. 2E. Dl+ past-EE cell number declined at 14 and 18 h after IR and because cell sized increased, the authors conclude that EE cells that de-differentiated into ISCs subsequently re-differentiated into EC’s. To reach this conclusion, the authors should count past-EEs that are positive for EC markers. Cell size alone is insufficient evidence.
3.8. Author response:
We have added data quantifying the proportion of past EEs that are positive for the EC marker Pdm1, showing that past EEs were more likely to be ECs in guts examined 14 h after radiation (see Fig. 2F - 2G, Page 9, Line 189).
- Fig. 6. Where are the % numbers for ISC, EB and EE’s coming from? And wouldn’t these change with time after IR, etc?
3.9. Author response:
The numbers came from the calculation of the percentage of the absolute values of control and 14 h post-IR conditions from Fig. 2E. These numbers changed with time after radiation. We realized that the precise numbers were misleading. We therefore have removed such illustration and instead added phrases “more current EEs → past EEs, more past EEs being ISCs → past EEs being ECs” to describe the increase in past EE cell number and the shift in the composition of past EEs (see Fig. 6).
- Improve Figure 1B: Pros and Dl are shown in the same color, creating confusion. If both are stained together, different colors or clearer labeling should be used. Clarify how cells are identified as Pros+ vs Dl+.
3.10. Author response:
Anti-Pros and anti-Dl antibodies were produced from the same host species and were detected with the same secondary antibody, so they were in the same color. We have stated that solid nuclear staining indicates Pros, whereas punctate cytoplasmic staining indicates Dl (see Page 5, Line 100, 102, and 103). Such staining has been reported in previous studies (for example, Fig. 2A - 2B, Veneti et al. Nat. Commun. 2024 DOI: 10.1038/s41467-024-46119-9).
- Why is Dl (supposed to be cytoplasmic) overlapping with nuclear GFP in cells #3 and 4 in Fig. 1B?
3.11. Author response:
Because Dl signals were located apically to DAPI/GFP signals, the overlap was likely due to Z-projection from stacked slices. We present below orthogonal slices along the z-axis, from top to bottom by row, and composite and individual color channels, from left to right by columns, for cell #3 (left) and cell #4 (right).
For cell #3, Dl signals were present in slices 1/8 and 2/8 and disappeared in slice 3/8, whereas DAPI signals appeared from slice 2/8. For cell #4, Dl signals surrounded DAPI signals when viewed separately. In addition, we realized that nuclear GFP signals slightly outgrew DAPI signals, despite our confirmation that the GFP channel was not saturated.
We have included separate color channels for DAPI signals and Pros, Dl and DAPI merged channels, showing that Dl signals were absent from the nucleus. For cell #3, in which the nuclear DAPI and cytoplasmic Dl cannot be distinguished in the stacked view, we show the images from a single orthogonal slice in the main panel, and the image from stacked slices as insets (see Fig. 1C).
- Fig. S1E and F. Very hard to see what the authors describe about Arm and Cora. One problem is that cell boundaries are not visible, just the nuclei, so it is hard to know whether cell-cell interactions the authors describe as normal are really normal. Another problem is the overlap of Arm (supposed to be cytoplasmic) with the nuclear GFP signal. What is that?
3.12. Author response:
Regarding the invisibility of cell boundaries, we have improved the image of anti-Cora staining and added anti-Mesh staining and a separate color channel for DAPI signals to reinforce junction integrity (see Fig. S1H - S1I).
Regarding the overlap of Arm signals with nuclear GFP signals, we realized similar problems as those noted in 3.11. Author response. We present below orthogonal slices along the z-axis and combined and individual color channels, for cell #2 (left) and cell #3 (right). For both cells, Arm signals did not overlap with DAPI signals. We have adjusted the maximum intensity projection to include slices 1-4 instead of 1-8 and added a separate color channel for DAPI signals to avoid the signals appearing to overlap (see Fig. S1G).
- Include a simple schematic of ISC to EE/EC lineages for readers unfamiliar with Drosophila gut biology.
3.13. Author response:
We have included a schematic (see Fig. 1A). Although not requested, we have also improved Fig. 1B to enhance clarity.
- Discuss the regional difference in Xrp1 efficacy (R2a vs R2b). Is there something known about gene expression differences in different gut regions that can explain the results?
3.14. Author response:
At present, we do not have an explanation for these results. We have refined our discussion regarding such regional differences (see Page 16 - 17, Line 381 - 390).
- Consider moving scRNAseq (Fig. S1G) into main paper: this is a central part of the conclusion.
3.15. Author response:
We have moved Fig. S1G, as well as Fig. S1H and S1I, into the main figure (see Fig. 1G - 1I).
- Xrp1 is existing as a short of long isoforms. The short form has been recently proposed to be required for cell competition (https://doi.org/10.1101/2025.06.15.659587) whereas Xrp1 long isoform may be responsible for reduced cell growth. Could the authors test which isoform is induced in the gut after irradiation? Is the overexpression of Xrp1 long isoform having the same effect that the short isoform used by the authors.
4.1. Author response:
We prefer not to distinguish whether the long or short Xrp1 isoform is induced in the gut after radiation. This presents technical challenges and falls outside the scope of the present study. As noted in 3.2. Author response, we instead report in the revised manuscript that both isoforms similarly promote EE plasticity.
- Xrp1 is known for its role in cell competition and elimination of looser cells by induction of apoptosis. It would be interesting to check for induction of cell death and/or caspase activation in the fly gut after irradiation and verify a non apoptotic role of DRONC activation in this context using a Dronc RNAi (as proposed by Bergmann lab (https://doi.org/10.1038/s41598-021-81261-0) or Baena-Lopez lab (DOI: 10.15252/embr.201948892)). Overexpression of Xrp1 could be combined with UAS-p35.
4.2. Author response:
We prefer not to perform Dronc-RNAi, because we did not observe Dronc activation downstream to Xrp1, as shown in 2.4. Author response.
- They authors interpret fate-conversion as beneficial for tissue repair but never test whether blocking this process impairs recovery or organismal survival or whether promoting it improves outcomes.
4.3. Author response:
We prefer not to examine organismal survival. We agree that organismal survival would be informative, but our study focuses on epithelial cell number, which will be tested as noted in 2.7. Author response. We will not mention broad claims at the organismal level.
- Related to the above, it would be helpful to know if fate-converted cells function as true ISCs or ECs (e.g., through proliferation or absorption assays).
4.4. Author response:
We prefer not to perform absorptive assays due to technical challenges. We will instead test proliferation, as noted in 2.8. Author response, and note our limitations.
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DOI: 10.4103/NRR.NRR-D-23-01965
Resource: (Thermo Fisher Scientific Cat# A-11008, RRID:AB_143165)
Curator: @scibot
SciCrunch record: RRID:AB_143165
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DOI: 10.1242/jcs.263779
Resource: RRID:Addgene_12260
Curator: @olekpark
SciCrunch record: RRID:Addgene_12260
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DOI: 10.1172/jci.insight.191245
Resource: None
Curator: @olekpark
SciCrunch record: RRID:Addgene_19044
RRID:AB_2138147
DOI: 10.1016/j.isci.2025.113246
Resource: (Abcam Cat# ab92434, RRID:AB_2138147)
Curator: @scibot
SciCrunch record: RRID:AB_2138147
Gregory, o' my word, we'll not carry coals.
What does this mean? carrying coals during this time period signified a person's low status, equaling little or giving no respect. Sampson was essentially saying that they will tolerate no disrespect from the house of Montague. Right out the gate, you certainly sense a high level of disregard from Sampson for house of Montague.
Valenzuela et al. (2020) llegaron a la conclusión de que el 90% está satisfecho con la COI y que al tener una percepción favorable produce en ellos SL.También se determinó el nivel de SL y sus dimensiones, de los cuales tenemos que: la SL está en un nivel alto con un 94.2%; las condiciones físicas y/o materiales, en nivel alto, con un 71.7%; los beneficios laborales y/o remunerativos, en un nivel alto, con un 67.5%; las políticas administrativas, en un nivel alto, con un 80%; las relaciones sociales, en nivel alto, con un 82.5%; el desarrollo personal, en nivel alto, con un 76.7%; el desempeño de tareas, en nivel alto, con un 80.8%; finalmente, la relación con la autoridad está en un nivel alto, con un 82.5%
Es revelador que, aunque el salario es importante (67.5%), la satisfacción laboral en la municipalidad es alta (94.2%) gracias a factores como la buena relación con los jefes (82.5%) y un ambiente de trabajo positivo (82.5%). Esto demuestra que un buen clima laboral y sentirse valorado son tan o más cruciales que los beneficios económicos para la felicidad de los empleados.
Para el procesamiento de los datos de esta investigación se hizo uso del paquete estadístico SPSS 26 y del programa Excel. Para determinar el uso del coeficiente adecuado, se realizó la prueba de normalidad Kolmogorov-Smirnov, de acuerdo a los resultados que dieron un nivel de sig. Menor a 0.05 se tomó la decisión de utilizar el coeficiente Rho Spearman. En cuanto a la regla de para determinar la existencia o no de relación se consideró que si p-valor es mayor >0.05 no existe relación, y si el p-valor es menor <0.05 existe relación; con respecto a los niveles de relación, se consideró los parámetros propuestos por Pallant (2011) que para esta investigación consideró los parámetros de 0,4 a 0,69 que indica una correlación positiva.
Este apartado es crucial porque demuestra el rigor metodológico del estudio. La elección de la prueba estadística no fue arbitraria, sino que se justificó con una prueba de normalidad . La clara explicación de los criterios hace que el proceso sea transparente y confiable, transformando los datos en hallazgos sólidos.
“Comunicación organizacional en las empresas del sur de Veracruz en tiempos de Covid-19”, la cual contó con la participación de 382 participantes. En ella se determinó que la percepción es favorable en un 43%; además, en esa investigación, se puso de manifiesto la enorme importancia de este constructo en tiempos complicados
Esta investigación podría o no ser representativa de la población estudiada ya que fue realizada en tiempos de pandemia lo cual bien podría haber afectado la percepción de los trabajadores sobre su ambiente laboral.
Creo que este tipo de estudios ayudan a que las instituciones se den cuenta de que no todo es salario o beneficios. La manera en que se comunican con los trabajadores influye mucho en cómo se sienten y en su motivación para trabajar.
Al no realizar una buena COI, se torna muy difícil el logro de los objetivos organizacionales; las organizaciones sin COI son calificadas como inertes, esto debido a que sin información no habría coordinación ni interacción entre las personas.
Esto implica en todos los ámbitos organizacionales, donde si no mantienes una buena COI, los objetivos grupales, individuales y en general, se ven inconclusos o con demasiadas dificultades para lograr los objetivos.
Tanto Silva (2011) como Sánchez y Nava (2007) coinciden en que los elementos de la COI son la “comunicación ascendente”, que es la que va desde los trabajadores del nivel operativo hacia los directivos; la “descendente”, que va desde el nivel directivo hasta los trabajadores del nivel operativo; y la “comunicación horizontal”, que se desarrolla entre los trabajadores del mismo nivel jerárquico.La SL es un sentimiento placentero o positivo con respecto al lugar de trabajo o por las actividades laborales que realiza una persona.
Me parece el punto más crítico dentro de una empresa ya que la manipulación o malinterpretacion de la información puede ocasionar problemas muy críticos
Es necesario definir los términos de inclusión y exclusión para determinar la muestra:Inclusión: Trabajadores que al momento de realizada la encuesta estaban realizando sus labores cotidianas.Exclusión: Trabajadores gozando de vacaciones o algún tipo de licencia.
Excluir a los trabajadores que se encuentran en su descanso puede mermar la obtención de resultados en el análisis estadístico, así mismo, un buen descanso puede significar también una mejor perspectiva o una visión más clara del ambiente en el que se trabaja, a comparación del veredicto que pudieras tener mientras realizas tus labores, podría afectar positiva o negativamente tu respuesta en función de las condiciones momentáneas en las que te encuentres.
Se resalta la importancia de que la COI que no es una opción el realizarla, sino una obligación, debido a que las organizaciones (indistintamente del rubro en que se desenvuelve, si son públicas o privadas...) deben realizar la información de sus actividades. Esto ayuda a modernizar el desarrollo de la gestión y su importancia radica en que al implementarla garantiza el cumplimiento de los objetivos y fomenta la vida organización
deja claro que la COI no es algo accesorio, sino esencial para cualquier organización dándonos una idea del por qué es el sistema a seguir dentro de las insdustrias y/o empresas
una población son individuos con características comunes y de interés para el investigador. Quienes participaron en esta investigación fueron los colaboradores de la municipalidad provincial de Hualgayoc, que, según el área de personal, son un total de 120 personas en los diferentes áreas y modalidades de contrato.
Indagar más sobre la población estudiada podría darnos más detalles acerca de sus resultados, la comunicación entre individuos depende directamente de las cualidades de la población, esta información podría o no ser acorde a la realidad de diversas poblaciones.
Se resalta la importancia de que la COI que no es una opción el realizarla, sino una obligación, debido a que las organizaciones (indistintamente del rubro en que se desenvuelve, si son públicas o privadas...) deben realizar la información de sus actividades. Esto ayuda a modernizar el desarrollo de la gestión y su importancia radica en que al implementarla garantiza el cumplimiento de los objetivos y fomenta la vida organización.
A pesar de que se resalte la importancia de la SL y COI dentro de las organizaciones es de notar que en los campos reales es difícil de lograr y requiere mucho esfuerzo y dinero para capacitar a gente que pueda fomentar dichas prácticas de dicha cualidad.
Aunque no lo parezca, es de gran dificultad implementar este tipo de prácticas en un ambiente laboral por la cantidad de recursos requeridos para fomentar la SL y COI.
Esta investigación se desarrolló bajo una metodología de tipo básica, ya que se enfocó en un tema de estudio específico y en hacen aportes al respecto (Lam, 2016). La investigación se realizó bajo un enfoque cuantitativo, ya que se basó en la valoración numérica y tomó como medio la estadística (Juárez et al., 2015). El nivel es descriptivo-correlacional; esto es así porque, por un lado, como explicaron Martínez et al. (2016), las investigaciones descriptivas, presentan características estudiadas de un determinado tema en un momento determinado, y, por otro, es correlacional porque buscan la relación de dos o más constructos. El diseño de la investigación es no experimental y de corte transversal, porque el recojo de la información se realizó en un momento determinado y el resultado es tal y como se encontró en la realidad
Algo que me gusta del documento presente es que te desglosan la manera en la que se realizó el trabajo, menciona su metodología, mencionan que la investigación es no experimental, es una buena manera de facilitar la investigación a alguna persona y si encuentra lo que está buscando
INTRODUCCIÓNContar con colaboradores que sientan satisfacción por el trabajo es fundamental para cualquier organización, independientemente del rubro en el que se desempeñen. Según Pujol y Dabos (2018), los colaboradores que sienten satisfacción laboral (SL) muestran afecto por el trabajo y concentran sus energías en el desarrollo de las actividades encomendadas.
Sin duda es algo fundamental que los trabajadores o colaboradores tengan algo de cariño por algún proyecto o trabajo ya que de esta manera se puede llegar a una meta mejor hecha o incluso con tendencia a ser perfecta, ya que las cosas hechas con cariño y dedicación siempre son lo que debemos realizar
debemos avanzar hacia un sistema centralizado de pagos para publicación en revistas de acceso abierto, a través de convenios nacionales con editoriales o fondos específicos administrados por agencias públicas.
Aquí hay que tener mucho cuidado: esos “convenios nacionales” no son otra cosa que los Acuerdos Transformativos que están firmando nuestras universidades por presión de las editoriales comerciales que les hacen "cuentas alegres" a los responsables de las políticas de las universidades, haciéndoles creer que nos hacen un favor. Y no, la verdad es que experiencias en Europa muestran que no reducen desigualdades, solo negocian precios globales para un mismo modelo corporativo (Baldwin & Cavanagh, 2024). ¿Cuál es tu fuente para hablar de "experiencias exitosas en Europa?
Estos costos pueden superar en promedio fácilmente los 2.000 o 3.000 dólares por artículo, pero las tarifas para las revistas más reputadas están arriba de los 10.000 dólares.
Esos montos se han disparado en parte por la lógica inflacionaria de los APCs tras la firma de los Acuerdos Transformativos. Entre 2013 y 2016 subieron 16% (Meagher, 2021) y la tendencia no se ha detenido.
No me parece adecuado que se quiera tratar el tema de desarrollo de comunicación con datos cuantitativos, no veo mucho sentido o como demostrar algo. Tal vez con gráficas sobre una pregunta o un punto de vista específico pero,con tablas no lo siento tan apropiado.
En parte no estoy de acuerdo en confiarle a un solo sitio o institución las características que no hacen correctas una buena vinculación al momento de trabajar, no todos los lugares serán iguales desde el simple hecho de que no se cuentan con las mismas características.
La SL resulta ser algo útil cuando bien sabemos que el puesto de jefe es algo muy importante, el como sea visto el habla mucho sobre como serán sus empleados y como a su vez ellos se sentirán motivados por él. Poder darle una idea a los jefes sobre aquellas cosas que desmotivan o perjudican a los empleados, ayuda a una mejor comunicación laboral, donde se podrá tener mayor confianza al momento de dudas, nuevas ideas o hasta para consultar sobre algún error. Un buen jefe es capaz de guiar a todo su equipo al éxito.
Laport et (2010) nos dice algo muy interesante, la imagen que generamos de nuestro espacio de trabajo habla mucho de nosotros, le permite a las demás personas darse una opinión de nosotros, sea buena o mala, donde podemos obtener un reconocimiento de esfuerzo de parte de ellos,atribuyendo un deseo de que gusten colaborar con nosotros.
Citando lo dicho por Nazar (2012), donde menciona que la SL ayuda a las personas, me parece muy relevante mencionar como desde siempre, parte de nuestra comunicación al momento de socializar con los demás, siempre biuscamos de alguna manera la aprobación al estar en un grupo o entorno de trabajo, un espacio en donde al sentir esa validación nos notivamos a realizar mejor nuestros trabajos laborales, generando un sentimiento tal vez de que si somos de ayuda para la empresa.
Tema 2 Básicamente, la COI es la manera en que fluye la info dentro de una empresa o organizacion Sirve para que la gente trabaje en equipo, se coordine mejor y se logren los objetivos. Si no existe la organización se estanca o hasta puede venirse abajo.
cada expansión publicada en las plataformas respondió a la capacidad demejora continua e instantánea que tiene la noticia transmedia. Cada ampliación,contracción o modificación de la información publicada en Facebook, Twitter oYouTube permitió editar la serialidad del sentido estipulada por la publicaciónperiódica y cerrada del medio. Cada fragmento expansivo dio nueva forma,reenmarcó o remezcló la información ya antes entregada por la misma franquicia.
Noticia siempre en desarrollo en lo transmedia.
en una primera fase, se compone de fragmentos diferentesde un mismo acontecimiento que se introducen a través del canal principal yse expanden a través de más medios o plataformas para luego ser exploradosy experimentados en sus diferentes repositorios; en el segundo momento, quepuede ser simultáneo, logra, además de ser compartida y comentada, que unaparte de la audiencia intervenga, modifique o resignifique por lo menos algunaporción la información propuesta por el productor seminal (Jenkins, 2003).
Fases de las noticias transmedia
La noticia transmedia, a diferencia de la multiplataforma y la crossmedia, genera“experiencias en el público, con el fin de motivar y hacer que participe, asumiendoun rol activo en la expansión” (Scolari, 2013, en Larrondo Ureta, 2016, p. 37). Así,los proyectos transmedia promueven que el usuario y los grupos de coproducción(agencias de noticias, ONG, fans, entre otros) puedan completar, ajustar, mostrarotro punto de vista y contradecir la información, sin alterar o trastocar la noticiaoriginal (Mendieta Briceño & Garcés, 2022).
Transmedia
la prensa multiplataforma adapta o traduce lamisma noticia a diversos soportes, el crossmedia journalism despliega fragmentoscomplementarios y dependientes que se constituyen como partes de un únicoacontecimiento, y el periodismo transmedia introduce por un canal y expandemediante diferentes medios y plataformas información interrelacionada –queguarda autonomía narrativa y sentido completo– pero que a su vez aporta detallesinéditos a una historia periodística global.
Diferencias entre multiplataforma, crossmedia y transmedia
sinistro
Súmula vinculante 32/STF
Trecho do voto e da ementa do RE 588.149/SP (Rel. Min. Gilmar Mendes, Plenário, julgado em 16/02/2011):
O eminente Ministro Sydney Sanches concluiu que assim como não compete aos estados fazer incidir o ICMS sobre a circulação de dinheiro, que na operação de câmbio é a mercadoria, também a eles não compete tributar a alienação de salvados, a qual se integra à operação de seguros.
Registro que por vedação legal ‘As Sociedades Seguradoras não poderão explorar qualquer outro ramo de comércio ou indústria’ (art. 73 do Decreto-Lei n. 73, de 21.11.1966), de maneira que elas não são e nem poderiam ser ‘comerciantes de ferro velho’.
O que ocorre é que por disposição contratual as seguradoras recebem por ato unilateral a propriedade do bem nas hipóteses em que, em razão de sinistro, tenha perdido mais de 75% do valor segurado.
Ressalto que as companhias de seguro são obrigadas a pagar ao segurado 100% do valor do bem. A posterior alienação dos salvados, pelas seguradoras, tem, quando muito, o condão de recuperar parcela da indenização que haja superado o dano ocorrido. Não há, dessa forma, finalidade de obter lucro, não havendo, portanto, intenção comercial.
Este é o sentido da jurisprudência do Supremo Tribunal Federal, conforme se depreende do Enunciado n. 541 da Súmula do Tribunal: ‘O imposto sobre vendas e consignações não incide sobre a venda ocasional de veículos e equipamentos usados, que não se insere na atividade profissional do vendedor, e não é realizada com o fim de lucro, sem caráter, pois, de comercialidade’.
O objeto das operações das seguradoras é o seguro. A eventual alienação dos salvados não os torna mercadorias, visto que as companhias seguradoras não possuem por objeto social a circulação de mercadorias, constituindo a referida alienação um elemento da própria operação de seguro, consoante exposto acima e de forma clara no voto do Ministro Sydney Sanches, relator da ADI-MC n. 1.332/RJ.”
VIII
ICMS e “leasing” internacional - 6
Tese fixada - Não incide o ICMS na operação de arrendamento mercantil internacional, salvo na hipótese de antecipação da opção de compra, <u>quando configurada a transferência da titularidade do bem</u>.
Resumo - Não incide o ICMS importação na operação de arrendamento mercantil internacional, salvo na hipótese de antecipação da opção de compra na medida em que o arrendamento mercantil não implica, necessariamente, transferência de titularidade sobre o bem.
Com base nessa orientação, o Plenário, por maioria e em conclusão de julgamento, desproveu recurso extraordinário se discutia a constitucionalidade da incidência de ICMS sobre operações de importação de mercadorias, sob o regime de arrendamento mercantil internacional — v. Informativos 629 e 729. O Tribunal assinalou que a incidência do ICMS pressuporia operação de circulação de mercadoria.
Assim, se não houver aquisição de mercadoria, mas <u>mera posse</u> decorrente do arrendamento, não se poderia cogitar de circulação econômica. Dessa forma, sublinhou que caberia à Fazenda Pública examinar o contrato de arrendamento para verificar a incidência de ICMS. Frisou que não haveria a aludida incidência sobre a operação de arrendamento mercantil sempre que a mercadoria fosse passível de restituição ao proprietário e enquanto não fosse efetivada a opção de compra.
Por outro lado, afirmou que sobre a operação de arrendamento a envolver bem insuscetível de devolução, fosse por circunstâncias naturais ou físicas ou por se tratar de insumo, incidiria ICMS, porque nessa hipótese o contrato teria apenas a forma de arrendamento, mas conteúdo de compra e venda. Apontou que, nos termos do acórdão recorrido, o caso dos autos seria de contrato de arrendamento mercantil internacional de bem suscetível de devolução, sem opção de compra.
Ademais, enfatizou que o entendimento de que o ICMS incidiria sobre toda e qualquer entrada de mercadoria importada poderia resultar em situações configuradoras de afronta ao princípio constitucional da vedação de confisco (CF, art. 150, IV). Isso porque, no caso de mercadoria que não constitua o patrimônio do arrendatário, o tributo, ao invés de integrar o valor da mercadoria, como seria da natureza do ICMS, expropriaria parcela do efetivo patrimônio da empresa. Salientou que os conceitos de direito privado não poderiam ser desnaturados pelo direito tributário.
Vencidos os Ministros Gilmar Mendes (relator) e Teori Zavascki, que davam provimento ao recurso. O relator aplicava o precedente firmado no RE 206.069/SP (DJU de 1º.9.2006), de modo a garantir a incidência do ICMS na importação de bem ou mercadoria provenientes do exterior, independentemente da natureza do contrato internacional celebrado. O Ministro Teori Zavascki, em acréscimo, pontuava que a natureza e o conteúdo do contrato celebrado no exterior não poderia comprometer a ocorrência do fato gerador do ICMS.
Legislação: CF: art. 150, IV
Precedentes: RE 206.069/SP (DJU de 1º.9.2006)
ck o
use of the metaphor as sickness is important alos, is being woc beingsick when u are in the world, becaue like either ur crazy and everyone else is normal or they're crazy and ur the only one who is normal, also black women becoming an object of learning for the world, and the decentering of herself in the poem, but it is the larger experience for being woc
seguridad se percibe
creo que puede confundir hablar de percepción en seguridad objetiva, pues más que relacionarse a la sensación de seguridad, es la vivencia de hechos que vulneran o resguardan la seguridad (peleas callejeras, asaltos, etc).
Assistant linked to federal registration and award history for Arayna Technology Solutions (UEI LQKTFZMD19B3), update here. Upload a capability statement or catalogue for your company here or upload an opportunity-specific document here Some suggestions:Please summarize the work to be completed under this opportunityDo the documents mention an incumbent contractor?Does this contract have any security clearance requirements?I'd like to anonymously submit a question to the procurement officer(s) Do they need certified people or something similar Loading Yes, the solicitation explicitly requires that all primary technical services be performed by individuals who are Mitel/Unify certified, qualified, and experienced in performing maintenance and repair of the specified equipment. Specifically, the contractor must:- Be a Mitel/Unify Systems certified partner authorized to perform on-network installations and maintenance.- Ensure that technicians have a complete understanding of system design, installation diagrams, and maintenance records for the equipment listed in the PWS.- Meet the following Air Force requirements for access: - Air Force Cyber Readiness Training (Annual) - CompTIA Security+ CertificationThese requirements are detailed in Section 4.0 of the [Performance Work Statement (PWS)](https://www.highergov.com/document/attachment-1-ellsworth-28th-bw-unify-sustainment-pws-pdf-4c2c3b/). Submit × Opportunity Assistant Document Library Capability Statements Opportunity Documents Drop or click to upload a Capability Statement (.pdf, .docx, or .xlsx) Close Select Document for Processing Select a document for analysis. Submit Match Score Factors Contributing to Match Registered with primary NAICS 541519 matches this opportunity. Registered for NAICS 541519 - Other Computer Related Services This opportunity is open to competition Potential Issues or Gaps Minimal or no matching keywords found for your capabilities May have limited or no recent experience with NAICS 541519 - Other Computer Related Services Registration not found for PSC DG10 - IT And Telecom - Network As A Service May have limited or no recent experience with PSC DG10 - IT And Telecom - Network As A Service Little or no prior performance with Air Combat Command This opportunity was marked No Bid by: Arayna Description Original Summary Original Summary The 28th Contracting Squadron, Ellsworth AFB, South Dakota, has issued this Solicitation, FA469025Q0062, to compete and award a Firm-Fixed Price Contract for sustainment and repair for the Mitel/Unify OpenScape Systems located at Ellsworth Air Force Base, South Dakota. This requriement is being solicited Full & Open (No Small Business Set-Aside), limited to Unify OpenScape Brand Name items and certified Technicians. Attached to this Solicitation Notice are: Standard Form 1449 Solicitation Attachment 1 - Performance Work Statement (PWS) Attachment 2 Wage Determination All questions and comments on this solicitation must be submitted in writing to joshua.johnson.233@us.af.mil no later than Wednesday, 10 September 2025 by 12:00pm MDT. Please title all emails with questions with the following subject line: FA469025Q0062 - Unify Mainteance. All interested and responsible entities are invited to submit a quote that will be considered by the 28th Contracting Office at Ellsworth AFB, South Dakota. The award will be based on the criteria established in the solicitation. Vendor quotes and all items required as listed within the Addendum to 52.212-1, Instruction to Offerors, are due to be submitted no later than Wednesday, 17 September 2025 by 3:00pm MDT to joshua.johnson.233@us.af.mil. Please title all submissions with the following subject line: FA469025Q0062 - Unify Maintenance Contractors submitting a quote must have and list within the quote their assigned Cage Code and be registered and ACTIVE in the System of Award Management (SAM) at www.sam.gov to be eligible for award. Auto-generated summaries available on select opportunities Background The 28th Contracting Squadron at Ellsworth Air Force Base, South Dakota, is issuing Solicitation FA469025Q0062 for a Firm-Fixed Price Contract aimed at the sustainment and repair of Mitel/Unify OpenScape Systems. This requirement is open to all vendors (Full & Open) but is limited to Unify OpenScape Brand Name items and certified technicians. The goal of this contract is to ensure the operational integrity of the telecommunications systems at the base.Work Details The contractor shall provide all personnel, equipment, tools, materials, supervision, and any other items and services necessary to ensure that the Unify system is operational. Key tasks include: - Performing maintenance and repair on Mitel/Unify telecommunications hardware and software installed throughout Ellsworth AFB. - Diagnosing and resolving system issues for applications such as OpenScape Voice, OpenScape Xpert, iNemsoft radio interface, and ASC voice. - Ensuring that all technical services are performed by individuals who are certified, qualified, and experienced in Mitel/Unify systems. - The scope of work includes sustainment and repair of specific equipment listed in Section 6.0 Equipment List of the Performance Work Statement (PWS).Period of Performance The contract will have a base period of 12 months with four additional option years, each lasting 12 months.Place of Performance Ellsworth Air Force Base, South Dakota. Show Less List Text Overview Agency Air Combat Command (ACC) [DoD - USAF] Response Deadline Sept. 17, 2025, 5:00 p.m. EDT Due in 12 Days Posted Sept. 4, 2025, 12:14 p.m. EDT Set Aside None NAICS 541519 - Other Computer Related Services PSC DG10 - IT And Telecom - Network As A Service Place of Performance Ellsworth AFB, SD 57706 United States Source Open Current SBA Size Standard $34 Million Pricing Fixed Price Est. Level of Competition Average Est. Value Range Experimental $50,000 - $150,000 (AI estimate) On 9/4/25 Air Combat Command issued Solicitation FA469025Q0062 for Unify OpenScape Maintenance due 9/17/25. The opportunity was issued full & open with NAICS 541519 and PSC DG10. Primary Contact Name Marc L Bellucci Profile Email marc.bellucci.1@us.af.mil Phone (605) 385-1782 Secondary Contact Name Joshua Johnson Profile Email joshua.johnson.233@us.af.mil Phone (605) 385-1734 Download All Explore Documents Posted documents for Solicitation FA469025Q0062 5102050100 ShownDocumentAgencyPosted DateSourceDownloadDocumentAgencyPosted DateSourceDownload Attachment 1 - Ellsworth 28th BW Unify Sustainment PWS.pdf Air Combat Command 09/04/25Contract Opportunity Text Snapshot This performance work statement (PWS) outlines the requirements for the sustainment and repair of Mitel/Unify systems at Ellsworth Air Force Base, specifically for the 28th Bomb Wing (BW). The contractor is tasked with providing all necessary personnel, equipment, tools, materials, and supervision to ensure the operational status of the Unify system. Key services include maintenance and repair... Attachment 3 - single source justification Redacted.pdf Air Combat Command 09/04/25Contract Opportunity Text Snapshot This single source justification is for a simplified acquisition related to the unify maintenance contract at ellsworth air force base (afb). the contracting activity is managed by the 28th contracting squadron, and the justification outlines the necessity for continuous maintenance, warranty support, software licenses, and timely updates for the existing unify openscape voice communication... Solicitation - FA469025Q0062.pdf Air Combat Command 09/04/25Contract Opportunity Text Snapshot This solicitation (FA469025Q0062) is for commercial products and services, specifically focused on Unify maintenance services. The solicitation outlines the requirements for a 12-month maintenance contract with options for four additional 12-month periods. The pricing arrangement is firm fixed price, and the total quantity required is specified as one unit for each period. The solicitation is... Attachment 2 - WD 15-5367 (Rev 29) dated 08jul25 (1).pdf Air Combat Command 09/04/25Contract Opportunity Text Snapshot This wage determination is issued by the U.S. Department of Labor, specifically under the Service Contract Act, with wage determination no. 2015-5367 and revision no. 29, dated July 8, 2025. It outlines the minimum wage rates and fringe benefits that contractors must pay to workers performing on federal service contracts in South Dakota, particularly in the counties of Custer, Meade, and... Question & Answer The AI Q&A Assistant has moved to the bottom right of the page Export Visible Records Clipboard CSV Excel All Records CSV Excel Opportunity Lifecycle Procurement notices related to Solicitation FA469025Q0062 5102050100 ShownTitleTypeAgencySet AsidePostedDeadlineDescriptionTitleTypeAgencySet AsidePostedDeadlineDescription Unify OpenScape Maintenance 25%Solicitation Air Combat Command None09/04/2509/17/25Description The 28th Contracting Squadron, Ellsworth AFB, South Dakota, has issued this Solicitation, FA469025Q0062, to compete and award a Firm-Fixed Price Contract for sustainment and repair for the Mitel/Unify OpenScape Systems located at Ellsworth Air Force Base, South Dakota. This requriement is being solicited Full & Open (No Small Business Set-Aside), limited to Unify OpenScape Brand Name items and certified Technicians. Attached to this Solicitation Notice are: Standard Form 1449 Solicitation Attachment 1 - Performance Work Statement (PWS) Attachment 2 Wage Determination All questions ...show moreThe 28th Contracting Squadron, Ellsworth AFB, South Dakota, has issued this Solicitation, FA469025Q0062, to compete and award a Firm-Fixed Price Contract for sustainment and repair for the Mitel/Unify OpenScape Systems located at Ellsworth Air Force Base, South Dakota. This requriement is being solicited Full & Open (No Small Business Set-Aside), limited to Unify OpenScape Brand Name items and certified Technicians. Attached to this Solicitation Notice are: Standard Form 1449 Solicitation Attachment 1 - Performance Work Statement (PWS) Attachment 2 Wage Determination All questions and comments on this solicitation must be submitted in writing to joshua.johnson.233@us.af.mil no later than Wednesday, 10 September 2025 by 12:00pm MDT. Please title all emails with questions with the following subject line: FA469025Q0062 - Unify Mainteance. All interested and responsible entities are invited to submit a quote that will be considered by the 28th Contracting Office at Ellsworth AFB, South Dakota. The award will be based on the criteria established in the solicitation. Vendor quotes and all items required as listed within the Addendum to 52.212-1, Instruction to Offerors, are due to be submitted no later than Wednesday, 17 September 2025 by 3:00pm MDT to joshua.johnson.233@us.af.mil. Please title all submissions with the following subject line: FA469025Q0062 - Unify Maintenance Contractors submitting a quote must have and list within the quote their assigned Cage Code and be registered and ACTIVE in the System of Award Management (SAM) at www.sam.gov to be eligible for award. Unify OpenScape Maintenance 0%Sources Sought Air Combat Command None04/30/2505/09/25Description This Sources Sought / RFI is issued solely for market research purposes in accordance with Federal Acquisition Regulation (FAR) Part 10 and is not a solicitation for proposals. This notice does not obligate the Government to award a contract or otherwise pay for the information provided in response. The Government will use information received in response to this notice to determine the appropriate acquisition strategy for the requirement. The 28th Maintenance Group (28 MXG) at Ellsworth AFB, South Dakota requires the contractor to provide all personnel, equipment, tools, materials, ...show moreThis Sources Sought / RFI is issued solely for market research purposes in accordance with Federal Acquisition Regulation (FAR) Part 10 and is not a solicitation for proposals. This notice does not obligate the Government to award a contract or otherwise pay for the information provided in response. The Government will use information received in response to this notice to determine the appropriate acquisition strategy for the requirement. The 28th Maintenance Group (28 MXG) at Ellsworth AFB, South Dakota requires the contractor to provide all personnel, equipment, tools, materials, supervision and any other items and services necessary to accomplish maintenance required. The primary technical services shall be performed by individuals who are Mitel/Unify certified, qualified, and experienced in performing maintenance and repair of equipment, crisis management, dispatch consoles, and all associated Unify OpenScape telecommunications hardware and software installed throughout Ellsworth 28 BW. PWS is attached to this RFI. NOTE: IF YOU DO NOT INTEND TO SUBMIT A PROPOSAL FOR THIS PROJECT WHEN IT IS FORMALLY ADVERTISED, PLEASE DO NOT SUBMIT A RESPONSE TO THIS SOURCES SOUGHT / RFI. Information requested: All interested parties are invited to provide information about your company/institution, or any teaming or joint venture partners. Interested vendors are requested to submit the following information, clearly indicating whether you are providing information: Company name, address, point of contact with phone number and email address, CAGE code, business size status (e.g., small business, large business), and website (if applicable). Manufacturer and model number. Detailed product specifications and brochures. Maintenance requirements and service support Availability and lead time. The 28 MXG will review all vendors who respond to this sources sought to determine if other companies can perform the required repairs; and if aftermarket parts can meet the government’s needs. Submission Instructions: All responses must be submitted electronically to marc.bellucci.1@us.af.mil and joshua.johnson.233@us.af.mil no later than 4:00 PM MDT, Thursday, 09 May 2025. Please include "Sources Sought / Mitel/Unify Annual Sustainment. Questions relevant to this notice shall be sent electronically to the above email address. NO PHONE INQUIRIES WILL BE ACCEPTED. All communication shall be in writing and submitted electronically with reference " Mitel/Unify Annual Sustainment.” Disclaimer: This Sources Sought / RFI is issued solely for information and planning purposes only and does not constitute a solicitation. The Government is not obligated to award a contract as a result of this announcement. No reimbursement will be made for any costs associated with providing information in response to this announcement or any follow-up requests. The Government shall not be liable for or suffer any consequential damages for any improperly identified information. Incumbent or Similar Awards Contracts Similar to Solicitation FA469025Q0062 510 ShownAward IDAwardeeAwarding AgencyPotential ValueSet AsideStartEndSimilarityDescriptionAward IDAwardeeAwarding AgencyPotential ValueSet AsideStartEndSimilarityDescription FA469022C0004Advancia Aeronautics Air Combat Command $370.2K8AN07/26/2208/09/25 Description 1 FTE MEDICAL IT SUPPORT TECHNICIAN47QTCA21A001G-FA469025FG018Impres Technology Solutions Air Combat Command $10.8KNone10/01/2409/30/25 Description IT AND TELECOM -NETWORK: DIGITAL NETWORK PRODUCTSNNG15SC41B-FA486123F0261Iron Bow Technologies Air Combat Command $4.0MNone07/11/2309/30/25 Description 805 COMBAT TRAINING SQUADRON SPECIAL ACCESS PROGRAM DIGITAL ENVIRONMENT (SAP DE) EQUIPMENT, CONFIGURATION AND INSTALL. Potential Bidders and Partners Awardees that have won contracts similar to Solicitation FA469025Q0062 Explore in Partner Finder Advancia Aeronautics 2024 Obligations: $35.4 million Microtechnologies 2024 Obligations: $142.9 million ENSCO 2024 Obligations: $98.9 million CDO Technologies 2024 Obligations: $13.6 million Referentia Systems 2024 Obligations: $9.1 million UIC Government Services 2024 Obligations: $369.6 million RTX 2024 Obligations: $30.4 billion World Wide Technology 2024 Obligations: $733.9 million Similar Active Opportunities Open contract opportunities similar to Solicitation FA469025Q0062 Experiments, Prototypes, Research, and Evaluation Supporting Systems (EXPRESS) Agency: Air Force Research Laboratory (AFRL) [DoD - USAF - AFMC] Deadline: Feb. 14, 2027, 5:00 p.m. EST Type: Solicitation Set Aside: None NAICS: 541715 - Research and Development in the Physical, Engineering, and Life Sciences (except Nanotechnology and Biotechnology) Hush House Inspection Agency: Pacific Air Forces (PACAF) [DoD - USAF] Deadline: Sept. 23, 2025, 10:00 p.m. EDT Type: Synopsis Solicitation Set Aside: None NAICS: 541350 - Building Inspection Services Combat Identification Automated Target Recognition Technology (CATCH) Call 03 Agency: Department of the Air Force (USAF) [DoD] Deadline: Sept. 18, 2025, 5:00 p.m. EDT Type: Solicitation Set Aside: None NAICS: 541715 - Research and Development in the Physical, Engineering, and Life Sciences (except Nanotechnology and Biotechnology) JRE Help Desk Agency: Air Combat Command (ACC) [DoD - USAF] Deadline: Sept. 9, 2026, 4:00 p.m. EDT Type: Solicitation Set Aside: None NAICS: 541512 - Computer Systems Design Services DATA CENTER MONITORING MODERNIZATION Agency: U.S. Air Forces Europe and Africa (USAFE) [DoD - USAF] Deadline: Sept. 15, 2025, 11:00 a.m. EDT Type: Synopsis Solicitation Set Aside: None NAICS: 541513 - Computer Facilities Management Services THUNDER COMMERCIALLY AUGMENTED MISSION PLATFORM (CAMP): DEVSECOPS SOFTWARE LICENSE Agency: Air Force Sustainment Center (AFSC) [DoD - USAF - AFMC] Deadline: Sept. 20, 2025, 4:00 p.m. EDT Type: Sources Sought Set Aside: None NAICS: 541519 - Other Computer Related Services Additional Details Source Agency Hierarchy DEPT OF DEFENSE > DEPT OF THE AIR FORCE > AFGSC > FA4690 28 CONS PKC FPDS Organization Code 5700-FA4690 Source Organization Code 500022516 Last Updated Sept. 4, 2025 Last Updated By joshua.johnson.233@us.af.mil Archive Date Oct. 2, 2025 search_params = {} table = 'contract_opportunity' key = 'FA469025Q0062-Solicitation-62400' sol = "FA469025Q0062" sol_clean = "FA469025Q0062" key3 = 'c28f08e1161a4ddfb1151beb3b162400' path_key = 'FA469025Q0062-Solicitation-62400' display = 'Solicitation - Unify OpenScape Maintenance' download_params = {"fed_opportunity": {"code": [{"key": `${key}`, "display": `${display}`}], "include": "Include"}, "id": "download"} show_bidders = true award_flag = false contract_flag = false idv_flag = false incumbent_flag = 'Exists' //true award_notification_threshold = 0 type_code = 'o' dibbs_flag = false store_recent_flag = true enable_expander = true track_key = "FA469025Q0062" //defining a separate key to track here than the default key transaction_key = 'c28f08e1161a4ddfb1151beb3b162400' api_transaction_key = 'c28f08e1161a4ddfb1151beb3b162400' opp_source = 'sam' solicitation_year = 'None' topic_code = 'None' nsn = 'None' no_link_flag = 'false' show_pricing = false show_supplier = false About Contact Terms Privacy © 2025 HigherGov
Specifically, the contractor must: