... signalling in the shade avoidance response

Also cyanobacteria can sense UV-B radiation. I'm quite sure that we have not yet found all possible UV-B receptors. I think we need to do some more literature search and write something about the potential existrance of other UV-B receptors.

Since we don't talk about the consequences of light reception thorugh the other receptors, I would not include the generation of ROS by light reception here. There are two more figures for signal transduction below. Should we include another one on ROS? Since this is a somewhat uncertain topic, I would not do that but rather add a little paragraph (at the time of writing this I have not yet read the whole chapter).

At some point we need to update all citations. I won't do that now, but in a separate session.

insert "UV radiation" after "of"

This is double

For Japanese bidding? Who were these residents?

There are no Z variables in the equation given above

Typo. It should be "k+r"

Операционная система - это многокомплексная программа, которая может пользоваться всеми ресурсами устройства, на котором она установлена. она некоторый посредник между пользователем, который хочет что-то делать на этом устройстве, и hardware частью компьютера

for - article - The End of Scarcity? From ‘Polycrisis’ to Planetary Phase Shift - Nafeez Ahmed - gene-culture coevolution - adjacency - indyweb dev - individual / collective evolutionary learning - provenance - tracing the evolution of ideas - gene-culture coevolution

adjacency - between - indyweb dev - individual / collective evolutionary learning - provenance - tracing the evolution of ideas - gene-culture coevolution - adjacency relationship - As DNA and epigenetics plays the role of transmitting biological adaptations, language and symmathesy play the role of transmitting cultural adaptations

quietly cleaning a quiet deluxe by [[Just My Typewriter]]

Cleaning the case, exterior and some of interior of a Royal Quiet De Luxe typewriter. She does a somewhat minimal job here.

She could have disassembled a bit more and done a better job with a toothbrush and mineral spirits on the inside.

Not a horrible recommendation for a beginner, but could have gone further and been a bit more comprehensive.

intention vectors in mindscape

converse

• makes my weekly reviews a breeze

meme put things in fornot of me - so that I do not forget

help 1 to recollect - what you need = for whatever purpoe - anytime - anywher - any how

intntional contextual reall

the information I have bothered to save and organise will be unused.

Royal Quiet De Luxe Typewriter Adjustment Print Quality Height Balance On-Feet Shift Motion by [[Phoenix Typewriter]]

He made sure the carriage isn't out of alignment which can cause on feet issues as well.

Adjust the basket stops higher or lower as necessary. Try 1/2 to full turn and test each

The adjustment points are between the body and the carriage about an inch inside the body shell.

Do upper case first. The first set of screws/nuts just next to the outside of the typewriter are for lower case and the second set just inside of those are for upper case.

Turning the adjustment screws clockwise should push the carriage stops down just a bit.

Some good characters to check are H, h, p, y, and 8.

When doing type alignment, Duane Jensen was taught to use an old/used ribbon instead of a new, wet/dark ribbon for better performance in testing. New ribbons don't show the differences as well.

He's noticed that ribbon from Around the Office are dreadful.

Surprising how low the percentage of "cases filed in California's trial courts result in an appellate disposition" are. Personally I thought it was a low number but something like 2-3%, not in the hundredths of a decimal. Given how low the percentage is it helps me get another reason why casebooks focus so much appellate rather than trial court designs. If a case is heard in appellate court that automatically gives it some importance in the legal world.

Sounds very similar to the phrase "when you're a hammer, everything looks like a nail." With very little understanding of the law these unimportant surface-level details are viewed as important, these "post-literate" individuals (as the paper calls them) trying to make these unrelated cases connect to what they are researching. I presume that only through a good understanding of how to properly do legal research that this issue is resolved.

Books are about large blocks of uninterrupted time...

( ~13:00)

Perhaps. I don't think so. With a Zettelkasten I believe you can write without 4-hour deep work blocks... However, maybe he is right... I haven't really written yet so I can't be certain.

Ryan Holiday says that our society struggles with accepting that we owe things to other people...

This reminds me of Simone Weil's notion of "no rights, only responsibilities"... A right by itself has no power, only obligation has. A right is an obligation toward us fulfilled. Only other people have rights, and we have obligations.

Getting into this frame of mind allows one to live a far more righteous and fulfilled as well as calm life. Once you acknowledge that you have no rights, you can not cling to them, and thus you don't view things as unfair to you.

"The Stoic Practice is a Dialogue With The Self" -- Ryan Holiday (~7:58)

I think this is also true for Zettelkasten. You write for yourself. Only you need to understand your notes, nobody else.

Stoicism is about taking the thorn out of your own eye before throwing stones at others.

( ~5:58)

"You get surprised even by your own notes."

Yes, that's exactly what Niklas Luhmann mentioned as the prerequisite for effective communication (with a Zettelkasten).

"You can see I have quite a lot notes I have to make."

This is a difference in mentality between Ryan Holiday and me (as well as Muhammed Ali Kilic)

@M.AKilic50

Our mentality (inspired by GTD and other standard productivity stuff, mostly Flow) is to avoid creating homework.

You don't HAVE to make notes on something. You select what you deem valuable and are interested in working with at the moment.

Because of the marginal gains effect I wrote about earlier, it doesn't matter if you don't make a lot of notes. Besides, you can always return later--especially with a proper bib card and potentially a custom index/ToC for a book.

A Zettelkasten is the lazy man's path to excellence.

(this is an ironic statement of mine because a Zettelkasten asks a lot of work over time. However, it doesn't have to be on a day to day basis. Plus you work only on what you want, hence it doesn't require that much discipline.)

"If you do one or two positive contributions a day, it adds up." - Ryan Holiday

Perhaps this is the essence of both Zettelkasten and Commonplace books; Marginal Gains.

Exponentional Increase over time. Upon first glance, it seems linear (1+1 = 2)... However, the formula is different because, at least in Zettelkasten, a new note means N new possible connections as this new note can virtually be connected to all other notes. In a Zettelkasten this is explicit, in a commonplace book connections are implicit.

Removing Feet from Royal Quiet DeLuxe Typewriter. by [[DC Types]]

Not what I was hoping for in terms of removing the screws holding the feet in.

https://www.theguardian.com/world/2024/oct/20/starmer-faces-slavery-reparations-demands-at-commonwealth-summit

AWSの試験のこと

難しいから理屈は覚えなくてもいいかも 一応リンクだけ...

XGBoost、LightGBM、CatBoostはライブラリやフレームワークの名前

クラスタリング（グループ化）

分類・回帰タスク

学習データ→ミニバッチ ミニバッチの中で予測されたデータと正解の誤差を計算 各パラメータの微分に負の符合( - )と学習関数をかける この更新方法を確率的勾配降下法という

並列処理をするからGPUなどのハードウェアに負担がかかるため高性能なマシンが必要

常に監視して検証をすることで過学習や余分な処理をなくす

Résumé de la vidéo [00:00:00][^1^][1] - [00:13:39][^2^][2]:

Cette vidéo explore KidZania, un parc d'attractions pour enfants qui simule le monde du travail et de la consommation. Elle examine les aspects éducatifs et publicitaires du parc, ainsi que les implications idéologiques.

Points forts : + [00:00:00][^3^][3] Introduction à KidZania * Parc d'attractions éducatif pour enfants * Présent dans une vingtaine de pays * Conçu pour initier les enfants au monde du travail + [00:01:20][^4^][4] Fonctionnement du parc * Mini-ville à taille d'enfant * Enfants reçoivent de l'argent fictif (Kidzos) * Activités de jeu de rôle pour gagner et dépenser des Kidzos + [00:02:28][^5^][5] Partenariats avec des entreprises * Activités gérées par de vraies entreprises * Exemples : Nivea, McDonald's, Pizza Hut * Entreprises paient un loyer pour être présentes + [00:05:00][^6^][6] Critiques et analyses * Mélange de communication éducative et publicitaire * Idéalisation du marché et du travail salarié * Absence de secteurs publics et de concurrence + [00:10:42][^7^][7] Implications idéologiques * Présentation du capitalisme comme une évidence * Fantasmagorie du capital dans l'architecture du parc * Différence entre la perception des enfants et celle des adultes

Résumé de la vidéo [00:00:00][^1^][1] - [00:14:47][^2^][2]:

Cette vidéo explore la mesure de la pauvreté en France, en comparant les indicateurs statistiques avec les expériences subjectives des individus. Elle se base sur un article d'Eléonore Richard publié en 2022.

Temps forts: + [00:00:00][^3^][3] Définition de la pauvreté * Difficulté à définir objectivement * Importance des indicateurs objectifs * Approches de l'Insee : pauvreté monétaire et en conditions de vie + [00:01:00][^4^][4] Article d'Eléonore Richard * Étude sur le ressenti de la pauvreté * Importance de relever certains seuils * Utilisation de données de l'Insee + [00:03:00][^5^][5] Calcul du niveau de vie * Définition du revenu disponible * Calcul du niveau de vie médian * Seuil de pauvreté à 60 % du niveau de vie médian + [00:06:00][^6^][6] Indicateurs de pauvreté * Comparaison internationale * Limites des indicateurs monétaires * Pauvreté en conditions de vie + [00:09:00][^7^][7] Analyse des données * Régression logistique * Impact du seuil de pauvreté sur le bien-être * Importance du seuil à 80 % du revenu médian

Résumé de la vidéo [00:00:00][^1^][1] - [00:15:16][^2^][2]:

Cette vidéo explore pourquoi les filles sont moins présentes dans les filières scientifiques malgré leur réussite académique. Elle examine les facteurs sociaux et les stéréotypes influençant leurs choix d'orientation.

Temps forts: + [00:00:00][^3^][3] Sous-représentation des filles en sciences * Les filles sont moins nombreuses dans les filières scientifiques prestigieuses * Étude de 2018 sur les choix d'orientation des élèves * Importance des choix d'orientation sur les écarts de salaire + [00:01:01][^4^][4] Étude sur les choix d'orientation * Enquête auprès de 8500 élèves en Île-de-France * Financement par L'Oréal pour sensibiliser les élèves * Analyse des données sans intervention de sensibilisation + [00:07:20][^5^][5] Différences de goût pour les sciences * Moins de filles aiment les sciences et envisagent une carrière scientifique * Confiance en soi plus faible chez les filles * Influence des stéréotypes de genre + [00:09:23][^6^][6] Impact des stéréotypes * Stéréotypes sur les capacités en mathématiques et les carrières scientifiques * Moins de filles pensent pouvoir concilier carrière scientifique et vie de famille * Importance de l'information sur les perspectives de salaire + [00:12:04][^7^][7] Analyse des choix d'orientation * Variables expliquant les écarts de choix d'orientation * Goûts et confiance en soi comme facteurs principaux * Rôle marginal des stéréotypes sur les choix d'orientation

New 3D-printed elements for the IBM Selectric by [[Richard Polt]]

Selectric Service Foot<br /> https://www.printables.com/model/895168-selectric-service-foot

New Selectric Type Elements by [[Joe Van Cleave]]

JVC tries out a new 3D printed type element in an IBM Selectric he got from David Hayden

David Hayden <br /> Austin Selectric Rescue<br /> https://selectricrescue.org/

Custom type elements for the IBM Selectric

ᔥ[[Joe Van Cleave]] in New Selectric Type Elements<br /> (accessed:: 2024-10-19 11:42:15)

Custom 3D Printed Selectric Elements are HERE! (Vogue! Papyrus!) by [[Theodore Munk]]

Engaging in a Zettelkasten/Commonplace book in this way is equal to inherent spaced repetition and recall perhaps?

Especially if you allow some time of rumination... Read book, wait a few days to a few weeks before processing it. The book's contents remain in the back of your mind.

Then when processing you get engaged with the substance again and therefore interrupt the ebbinghaus curve.

Perhaps I need to argue more with the authors and the content, as Adler & van Doren also recommend.

This might be a limitation in (the way I do) Zettelkasten. Because I am not writing in the margins and not engage in "tearing up" the book, I am less inclined to argue against/with the work.

Maybe I need to do this more using bib-card. Further thought on implementation necessary...

Perhaps a different reason is that I like to get through most books quickly rather than slowly. Sometimes I do the arguing afterward, within my ZK.

I need to reflect on this at some point (in the near future) and optimize my processes.

Watching this now, I am reminded that I really want to read more. To become erudite.

什么玩意这么热闹

↓typo

作っていたものを

「始める」のほうがよさそうです。

ここから下は特徴の説明をしているようなので、URLの掲載とは別に、「Streamlit Cloudには以下の特徴があります。」のような説明を先に書いてから列挙するのがいいと思います。

クリッカブルなリンクにしたほうがよさそうです。

「、」が多いのでここはトルでよさそうです。

たぶん「で」が正しいのでは？

後ろに()をつけた方がよさそうです。

後ろに()をつけた方がよさそうです。

後ろに()をつけた方がよさそうです。

後ろに()をつける表記と混ざっているので、()をつける方に統一したほうがよさそうです。

何を説明するのか書いたほうがよさそうです（おそらくst.writeの用途？）

トル

「、」が多いのでどちらかをトルでよさそうです。

search.google =notion+daily+notes

goole-search.url: https://www.google.co.uk/search?q=notion+daily+notes

plex - self.link - to

to:

google-search - notion daily notes

humans affecting underlying genomic basis of fisheries

*then

This is true due to the risk of getting robbed.

It is a good idea to rebuild trust within the community.

Its smart to play into what people want.

This makes sense, as people were in a big panic then.

This was after Elliot had laid into Tatiana about using the dragon for show and not for reasons that may somewhat be valuable for her in the long run.

This is a way that Elliot shows even though Tatiana can be worrisome he still felt some type of empathy for her in this situation.

I think it's very smart to focus on rebuilding the countrys economy.

I think this is only semi true, as another form of happiness is due to the relief having money brings.

I find it interesting that hes making it out, to seem like a problem he too faces.

I like the fact his trying to be honest.

How many things had to go perfectly for me to exist in the flesh?

It's interesting to see the major effect the Great Depression had on America.

It's crazy the difference in salary between farmers on the West Coast and farmers in the South.

I think it's interesting that his presidency had such a devastating effect.

I think it's so crazy how so much money can disappear in a blink of an eye.

I found this passage really shocking. It’s surprising that many people thought the economy would bounce back quickly when things were so bad. Hoover saying “the depression is over” feels almost unreal. The Smoot-Hawley Tariff made things worse, causing trade to drop a lot. This shows how quickly hope can turn into trouble, especially with bad decisions.

I found this information really surprising. It’s shocking that while the stock market was doing well, most Americans weren’t getting richer. The gap between the rich and poor was huge, with the wealthiest seeing their incomes rise a lot while everyday people struggled. It’s hard to believe that the government supported policies that helped the rich even more. This shows that a strong economy doesn’t mean everyone is doing well, and we need to pay attention to these inequalities to prevent future problems.

I found it surprising how easily people could buy stocks with borrowed money during the 1920s. They only needed to put down a small percentage, like $50 to buy $1,000 worth of shares. It worked well while prices went up, but when the market dropped, they couldn’t pay back their loans, which helped cause the big crash. It’s shocking how this risky system led to the stock market collapse and eventually the Great Depression.

I found this pretty funny! The idea of using stock market terms for love is clever, but it’s ironic that the song came out just before the stock market crashed. It’s surprising how quickly the mood changed from excitement to the Great Depression.

Roosevelt's use of presidential authority to bring about long-lasting change is interesting.

I'm glad president franklin got majority voted because i felt like Herbert hoover would've done worse for the country.

I wonder when this whole time took place, I feel like it was the time when hoover was president, and I heard that he was a selfish corrupt president since he was not helping his people.

That's super crazy after how the old soldiers sacrificed their own life to help the country in war, just for them to get treated like this.

I wonder what the difference from before and after the making of the movie was towards the sympathy for migrants. Was it the increased awareness of their struggles or the similarities between their struggles and US citizen's struggles of the time?

Seems kind of obvious that other countries would want to retaliate if the US was charging so much for imports, while not having to pay nearly as much in exports.

This is such a horrible response to those in need. The veterans worked for the US to help it cause, and they're just being tossed aside.

I'm confused on how this wasn't seen as a problem. To me, it seems like people are getting a large sum of money from just a small buy in. How did 1000 dollars worth of shares come from just 50 to 100 dollars?

The light could be a symbol for the afterlife and by continuing on the group could be ultimately racing towards their end.

could be a demonstration of natural selection by showing how despite the groups best efforts, they still fall short of safety

Tragedy will often bring an unlikely group together and allow them to bond.

Creates a dark and cynical tone for the story and may foreshadow tragedy later on since they are all aware that there is a chance they will die at sea.

The rough sea environment coincides with the naturalism tendency to place the story in a harsh environment..

or leave the co

need to have a bias for action and realize when there's a sunk cost fallacy

Changing numbering to reflect list of options would be more clear. In this case, "2. cout << h->key << endl;"

All theory is a learning in the development of a child, therefore it helps us in the social, cultural, emotional, physical, cognitive growth, etc. For me, Vygolsky's theory is essential since through the cognitive we learn to develop within society also through our own experiences

If all members of the household collaborated in household chores, it would be of great help to everyone because we are all putting in the time, effort and attitude to have a clean healthy home and work together.

The main role I have in my family is to be a mother and a provider. I take care of my kids , help around the house, and bring income to the house. Some boundaries I wish I had is spending more time with my family, because communication and relationships is important to me, but I am always working, and my kids are always busy.

This last line is very powerful. It affirms the rootedness of Native Americans in Californian lands as their beliefs and myths themselves were centered around the mundane things that were "the body, the village, the earth".

Often rare to find myths resemble more human, tangible features that we can relate to more.

I really love the idea of just "going to be." Like others have touched upon, this creation story is distinct from others I know about in that its intentions are plain and simple: they want the people to live freely and happily--as compared to having to earn happiness and basic necessities like food via strict customs and/or values.

This is what basic principle we have learned in science class or experiment. The variable in the experiment should be seriously asked. No other factors are allowed to create chaotic results or bias. People's attention sometimes are attracted by absurd or strange news, so they are easy to spread those wrong messages.

This is what we have learned in science class and experiments. We should always make sure of the accuracy of the setting. No other elements or factors affect the result with errors. And this is also related to the content of the class I am taking this quarter: Calling Bullshit.

Most of the time, social media always wanna gain information and track people's actions. The registration not only asks for basic contact information like email and phone number. Gender, interests, invite friends are the purpose of using social media and are included as unskippable steps.

What does realism mean here?

Wow, I didn't know that Romeo and Juliet is from some other story!

What does antiquity mean?

The Annual Energy Outlook (AEO) report by the US Energy Information Agency surveys long-term energy trends and is widely respected by traditional energy producers and analysts across many fields. The 2023 AEO concluded that "we project that the United States will remain a net exporter of petroleum products and natural gas through 2050 in all AEO2023 cases." Further, "As a result, in AEO2023, we see renewable generating capacity growing in all regions of the United States in all cases. Across all cases, compared with 2022, solar generating capacity grows by about 325% to 1019% by 2050, and wind generating capacity grows by about 138% to 235%. We see growth in installed battery capacity in all cases to support this growth in renewables. Across the span of AEO cases, relative to 2022, natural gas generating capacity ranges from an increase of between 20% to 87% through 2050." https://www.eia.gov/outlooks/aeo/narrative/index.php#ExecutiveSummary

This chapter provides no evidence that the US is experiencing an energy crisis. To wit: much evidence such as of record oil production suggests the opposite: https://www.forbes.com/sites/rrapier/2024/03/12/eia-confirms-historic-us-oil-production-record/

I believe only the numerator needs to be multiplied here.

B

Noematta

Sensory and the noetic

The advent of

Fallen material world

Complex imperfection

Axial age

Well said

Dato

Test @

Após a leitura dos materiais partilhados, deixo aqui um contributo do que me parece essencial: A seleção das atividades eletrónicas é crucial para um ensino e uma aprendizagem eficazes em ambientes virtuais. Almenara, Osuna & Cejudo (2014) sugerem critérios para a seleção da melhor atividade eletrónica, incluindo a consideração das caraterísticas dos alunos, a mobilização de diferentes aptidões, a promoção de diversas competências e a maior motivação. As actividades electrónicas são cruciais para a aprendizagem dos alunos, apoiando os objectivos e integrando-se no currículo. A motivação é essencial para reforçar as necessidades de aprendizagem. O feedback imediato é crucial para compreender os progressos e identificar as áreas a melhorar. As actividades electrónicas permitem uma aprendizagem online ativa e participativa, utilizando princípios e tecnologias apropriados. A escolha de uma estrutura de atividade eletrónica depende das opções pedagógicas, didácticas e metodológicas.

I love how this blogpost creates a highlighted link to the original post which they're quoting along with the commanding words "View in context at www.fromjason.xyz".

Skal være \(u\)

Hallo, ich hab gerade versucht die erste Uebung zu testen, aber hab leider folgende Fehler bekommen:

Tests konnten nicht ausgeführt werden Jobe Serveranfrage fehlgeschlagen.HTTP response from Jobe was 502: null

Haben alle auch das gleiche Problem?

reply to u/General-Writing1764 at https://old.reddit.com/r/typewriters/comments/1g7a8y5/does_anyone_know_how_do_they_make_new_platens/

I'm guessing that JJ Short is taking the original, removing the rubber. Placing the core into a mold and pouring in new material which hardens. Once done they put it on a lathe and turn it down to the appropriate (original) diameter. Potentially they're sanding the final couple of thousands of an inch for finish.

I'd imagine that if you asked them, they could/would confirm this general process.

The only other shop I've heard doing platen work is Bob at Typewriter Muse, but I haven't gone through his YouTube videos to see what his process looks like. (I'm pretty sure he documents some of it there.)

The assertion that these bones "can harm no one" introduces a paradox. While they signify death and the end of vitality, their inertness suggests that the past cannot actively disrupt the present. This resonates with the broader themes of the poem, which often grapple with the weight of history and the haunting presence of memory. In a world characterized by despair, the line hints at a resigned acceptance of the past’s inability to inflict further harm, positioning it as a relic rather than an active force.

The repetition of "Shantih shantih shantih" in the final lines of "The Waste Land" functions as a continuum rather than a concrete ending, embodying a search for peace amidst chaos. The term, meaning "the peace that passeth understanding," resonates deeply with the poem’s overarching themes of fragmentation and despair.

This triplet of peace is both a culmination and an invitation, suggesting that true tranquility may lie not in resolution but in the ongoing quest for harmony. Instead of providing a definitive conclusion, the repetition creates a rhythmic pulse that echoes throughout the poem, reminiscent of a mantra. It invites the reader to contemplate the cyclical nature of existence—where endings lead to new beginnings.

The line "I do not know whether a man or a woman" emerges from the section "A Game of Chess," which delves into the disintegration of human connection. This line signifies the speaker’s existential uncertainty, reflecting a world where traditional gender roles and identities have become muddied and irrelevant.

Here, the ambiguity echoes the broader themes of alienation and fragmentation, as the characters struggle to communicate and connect. The uncertainty of gender mirrors the breakdown of personal relationships, suggesting that in a chaotic, post-war landscape, even the most fundamental aspects of identity are in flux. This disorientation emphasizes the emotional paralysis faced by individuals, reinforcing the haunting sense of isolation pervading the poem.

In T.S. Eliot's "The Waste Land," the line "Who is the third who walks always beside you?" evokes a haunting presence, layering the poem with existential uncertainty and a sense of companionship laced with disquiet. From the speaker's point of view, this line captures an intimate yet unsettling inquiry that transcends the immediate relationships, hinting at a spiritual or existential specter that accompanies the living.

The phrase suggests an ambiguous, almost spectral companionship—one that suggests both intimacy and alienation. The “third” figure implies a triangular relationship, where the speaker and another are not alone; instead, they are shadowed by an elusive presence. This presence is not just a literal figure but symbolizes collective trauma, history, or perhaps the weight of modern disillusionment. It evokes a sense of haunting, as if the past—whether personal or cultural—lingers ominously, shaping the present.

Moreover, the use of “who” implies a search for identity, suggesting that the speaker grapples with understanding not just the presence of this third entity, but also their own place within the existential landscape of the poem. The question is both a plea and a probe, inviting readers to ponder the nature of companionship in a fractured world. The haunting nature of this inquiry lies in its open-endedness; it suggests that the answer may elude the speaker, reinforcing the poem's overarching themes of fragmentation and despair.

Ultimately, this line encapsulates the haunting complexity of human experience—where the past, the present, and the metaphysical intertwine, leaving the speaker (and the reader) in a state of reflective disquiet. The third figure symbolizes both loss and the ongoing search for meaning, a companion that walks with us, whether we acknowledge it or not.

Das Amazonasgebiet leidet im zweiten Jahr unter extremer Dürre. Es gab noch nie so wenig Niederschläge, die Dürre dauerte noch nie so lang und betraf noch nie ein so großes Gebiet. Waldbrände machten Sao Paolo zur Stadt mit der schlechtesten Luftqualität der Welt. Auch die meisten übrigen Staaten Südamerikas sind betroffen. Ursachen sind ein durch die globale Erhitzung verstärkter El Niño und die Rekordtemperaturen des Nordatlantik. Ausführlicher Bericht https://www.nytimes.com/2024/10/19/world/americas/south-america-drought-amazon-river.html

https://www.nytimes.com/2024/10/19/world/americas/south-america-drought-amazon-river.html

moleculas del sistema inmune en tortugas: Lisozymas con actividad antibacteriana y otrs moleculas son catelicidinas con actividad antifungica yactividad antibacteriana, incluso mas potente que farmacos ampicilina y bencilpenicilina. En el cocodrilo siames se aislo una pequeña proteina cationica 2008). A small cationic protein was isolated from the Siamese crocodile (Crocodylus siamensis), which demonstrated antibacterial activity against S. typhi, E. coli, S. aureus, Staphylococcus epidermidis, K. pneumoniae, P. aeruginosa and Vibrio chorelae (Preecharram et al., 2008). These antimicrobial peptides offer potent protection for reptiles against infection as well as provide exciting opportunities in the search for new clinical or agricultural antibiotics.

Las defensinas son: proteins that have a characteristic -sheet-rich fold as well as six disulphide-linked cysteines and have been found in all mammals that have been examined as well as in birds

Se han descrito defensinas con actividad antibacteriana against Escherichia coli and Salmonella typhimurium as well as antiviral activity against the Chandipura virus (Chattopadhyay et al., 2006). The first - defensin from reptilian leukocytes was recently isolated from the European pond turtle Emys orbicularis. Known as TBD-1, the peptide demonstrated strong activity against E. coli, Listeria monocytogenes, Candida albicans and methicillin-resistant Staphylococcus aureus (Stegemann, 2009).

El timo y el bazo se mantienen activos durante el verano e involucionan en en invierno cuando se encuentran en hibernación

Reptiles are the only ectothermic amniotes, and therefore become a pivotal group to study in order to provide important insights into both the evolution of the immune system as well as the functioning of the immune system in an ecological setting.

Ein „mediterraner Zyklon“ verursacht an diesem Wochenende überall in Italien extreme Unwetter. In der vergangenen Woche kam es in Ligurien stellenweise innerhalb weniger Stunden zu Niederschlägen von 300 mm (an einigen Punkten Frankreichs zu 600mm). Bei Piombino fielen in einer Stunde 120 mm Regen. Diese „tropischen“ Niederschlagsmengen weichen deutlich vom gewohnten mediterranen Klima ab. https://www.repubblica.it/italia/2024/10/19/news/previsioni_meteo_weekend_maltempo_nubifragi_allerta_rossa-423564680/

for - book - Clash of the Cartels: Unmasking the global drug kingpins stalking South Africa - Caryn Dolley - Columbia drug trafficking in South Africa

for - quote - organized crime in Cape Town

quote - organized crime in Cape Town - Andre Lincoln - Caryn Dolley - (see below) - Why you don’t see it is because it’s subtle, very sophisticated and it is a massive business. - How many restaurants and clubs on these famous streets are paying protection money to criminals? It's pretty startling - And what about construction shakedowns? 63 billion Rand of projects impacted in 2019 - https://hyp.is/Smjb3I5CEe-fXHsx-Sy8kQ/www.inclusivesociety.org.za/post/overview-of-the-construction-mafia-crisis-in-south-africa

for - organized crime - Cape Town - hidden protection scheme - Andre Lincoln

for - polycrisis - organized crime - Daily Maverick article - organized crime - Cape Town - How the state colludes with SA’s underworld in hidden web of organised crime – an expert view - Victoria O’Regan - 2024, Oct 18 - book - Man Alone: Mandela’s Top Cop – Exposing South Africa’s Ceaseless Sabotage - Daily Maverick journalist Caryn Dolley - 2024 - https://viahtml.hypothes.is/proxy/https://shop.dailymaverick.co.za/product/man-alone-mandelas-top-cop-exposing-south-africas-ceaseless-sabotage/?_gl=11mkyl5s_gcl_auODI2MTMxODEuMTcyNjI0MDAwMg.._gaNzQ5NDM3NzE0LjE3MjMxODY0NzY._ga_Y7XD5FHQVG*MTcyOTM1MjgwOS4xLjAuMTcyOTM1MjgxOS41MC4wLjkyNTE5MDk2OA..

summary - This article revolves around the research of South African crime reporter Caryn Dolley on the organized web of crime in South Africa - She discusses the nexus of - trans-national drug cartels - local Cape Town gangs - South African state collusion with gangs - in her new book: Man Alone: Mandela's Top Cop - Exposing South Africa's Ceaseless Sabotage - It illustrates how on-the-ground efforts to fight crime are failing because they do not effectively address this criminal nexus - The book follows the life of retired top police investigator Andre Lincoln whose expose paints the deep level of criminal activity spanning government, trans-national criminal networks and local gangs - Such organized crime takes a huge toll on society and is an important contributor to the polycrisis. - Non-linear approaches are necessary to tackle this systemic problem - One possibility is a trans-national citizen-led effort

for - stats - construction mafia impacts - South Africa - 2019 - R63 billion - Overview of the Construction Mafia Crisis in South Africa - Inclusive Society Institute - 2023

eLife Assessment

This important work advances our understanding of parabrachial CGRP threat function. The evidence supporting CGRP aversive outcome signaling is solid, while the evidence for cue signaling and fear behavior generation is incomplete. The work will be of interest to neuroscientists studying defensive behaviors.

Reviewer #1 (Public Review):

Summary

The authors asked if parabrachial CGRP neurons were only necessary for a threat alarm to promote freezing or were necessary for a threat alarm to promote a wider range of defensive behaviors, most prominently flight.

Major Strengths of Methods and Results

The authors performed careful single-unit recording and applied rigorous methodologies to optogenetically tag CGRP neurons within the PBN. Careful analyses show that single-units and the wider CGRP neuron population increases firing to a range of unconditioned stimuli. The optogenetic stimulation of experiment 2 was comparatively simpler but achieved its aim of determining the consequence of activating CGRP neurons in the absence of other stimuli. Experiment 3 used a very clever behavioral approach to reveal a setting in which both cue-evoked freezing and flight could be observed. This was done by having the unconditioned stimulus be a "robot" traveling along a circular path at a given speed. Subsequent cue presentation elicited mild flight in controls and optogenetic activation of CGRP neurons significantly boosted this flight response. This demonstrated for the first time that CGRP neuron activation does more than promote freezing. The authors conclude by demonstrating that bidirectional modulation of CGRP neuron activity bidirectionally affects freezing in a traditional fear conditioning setting and affects both freezing and flight in a setting in which the robot served as the unconditioned stimulus. Altogether, this is a very strong set of experiments that greatly expand the role of parabrachial CGRP neurons in threat alarm.

Weaknesses

In all of their conditioning studies the authors did not include a control cue. For example, a sound presented the same number of times but unrelated to US (shock or robot) presentation. This does not detract from their behavioral findings. However, it means the authors do not know if the observed behavior is a consequence of pairing. Or is a behavior that would be observed to any cue played in the setting? This is particularly important for the experiments using the robot US.

The authors make claims about the contribution of CGRP neurons to freezing and fleeing behavior, however, all of the optogenetic manipulations are centered on the US presentation period. Presently, the experiments show a role for these neurons in processing aversive outcomes but show little role for these neurons in cue responding or behavior organizing. Claims of contributions to behavior should be substantiated by manipulations targeting the cue period.

Appraisal

The authors achieved their aims and have revealed a much greater role for parabrachial CGRP neurons in threat alarm.

Discussion

Understanding neural circuits for threat requires us (as a field) to examine diverse threat settings and behavioral outcomes. A commendable and rigorous aspect of this manuscript was the authors decision to use a new behavioral paradigm and measure multiple behavioral outcomes. Indeed, this manuscript would not have been nearly as impactful had they not done that. This novel behavior was combined with excellent recording and optogenetic manipulations - a standard the field should aspire to. Studies like this are the only way that we as a field will map complete neural circuits for threat.

Reviewer #2 (Public Review):

-Summary of the Authors' Aims:<br /> The authors aimed to investigate the role of calcitonin gene-related peptide (CGRP) neurons in the parabrachial nucleus (PBN) in modulating defensive behaviors in response to threats. They sought to determine whether these neurons, previously shown to be involved in passive freezing behavior, also play a role in active defensive behaviors, such as fleeing, when faced with imminent threats.

-Major Strengths and Weaknesses of the Methods and Results:<br /> The authors utilized an innovative approach by employing a predator-like robot to create a naturalistic threat scenario. This method allowed for a detailed observation of both passive and active defensive behaviors in mice. The combination of electrophysiology, optogenetics, and behavioral analysis provided a comprehensive examination of CGRP neuron activity and its influence on defensive behaviors. The study's strengths lie in its robust methodology, clear results, and the multi-faceted approach that enhances the validity of the findings.

No notable weakness found.

-Appraisal of Aims and Results:<br /> The authors successfully achieved their aims by demonstrating that CGRP neurons in the PBN modulate both passive and active defensive behaviors. The results clearly show that activation of these neurons enhances fear memory and promotes conditioned fleeing behavior, while inhibition reduces these responses. The study provides strong evidence supporting the hypothesis that CGRP neurons act as a comprehensive alarm system in the brain.

-Impact on the Field and Utility of Methods and Data:<br /> This work has significant implications for the field of neuroscience, particularly in understanding the neural mechanisms underlying adaptive defensive behaviors. The innovative use of a predator-like robot to simulate naturalistic threats adds ecological validity to the findings and may inspire future studies to adopt similar approaches. The comprehensive analysis of CGRP neuron activity and its role in defensive behaviors provides valuable data that could be useful for researchers studying fear conditioning, neural circuitry, and behavior modulation.

-Additional Context:<br /> The study builds on previous research that primarily focused on the role of CGRP neurons in passive defensive responses, such as freezing. By extending this research to include active responses, the authors have provided a more complete picture of the role of these neurons in threat detection and response. The findings highlight the versatility of CGRP neurons in modulating different types of defensive behaviors based on the perceived intensity and immediacy of threats.

Overall, this manuscript makes a significant contribution to our understanding of the neural basis of defensive behaviors and offers valuable methodological insights for future research in the field.

Reviewer #3 (Public Review):

Strengths:<br /> The study used optogenetics together with in vivo electrophysiology to monitor CGRP neuron activity in response to various aversive stimuli including robot chasing to determine whether they encode noxious stimuli differentially. The study used an interesting conditioning paradigm to investigate the role of CGRP neurons in the PBN in both freezing and flight behaviors.

Weakness:<br /> The major weakness of this study is that the chasing robot threat conditioning model elicits weak unconditioned and conditioned flight responses, making it difficult to interpret the robustness of the findings. Furthermore, the conclusion that the CGRP neurons are capable of inducing flight is not substantiated by the data. No manipulations are made to influence the flight behavior of the mouse. Instead, the manipulations are designed to alter the intensity of the unconditioned stimulus.

eLife Assessment

This study presents a valuable finding on the identification of a complex consisting of NHE1, hERG1, β1/integrin and NaV1.5 on the membrane of breast cancer cells. The evidence supporting the claims of the authors is somewhat incomplete. The inclusion of clarification of some experimental design and the amendment of cropping Western blot data would have strengthened the study. The work will be of interest to scientists working on breast cancer.

Reviewer #1 (Public Review):

This manuscript by Capitani et al. extends previous studies of ion channel expression in triple-negative breast cancer cell lines. Probing four phenotypically different breast cancer cell lines, they used co-IP and confocal immunofluorescence (IF) colocalization to reveal that beta1 integrin forms a complex with the neonatal form of the Na+ channel NaV1.5 (nNaV1.5) and the Na+/H+ antiporter NHE1 in addition to previously reported hERG1. They used siRNA to show that silencing beta1 results in a co-depletion of hERG and Nav1.5, further supporting the conclusion that they form a complex; a complementary enhancement of Na current with increased hERG expression was also demonstrated. These data compellingly describe a complex of membrane proteins unregulated in breast cancer and thus present novel potential targets for treatment.

There are several concerns with experimental approaches. How fluorescence measurements were compared and controlled among experiments was not described, and masks drawn to define membrane expression seemed arbitrary, and included in some cases large sections of cytoplasm. There are issues associated with the use of channel blocking agents and a bifunctional small-chain antibody that are not well rationalized. Why are they being used, to test what hypotheses or disrupt what processes? The extremely high concentrations of E-4031 (4000x IC50 for block), e.g., are not expected to have selective actions. The effects of E-4031 at high concentrations altering cytoskeleton properties associated with invasiveness (and thus cancer progression) are questionable. There are numerous problems with co-IPs together carried out together with knock-down, which in one case depleted the protein targeted by the primary IP antibody. Western blots (WB) were quantified by comparing treatment to control, which does not control for loading errors. The control and treated signals should be divided by the respective tubulin signals to control for loading errors. Then the treated value can be compared with the control.

Reviewer #2 (Public Review):

The manuscript by Chiara Capitani and Annarosa Arcangeli reports the identification of a complex comprising NHE1,hERG1, β1 integrin, and NaV1.5 on the plasma membrane of breast cancer cells. The authors further investigated the mutual regulatory interactions among these proteins using Western blotting and co-immunoprecipitation assays. They also examined the downstream signaling pathways associated with this complex and assessed its impact on the malignant behavior of breast cancer cells.

Strengths

The manuscript used different breast cancer cell lines and combined Western blot, immunostaining, and electrophysiology to provide evidence for the proposed complex. The inhibitors are also used to test the requirement of channel activity to function in the development of breast cancer cells with in-vitro studies.

Weaknesses

The data shown in this manuscript include the western blots that are cropped and imaged separately to draw conclusions about protein levels and changes in immunoprecipitation. These cannot be done on separate, cropped blots but must be imaged together to make these comparisons.

Antibodies used for hERG, NaV1.5 and β1 integrin must be validated to work for IP using KO or KD cell lines for the respective proteins to demonstrate specificity. The same goes for all the immunofluorescence imaging shown in the manuscript as these are all key pieces of data to support the conclusions.

eLife Assessment

SGLT2 inhibitors (SGLT2i) have assumed important roles in reducing cardiovascular risk, particularly in those with diabetes. It has become appreciated that its protective effects are likely beyond their ability to lower blood sugar levels. This research presents a novel approach to studying the SGLT2i mechanism of action which is yet to be fully elucidated.

Reviewer #1 (Public Review):

The authors examined the hypothesis that plasma ApoM, which carries sphingosine-1-phosphate (S1P) and activates vascular S1P receptors to inhibit vascular leakage, is modulated by SGLT2 inhibitors (SGLTi) during endotoxemia. They also propose that this mechanism is mediated by SGLTi regulation of LRP2/ megalin in the kidney and that this mechanism is critical for endotoxin-induced vascular leak and myocardial dysfunction. The hypothesis is novel and potentially exciting. However, the author's experiments lack critical controls, lack rigor in multiple aspects, and overall does not support the conclusions.

Reviewer #2 (Public Review):

Apolipoprotein M (ApoM) is a plasma carrier for the vascular protective lipid mediator sphingosine 1-phospate (S1P). The plasma levels of S1P and its chaperones ApoM and albumin rapidly decline in patients with severe sepsis, but the mechanisms for such reductions and their consequences for cardiovascular health remain elusive. In this study, Ripoll and colleagues demonstrate that the sodium-glucose co-transporter inhibitor dapagliflizin (Dapa) can preserve serum ApoM levels as well as cardiac function after LPS treatment of mice with diet-induced obesity. They further provide data to suggest that Dapa preserves serum ApoM by increasing megalin-mediated reabsorption of ApoM in renal proximal tubules and that ApoM improves vascular integrity in LPS treated mice. These observations put forward a potential therapeutic approach to sustain vascular protective S1P signaling that could be relevant to other conditions of systemic inflammation where plasma levels of S1P decrease. However, although the authors are careful with their statements, the study falls short of directly implicating megalin in ApoM reabsorption and of ApoM/S1P depletion in LPS-induced cardiac dysfunction and the protective effects of Dapa.

The observations reported in this study are exciting and potentially of broad interest. The paper is well written and concise, and the statements made are mostly supported by the data presented. However, the mechanism proposed and implied is mostly based on circumstantial evidence, and the paper could be substantially improved by directly addressing the role of megalin in ApoM reabsorption and serum ApoM and S1P levels and the importance of ApoM for the preservation for cardiac function during endotoxemia. Some observations that are not necessarily in line with the model proposed should also be discussed.

The authors show that Dapa preserves serum ApoM and cardiac function in LPS-treated obese mice. However, the evidence they provide to suggest that ApoM may be implicated in the protective effect of Dapa on cardiac function is indirect. Direct evidence could be sought by addressing the effect of Dapa on cardiac function in LPS treated ApoM deficient and littermate control mice (with DIO if necessary).

The authors also suggest that higher ApoM levels in mice treated with Dapa and LPS reflect increased megalin-mediated ApoM reabsorption and that this preserves S1PR signaling. This could be addressed more directly by assessing the clearance of labelled ApoM, by addressing the impact of megalin inhibition or deficiency on ApoM clearance in this context, and by measuring S1P as well as ApoM in serum samples.

Methods: More details should be provided in the manuscript for how ApoM deficient and transgenic mice were generated, on sex and strain background, and on whether or not littermate controls were used. For intravital microscopy, more precision is needed on how vessel borders were outland and if this was done with or without regard for FITC-dextran. Please also specify the type of vessel chosen and considerations made with regard to blood flow and patency of the vessels analyzed. For statistical analyses, data from each mouse should be pooled before performing statistical comparisons. The criteria used for choice of test should be outlined as different statistical tests are used for similar datasets. For all data, please be consistent in the use of post-tests and in the presentation of comparisons. In other words, if the authors choose to only display test results for groups that are significantly different, this should be done in all cases. And if comparisons are made between all groups, this should be done in all cases for similar sets of data.

Reviewer #3 (Public Review):

The authors have performed well designed experiments that elucidate the protective role of Dapa in sepsis model of LPS. This model shows that Dapa works, in part, by increasing expression of the receptor LRP2 in the kidney, that maintains circulating ApoM levels. ApoM binds to S1P which then interacts with the S1P receptor stimulating cardiac function, epithelial and endothelial barrier function, thereby maintaining intravascular volume and cardiac output in the setting of severe inflammation. The authors used many experimental models, including transgenic mice, as well as several rigorous and reproducible techniques to measure the relevant parameters of cardiac, renal, vascular, and immune function. Furthermore, they employ a useful inhibitor of S1P function to show pharmacologically the essential role for this agonist in most but not all the benefits of Dapa. A strength of the paper is the identification of the pathway responsible for the cardioprotective effects of SGLT2is that may yield additional therapeutic targets. There are some weaknesses in the paper, such as, studying only male mice, as well as providing a power analysis to justify the number of animals used throughout their experimentation. Overall, the paper should have a significant impact on the scientific community because the SGLT2i drugs are likely to find many uses in inflammatory diseases and metabolic diseases. This paper provides support for an important mechanism by which they work in conditions of severe sepsis and hemodynamic compromise.

eLife Assessment

This study provides a valuable new perspective on how motor learning occurring in one state generalizes to new states (for example, a different limb posture). The proposed model improves upon previous theories in its ability to predict patterns of generalization, but evidence supporting this specific proposed model over possible alternatives is incomplete. The newly proposed theory appears promising but would be more convincing if its conceptual and theoretical basis were clearer and more rigorously derived.

Reviewer #1 (Public Review):

This paper proposes a novel framework for explaining patterns of generalization of force field learning to novel limb configurations. The paper considers three potential coordinate systems: cartesian, joint-based, and object-based. The authors propose a model in which the forces predicted under these different coordinate frames are combined according to the expected variability of produced forces. The authors show, across a range of changes in arm configurations, that the generalization of a specific force field is quite well accounted for by the model.

The paper is well-written and the experimental data are very clear. The patterns of generalization exhibited by participants - the key aspect of the behavior that the model seeks to explain - are clear and consistent across participants. The paper clearly illustrates the importance of considering multiple coordinate frames for generalization, building on previous work by Berniker and colleagues (JNeurophys, 2014). The specific model proposed in this paper is parsimonious, but there remain a number of questions about its conceptual premises and the extent to which its predictions improve upon alternative models.

A major concern is with the model's premise. It is loosely inspired by cue integration theory but is really proposed in a fairly ad hoc manner, and not really concretely founded on firm underlying principles. It's by no means clear that the logic from cue integration can be extrapolated to the case of combining different possible patterns of generalization. I think there may in fact be a fundamental problem in treating this control problem as a cue-integration problem. In classic cue integration theory, the various cues are assumed to be independent observations of a single underlying variable. In this generalization setting, however, the different generalization patterns are NOT independent; if one is true, then the others must inevitably not be. For this reason, I don't believe that the proposed model can really be thought of as a normative or rational model (hence why I describe it as 'ad hoc'). That's not to say it may not ultimately be correct, but I think the conceptual justification for the model needs to be laid out much more clearly, rather than simply by alluding to cue-integration theory and using terms like 'reliability' throughout.

A more rational model might be based on Bayesian decision theory. Under such a model, the motor system would select motor commands that minimize some expected loss, averaging over the various possible underlying 'true' coordinate systems in which to generalize. It's not entirely clear without developing the theory a bit exactly how the proposed noise-based theory might deviate from such a Bayesian model. But the paper should more clearly explain the principles/assumptions of the proposed noise-based model and should emphasize how the model parallels (or deviates from) Bayesian-decision-theory-type models.

Another significant weakness is that it's not clear how closely the weighting of the different coordinate frames needs to match the model predictions in order to recover the observed generalization patterns. Given that the weighting for a given movement direction is over-parametrized (i.e. there are 3 variable weights (allowing for decay) predicting a single observed force level, it seems that a broad range of models could generate a reasonable prediction. It would be helpful to compare the predictions using the weighting suggested by the model with the predictions using alternative weightings, e.g. a uniform weighting, or the weighting for a different posture. In fact, Fig. 7 shows that uniform weighting accounts for the data just as well as the noise-based model in which the weighting varies substantially across directions. A more comprehensive analysis comparing the proposed noise-based weightings to alternative weightings would be helpful to more convincingly argue for the specificity of the noise-based predictions being necessary. The analysis in the appendix was not that clearly described, but seemed to compare various potential fitted mixtures of coordinate frames, but did not compare these to the noise-based model predictions.

Reviewer #2 (Public Review):

Leib & Franklin assessed how the adaptation of intersegmental dynamics of the arm generalizes to changes in different factors: areas of extrinsic space, limb configurations, and 'object-based' coordinates. Participants reached in many different directions around 360{degree sign}, adapting to velocity-dependent curl fields that varied depending on the reach angle. This learning was measured via the pattern of forces expressed in upon the channel wall of "error clamps" that were randomly sampled from each of these different directions. The authors employed a clever method to predict how this pattern of forces should change if the set of targets was moved around the workspace. Some sets of locations resulted in a large change in joint angles or object-based coordinates, but Cartesian coordinates were always the same. Across three separate experiments, the observed shifts in the generalized force pattern never corresponded to a change that was made relative to any one reference frame. Instead, the authors found that the observed pattern of forces could be explained by a weighted combination of the change in Cartesian, joint, and object-based coordinates across test and training contexts.

In general, I believe the authors make a good argument for this specific mixed weighting of different contexts. I have a few questions that I hope are easily addressed.

Movements show different biases relative to the reach direction. Although very similar across people, this function of biases shifts when the arm is moved around the workspace (Ghilardi, Gordon, and Ghez, 1995). The origin of these biases is thought to arise from several factors that would change across the different test and training workspaces employed here (Vindras & Viviani, 2005). My concern is that the baseline biases in these different contexts are different and that rather the observed change in the force pattern across contexts isn't a function of generalization, but a change in underlying biases. Baseline force channel measurements were taken in the different workspace locations and conditions, so these could be used to show whether such biases are meaningfully affecting the results.

Experiment 3, Test 1 has data that seems the worst fit with the overall story. I thought this might be an issue, but this is also the test set for a potentially awkwardly long arm. My understanding of the object-based coordinate system is that it's primarily a function of the wrist angle, or perceived angle, so I am a little confused why the length of this stick is also different across the conditions instead of just a different angle. Could the length be why this data looks a little odd?

The manuscript is written and organized in a way that focuses heavily on the noise element of the model. Other than it being reasonable to add noise to a model, it's not clear to me that the noise is adding anything specific. It seems like the model makes predictions based on how many specific components have been rotated in the different test conditions. I fear I'm just being dense, but it would be helpful to clarify whether the noise itself (and inverse variance estimation) are critical to why the model weights each reference frame how it does or whether this is just a method for scaling the weight by how much the joints or whatever have changed. It seems clear that this noise model is better than weighting by energy and smoothness.

Are there any force profiles for individual directions that are predicted to change shape substantially across some of these assorted changes in training and test locations (rather than merely being scaled)? If so, this might provide another test of the hypotheses.

I don't believe the decay factor that was used to scale the test functions was specified in the text, although I may have just missed this. It would be a good idea to state what this factor is where relevant in the text.

Reviewer #3 (Public Review):

The author proposed the minimum variance principle in the memory representation in addition to two alternative theories of the minimum energy and the maximum smoothness. The strength of this paper is the matching between the prediction data computed from the explicit equation and the behavioral data taken in different conditions. The idea of the weighting of multiple coordinate systems is novel and is also able to reconcile a debate in previous literature.

The weakness is that although each model is based on an optimization principle, but the derivation process is not written in the method section. The authors did not write about how they can derive these weighting factors from these computational principles. Thus, it is not clear whether these weighting factors are relevant to these theories or just hacking methods. Suppose the author argues that this is the result of the minimum variance principle. In that case, the authors should show a process of how to derive these weighting factors as a result of the optimization process to minimize these cost functions.

In addition, I am concerned that the proposed model can cancel the property of the coordinate system by the predicted variance, and it can work for any coordinate system, even one that is not used in the human brain. When the applied force is given in Cartesian coordinates, the directionality in the generalization ability of the memory of the force field is characterized by the kinematic relationship (Jacobian) between the Cartesian coordinate and the coordinate of interest (Cartesian, joint, and object) as shown in Equation 3. At the same time, when a displacement (epsilon) is considered in a space and a corresponding displacement is linked with kinematic equations (e.g., joint displacement and hand displacement in 2 joint arms in this paper), the generated variances in different coordinate systems are linked with the kinematic equation each other (Jacobian). Thus, how a small noise in a certain coordinate system generates the hand force noise (sigma_x, sigma_j, sigma_o) is also characterized by the kinematics (Jacobian). Thus, when the predicted forcefield (F_c, F_j, F_o) was divided by the variance (F_c/sigma_c^2, F_j/sigma_j^2, F_o/sigma_o^2, ), the directionality of the generalization force which is characterized by the Jacobian is canceled by the directionality of the sigmas which is characterized by the Jacobian. Thus, as it has been read out from Fig*D and E top, the weight in E-top of each coordinate system is always the inverse of the shift of force from the test force by which the directionality of the generalization is always canceled. Once this directionality is canceled, no matter how to compute the weighted sum, it can replicate the memorized force. Thus, this model always works to replicate the test force no matter which coordinate system is assumed. Thus, I am suspicious of the falsifiability of this computational model. This model is always true no matter which coordinate system is assumed. Even though they use, for instance, the robot coordinate system, which is directly linked to the participant's hand with the kinematic equation (Jacobian), they can replicate this result. But in this case, the model would be nonsense. The falsifiability of this model was not explicitly written.

eLife Assessment

This important work advances our understanding of factors influencing early childhood development. The large sample size and methodology applied make the findings of this study convincing; however, support for some of the claims made by the authors is incomplete. The work will be of interest to researchers in developmental science and early childhood pediatrics.

Reviewer #1 (Public Review):

Padilha et al. aimed to find prospective metabolite biomarkers in serum of children aged 6-59 months that were indicative of neurodevelopmental outcomes. The authors leveraged data and samples from the cross-sectional Brazilian National Survey on Child Nutrition (ENANI-2019), and an untargeted multisegment injection-capillary electrophoresis-mass spectrometry (MSI-CE-MS) approach was used to measure metabolites in serum samples (n=5004) which were identified via a large library of standards. After correlating the metabolite levels against the developmental quotient (DQ), or the degree of which age-appropriate developmental milestones were achieved as evaluated by the Survey of Well-being of Young Children, serum concentrations of phenylacetylglutamine (PAG), cresol sulfate (CS), hippuric acid (HA) and trimethylamine-N-oxide (TMAO) were significantly negatively associated with DQ. Examination of the covariates revealed that the negative associations of PAG, HA, TMAO and valine (Val) with DQ were specific to younger children (-1 SD or 19 months old), whereas creatinine (Crtn) and methylhistidine (MeHis) had significant associations with DQ that changed direction with age (negative at -1 SD or 19 months old, and positive at +1 SD or 49 months old). Further, mediation analysis demonstrated that PAG was a significant mediator for the relationship of delivery mode, child's diet quality and child fiber intake with DQ. HA and TMAO were additional significant mediators of the relationship of child fiber intake with DQ.

Strengths of this study include the large cohort size and study design allowing for sampling at multiple time points along with neurodevelopmental assessment and a relatively detailed collection of potential confounding factors including diet. The untargeted metabolomics approach was also robust and comprehensive allowing for level 1 identification of a wide breadth of potential biomarkers. Given their methodology, the authors should be able to achieve their aim of identifying candidate serum biomarkers of neurodevelopment for early childhood. The results of this work would be of broad interest to researchers who are interested in understanding the biological underpinnings of development and also for tracking development in pediatric populations, as it provides insight for putative mechanisms and targets from a relevant human cohort that can be probed in future studies. Such putative mechanisms and targets are currently lacking in the field due to challenges in conducting these kind of studies, so this work is important.

However, in the manuscript's current state, the presentation and analysis of data impede the reader from fully understanding and interpreting the study's findings. Particularly, the handling of confounding variables is incomplete. There is a different set of confounders listed in Table 1 versus Supplementary Table 1 versus Methods section Covariates versus Figure 4. For example, Region is listed in Supplementary Table 1 but not in Table 1, and Mode of Delivery is listed in Table 1 but not in Supplementary Table 1. Many factors are listed in Figure 4 that aren't mentioned anywhere else in the paper, such as gestational age at birth or maternal pre-pregnancy obesity.

The authors utilize the directed acrylic graph (DAG) in Figure 4 to justify the further investigation of certain covariates over others. However, the lack of inclusion of the microbiome in the DAG, especially considering that most of the study findings were microbial-derived metabolite biomarkers, appears to be a fundamental flaw. Sanitation and micronutrients are proposed by the authors to have no effect on the host metabolome, yet sanitation and micronutrients have both been demonstrated in the literature to affect microbiome composition which can in turn affect the host metabolome.

Additionally, the authors emphasized as part of the study selection criteria the following,<br /> "Due to the costs involved in the metabolome analysis, it was necessary to further reduce the sample size. Then, samples were stratified by age groups (6 to 11, 12 to 23, and 24 to 59 months) and health conditions related to iron metabolism, such as anemia and nutrient deficiencies. The selection process aimed to represent diverse health statuses, including those with no conditions, with specific deficiencies, or with combinations of conditions. Ultimately, through a randomized process that ensured a balanced representation across these groups, a total of 5,004 children were selected for the final sample (Figure 1)."

Therefore, anemia and nutrient deficiencies are assumed by the reader to be important covariates, yet, the data on the final distribution of these covariates in the study cohort is not presented, nor are these covariates examined further.

The inclusion of specific covariates in Table 1, Supplementary Table 1, the statistical models, and the mediation analysis is thus currently biased as it is not well justified.

Finally, it is unclear what the partial-least squares regression adds to the paper, other than to discard potentially interesting metabolites found by the initial correlation analysis.

Reviewer #2 (Public Review):

A strength of the work lies in the number of children Padilha et al. were able to assess (5,004 children aged 6-59 months) and in the extensive screening that the Authors performed for each participant. This type of large-scale study is uncommon in low-to-middle-income countries such as Brazil.<br /> The Authors employ several approaches to narrow down the number of potentially causally associated metabolites.<br /> Could the Authors justify on what basis the minimum dietary diversity score was dichotomized? Were sensitivity analyses undertaken to assess the effect of this dichotomization on associations reported by the article? Consumption of each food group may have a differential effect that is obscured by this dichotomization.<br /> Could the Authors specify the statistical power associated with each analysis?<br /> Could the Authors describe in detail which metric they used to measure how predictive PLSR models are, and how they determined what the "optimal" number of components were?<br /> The Authors use directed acyclic graphs (DAG) to identify confounding variables of the association between metabolites and DQ. Could the dataset generated by the Authors have been used instead? Not all confounding variables identified in the literature may be relevant to the dataset generated by the Authors.<br /> Were the systematic reviews or meta-analyses used in the DAG performed by the Authors, or were they based on previous studies? If so, more information about the methodology employed and the studies included should be provided by the Authors.<br /> Approximately 72% of children included in the analyses lived in households with a monthly income superior to the Brazilian minimum wage. The cohort is also biased towards households with a higher level of education. Both of these measures correlate with developmental quotient. Could the Authors discuss how this may have affected their results and how generalizable they are?<br /> Further to this, could the Authors describe how inequalities in access to care in the Brazilian population may have affected their results? Could they have included a measure of this possible discrepancy in their analyses?<br /> The Authors state that the results of their study may be used to track children at risk for developmental delays. Could they discuss the potential for influencing policies and guidelines to address delayed development due to malnutrition and/or limited access to certain essential foods?

Reviewer #3 (Public Review):

The ENANI-2019 study provides valuable insights into child nutrition, development, and metabolomics in Brazil, highlighting both challenges and opportunities for improving child health outcomes through targeted interventions and further research.

Strengths of the methods and results:<br /> (1) The study utilizes data from the ENANI-2019 cohort, which was already existing. This cohort choice allows for longitudinal assessments and exploration of associations between metabolites and developmental outcomes. In addition, there was conservation of resources which are scanty in all settings in the current scenario.<br /> (2) The study aims to investigate the relationship between circulating metabolites (exposure) and early childhood development (outcome), specifically developmental quotient (DQ). The objectives are clearly stated, which facilitates focused research questions and hypotheses. The population that is studied is clearly mentioned.<br /> (3) The study accessed a large number of children under five years, with blood collected from a final sample size of 5,004 children. The exclusion of infants under six months due to venipuncture challenges and lack of reference values highlights practical considerations in research design.<br /> The study sample reflects a diverse range of children in terms of age, sex distribution, weight status, maternal education, and monthly family income. This diversity enhances the generalizability of findings across different sociodemographic groups within Brazil.<br /> (4) The study uses standardized measures (e.g., DQ assessments) and chronological age. Confounding variables, such as child's age, diet quality, and nutritional status, are carefully considered and incorporated into analyses through a Directed Acyclic Graph (DAG). The mean DQ of 0.98 indicates overall developmental norms among the studied children, with variations noted across different demographic factors such as age, region, and maternal education. The prevalence of Minimum Dietary Diversity (MDD) being met by 59.3% of children underscores dietary patterns and their potential impact on health outcomes. The association between nutritional status (weight-for-height z-scores) and developmental outcomes (DQ) provides insights into the interplay between nutrition and child development.<br /> The study identified key metabolites associated with developmental quotient (DQ):<br /> Component 1: Branched-chain amino acids (Leucine, Isoleucine, Valine).<br /> Component 2: Uremic toxins (Cresol sulfate, Phenylacetylglutamine).<br /> Component 3: Betaine and amino acids (Glutamine, Asparagine).<br /> The study focused on several serum metabolites like PAG (phenylacetylglutamine), CS (p-cresyl sulfate), HA (hippuric acid), TMAO (trimethylamine-N-oxide), MeHis (methylhistidine), and Crtn (creatinine). These metabolites are implicated in various metabolic pathways linked to gut microbiota activity, amino acid metabolism, and dietary factors.<br /> These metabolites explained a significant portion of both metabolite variance (39.8%) and DQ variance (4.3%). The study suggests that these metabolites can be used as proxy measures of the gut microbiome in children.<br /> (5) The use of partial least square regression (PLSR) with cross-validation (80% training, 20% testing) which is a robust approach to identify metabolites predictive of DQ, which minimizes overfitting. This model allows for outliers to remain outliers for transparency.<br /> The Directed Acyclic Graph (DAG) identifies and adjusts for confounding variables (e.g., child's diet quality, nutritional status) and strengthens the validity of findings by controlling for potential biases. Developmental and gender differences were studied by testing interactions with the age of the child and the sex.<br /> Mediation analysis exploring metabolites as potential mediators provides insights into underlying pathways linking exposures (e.g., diet, microbiome) with DQ.<br /> The use of Benjamini-Hochberg correction for multiple comparisons and bootstrap tests (5,000 iterations) enhances the reliability of results by controlling false discovery rates and assessing significance robustly.

Significant correlations between serum metabolites and DQ, particularly negative associations with certain metabolites like PAG and CS, suggest potential biomarkers or pathways influencing developmental outcomes. Notably, these associations varied with age, suggesting different metabolic impacts during early childhood development.

Weaknesses:<br /> (1) The data collected was incomplete especially those related to breastfeeding history and birth weight. These have been mentioned in the limitations of the study but yet might have been potential confounders or even factors leading to the particular identified metabolite state of the population.<br /> (2) Other tests than mediation analysis might have been used to ensure reliability and robustness of the data. How data was processed, data cleaning methods, how outliers were handled and sensitivity analyses would ensure robustness of the findings.<br /> (3) The generalizability of the data is not sound especially considering the children mostly belonged to a higher socioeconomic group in Brazil with mother or caregiver education being above a certain level. Comparative studies with children from other socio-economic groups and other cohorts might have been useful. Consideration of sample size adequacy and power analysis might have helped in generalizing the findings.<br /> (4) Caution is needed in interpreting causality from this data because of the nature of the study design Discussing alternative explanations and potential confounding factors in more depth could strengthen the conclusions.

Appraisal<br /> (1) The aims of the study were to identify associations between children's serum metabolome and Early Childhood development. This aim was met. The results do confirm their conclusions.<br /> Impact of the work on the field

(1) Unless actual gut microbiome of children in this age group from gut bacteria examination or gastrointestinal examination of the gut of children, the causality of gut metabolome on early childhood development cannot be established with certainty. Because this may not be possible in every situation, proxy methods such as the one elucidated here might be useful, considering the risk-benefit ratio.<br /> (2) More research is needed on this theme through longitudinal studies to validate these findings and explore underlying pathways involving gut-brain interactions and metabolic dysregulation.<br /> Other readings: Readers are advised to read other research from other countries and other languages to understand the connection between gut microbiome, metabolite spectra, and child development. In addition to study the effect of these factors on child mental development too.

Readers might consider the following questions:<br /> (1) Should investigators study the families through direct observation of diet and other factors to look for a connection between food taken in and gut microbiome and child development?<br /> (2) Can an examination of the mother's gut microbiome influence the child's microbiome? Can the mother or caregiver's microbiome influence early childhood development?<br /> (3) Is developmental quotient enough to study early childhood development? Is it comprehensive enough?

eLife Assessment

This important work addresses the role of Marcks/Markcksl during spinal cord development and regeneration. The study is exceptional in combining molecular approaches to understand the mechanisms of tissue regeneration with behavioural assays, which is not commonly employed in the field. The data presented is convincing and comprehensive, using many complementary methodologies.

Reviewer #1 (Public Review):

In this manuscript, El Amri et al. are exploring the role of Marcks and Marcksl1 proteins during spinal cord development and regeneration in Xenopus. Using two different techniques to knockdown their expressions, they argue that these proteins are important for neural progenitors proliferation and neurites outgrowth in both contexts. Finally, using a pharmalogical approach, they suggest that Marcks and Marcksl1 work by modulating the activity of PLD and the levels of PIP2 whilst PKC could modulate Marcks activity.<br /> The strength of this manuscript resides in the ability of the authors to knockdown the expression of 4 different genes using 2 different methods to assess the role of this protein family during early development and regeneration at the late tadpole stage. This has always been a limiting factor in the field as the tools to perform conditional knockouts in Xenopus are very limited. However, this will not really be applicable to essential genes as it relies on the general knockdown of protein expression. The generation of antibodies able to detect endogenous Marcks/Marcksl1 is also a powerful tool to assess the extent to which the expression of these proteins is down-regulated.<br /> Whilst there is a great amount of data provided in this manuscript and there is strong evidence to show that Marcks are important for spinal cord development and regeneration, their roles in both contexts is not explored fully. The description of the effect of knocking down Marcks/Marcksl1 on neurons and progenitors is rather superficial and the evidence for the underlying mechanism underpinning their roles is not very convincing.

Reviewer #2 (Public Review):

M. El Amri et al., investigated the functions of Marcks and Marcks like 1 during spinal cord (SC) development and regeneration in Xenopus laevis. The authors rigorously performed loss of function with morpholino knock-down and CRISPR knock-out combining rescue experiments in developing spinal cord in embryo and regeneration in tadpole stage.

For the assays in the developing spinal cord, a unilateral approach (knock-down/out only one side of the embryo) allowed the authors to assess the gene functions by direct comparing one-side (e.g. mutated SC) to the other (e.g. wild type SC on the other side). For the assays in regenerating SC, the authors microinject CRISPR reagents into 1-cell stage embryo. When the embryo (F0 crispants) grew up to tadpole (stage 50), the SC was transected. They then assessed neurite outgrowth and progenitor cell proliferation. The validation of the phenotypes was mostly based on the quantification of immunostaining images (neurite outgrowth: acetylated tubulin, neural progenitor: sox2, sox3, proliferation: EdU, PH3), that are simple but robust enough to support their conclusions. In both SC development and regeneration, the authors found that Marcks and Marcksl1 were necessary for neurite outgrowth and neural progenitor cell proliferation.<br /> The authors performed rescue experiments on morpholino knock-down and CRISPR knock-out conditions by Marcks and Marcksl1 mRNA injection for SC development and pharmacological treatments for SC development and regeneration. The unilateral mRNA injection rescued the loss-of-function phenotype in the developing SC. To explore the signalling role of these molecules, they rescued the loss-of-function animals by pharmacological reagents They used S1P: PLD activator, FIPI: PLD inhibitor, NMI: PIP2 synthesis activator and ISA-2011B: PIP2 synthesis inhibitor. The authors found the activator treatment rescued neurite outgrowth and progenitor cell proliferation in loss of function conditions. From these results, the authors proposed PIP2 and PLD are the mediators of Marcks and Marcksl1 for neurite outgrowth and progenitor cell proliferation during SC development and regeneration. The results of the rescue experiments are particularly important to assess gene functions in loss of function assays, therefore, the conclusions are solid. In addition, they performed gain-of-function assays by unilateral Marcks or Marcksl1 mRNA injection showing that the injected side of the SC had more neurite outgrowth and proliferative progenitors. The conclusions are consistent with the loss-of-function phenotypes and the rescue results. Importantly, the authors showed the linkage of the phenotype and functional recovery by behavioral testing, that clearly showed the crispants with SC injury swam less distance than wild types with SC injury at 10-day post surgery.<br /> Prior to the functional assays, the authors analyzed the expression pattern of the genes by in situ hybridization and immunostaining in developing embryo and regenerating SC. They confirmed that the amount of protein expression was significantly reduced in the loss of function samples by immunostaining with the specific antibodies that they made for Marcks and Marcksl1. Although the expression patterns are mostly known in previous works during embryo genesis, the data provided appropriate information to readers about the expression and showed efficiency of the knock-out as well.

MARCKS family genes have been known to be expressed in the nervous system. However, few studies focus on the function in nerves. This research introduced these genes as new players during SC development and regeneration. These findings could attract broader interests from the people in nervous disease model and medical field. Although it is a typical requirement for loss of function assays in Xenopus laevis, I believe that the efficient knock-out for four genes by CRISPR/Cas9 was derived from their dedication of designing, testing and validation of the gRNAs and is exemplary.

Weaknesses,<br /> 1) Why did the authors choose Marcks and Marcksl1?<br /> The authors mentioned that these genes were identified with a recent proteomic analysis of comparing SC regenerative tadpole and non-regenerative froglet (Line (L) 54-57). However, although it seems the proteomic analysis was their own dataset, the authors did not mention any details to select promising genes for the functional assays (this article). In the proteomic analysis, there must be other candidate genes that might be more likely factors related to SC development and regeneration based on previous studies, but it was unclear what the criteria to select Marcks and Marcksl1 was.

2) Gene knock-out experiments with F0 crispants,<br /> The authors described that they designed and tested 18 sgRNAs to find the most efficient and consistent gRNA (L191-195). However, it cannot guarantee the same phenotypes practically, due to, for example, different injection timing, different strains of Xenopus laevis, etc. Although the authors mentioned the concerns of mosaicism by themselves (L180-181, L289-292) and immunostaining results nicely showed uniformly reduced Marcks and Marcksl1 expression in the crispants, they did not refer to this issue explicitly.

3) Limitations of pharmacological compound rescue<br /> In the methods part, the authors describe that they performed titration experiments for the drugs (L702-704), that is a minimal requirement for this type of assay. However, it is known that a well characterized drug is applied, if it is used in different concentrations, the drug could target different molecules (Gujral TS et al., 2014 PNAS). Therefore, it is difficult to eliminate possibilities of side effects and off targets by testing only a few compounds.

Reviewer #3 (Public Review):

El Amri et al conducted an analysis on the function of marcks and marcksl in Xenopus spinal cord development and regeneration. Their study revealed these proteins are crucial for neurite outgrowth and cell proliferation, including Sox2+ progenitors. Furthermore, they suggested these genes may act through the PLD pathway. The study is well-executed with appropriate controls and validation experiments, distinguishing it from typical regeneration research by including behavioral assays. The manuscript is commendable for its quantifications, literature referencing, careful conclusions, and detailed methods. Conclusions are well-supported by the experiments performed in this study. Overall, this manuscript contributes to the field of spinal cord regeneration and sets a good example for future research in this area.

Welcome back and in this demo lesson you're going to learn how to install the Docker engine inside an EC2 instance and then use that to create a Docker image.

Now this Docker image is going to be running a simple application and we'll be using this Docker image later in this section of the course to demonstrate the Elastic Container service.

So this is going to be a really useful demo where you're going to gain the experience of how to create a Docker image.

Now there are a few things that you need to do before we get started.

First as always make sure that you're logged in to the I am admin user of the general AWS account and you'll also need the Northern Virginia region selected.

Now attached to this lesson is a one-click deployment link so go ahead and click that now.

This is going to deploy an EC2 instance with some files pre downloaded that you'll use during the demo lesson.

Now everything's pre-configured you just need to check this box at the bottom and click on create stack.

Now that's going to take a few minutes to create and we need this to be in a create complete state.

So go ahead and pause the video wait for your stack to move into create complete and then we're good to continue.

So now this stack is in a create complete state and we're good to continue.

Now if you're following along with this demo within your own environment there's another link attached to this lesson called the lesson commands document and that will include all of the commands that you'll need to type as you move through the demo.

Now I'm a fan of typing all commands in manually because I personally think that it helps you learn but if you are the type of person who has a habit of making mistakes when typing along commands out then you can copy and paste from this document to avoid any typos.

Now one final thing before we finish at the end of this demo lesson you'll have the opportunity to upload the Docker image that you create to Docker Hub.

If you're going to do that then you should pre sign up for a Docker Hub account if you don't already have one and the link for this is included attached to this lesson.

If you already have a Docker Hub account then you're good to continue.

Now at this point what we need to do is to click on the resources tab of this stack and locate the public EC2 resource.

Now this is a normal EC2 instance that's been provisioned on your behalf and it has some files which have been pre downloaded to it.

So just go ahead and click on the physical ID next to public EC2 and that will move you to the EC2 console.

Now this machine is set up and ready to connect to and I've configured it so that we can connect to it using Session Manager and this avoids the need to use SSH keys.

So to do that just right-click and then select connect.

You need to pick Session Manager from the tabs across the top here and then just click on connect.

Now that will take a few minutes but once connected you should see this prompt.

So it should say SH- and then a version number and then dollar.

Now the first thing that we need to do as part of this demo lesson is to install the Docker engine.

The Docker engine is the thing that allows Docker containers to run on this EC2 instance.

So we need to install the Docker engine package and we'll do that using this command.

So we're using shudu to get admin permissions then the package manager DNF then install then Docker.

So go ahead and run that and that will begin the installation of Docker.

It might take a few moments to complete it might have to download some prerequisites and you might have to answer that you're okay with the install.

So press Y for yes and then press enter.

Now we need to wait a few moments for this install process to complete and once it has completed then we need to start the Docker service and we do that using this command.

So shudu again to get admin permissions and then service and then the Docker service and then start.

So type that and press enter and that starts the Docker service.

Now I'm going to type clear and then press enter to make this easier to see and now we need to test that we can interact with the Docker engine.

So the most simple way to do that is to type Docker space and then PS and press enter.

Now you're going to get an error.

This error is because not every user of this EC2 instance has the permissions to interact with the Docker engine.

We need to grant permissions for this user or any other users of this EC2 instance to be able to interact with the Docker engine and we're going to do that by adding these users to a group and we do that using this command.

So shudu for admin permissions and then user mod -a -g for group and then the Docker group and then EC2 -user.

Now that will allow a local user of this system, specifically EC2 -user, to be able to interact with the Docker engine.

Okay so I've cleared the screen to make it slightly easier to see now that we've added EC2 -user the ability to interact with Docker.

So the next thing is we need to log out and log back in of this instance.

So I'm going to go ahead and type exit just to disconnect from session manager and then click on close and then I'm going to reconnect to this instance and you need to do the same.

So connect back in to this EC2 instance.

Now once you're connected back into this EC2 instance we need to run another command which moves us into EC2 user so it basically logs us in as EC2 -user.

So that's this command and the result of this would be the same as if you directly logged in to EC2 -user.

Now the reason we're doing it this way is because we're using session manager so that we don't need a local SSH client or to worry about SSH keys.

We can directly log in via the console UI we just then need to switch to EC2 -user.

So run this command and press enter and we're now logged into the instance using EC2 -user and to test everything's okay we need to use a command with the Docker engine and that command is Docker space ps and if everything's okay you shouldn't see any output beyond this list of headers.

What we've essentially done is told the Docker engine to give us a list of any running containers and even though we don't have any it's not erred it's simply displayed this empty list and that means everything's okay.

So good job.

Now what I've done to speed things up if you just run an LS and press enter the instance has been configured to download the sample application that we're going to be using and that's what the file container.zip is within this folder.

I've configured the instance to automatically extract that zip file which has created the folder container.

So at this point I want you to go ahead and type cd space container and press enter and that's going to move you inside this container folder.

Then I want you to clear the screen by typing clear and press enter and then type ls space -l and press enter.

Now this is the web application which I've configured to be automatically downloaded to the EC2 instance.

It's a simple web page we've got index.html which is the index we have a number of images which this index.html contains and then we have a docker file.

Now this docker file is the thing that the docker engine will use to create our docker image.

I want to spend a couple of moments just stepping you through exactly what's within this docker file.

So I'm going to move across to my text editor and this is the docker file that's been automatically downloaded to your EC2 instance.

Each of these lines is a directive to the docker engine to perform a specific task and remember we're using this to create a docker image.

This first line tells the docker engine that we want to use version 8 of the Red Hat Universal base image as the base component for our docker image.

This next line sets the maintainer label it's essentially a brief description of what the image is and who's maintaining it in this case it's just a placeholder of animals for life.

This next line runs a command specifically the yum command to install some software specifically the Apache web server.

This next command copy copies files from the local directory when you use the docker command to create an image so it's copying that index.html file from this local folder that I've just been talking about and it's going to put it inside the docker image in this path so it's going to copy index.html to /var/www/html and this is where an Apache web server expects this index.html to be located.

This next command is going to do the same process for all of the jpegs in this folder so we've got a total of six jpegs and they're going to be copied into this folder inside the docker image.

This line sets the entry point and this essentially determines what is first run when this docker image is used to create a docker container.

In this example it's going to run the Apache web server and finally this expose command can be used for a docker image to tell the docker engine which services should be exposed.

Now this doesn't actually perform any configuration it simply tells the docker engine what port is exposed in this case port 80 which is HTTP.

Now this docker file is going to be used when we run the next command which is to create a docker image.

So essentially this file is the same docker file that's been downloaded to your EC2 instance and that's what we're going to run next.

So this is the next command within the lesson commands document and this command builds a container image.

What we're essentially doing is giving it the location of the docker file.

This dot at the end contains the working directory so it's here where we're going to find the docker file and any associated files that that docker file uses.

So we're going to run this command and this is going to create our docker image.

So let's go ahead and run this command.

It's going to download version 8 of UBI which it will use as a starting point and then it's going to run through every line in the docker file performing each of the directives and each of those directives is going to create another layer within the docker image.

Remember from the theory lesson each line within the docker file generally creates a new file system layer so a new layer of a docker image and that's how docker images are efficient because you can reuse those layers.

Now in this case this has been successful.

We've successfully built a docker image with this ID so it's giving it a unique ID and it's tagged this docker image with this tag colon latest.

So this means that we have a docker image that's now stored on this EC2 instance.

Now I'll go ahead and clear the screen to make it easier to see and let's go ahead and run the next command which is within the lesson commands document and this is going to show us a list of images that are on this EC2 instance but we're going to filter based on the name container of cats and this will show us the docker image which we've just created.

So the next thing that we need to do is to use the docker run command which is going to take the image that we've just created and use it to create a running container and it's that container that we're going to be able to interact with.

So this is the command that we're going to use it's the next one within the lesson commands document.

It's docker run and then it's telling it to map port 80 on the container with port 80 on the EC2 instance and it's telling it to use the container of cats image and if we run that command docker is going to take the docker image that we've got on this EC2 instance run it to create a running container and we should be able to interact with that container.

So if you go back to the AWS console if we click on instances so look for a4l-public EC2 that's in the running state.

I'm just going to go ahead and select this instance so that we can see the information and we need the public IP address of this instance.

Go ahead and click on this icon to copy the public IP address into your clipboard and then open that in a new tab.

Now be sure not to use this link to the right because that's got a tendency to open the HTTPS version.

We just need to use the IP address directly.

So copy that into your clipboard open a new tab and then open that IP address and now we can see the amazing application if it fits i sits in a container in a container and this amazing looking enterprise application is what's contained in the docker image that you just created and it's now running inside a container based off that image.

So that's great everything's working as expected and that's running locally on the EC2 instance.

Now in the demo lesson for the elastic container service that's coming up later in this section of the course you have two options.

You can either use my docker image which is this image that I've just created or you can use your own docker image.

If you're going to use my docker image then you can skip this next step.

You don't need a docker hub account and you don't need to upload your image.

If you want to use your own image then you do need to follow these next few steps and I need to follow them anyway because I need to upload this image to docker hub so that you can potentially use it rather than your own image.

So I'm going to move back to the session manager tab and I'm going to control C to exit out of this running container and I'm going to type clear to clear the screen and make it easier to see.

Now to upload this to docker hub first you need to log in to docker hub using your credentials and you can do that using this command.

So it's docker space login space double hyphen username equals and then your username.

So if you're doing this in your own environment you need to delete this placeholder and type your username.

I'm going to type my username because I'll be uploading this image to my docker hub.

So this is my docker hub username and then press enter and it's going to ask for the corresponding password to this username.

So I'm going to paste in my password if you're logging into your docker hub you should use your password.

Once you've pasted in the password go ahead and press enter and that will log you in to docker hub.

Now you don't have to worry about the security message because whilst your docker hub password is going to be stored on the EC2 instance shortly we're going to terminate this instance which will remove all traces of this password from this machine.

Okay so again we're going to upload our docker image to docker hub so let's run this command again and you'll see because we're just using the docker images command we can see the base image as well as our image.

So we can see red hat UBI 8.

We want the container of cats latest though so what you need to do is copy down the image ID of the container of cats image.

So this is the top line in my case container of cats latest and then the image ID.

So then we need to run this command so docker space tag and then the image ID that you've just copied into your clipboard and then a space and then your docker hub username.

In my case it's actrl with 1L if you're following along you need to use your own username and then forward slash and then the name of the image that you want this to be stored as on docker hub so I'm going to use container of cats.

So that's the command you need to use so docker tag and then your image ID for container of cats and then your username forward slash container of cats and press enter and that's everything we need to do to prepare to upload this image to docker hub.

So the last command that we need to run is the command to actually upload the image to docker hub and that command is docker space push so we're going to push the image to docker hub then we need to specify the docker hub username so again this is my username but if you're doing this in your environment it needs to be your username and then forward slash and then the image name in my case container of cats and then colon latest and once you've got all that go ahead and press enter and that's going to push the docker image that you've just created up to your docker hub account and once it's up there it means that we can deploy from that docker image to other EC2 instances and even ECS and we're going to do that in a later demo in this section of the course.

Now that's everything that you need to do in this demo lesson you've essentially installed and configured the docker engine you've used a docker file to create a docker image from some local assets you've tested that docker image by running a container using that image and then you've uploaded that image to docker hub and as I mentioned before we're going to use that in a future demo lesson in this section of the course.

Now the only thing that remains to do is to clear up the infrastructure that we've used in this demo lesson so go ahead and close down all of these extra tabs and go back to the cloud formation console this is the stack that's been created by the one click deployment link so all you need to do is select this stack it should be called EC2 docker and then click on delete and confirm that deletion and that will return the account into the same state as it was at the start of this demo lesson.

Now that is everything you need to do in this demo lesson I hope it's been useful and I hope you've enjoyed it so go ahead and complete the video and when you're ready I look forward to you joining me in the next.

That is a very valid concern. AI-generated content, such as from ChatGPT, tends to spam online platforms like email and Wikipedia with misinformation, making people not trust the platforms. Because Wikipedia, for example, enables users to edit entries, it is highly susceptible to the addition of false information. There are systems in place for moderation, but it's tough to keep up with how quickly AI can generate content. It requires stronger editorial controls and awareness on the part of users to maintain the reliability of such platforms.

Does anyone know the original Italian word for "work"?

What does the Italian word "work" convey in Montessori's time?

Language evolution! Historical discussion.

[MAPS 2024 conversation] Italian translations of the term "work": * "meaningful activity" * "play" (lavora) ... i.e., "meaningful play Context: English translations of Montessori's original writing. Italian has different meanings than English translations Historical context matters as it relates to the meaning of terms.

https://www.nytimes.com/2024/08/28/climate/canada-wildfires-emissions-carbon.html

Studien: https://www.nature.com/articles/s41586-024-07878-z

I completely agree with this. As TikTok gained popularity with its short videos, many other platforms quickly adopted this feature for creating and sharing short-form content. Instagram introduced Reels, and YouTube launched Shorts, both experiencing significant growth as a result. Even Spotify has now incorporated a similar short video format.

Discipline

Related to agency

This is related to normalization

Broader /related Term: Third Plane of Development; Adolescence

what is quarks?

Die aktuelle weltweite Korallenbleiche ist bereits die vierte in 25 Jahren. Die Temperaturen in einem großen Teil der tropischen Meere lagen in diesem Sommer 3° über dem Durchschnitt. Im Gespräch mit der Repubblica erklärt der Korallenexperte Roberto Danovaro, dass ein Viertel der globalen Korallenbestände bereits verloren ist. Korallenriffe, die Ökosysteme mit der größten Biodiversität, sind durch die globale Erhitzung besonders verwundbar https://www.repubblica.it/green-and-blue/dossier/negazionisti-climatici/2024/10/07/news/barriera_corallina_sbiancamento_crisi_clima_roberto_danovaro-423531902/

https://pierce.instructure.com/courses/2517205/pages/week-6-required-reading?module_item_id=87293778

https://pierce.instructure.com/courses/2517205/pages/week-5-required-reading?module_item_id=87293770

https://pierce.instructure.com/courses/2517205/pages/week-4-required-viewing?module_item_id=87293761

https://pierce.instructure.com/courses/2517205/pages/week-2-required-reading-2?module_item_id=87293741

https://pierce.instructure.com/courses/2517205/pages/week-1-required-readings?module_item_id=87293729

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Extreme Regenfälle im Südosten und Zentrum Frankreichs im September gehören zu einer sogenannten „Mittelmeerepisode“ (épisode méditerranéen). Diese Ereignisse intensivieren sich durch die globale.Erhitzung. Die Libération hat dazu Klimaforschende befragt, und sie verweist auf eine aktualisierte Studie zur Erwärmung in Frankreich https://www.liberation.fr/environnement/climat/les-intemperies-dans-le-sud-est-de-la-france-enieme-illustration-des-effets-devastateurs-du-rechauffement-climatique-20241018_HCAHWSJACRADZNAVD5G7IZE66M/

Studie: https://esd.copernicus.org/articles/13/1397/2022/