I think that is also help me, it really teaches me how to deal with those negative feelings.

I can understand his feelings, because I always has same feeling during my childhood. Asian family is strict to the grades, we must do our best to get the A. If not, family members will be disappointed to you.

eLife Assessment

This important study presents a novel approach to enhance the therapeutic potential of mesenchymal stromal cells (MSCs) by genetically modifying their glycogen synthesis pathway, resulting in increased glycogen accumulation and improved cell survival under starvation conditions, particularly in the context of experimental pulmonary fibrosis. The methods and findings are generally solid and could be strengthened in the future by investigating the kinetics of persistence, the immunomodulatory effects, and the underlying improved mechanism of action of MSCs in this pulmonary fibrosis model. If confirmed, this approach could suggest potential methods to improve the therapeutic functionality of MSCs in cell therapy strategies.

Reviewer #1 (Public review):

Summary:

This study provides the first evidence that glucose availability, previously shown to support cell survival in other models, is also a key determinant for post-implantation MSC survival in the specific context of pulmonary fibrosis. To address glucose depletion in this context, the authors propose an original, elegant, and rational strategy: enhancing intracellular glycogen stores to provide transplanted MSCs with an internal energy reserve. This approach aims to prolong their viability and therapeutic functionality after implantation.

Strengths:

The efficacy of this metabolic engineering strategy is robustly demonstrated both in vitro and in an orthotopic mouse model of pulmonary fibrosis.

Reviewer #2 (Public review):

Summary:

In this article, the authors investigate enhancing the therapeutic and regenerative properties of mesenchymal stem cells (MSCs) through genetic modification, specifically by overexpressing genes involved in the glycogen synthesis pathway. By creating a non-phosphorylatable mutant form of glycogen synthase (GYSmut), the authors successfully increased glycogen accumulation in MSCs, leading to significantly improved cell survival under starvation conditions. The study highlights the potential of glycogen engineering to improve MSC function, especially in inflammatory or energy-deficient environments. However, critical gaps in the study's design, including the lack of validation of key findings, limited differentiation assessments, and missing data on MSC-GYSmut resistance to reactive oxygen species (ROS), necessitate further exploration.

Strengths:

(1) Novel Approach: The study introduces an innovative method of enhancing MSC function by manipulating glycogen metabolism.

(2) Increased Glycogen Storage: The genetic modification of GYS1, resulting in GYSmut, significantly increased glycogen accumulation, leading to improved MSC survival under starvation, which has strong implications for enhancing MSC therapeutic properties in energy-deficient environments.

(3) Potential Therapeutic Impact: The findings suggest significant therapeutic potential for MSCs in conditions that require improved survival, persistence, and immunomodulation, especially in inflammatory or energy-limited settings.

(4) In Vivo Validation: The in vivo murine model of pulmonary fibrosis demonstrated the improved survival and persistence of MSC-GYSmut, supporting the translational potential of the approach.

Weaknesses:

(1) Lack of Differentiation Assessments: The study did not evaluate key MSC differentiation pathways, including chondrogenic and osteogenic differentiation. The absence of analysis of classical MSC surface markers and multipotency limits the understanding of the full potential of MSC-GYSmut.

(2) Missing Validation of RNA Sequencing Data: Although RNA sequencing data revealed promising transcriptomic changes in chondrogenesis and metabolic pathways, these findings were not experimentally validated, limiting confidence.

(3) Lack of ROS Resistance Analysis: Resistance to reactive oxygen species (ROS), an important feature for MSCs under regenerative conditions, was not assessed, leaving out a critical aspect of MSC function.

(4) Limited Exploration of Immunosuppressive Properties: The study did not address the immunosuppressive functions of MSC-GYSmut, which are critical for MSC-based therapies in clinical settings.

Conclusion:

The study presents an exciting new direction for enhancing MSC function through glycogen metabolism engineering. While the results show promise, key experiments and validations are missing, and several areas, such as differentiation capacity, ROS resistance, and immunosuppressive properties, require further investigation. Addressing these gaps would solidify the conclusions and strengthen the potential clinical applications of MSC-GYSmut in regenerative medicine.

Author response:

The following is the authors’ response to the original reviews.

Reviewer #1 (Public Review)：

(1) Glycogen biosynthesis typically involves several enzymes. In this context, could the authors comment on the effect of overexpressing a single enzyme - especially a mutant version - on the structure or quality of the glycogen synthesized?

While quantitative molecular weight analysis of synthesized glycogen was not performed, we documented changes in glycogen particle morphology. GYSmut overexpression resulted in significantly enlarged singular glycogen granules, suggesting potential high molecular mass, while GYS-GYG co-overexpression in MSCs (GYG being the essential enzyme for glycogen synthesis initiation) produced a diffuse glycogen distribution pattern rather than particulate structures. We have incorporated this result as new Figure S2C.

These results suggest that overexpression of specific glycogen-metabolizing enzymes significantly influences glycogen structure. Consequently, targeted modulation of glycogen architecture and properties through key enzymes represents a potential avenue for future investigation.

(2) Regarding the in vitro starvation experiments (Figure 2C), what oxygen conditions (pO₂) were used? Are these conditions physiologically relevant and representative of the in vivo lung microenvironment?

Our in vitro starvation experiments (Figure 3C) were conducted under normoxic (21%). The oxygen concentration in human lungs is physiologically lower than atmospheric levels, with healthy individuals exhaling air containing approximately 16% oxygen (Thalakkotur Lazar Mathew, Diagnostics 2015). To our knowledge, direct measurements of alveolar oxygen concentration in pulmonary fibrosis are rare. Therefore, to evaluate the performance of GYSmut under hypoxic conditions, in the revised manuscript, Figure S2 has been augmented to include assessment of cell performance under combined hypoxia （oxygen concentration < 5%）and nutrient deprivation stress, which further corroborate the superiority of the GYSmut group over the control under different oxygen concentrations. 

(3) In the in vitro model, how many hours does it take for the intracellular glycogen reserve to be completely depleted under starvation conditions?

While quantitative cell viability data were recorded up to 72 hours post-implantation (Fig 3C), we observed cell viability at approximately 96 hours. We noticed that the presence of glycogen particles exhibited a correlation with sustained cell viability. However, reliable quantitative assessment of glycogen became increasingly challenging upon significant depletion of viable cells, thereby limiting our measurements during later time points.

(4) For the in vivo model, is there a quantitative analysis of the survival kinetics of the transplanted cells over time for each group? This would help to better assess the role and duration of glycogen stores as an energy buffer after implantation.

We tracked the in vivo distribution and persistence of implanted MSCs using enzymatic activity quantification assays (using Gluc luciferase assay) and live animal imaging (using Akaluc luciferase). The revised manuscript includes quantitative analysis of the in vivo fluorescence imaging data, which has been supplemented as Figure S4. Glycogen-engineered MSCs and control cells were quantitatively assessed at three discrete time points post-implantation. This quantification revealed a transient divergence in cell viability between the experimental and control groups around day 7. However, fluorescence in both cohorts subsequently declined to similar levels over the extended observation period.

(5) Finally, the study was performed in male mice only. Could sex differences exist in the efficacy or metabolism of the engineered MSCs? It would be helpful to discuss whether the approach could be expected to be similarly effective in female subjects.

We appreciate the reviewer’s important question regarding potential sex differences. Our study used male mice based on three key considerations: 1) Clinical Relevance: Idiopathic pulmonary fibrosis (IPF) shows significant male predominance, with diagnosis rates 3.5-fold higher in men (37.8% vs 10.6%, p<0.0001) and greater diagnostic confidence (Assayag et al., Thorax 2020). 2) Model Consistency: The bleomycin model (our chosen method) demonstrates more consistent fibrotic responses in male mice (Gul et al., BMC Pulm Med 2023). 3) Biological Rationale:

Estrogen’s protective effects in females may confound therapeutic assessments (cited in Assayag et al.).

We fully acknowledge this limitation and will include female subjects in subsequent translational studies. The therapeutic principle should theoretically apply to both sexes, but we agree this requires experimental validation.

(6) The number of mice for each group and time point should be specified.

The manuscript text has been revised to enhance clarity, and the number of mice for each group and time point has been specified (line 170 to 182).

Reviewer #2 (Public Review):

(4) Inconsistencies in In Vivo Data: There is a discrepancy between the number of animals shown in the figures and the graph (three individuals vs. five animals), as well as missing details on how luciferase signal intensity was quantified, requiring further clarification.

To assess MSC survival in vivo, we employed two strategies utilizing distinct luciferases optimized for specific detection modalities. MSC viability was quantified ex vivo through Gaussia luciferase (Gluc) activity, leveraging its high sensitivity and established commercial assay kits (n = 3 mice per group per time point). For non-invasive longitudinal tracking within living animals, MSC distribution and viability were monitored via in vivo bioluminescence imaging using Akaluc luciferase, selected for its superior tissue penetration and sensitivity in situ (n = 5 mice per group).The manuscript text has been revised to enhance clarity, and the experiment protocols for luciferase signal detection and quantification has been added into Methods.

（1) (2) (3) (5):

We fully agree that further investigation into the functional consequences of glycogen engineering in MSCs – encompassing core cellular functions, immunomodulatory properties, and associated signaling pathways – is important to fully elucidate the underlying mechanisms. Cellular metabolism is intrinsically intertwined with diverse physiological processes. Consequently, we believe that glycogen engineering exerts multifaceted effects on MSCs, likely extending beyond the modulation of any single specific pathway. Studying the metabolic perturbation induced by such engineering approaches in mammalian cells represents an interesting field. The exploration of these aspects remains an long-term research objective within our group.

Reviewer #2 (Recommendations for the authors):

(6) Clarification of Data in the Murine Model:

In Figure 4B, there is a discrepancy between the number of animals shown in the image (five) and those represented in the graph (three). This discrepancy needs clarification. Additionally, the study lacks information regarding the intensity of the signal in the luciferase assays. It is unclear how luciferase expression in the mice was quantified, and providing this detail would enhance the understanding of the data presented.

We sincerely appreciate these valuable suggestions. We have revised the relevant text for greater clarity. Figure 4B and Figure 4C present results from two distinct experimental approaches, each employing different luciferase reporters and measurement methodologies, and different num of mice were used in these two experiments.

Quantitative data derived from the in vivo bioluminescence imaging has been supplemented as Figure S4. The experiment protocols for luciferase signal detection and quantification has been added into Methods.

To other recommendations of reviewer 2：

We sincerely appreciate your valuable insights, which demonstrate your deep expertise. We fully agree that beyond nutrient availability, factors such as reactive oxygen species (ROS) and the immune microenvironment are also critical limitations affecting the survival and therapeutic efficacy of implanted MSCs.

We propose that glycogen engineering exerts broad effects on MSCs. These effects manifest as changes in multiple cellular characteristics, including proliferation, differentiation, surface marker expression, antioxidant capacity, and immunomodulatory activity – all crucial factors for the therapeutic purpose of MSCs.

We believe these changes likely involve complex networks of interconnected regulatory factors. The underlying mechanisms might be clarified through proteomic and metabolomic profiling.

However, comprehensively investigating these interconnected aspects requires significant time and resources. Some components of this research extend beyond the current scope of our project. Nevertheless, exploring these mechanisms remains an important objective, and we will actively work to investigate them further in our ongoing studies.

There are racial and ethnic factors that play a big role in this question. Generally, POC spend more time with extended family than white people.

There are limits to this study. The responses weren't collected in real time, there's no results for long-term responses, and there's a need to better measure the quality of social interactions. Compare the results with caution

Extended family benefits are more compensatory

older siblings play a more valuable role than younger siblings, but they still both can improve well-being

I wonder if the long-term stressors are because of this factor, or other factors?

this study reveals benefits that add to the study of long-term consequences of stepparents on adolescents.

strong stepparent relationships depend on each individual child and stepparent.

is this because they didn't feel inclined to spend time with the stepparent? Or both parents because the parents are spending more time together?

there are generally more benefits to interacting with siblings

while there are benefits to interacting with other family members, the most important members are still the parents, which is why most studies are sone on parent-child relationships.

While parental relationships are most important, studies revealed that interacting with both parents was more beneficial, suggesting benefits to interacting with multiple family members.

This study was important in expanding our knowledge of adolescent development because it studied more than just immediate and parent-child relationships. It revealed the importance of interacting with other family members and how each member affcets adolescents

The American Time Use Survey- conducted annually to gather info about how people allocate their time. (Work, Family, Leisure, etc.)

compensation theory- when individuals adjust their behavior in response to perceived disadvantages or transgressions

congruence theory- model of attitude adjustments made after a person receives new info. (initially separate attitudes become connected after learning new info)

why is it most beneficial for them? I'm guessing it has something to do with how much time the parent spends trying to fulfill both parental roles.

Interacting with younger sibling is common, but the effect on adolescents isn't studied enough. We mostly have results of affects older siblings have

The role resident parents play during interactions with nonresident parents can affect the emotions adolescents have when interacting with nonresident parents

social capital- the value (advancing the goals of individuals and groups) from positive connections and relationships between individuals

Other family relationships are affected by parent-child relationships and any factor that affects a parent-child relationship, affects all family relationships (interactions, alliances, and boundaries)

Social learning theory- learning that occurs through observation, imitation, and modeling ex: kids will learn how to positively interact with other family members because they learned out to socially interact w their parents.

Parent-child interactions in a nonnuclear home (such as single parents or stepparents) can have resulting consequences for other family interactions. Both examples showed benefit of interacting with other family members. One by seeking out positive interactions with other members, and the other by a social learning theory.

exploring the different moods and responses children have when interacting with different family members (not just nuclear) provides us with a more in depth and nuanced view of adolescent core-affect

core affect- influence reflexes, cognition, perception, and behavior

The research isn't focused on the effects of non-parent- child relationships on adolescent development, meaning there's missing factors in current studies that could be crucial to understanding heterogeneity in adolescents.

Heterogeneity- Diversity in character/content

Within-Group analysis- statistical method that compares same subjects in different conditions or at different points in time

Heterogeneity isn't commonly studied in relation to this theory, so they used a large, nationally representative data sample of adolescents to separately study adolescent feelings in different home contexts by doing a within- group analysis. Adolescents had benefits from interacting with whoever they lived with.

This study helps with designing interventions for adolescents because it conveys more precise info about which family members positively influence emotional responses in adolescents.

The Family systems theory suggests that variation in home contexts can shape emotional functioning and social developments in adolescents.

Family structure is an important role in the diversity of character in adolescents. their responses and mood with certain family members member can affect their relationships

The study focuses on how often adolescents interact w family members and how they feel when spending time with certain members.

Another example of how important representation and how detrimental a lack of it is.

I imagine that introducing renewable assignments would require a ton of set up and building a foundation for the students so they understand the concept, reasoning etc. before getting started.

Giving student options so that they can create something meaningful to them is so important!

This goes back to the theme and discussion from last week on centering care and creating a brave space.

C'est à cause de l'être humain qui laisse le numérique intègre dans tous les domaines de la vie, qui nous aboutis à perde de plus en plus notre identité

Je ne vois pas "l'évaluation" comme il est indiqué dans le texte, nous évaluons quelque trucs mais pas notre utilisation est tout le temps pour évaluer les choses; mais nous sommes évaluer par les inventeurs de numérique et l'algorithme

on ne peut plus être indépendant d'eux

Les avantages du numérique

Utilisation de la psychologie pour nous faire attacher au numérique

L'importance de l'interface dans le numérique

La naturalisation de numérique qui devient une partie essentiel dans notre quotidien

This entire section can be useful for educators working on developing Guidance Policies. Preventative measures are key in, well, preventing any unwanted situations. I think it's important for educators to self-reflect and make sure they are promoting a safe, kind, and appropriate environment before introducing the children to it.

This reminds me of a time last summer when a child climbed to the top of the monkey bars, and was incapable of getting down to try to get down. She cried as everyone looked on. I offered suggestions such as scooting across to the lowest part where she could safely get down onto the connected playground structure. As well as holding onto the monkey bars and slipping through the middle, and safely dropping on the floor. However, she was too scared to try anything, remaining frozen in fear at the top.

After about 15 minutes, the teacher and I decided to have me remain on the floor and lift her tiny hands from clenching onto the monkey bars. At the same time, the main teacher went behind the student, picked her up, and passed her to me to help set her down safely onto the playground structure. Although the student was scared, she was relieved to be out of the situation. It surprised me that there were regulations set for instances like this. This is really good to know for any future situations.

In some cases, children may come from a home where neither of these two key concepts of child development are present. What are ways that educators can mitigate an/or work our way around any possible negative results in school environment stemming from the lack of these two key concepts?

I feel these are great questions to get feedback from your colleagues, to make sure we have covered everything for the kids safety. I believe the last question is crucial because our job is to teach but more than that is to provide an environment for the kids to safe and for their health.

I believe this is crucial for us teacher to know and understand, and the reason why we should pay attention to every child observe how they react and act to certain situations so when cases like this comes up we can identify how to provide the right responsive step so the child can feel comfortable but not bring bad memories.

I was surprised to know that we only hold the child to remove them from the dangerous situation and how long is the appropriate holding time ?

This article describes how a comedian pretended to be the official FBI account on Twitter and posted a joke about Martin Luther King Jr's assassination. While the intention may have been humor, the impersonation clearly crossed ethical boundaries by exploiting both historical trauma and public trust. I think this example connects directly to Chapter 7's discussion of trolling and "bad faith". The comedian's defense that it was "just a joke" mirrors the Schrodinger's asshole concept, using humor as a shield to deny accountability when the audience reacts negatively. It raises an important question: when dose satire stop being social commentary and start becoming harm disguised as entertainment.

I find it fascinating that trolling began as a kind of "inside joke" among early Internet users. What started as playful testing of newcomers' knowledge later evolved into behaviors that intentionally cause harm or humiliation. Reading about the shift from lighthearted pranks to toxic online cultures like 4chan made me realize how easily community norms can slide when cruelty becomes entertainment. It also connects to Sartre's idea of "bad faith" mentioned earlier, the troll's refusal to take words seriously allows them to avoid responsibility. It makes me wonder whether trolling is less about anonymity and more about disengagement from empathy.

Un análisis con ELSOC

la conceptualizacion y medicion de cohesion social para AL del OCS

Relevaría aquí que, dada la relevancia de la cohesión social para mantener la estabilidad de la sociedad, el lazo etc, poder medirlo empiricamete es clave y ello es algo fragmentado a nivel internacional y no existe un esfuerzo como tal en Chile antes del OCS

Hi, I was interested in the full dataset but it looks like the data either have not yet been deposited or the placeholders have accidentally not been replaced here. I was also unable to also find them via an admittedly cursory search directly in the appropriate repos (recognizing that the US government shutdown may be delaying updating of NCBI).

Is this uppercased letters intentional?

Should be:

b. Call GET /1-click/verifications/{verificationUuid}

Might confuse the reader that he has to call GET /1-click/verifications

The varianceexplained.org analysis of Trump’s tweets is a great example of how data science can uncover patterns behind online personas — it really connects to this chapter’s discussion of authenticity. I found it fascinating how statistical language analysis can expose who’s “really” speaking behind a public account, showing that authenticity online can sometimes be measured rather than just perceived.

It feels like Schrodinger's Asshole's have become so much more prominent online in recent years and I think its a consequence of how cynical people have become. Its definitely a way for people to signal their actual beliefs and then attempt to bail out when they face social repercussions for those beliefs.

I think this video on Jennifer Lawrence is a perfect example of online personas, and people's parasocial relationships with celebrities. Jennifer Lawrence in the video is said the be the ultimate cool girl. Doing things like hanging out her her male co stars, casually eating junk food while maintaining a slim figure, and being unfiltered. A stark contrast from her often serious and tough roles-- such as Katniss Everdeen. This left a lot of people speculating if her "cool girl" personality was even real, or just something she contrived to seem more relatable. So as this shows, no matter how Jennifer Lawrence acts, she will always have people questioning the authenticity of what she choose to display to the public, despite them not even knowing her in person.

When reading this source, about COVID-19, one factor that I realized was the number of fatality's that were encountered for. It seems like this source actually undermines the actual number, perhaps to make people less in a panic. However, this could lead to the majority of the public to be untrusting of the sources giving them data, which in turn will lead to future reportings not being taken as serious.

I partially agree with this tweet's contents regarding mental health and relationships towards jobs. I also disagree with how large of a generalization this is of a huge group of people.

I remember back when this happened, when Twitter was still X. So many people cried out in anger when Trump was banned, claiming that free speech was being infringed upon. But Twitter was a private company that had the right to ban whoever it wanted. The reaction so many people, including Elon Musk and Trump himself, I feel was the writing on the walls to Musk eventually purchasing Twitter. But also looking into the question of authenticity, and seeing that Trump may have had a team specifically hired to post inflammatory rhetoric really speaks to how difficult moderation is on the internet, and how much the platform of Twitter has changed as Musk took over.

One of the things that I noticed in this source was the inauthenticity that was discussed about the COVID-19 death rates, showing that the true number of deaths was actually much higher than official reports said. Things like this can cause mistrust among societies, which can ultimately lead to their downfall.

That's a pretty interesting topic. Although people say people have the right to choose which type of phones they want, and there's no difference in using different operating systems regarding their perspective, education, and social status. However, sometimes we can see there's a difference between iOS and Android users; for example, Android users tend to know phones better, as Android is an open system. It's very interesting to see why people are posting different persepctive based on their device choice

I find it interesting how once the authenticity of the stories were called into question, people started to care less and got upset at the creator. If a story is entertaining, why should it matter that it's fictional?

Lonelygirl15's Wikipedia entry initially presented the creator as a real-life teenage vlogger, but was later revealed to be a scripted project. This example highlights that "authenticity" can be manufactured, and that the audience's trust can ultimately translate into a feeling of betrayal. It also shows that online persona is not just a matter of personal expression, but also involves power and control over narrative. When viewers discover the truth, the strong response reflects a view of authenticity as a social contract between creator and audience

This link talks about how anonymity online could be both harmful and beneficial for people. While it is hiding people from disrespecting or abusing others, it is protecting minorities from violence. For instance, women and LGBTQ individuals. Survivors of domestic violence rely on anonymity online to hide and heal from abusers. In the passage, it mentions that trans and queer women face threats online and offline, so anonymity protects their privacy from being invaded and attacked by others.  Survivors should also not be asked to leave the internet because it is not fair for them to get punished for someone else’s mistake. So if it is needed, use a fake name for your social media account to protect your privacy. 

This wikipedia page goes into an engraved tablet meant for Ea-nasir. It's a complaint from a customer about the bad quality of his goods. This was in the ancient mesipotamian culture and Ea-nasir was selling copper. They complained about the quality of the copper and the treatment they got from the delivery guy making it the oldest written complaint.

This story is absolutely wild. A woman named BethAnn pretended on Twitter to be a Native American queer woman who was a professor at ASU and who was a victim of sexual harassment. She used the account for her own personal gain. She used it to give credibility to herself (the fake account would post praise about BethAnn), lead MeTooSTEM, and even raise $75k through a gofundme. One day BethAnn announced that the woman behind the account had died of Covid and she even held a funeral for her. This level of deception is insane. It shows that of course, we can't believe everything we see on the Internet. It also serves as a warning to us of the level of deceit and manipulation that the Internet can be used to create.

This article analyzes how tweets posted from Donald Trump's Android device were significantly more negative and aggressive than those from his campaign's iPhone, which were more neutral and polished. The researchers used sentiment analysis to identify emotional differences between the two sets of tweet. I found this especially relevant to the discussion of inauthenticity in Chapter 6, because it shows that even when a social media account seems to belong to one person, multiple voices or personas may actually be behind it. This raises important ethical questions about authenticity and accountability online.

I think the story of Lonelygirl15 is particularly intriguing. At first, she shared her life on YouTube as an ordinary girl, and many people found her sincere and approachable, like a friend to confide in. But later, it was discovered that it was actually a performance, a web drama meticulously planned by actors and the production team. When the truth came out, the reaction of the audience was very strong - not only because they were deceived, but also because the feeling of "being understood" and "being connected" suddenly vanished. I can understand the emotion of betrayal because the trust we invest in the online world is, in essence, as precious as trust in real life. But looking back, I also think the phenomenon of Lonelygirl15 reminds us: Authenticity doesn't necessarily mean "complete truth". It can also be a more complex emotional experience. Even if it was a performance, the sense of companionship and resonance people gained while watching was also real. Perhaps this is the most contradictory and most human aspect of the internet age.

[f10] Regarding the “unidentified professor”: This report reveals how community skepticism (strange memorial service phone calls, no Arizona State University records) confirmed that @Sciencing_Bi was a McLaughlin scam and how the scam caused multiple harms: identity theft, community manipulation, and the discrediting of a genuine Title IX whistleblower. The name reference and institutional confirmation are powerful sources. Combined with BuzzFeed [f9] and The Verge [f13], this strengthens the timeline and mitigates the bias of a single source.

This is so valuable!

This refers to ads like the Jumbotron display.?

I think the parallels between the horses and racism are pretty strange.

Adding a bit more to the exploratory part of the work, by questioning why marriages with non-English-speaking spouses start off great, but later fail. The author then answers their own question.

This part initially surprised me when the topic was about talking to animals, then all of a sudden to started to talk about slavery.

The connection between humans and animals.

First glance at the author introducing communication as the topic of this work. Exploring how communication between animals and humans can be lost, but at the fault of the human.

I think authenticity as to who is actually making post on any social media is very important. With Donald Trump any tweet on his account is assumed to be his as it was his personal account, so having knowledge that another person was making tweets under his name is certainly frustrating.

I think it's interesting how normalized it is for a popular figure like the president to have their own social media manager. To me, it feels less personal knowing that every tweet a celebrity tweets is not actually their own thoughts and opinions.

Do you think it should be in quotes?

Vosotros

I was struck by the tension in the chapter between authenticity as honesty and authenticity as the matching of presentation and interaction. The idea that social media personas can “feel” authentic even when they’re partly performed makes me wonder: in platforms driven by algorithms and engagement metrics, do we discourage real authenticity because performative authenticity “wins” more often?

I think that it is super interesting that we can take artifacts that prove that being honest and being authentic is something that societies have relied on to stay successful for thousands of years.

Why does authenticity when watching something bother us? We sit down and watch fictional movies for hours at a time, watch hour long episodes of fictional TV shows weekly and give it our full attention, but what about watching someone online tell stories bothers us? I think it's likely associated with mistrust and dishonesty, like the passage says. Influences now lie all the time, but are much more careful and covert in how they do it, when as a result, when they're caught, the consequences are more dramatic in comparison to inauthenticity with low effort to cover it up.

This idea plays out often in short-form video content (such as TikTok, Youtube shorts, or Instagram reels). Creators like hayleyybaylee and janeinsane exemplify this idea. They made sarcastic and satirical videos that at first appear to be reality. Hayleyybaylee makes videos about a day in her life as a "billionaire's girlfriend," where she does absurd tasks around her fancy New York apartment dressed in full glam. Janeinsane makes videos about Utah moms. She exaggerates their stereotypical qualities (for instance, using Stanley cups). Both creators are playful and fun, and their audiences aren't mad that they aren't completely authentic. The purpose of their content isn't to inform or teach, it's simply to make others laugh. That is where it's excusable to let authenticity slide.

I can truly understand this statement. The feeling of being deceived is truly awful - not only because we were deceived, but also because we start to doubt our ability to make correct judgments. This reminds me of some "true stories" accounts I followed on social media earlier. Later, I discovered that they were actually fabricated. The sense of loss is deeper than just an information error. Perhaps the reason why we react so strongly to "falsehood" is that trust is an emotional investment for us. When others take advantage of this trust, we lose not only the authenticity of the information but also the sense of security between people.

Let’s answer this question: Performance itself does not compromise authenticity. As defined in this chapter, a connection is authentic when the service provided matches the actual service. If the audience can identify with a carefully crafted persona and feel the intimacy promised, the connection is authentic. If important facts are hidden (for example, through undisclosed sponsorships or deceptive practices), it becomes fake. In my research on the Bluesky bot, labeling posts as coming from a bot maintained trust and correlated the connection provided with reality.

Est-il usuel de désigner la péninsule par le seul adjectif « Indochinoise » ?

Pouvez-vous confirmer que le « fonds indo-chinois » est bien écrit avec cette orthographe et sans la majuscule dans le titre de ce document ?

Plus haut, à la note 9, la « Note sur le fonds Indochinois » est localisée sous la cote E317 ; dans la note précédente, la cote E178 est identifiée comme celle de la « Note à l'attention de Monsieur l'Administrateur général sur l'urgente nécessité ... ». La cote E178 est-elle correcte ?

Langetermijngeheugen = - grote capaciteit en duur - de items blijven inactief, totdat het STM ze ophaalt - PRIMING = activering van informatie in het LTM door sensorische input: gebeurt onbewust.

Processen om informatie te verplaatsen = - ophalen (nadenken over het verleden), tegenhanger coderen, van LMT naar STM. - coderen = iets proberen te onthouden voor de toekomst, dit is actief en bewust - aandacht = aandacht schenken aan een informatiebron, zodat zintuiglijke input naar ons werkgeheugen gaat.

belangrijke naam

GELEERDE RESPONS NEEMT AF

Parasocial relationships are so common nowadays, especially in pop culture. We see this behavior done with fans of artist, fans of movies, or social media influencers. I believe that there is a difference between "admiring" someones work and then "obsessing" over someones work. There should be a clear line between the two, and some people don't have that thinking. In this example, we see that this lady believes Mr. Rodgers knows of her. While this is true in someway, he doesn't really "know" her. He just acknowledges her. But to this girl she genuinely believes that she is a part of his life.

I personally wouldn't say that I am in any parasocial relationships with media figures or people in my life currently. I was however in a lot of parasocial relationships as a child with characters like SpongeBob, Finn from adventure time, and various other shows and cartoons.

This kind of relationship reminds me that most online celebrities or social media stars always create a feeling like this. With the development of communication technology, audiences can interact with the celebrities just using their own account online, eg. commenting under videos, making online face-to-face calls, etc. But no longer restricted to TVs, a one-way communication route. However, this may cause some of the fans to interrupt the normal life of these celebrities rudely, and their own life are actually occupied by chasing the celebrity, grabbing their attention at them no matter what will takes.

I see parasocial relationship on the internet all the time. Any tiny connection or feelings can create a parasocial relationship. Influencers and their fans or even just followers. Creating a parasocial relationship with viewers online can bring large profit to the influencers. Viewers are promoting influencers and celebrities based on the parasocial relationship that they have which make them feel connected.

Biologists have traditionally classified all living organisms into two major groups, the prokaryotes and the eukaryotes (Figure 2.1). A prokaryote is a unicellular organism with a relatively simple cell structure. **A eukaryote has a compartmentalized cell structure with components bounded by intracellular membranes; eukaryotes may be either unicellular or multicellular.

• let divide nu den =
• let rec aux nu den n =
• if (nu < den) then n else
• aux (nu-den) den (n+1)
• in aux nu den 0;;

let divide nu den = let rec aux nu den n = if (nu < den) then n else aux (nu-den) den (n+1) in aux nu den 0;;

tiger.

working vehicle.

healthy, delicious, food.

I would use words like wreckage, debris, rubble, smoke, sirens, ghastly.

it leaves you feeling like something is missing, just like how the speaker feels being haunted by their voice.

create lines instead of one big paragraph.

The final lines of Keats’ poem “What the Thrush Said” –“He who saddens/At thought of idleness cannot be idle,/And he's awake who thinks himself asleep” touches on, like many other past sources, a human concern with the senses. In my annotation on the “Fire Sermon Discourse" and Augustine’s confessions, I said that “to unite humanity, we must relinquish that which makes us so, embracing divinity and the word of our gods, though they may present differently across cultures.” taking the common thread between the two schools of thought as a divergence from our human-ness. Unlike these sources, however, Keats almost celebrates one's connection with their senses. Rather than an enforcing of separation (through fire sermons and such), where true spirituality can only be reached by purging oneself of all things that tie them to their humanity, Keats equates one's sense of purpose with emotions, with the lines “He who saddens/At thought of idleness cannot be idle,” suggesting that by existing, by living through one's emotions, that is proof enough of life and connection. Furthermore, he celebrates human knowledge in the lines “O fret not after knowledge -- I have none,/And yet the Evening listens.” In this line Keats finds himself celebrating that which makes us human. Though the speaker “I” doesn’t have all-encompassing knowledge and has not sacrificed themself to the divine, “the Evening listens.” The dual powerlessness and power held by humanity in this moment showcases the human’s place in existence. In a poem harkening the arrival of spring, after “the Winter's wind,” Keats doesn’t ask anything of humanity besides existence, allowing the world to revive itself while we sit and let the seasons change. Instead, taking direct inspiration from Keats in his own “What the Thunder Said,” Eliot places humanity back into a dead waste land, where there “is no water but only rock.” While humanity is passive, though they lack knowledge, they are celebrated with the arrival of spring in Keats’ work. Eliot’s world has no hope of spring, with not a drop of water in sight. Human agency is revoked, and they “can neither stand nor lie nor sit.” This “can” added in this line seems to be at odds with the “He who saddens/At thought of idleness cannot be idle,” of Keats. Though in both cases, there is the sense of inability to be idle. (with the definition of idle being dually “without purpose or effect; pointless” and “avoiding work; lazy). While in Keats’ case, he suggests that, by having emotion, even if lamenting on laziness, one can never truly be “pointless,” Eliot’s humans can never be idle, though perhaps lacking purpose, because they can never avoid the work of escaping or exploring the waste land.

The story of Pheblas in “Death by Water” mirrors that of Ulysses in Tennyson’s poem under the same name. In both stories, the men are drawn to the water, to a life of adventure despite their old age. Ulysses, through all of his experiences is “a part of all that [he has] met,” existing as all prior parts of himself. Furthermore, as he lives his last days, thinking about how little of him there is left, he sees salvation in the “eternal silence” and seeks to follow this like a “sinking star.” The idea of a “sinking star” inverts the typical shooting star – a symbol of hope and wishes. The “sinking star” then becomes a symbol of inevitability. Additionally, in a story connected to the sea in all ways, the “sinking” also acts as a reference to drowning – sinking and dying in the exact place Ulysses yearns to return to. Similarly, in “Death by Water” “Phlebas the Phoenician” (which is a direct reference back to Madame Sosostris’s “drowned Phoenician Sailor”) dies in the water. The lines “As he rose and fell/He passed the stages of his age and youth/Entering the whirlpool” showcase a similar phenomenon to Ulysses, passing through all previous versions to himself. Even his rising and falling harks back to Ulysses’ sinking. The parallel between the two men, who were both once young and handsome, sacrificed to the water that once sustained them is an interesting image.

This does not work for anyone trying to maintain anonymity online because you have to put your name thats on your ID or something really close to it. This could remove an aspect of authenticity for these people because if they feel exposed online they may be less authentic because they fear the consequences of that authenticity due to it being linked to their real name. This does however work for people who believe that accountability is the way to get authenticity this works because it holds people accountable for things they post by having their name tied to it.

instead of having gestures or body language to relay a story, the poetry can only use words to tell a story.

being a good listener means paying attention and focusing without interruptions and forcing your own feelings or perspectives on it.

a bad listener is someone who can focus only on the words and not the meaning behind them.

you must interpret it without applying your own feelings.

it seems like a lot of changing imagery.

the imagery seems like a circus. he is moving away from being foolish and moving onto the next day.

metaphor.

leonard put more effort to make the poetry into music vs making music into poetry.

bob didn't intend to be a poet but it flowed naturally within the music.

once he turned the writing into music it created massive success.

they used their music and their platforms to spread information about political injustices.

their different forms of poetry styles manifested into their music and shaped their careers.

music is a form of poetry in itself, and it exposes poetry to unsuspecting listeners.

Authenticity is not fixed but depends on context and community. For example, Obama’s different handshakes show how varied expressions can still be genuine. On the other hand, as Lindsay Ellis points out, creators often perform emotional labor to maintain their persona because it brings benefits. So, authenticity should be analyzed not just by what people do, but why they do it and what advantages they gain

Deciding whether or not to go to college and get my bachelors was a very difficult decision, and ultimately I was influenced by my parents. My dad especially encouraged me to go to residential college and get my degree, which is also encouraged by the master narrative.

Emerging adulthood

How would this study and its findings look if conducted in relatively poorer areas of the country? What does youth culture think about post-secondary education in Alabama, Mississippi, West Virginia?

This story about Dr. McLaughlin and the fake @Sciencing_Bi account really shocked me. It made me realize how easily emotional trust can be built, and then betrayed, on social media. As someone who often reads personal stories online, I've never thought deeply about whether those stories could be fabricated. What feels most unethical here is not only the lying itself, but how it exploited people's empathy for marginalized identities and real victims of harassment. It's frightening that authenticity online now requires skepticism, and that our compassion can be used as a tool for manipulation.

Adler, Mortimer J. 1940. “How to Mark a Book.” Saturday Review of Literature 6: 250–52. https://www.unz.com/print/SaturdayRev-1940jul06-00011/ (January 11, 2023).

Annotations: https://via.hypothes.is/https://docdrop.org/download_annotation_doc/Adler---1940---How-to-Mark-a-Book-fehef.pdf

Annotations alternate: https://jonudell.info/h/facet/?user=chrisaldrich&max=100&exactTagSearch=true&expanded=true&url=https%3A%2F%2Fdocdrop.org%2Fdownload_annotation_doc%2FAdler---1940---How-to-Mark-a-Book-fehef.pdf

Prior [.pdf copy]9https://stevenson.ucsc.edu/academics/stevenson-college-core-courses/how-to-mark-a-book-1.pdf): - Annotations https://hypothes.is/users/chrisaldrich?q=url%3Ahttps%3A%2F%2Fstevenson.ucsc.edu%2Facademics%2Fstevenson-college-core-courses%2Fhow-to-mark-a-book-1.pdf<br /> - Alternate annotation link https://jonudell.info/h/facet/?user=chrisaldrich&max=100&exactTagSearch=true&expanded=true&url=https%3A%2F%2Fstevenson.ucsc.edu%2Facademics%2Fstevenson-college-core-courses%2Fhow-to-mark-a-book-1.pdf

Summary

• Marking a book helps in increasing "the most efficient kind of reading."
• The marked (pun intended) difference between physical vs. intellectual ownership of books
• 3 types of book owners:
  • 1. collector of wood pulp and ink
  • 1. one whose read most and dipped into some
  • 1. one who's annotated and sucked the marrow out of them
• Active reading (annotating and staying awake) and engaging deeply, arguing with, and questioning the author is the point of reading.
• A historical record of your active reading allows you to continue the conversation you've had with the author and yourself. (p12)
• Adler's method of reading and marking:
  1. Underlining major points of importance
  2. Vertical lines for emphasis
  3. Marginal marks (stars, asterisks, etc.) (10-20 per book) to indicate the most important statements in conjunction with dogearing these pages for making it easier to find them subsequently
  4. Numbers in the margin to sequence arguments
  5. Page numbers in the margin for linking ideas across pages, ostensibly for juxtaposing them later
  6. Circling key words or phrases (unsaid here, but this is helpful for indexing as well as helping one to come to terms with the author)
  7. Marginal writing for synopsis of sections as well as questions raised by the text; use of endpapers for a personal index of ideas presented chronologically throughout the book
• Objections to marking books:
  • Using scratch pad (or index cards, which he doesn't mention specifically, but which could be implied) so as not to destroy a precious or rare physical copy (this is a repetition from earlier in the article)
  • Marking slows you down. This is part of the point! Slowing down makes you engage with the author and get more out of the text.
  • You can't loan books because they contain your important thoughts which you don't want to give away (and lose the historical record of your thinking). Solution: Simply require friends to buy their own copy.

This practice is not too dissimilar to that used by zettelkasten practitioners (including Niklas Luhmann) who broadly used his bibliographic cards this way.

By separating his index and ideas from the book and putting them into a physical index, it makes them easier to juxtapose with other ideas over time rather than having them anchored directly to the book itself. For academics and researchers, this will tend to help save time from having to constantly retrieve these portions from individual books.

This poke at "fancy" bookplates is a rhetorical call back to those who would attempt to weakly show only physical and not intellectual ownership by "pasting his bookplate inside the cover."

Was "fancy thinking" used in a pejorative sense prior to this article? Are subsequent uses of it poking fun of the parallelism (to fancy bookplates) which Adler sets up here?

The making of a personal index is a first step in building a mesh of knowledge. In just a few years, Vannevar Bush will speak of "associative trails" a phrase he uses twice in "As We May Think" (The Atlantic, July 1945), but of potentially more import is his phrase "associative indexing" which lays way to either juxtaposing or linking two ideas (either similar or disjoint) together. It bears asking the question of of whether it's more valuable to index and juxtapose similar ideas or disjoint ideas which may more frequently lead to better, more useful, and more relevant and rich future ideas.

It affords an immediate step, however, to associative indexing, the basic idea of which is a provision whereby any item may be caused at will to select immediately and automatically another. This is the essential feature of the memex. The process of tying two items together is the important thing. Bush, Vannevar. 1945. “As We May Think.” The Atlantic 176: 101–8. https://www.theatlantic.com/magazine/archive/1945/07/as-we-may-think/303881/ (October 22, 2022). #

Adler creates a very specific and subtle definition of ownership as it applies to books.

It's not too dissimilar to Antoine de Saint-Exupéry's idea of ownership or love of people. "Unique to me in all the world"

Many have spoken of "books as wallpaper" or "intellectual furniture", but here Mortimer J. Adler goes beyond owning them solely as material culture, but turning them into intellectual and personal culture.

When they sit upon the shelf after being intellectually owned, they can serve as a mnemonic touchstone, which is a method of supercharging their value as lowly "decorative wallpaper", and instead making them living active, intellectual wallpaper.

:

The frontal lobe controls decisions making emotion and behavior damage here can change who a person is.

After the accident, Gage's personality changed. he became impulsive and had trouble controlling emotions.

A railroad worker who survived when an iron rod went through his skills in 1848

It helps produce speech. damage here makes it hard to talk or form words.

Vision and visual processing

Hearing,memory, and understanding language

Touch,pressure, and spatial sense

Reasoning, planning, movement. and speech

Midbrain- helps process movement, hearing, and vision

controls thinking, emotions,memory, and decision making

they take this matter very seriously china has this document in it goverment website

in china playbook

I really agree with this idea that critique should be two-way. In many classroom or work settings, feedback feels one-sided — someone tells you what’s wrong, and you just listen. But when designers explain their rationale, it opens up a more meaningful conversation. I found Ko’s framing useful because it reminds me that critique is about growth and understanding, not just judgment. It changes my perspective on feedback — instead of feeling defensive, I can see it as a collaborative dialogue to refine ideas together.

I think this section nails something that goes way beyond design. It’s just good communication in general. I agree that giving balanced feedback forces you to slow down and actually see what’s working instead of jumping straight to criticism. The “shit sandwich” rule might sound funny, but it’s surprisingly useful, especially in group projects where people can get defensive fast. It made me realize that how feedback is delivered can decide whether an idea improves or dies.

Was this really yesterday?

/hyperpost/🌐/🎭/gyuri/do/web/snarf/📅/2025/10/2/Metaphysics.of.Adjacency.html

self

view

Its measure electrical activity. They use a array of electrodes around a person's head

Measures the blood flow and oxygen levels

Best showing for structure (anatomical detail)

once in the bloodstream the tracer's movement is monitored

a technique that takes a number of x-rays of a particular section of a person's body or brain

are we sure?

When Iago says "Call up her father," it is the beginning of his manipulation and how he gets others to be tools of his own cunning devices. On the surface, the line can be interpreted as simply an order, but in fact, it shows how cunning and devious Iago is. Instead of confronting Othello or Desdemona himself, he uses Roderigo to set things on fire for him. By waking Brabantio at midnight, Iago knows that he will be provoking anger, confusion, and fright, emotions that make humans vulnerable to manipulation. This is significant because it makes clear that Iago's evil does not result from aggression but from his ability to provoke reaction and manipulate feelings. He is aware of how to use timing and fright to make fighting break out in moments of peace, and this line alone is the start of that. It also shows one of the prevailing motifs of Othello: the idea that destruction is oftentimes started quietly in suggestion and manipulation and not in outright brutality. By having others "call up" the trouble for him, Iago gets to preserve his good name while the damage is being done, highlighting how risky lying can be when clothed in peaceful words.

See also the precursor of personal indexing which Mortimer J. Adler mentions in 1940: https://hypothes.is/a/cPcoAqhVEfC0rJOZ0Pm-8Q

academic brained? I don’t mean that too harshly but: social context matters!! people find formulas meaningful because they are literally performative not because of their novelty

Comment 2

Comment 1

Note 1

want it to be true

Is this working?

西班牙裔

eLife Assessment

This important manuscript evaluates how sample size and demographic balance of reference cohorts affect the reliability of normative models. The evidence supporting the conclusions is convincing, although some additional analysis and clarifications could improve the generalisability of the conclusions. This work will be of interest to clinicians and scientists working with normative models.

Reviewer #1 (Public review):

Summary:

Overall, this is a well-designed and carefully executed study that delivers clear and actionable guidance on the sample size and representative demographic requirements for robust normative modelling in neuroimaging. The central claims are convincingly supported.

Strengths:

The study has multiple strengths. First, it offers a comprehensive and methodologically rigorous analysis of sample size and age distribution, supported by multiple complementary fit indices. Second, the learning-curve results are compelling and reproducible and will be of immediate utility to researchers planning normative modelling projects. Third, the study includes both replication in an independent dataset and an adaptive transfer analysis from UK Biobank, highlighting both the robustness of the results and the practical advantages of transfer learning for smaller clinical cohorts. Finally, the clinical validation ties the methodological work back to clinical application.

Weaknesses:

There are two minor points for consideration:

(1) Calibration of percentile estimates could be shown for the main evaluation (similar to that done in Figure 4E). Because the clinical utility of normative models often hinges on identifying individuals outside the 5th or 95th percentiles, readers would benefit from visual overlays of model-derived percentile curves on the curves from the full training data and simple reporting of the proportion of healthy controls falling outside these bounds for the main analyses (i.e., 2.1. Model fit evaluation).

(2) The larger negative effect of left-skewed sampling likely reflects a mismatch between the younger training set and the older test set; accounting explicitly for this mismatch would make the conclusions more generalisable.

Reviewer #2 (Public review):

Summary:

The authors test how sample size and demographic balance of reference cohorts affect the reliability of normative models in ageing and Alzheimer's disease. Using OASIS-3 and replicating in AIBL, they change age and sex distributions and number of samples and show that age alignment is more important than overall sample size. They also demonstrate that models adapted from a large dataset (UK Biobank) can achieve stable performance with fewer samples. The results suggest that moderately sized but demographically well-balanced cohorts can provide robust performance.

Strengths:

The study is thorough and systematic, varying sample size, age, and sex distributions in a controlled way. Results are replicated in two independent datasets with relatively large sample sizes, thereby strengthening confidence in the findings. The analyses are clearly presented and use widely applied evaluation metrics. Clinical validation (outlier detection, classification) adds relevance beyond technical benchmarks. The comparison between within-cohort training and adaptation from a large dataset is valuable for real-world applications.

The work convincingly shows that age alignment is crucial and that adapted models can reach good performance with fewer samples. However, some dataset-specific patterns (noted above) should be acknowledged more directly, and the practical guidance could be sharper.

Weaknesses:

The paper uses a simple regression framework, which is understandable for scalability, but limits generalization to multi-site settings where a hierarchical approach could better account for site differences. This limitation is acknowledged; a brief sensitivity analysis (or a clearer discussion) would help readers weigh trade-offs. Other than that, there are some points that are not fully explained in the paper:

(1) The replication in AIBL does not fully match the OASIS results. In AIBL, left-skewed age sampling converges with other strategies as sample size grows, unlike in OASIS. This suggests that skew effects depend on where variability lies across the age span.

(2) Sex imbalance effects are difficult to interpret, since sex is included only as a fixed effect, and residual age differences may drive some errors.

(3) In Figure 3, performance drops around n≈300 across conditions. This consistent pattern raises the question of sensitivity to individual samples or sub-sampling strategy.

(4) The total outlier count (tOC) analysis is interesting but hard to generalize. For example, in AIBL, left-skew sometimes performs slightly better despite a weaker model fit. Clearer guidance on how to weigh model fit versus outlier detection would strengthen the practical message.

(5) The suggested plateau at n≈200 seems context-dependent. It may be better to frame sample size targets in relation to coverage across age bins rather than as an absolute number.

Author response

We would like to thank the editors and two reviewers for the assessment and the constructive feedback on our manuscript, “Toward Robust Neuroanatomical Normative Models: Influence of Sample Size and Covariates Distributions”. We appreciate the thorough reviews and believe the constructive suggestions will substantially strengthen the clarity and quality of our work. We plan to submit a revised version of the manuscript and a full point-by-point response addressing both the public reviews and the recommendations to the authors. 

Reviewer 1. 

In revision, we plan to address the reviewer’s comments by: (i) strengthen the interpretation of model fit through reporting the proportion of healthy controls within and outside the extreme percentile bounds; (ii) adding age-resolved overlays of model-derived percentile curves compared to those from the full reference cohort for key sample sizes and regions; (iii) quantifying age-distribution alignment between train and test set; and (iv) summarizing model performance as a joint function of age-distribution alignment and sample size.

Reviewer 2. 

In the revised manuscript, we will (i) expand the Discussion to more clearly outline the trade-offs between simple regression frameworks and hierarchical models for normative modeling (e.g., scalability, handling of multi-site variation, computational considerations), and discuss alternative approaches and harmonization as important directions for multi-site settings; (ii) contextualize OASIS-3 vs AIBL differences by quantifying train– test age-alignment across sampling strategies and emphasize that skewness should be interpreted relative to the target cohort’s alignment rather than absolute numbers. (iii) reassess sex-imbalance effects by reporting expected age distributions per condition and re-evaluate sex effects while controlling for age; (iv) investigate the apparent dip at n≈300 dip by increasing sub-sampling seeds, testing neighboring sample sizes, and using an alternative age-binning scheme to clarify the observed artifact; (v) clarify potential divergence between tOC separation and global fit under discrepancies in demographic distributions and relate tOC to age-alignment distance; (vi)  reframe the sample-size guidance in terms of distributional alignment rather than an absolute n.

ეს სტატია შეაჯამებს შვეიცარიაში მიმდინარე ბიომრავალფეროვნების კრიზისს და იმ გამოწვევებს, რომლებსაც ქვეყანა ხვდება დაცული ტერიტორიების გაფართოებისას. შვეიცარიამ გაამახვილა ყურადღება გარემოს დაცვის აუცილებლობაზე,თუმცა ქვეყნის დაცული ტერიტორიების ფართობი არ აკმაყოფილებს საერთაშორისო მიზნებს. სტატია მიანიშნებს ,რომ შვეიცარიის დაცული ტერიტორიების ფართობი მხოლოდ 6.6%-დან 13.4% პროცენტამდე მერყეობს ,რაც დაბალია 2020 წლისთვის დაწესებულ 17%იან მიზანთან შედარებით . სტატიიის ერთ-ერთი მთავარი თემააა 2016 წელს უშედეგოდ დასრულებული PARC ADULA პროექტი , რომელიც მიზნად ისახავდა ბიომრავალფეროვნების დაცვას და ადგილობრივი ეკონომიკის გაძლიერებას. თუმცა, ადგილობრივმა მოსახლეობამ დაგმო პროექტი , ტრადიციული საქმიანობის ხელშეშლის მიზეზით, როგორიცაა ნადირობა და თევზაობა. სტატია ხაზს უსვამს, რომ შვეიცარიის პოლიტიკა ადგილობრივ თვითმმართველობების ავტონომიას პრიორიტეტს აძლევს , რაც ართლებს ზოგად-ეროვნულ ინიციატივებს, როგორიცაა დაცული ტერიოტრიების გაფართოება. სტატიის ბოლო ნაწილი გვეუბნება , რომ შვეიცარიის მიზანია დაცული ტერიტორების გაფართოება, რისთვისაც საჭიროა ადგილობრივი თემების მხარდაჭერა, ცნობიერების ამაღლება და ეროვნული ინტერესესბის დაცვა.

eLife Assessment

This important paper reports the development of proteins and small molecules that induce degradation of a clinically-relevant oncogenic transcription factor, LMO2. The findings provide a proof of concept that PROTAC-type chemicals can be developed against intrinsically disordered proteins. The methods provide a blueprint for rational design of PROTACs starting from intracellular antibody paratopes. Overall, the paper is supported by solid evidence and will be of interest to chemical biologists and cancer pharmacologists.

Reviewer #2 (Public review):

Summary:

Sereesongsaeng et al. aimed to develop degraders for LMO2, an intrinsically disordered transcription factor activated by chromosomal translocation in T-ALL. The authors first focused on developing biodegraders, which are fusions of an anti-LMO2 intracellular domain antibody (iDAb) with cereblon. Following demonstrations of degradation and collateral degradation of associated proteins with biodegraders, the authors proceeded to develop PROTACs using antibody paratopes (Abd) that recruit VHL (Abd-VHL) or cereblon (Abd-CRBN). The authors show dose-dependent degradation of LMO2 in LMO2+ T-ALL cell lines, as well as concomitant dose-dependent degradation of associated bHLH proteins in the DNA-binding complex. LMO2 degradation via Abd-VHL was also determined to inhibit proliferation and induce apoptosis in LMO2+ T-ALL cell lines.

Strengths:

The topic of degrader development for intrinsically disordered proteins is of high interest and the authors aimed to tackle a difficult drug target. The authors evaluated methods including the development of biodegraders, as well as PROTACs that recruit two different E3 ligases. The study includes important chemical control experiments, as well as proteomic profiling to evaluate selectivity.

Weaknesses:

Several weaknesses remain in this study:

(1) The overall degradation achieved is not highly potent (although important proof-of-concept);

(2) The mechanism of collateral degradation is not completely addressed. The authors acknowledge possible explanations, which would require mutagenesis and structural studies to further dissect;

(3) The proteomics experiments do not detect LMO2, which the authors attribute to its size, making it difficult to interpret.

Author response:

The following is the authors’ response to the original reviews.

Reviewer #1 (Public review): 

Summary:

The authors describe the degradation of an intrinsically disordered transcription factor (LMO2) via PROTACs (VHL and CRBN) in T-ALL cells. Given the challenges of drugging transcription factors, I find the work solid and a significant scientific contribution to the field. 

Strengths: 

(1) Validation of LMO2 degradation by starting with biodegraders, then progressing to chemical degrades. 

(2)interrogation of the biology and downstream pathways upon LMO2 degradation (collateral degradation §

(3) Cell line models that are dependent/overexpression of LMO2 vs LMO2 null cell lines. 

(4) CRBN and VHL-derived PROTACs were synthesized and evaluated. 

Weaknesses: 

(1) The conventional method used to characterize PROTACs in the literature is to calculate the DC50 and Dmax of the degraders, I did not find this information in the manuscript. 

As noted in the reply to referee’s point 4 below, our first generation compounds are not highly potent. The DC₅₀ values have been computed specifically using Western blot reflected in the data shown in Fig. 2. The revised version Supplementary Fig. S3 shows these quantified Western blot data from a time course of treating KOPT-K1 cells with either Abd-CRBN and Abd-VHL, where the 24 hour blot data are shown in Figure 2, G and E, and the quantified data from each 24 hour treatment are quantified in Supplementary Fig. S3). With these data, the DC₅₀ values 9 μM for Abd-CRBN and 15 μM Abd-VHL), included in in the main text and the Supplementary Fig. S3 figure legend.

In addition, the loss of signal of the LMO2-Rluc reporter protein from PROTAC treated cells shown in Fig. 2M has been used to calculate a half-point of degradation; although strictly not DC₅₀, as it measures a reporter protein, this yielded values are 10 μM for Abd-CRBN and 9 μM Abd-VHL. 

(2) The proteomics data is not very convincing, and it is not clear why LMO2 does not show in the volcano plot (were higher concentrations of the PROTAC tested? and why only VHL was tested and not CRBN-based PROTAC?).

Due to the relatively small size of the LMO2 protein, it is challenging to produce enough unique peptides for reliable identification, especially to distinguish some proteins in the LMO2 complex.  

(3) The correlation between degradation potency and cell growth is not well-established (compare Figure 4C: P12-Ichikawa blots show great degradation at 24 and 48 hrs, but it is unclear if the cell growth in this cell line is any better than in PF-382 or MOLT-16) - Can the authors comment on the correlation between degradation and cell growth?  

In this study (Fig. 4) we did not aim to compare the effect of LMO2 loss on cell growth among LMO2 positive cells. Rather, we aimed to evaluate the LMO2 importance for cell growth in LMO2-expressing T-ALL cells compared to non-expressing cells and to correlate the loss of the protein with this effect on the cell growth. In addition, the treatment of cells with the LMO2 compounds did now show an effect to LMO2 negative cells until at least 48 hours of treatment indicating that low toxicity of our PROTAC compounds and providing correlation between LMO2 loss and cell growth. 

(4) The PROTACs are not very potent (double-digit micromolar range?) - can the authors elaborate on any challenges in the optimization of the degradation potency? 

The Abd methodology to use intracellular domain antibodies to screen for compounds that bind to intrinsically disordered proteins such as the LMO2 transcription factors offers a tractable approach to hard drug targets but, in so doing, creates challenging factors to improve the potency that are not the same as those targets for which structural data are available. LMO2 is an intrinsically disordered protein, for which soluble recombinant protein is not readily available to identify the binding pocket of compounds. The potency has so far been optimized solely based on the different moieties substituted in cell-based SAR studies (http://advances.sciencemag.org/cgi/content/full/7/15/eabg1950/DC1) and all new compounds were tested with BRET assays. Thus, currently optimization of the degradation potency (including properties such as improved solubility) for the LMO2-binding compounds relies on chemical modification the three areas of the compounds indicated in Fig. 2 B,C.  

(5) The authors mentioned trying six iDAb-E3 ligase proteins; I would recommend listing the E3 ligases tried and commenting on the results in the main text. 

The six chimaeric iDAb-E3 ligase proteins involved one anti-LMO2 iDAb and three different E3 ligase where either fused at the N- or the C-terminus of the VH (giving six protein formats). These six fusion proteins were described in the text referring to the degrader studies described in Supplementary Fig. 1. 

Reviewer #2 (Public review): 

Summary: 

Sereesongsaeng et al. aimed to develop degraders for LMO2, an intrinsically disordered transcription factor activated by chromosomal translocation in T-ALL. The authors first focused on developing biodegraders, which are fusions of an anti-LMO2 intracellular domain antibody (iDAb) with cereblon. Following demonstrations of degradation and collateral degradation of associated proteins with biodegraders, the authors proceeded to develop PROTACs using antibody paratopes (Abd) that recruit VHL (Abd-VHL) or cereblon (Abd-CRBN). The authors show dose-dependent degradation of LMO2 in LMO2+ T-ALL cell lines, as well as concomitant dose-dependent degradation of associated bHLH proteins in the DNA-binding complex. LMO2 degradation via Abd-VHL was also determined to inhibit proliferation and induce apoptosis in LMO2+ T-ALL cell lines. 

Strengths: 

The topic of degrader development for intrinsically disordered proteins is of high interest, and the authors aimed to tackle a difficult drug target. The authors evaluated methods, including the development of biodegraders, as well as PROTACs that recruit two different E3 ligases. The study includes important chemical control experiments, as well as proteomic profiling to evaluate selectivity. 

Weaknesses: 

The overall degradation is relatively weak, and the mechanism of potential collateral degradation is not thoroughly evaluated

The purpose of the study was to evaluate effects of LMO2 degraders. The mechanism of the observed collateral degradation could not be investigated directly within the scope of our study. In the main text, discussed two possible, not exclusive, explanations. One being that our work (and previously published, cited work) indicates that the DNA-binding bHLH proteins have relatively short half file (Supplementary Fig. S12) and may therefore be subject to normal turnover when the LMO2, which is in the complex, turns over. Further, the known structure of the LMO2-bHLH interactions (from Omari et al, doi: 10.1016/j.celrep.2013.06.008) was also examined for the location of lysines in the TAL1 & E47 partners (Supplementary Fig. S11). It is possible that their local association with the LMO2-E3-ligase complex created by the PROTAC interaction, could cause their concurrent degradation. Mutagenesis and structural analysis would be needed to establish this point.

In addition, experiments comparing the authors' prior work with their anti-LMO2 iDAb or Abl-L are lacking, which would improve our understanding of the potential advantages of a degrader strategy for LMO2.  

A major motivation behind developing the Antibody-derived (Abd) method to select compounds, which are surrogates of the antibody paratope, is because using iDAbs directly as inhibitors requires the development of delivery technologies for these macromolecules, as protein directly or as vectors or mRNA for their expression. Ultimately, high affinity anti-LMO2 iDAbs should directly be used as tractable inhibitors when delivery methods redeveloped. In the meantime, Abd compounds were envisaged as being surrogates suitable for development into reagents, and potentially drugs, by medicinal chemistry. We evaluated selected first generation LMO2-binding Abd compounds previously, finding their ability to interfere with LMO2-iDAb BRET signal to EC_max about 50% but these compounds do not have potency to have an effect on the interaction of LMO2 with a non-mutated iDAb (nM affinity). These data indicated that efficacy improvement for the PROTACs was needed. In addition, in the current study, we observed viability effects in T-ALL lines at high concentrations (20 μM) irrespective of LMO2 expression (Supplementary Fig. S 2A, B) These data indicated that efficacy improvement was needed and potentially converting the degraders (PROTACs) would add to in-cell potency. By adding the E3 ligase ligands, we found the toxicity of non-LMO2 expressing Jurkat was significantly reduced (Supplementary Fig. S 2E, F). 

Reviewer #2 (Recommendations for the authors): 

Suggestions for additional experiments: 

(1) The data presented is primarily focused on demonstrating targeted degradation of LMO2, with a focus on phenotypes such as proliferation and apoptosis. In this manuscript, there are limited comparative evaluations of anti-LMO2 iDAb or Abl-L to show the potential benefits of a degrader approach to their previously described work, as well as why targeted degradation is in fact, advantageous. For example, the authors' previous work has shown that anti-LMO2 iDAb inhibits tumor growth in a mouse transplantation model. Comparisons in vitro would be supportive of the importance of continued degrader optimization/development.  

we have previously shown that an anti-LMO2 scFv inhibits tumour growth in a mouse model but this work used an expressed scFv antibody that binds to LMO2 in nM range. The Abd compounds are much lower potency that the antibody and, because recombinant LMO2 is difficult to work with, we could only evaluate interactions of compounds with LMO2 in cell-based assays like BRET (LMO2-iDAb BRET). In this cell-based assay, the first generation Abd compounds do not have sufficient potency to block LMO2-iDAb interaction unless the affinity of the iDAb is reduced to sub-μM. The justification for proceeding on the degrader process rather than just using the protein-protein interaction (PPI) inhibition was based largely around the low potency of the first generation PPI compounds in cell assays and that incorporation protein degradation with PPI inhibition would enhance the efficacy.

In addition, the viability experiments are also very short-term; is there a reason why the authors did not carry out these experiments for 3-5 days to fully understand the impacts on proliferation? 

In Supplementary Fig. S5, we did show assays up to 3 days. In KOPT-K1 (LMO2+), the LMO2 levels were reduced during the time course of this assay (from a single compound dose at time zero) (Supplementary Fig S 5A, B). We also show CellTitreGlo assays up to 3 days and, with these second generation compounds, we observed sustained effects on KOPT-K1 (LMO2+) but low non-DMSO toxicity in Jurkat (LMO2-) (revised version Supplementary (Fig S5 C, D).

(2) The potential mechanism of collateral degradation is interesting and important in evaluating the on-target responses and consequences of degrading LMO2. At this time, the data supporting collateral degradation is limited and would be strengthened by showing that it is not due to a change in mRNA levels and not due to complex dissociation. Overall, the kinetics and depth of loss of complex members such as E47 in Figure 3 appear more substantial than LMO2 itself, and as presented, collateral degradation is not effectively demonstrated. In addition, to aid in the readers' assessments, additional background and references around the roles of TAL1 and E47 would be helpful. For example, structurally, where do they (and other associated proteins that are not degraded) fit in the complex? 

We have responded above in relation to the Public Review Comments and note that a structure of the complex was in submitted version (now revised version Supplementary Fig. S11). 

(3) In Figure 1A, the blots show decreased levels of endogenous CRBN with iDAB-CRBN. Is this a known consequence of this approach in these cell lines? Does the partial recovery of endogenous CRBN in KOPTK1 cells have any indication of iDAB-CRBN levels? 

We cannot be sure why the endogenous level of CRBN decreases in doxycycline treated cells. It has been shown (DOI:10.1371/journal.pone.0064561) that doxycycline used in the inducible expression system (and its derivatives), such as the lentivirus we used, has an effect to gene expression patterns, which can be increase or decrease expression. Although the published study did not examine CRBN expression, the effect might explain the CRBN expression decrease on doxycycline addition and remains the same level after that. 

(4) In Figure S7, the authors do not fully explain the results and why there is minimal rescue with epoxomicin (S7A) or MLN4924 (S7J). This could indicate an alternative mechanism of degradation and loss at play, given the lack of rescue. Can the authors comment on this discrepancy, and have they looked autophagy inhibitor or other agents to achieve the chemical rescue? 

In the experiments such as in revised version Supplementary Fig. S6, we used KOPT-K1 cells with a single concentration of the inhibitors and the cells may less susceptible to the epoxomicin (0.8 μM) but lenalidomide and free thalidomide restored the LMO2 levels fully. In the main text Fig. 3D, we also showed that including epoxomicin and thalidomide with the Abd-CRBN in KOPT-K1 and CCRF-CEM restore LMO2 levels, supporting the conclusion that the main mechanism of degradation is through ubiquitination proteosomal route.

(5) For the proteomics data, it would be helpful to have the proteins in yellow highlighted to have them noted in 5D and 5E. In addition, can the authors comment on why LMO2 or their collateral targets are not confirmed in the table? Furthermore, 5C is difficult to interpret; if there are no significantly changing proteins in the Jurkat cells, why are there pathways that are identified? 

As mentioned in reply to referee 1, due to the relatively small size of the LMO2 protein, it is challenging to produce enough unique peptides for reliable identification, especially to distinguish some proteins in the LMO2 complex where expression levels are low.

This confuses me because at the time they had slaves yet he is saying all men are originally equal, so then why do they have slaves if they are equal? I also wonder if he just included this because it would make him appeal to more people as to get as many people on his side as possible.

Paine's goal in saying this is to get the colonist to rally together and to unite as one against Britain. He is saying how the king is a tyrant and is slandering him pretty much so he can get people to join him.

He is telling the colonist to get ready and to be excited for the new world when they break free from Britain. He is saying that it will be a safe haven for freedom. The fugitive means freedom and he is telling the colonist to prepare to have freedom.

This paper is written for the people of the colonies to raise up against the Brits. His goal is also to give them a spark to push for independence. Through this paper he tries to give them some hope and reasons as to why they should break away.

Historical Context- He is talking to the colonist about how because of there being kings they caused wars by being too greedy. His goal in saying this is to turn the colonist against having a king and breaking away from the kings rule in Britain.

Not sure we need this separate page. The map can be embedded at the bottom of the Repository page.

Add 3 sections which link to other pages: 1. Publish Open Access 2. Share your Work in Purdue e-Pubs/ the Repository 3. Publish with Libraries

"ua maila" is translated "it is calm." Not sure how this was arrived at. To me, it looks like

"In the stillness of the birds, [because it has just] rained."

2.

Human Needs Theory directly conflicts with Survival of the fittest as it argues that aggression is not stemmed from innate human behavior, but rather, they stem from social circumstances that do not satisfy human needs.

eLife Assessment

This valuable study concerns a model for transgenerational epigenetic inheritance, the learned avoidance by C. elegans of the PA14 pathogenic strain of Pseudomonas aeruginosa. A recent study questioned whether transgenerational inheritance in this paradigm lacks robustness. The authors of this study have worked independently of the group that reported the original phenomenon and also independently of the group that challenged the original report. With solid data, this study independently validates findings previously reported by the Murphy group, confirming that the paradigm is reproducible elsewhere. The reviewers also appreciated the information on reagent sources used by different groups. The present study is therefore of broad interest to anyone studying genetics, epigenetics, or learned behavior.

Reviewer #1 (Public review):

Summary:

The manuscript addresses the discordant reports of the Murphy (Moore et al., 2019; Kaletsky et al., 2020; Sengupta et al., 2024) and Hunter (Gainey et al., 2025) groups on the existence (or robustness) of transgenerational epigenetic inheritance (TEI) controlling learned avoidance of C. elegans to Pseudomonas aeruginosa. Several papers from Colleen Murphy's group describe and characterize C. elegans transgenerational inheritance of avoidance behaviour. In the hands of the Murphy group, the learned avoidance is maintained for up to four generations, however, Gainey et al. (2025) reported an inability to observe inheritance of learned avoidance beyond the F1 generation. Of note, Gainey et al used a modified assay to measure avoidance, rather than the standard assay used by the Murphy lab. A response from the Murphy group suggested that procedural differences explained the inability of Gainey et al.(2025) to observe TEI. They found two sources of variability that could explain the discrepancy between studies: the modified avoidance assay and bacterial growth conditions (Kaletsky et al., 2025). The standard avoidance assay uses azide as a paralytic to capture worms in their initial decision, while the assay used by the Hunter group does not capture the worm's initial decision but rather uses cold to capture the location of the population at one point in time.

In this short report, Akinosho, Alexander, and colleagues provide independent validation of transgenerational epigenetic inheritance (TEI) of learned avoidance to P. aeruginosa as described by the Murphy group by demonstrating learned avoidance in the F2 generation. These experiments used the protocol described by the Murphy group, demonstrating reproducibility and robustness.

Strengths:

Despite the extensive analyses carried out by the Murphy lab, doubt may remain for those who have not read the publications or for those who are unfamiliar with the data, which is why this report from the Vidal-Gadea group is so important. The observation that learned avoidance was maintained in the F2 generation provides independent confirmation of transgenerational inheritance that is consistent with reports from the Murphy group. It is of note that Akinosho, Alexander et al. used the standard avoidance assay that incorporates azide, and followed the protocol described by the Murphy lab, demonstrating that the data from the Moore and Kaletsky publications are reproducible, in contrast to what has been asserted by the Hunter group.

Comments on revised version:

I am happy with the responses to reviews.

Reviewer #2 (Public review):

Summary:

The manuscript "Independent validation of transgenerational inheritance of learned pathogen avoidance in C. elegans" by Akinosho and Vidal-Gadea offers evidence that learned avoidance of the pathogen PA14 can be inherited for at least two generations. In spite of initial preference for the pathogen when exposed in a 'training session', 24 hours of feeding on this pathogen evoked avoidance. The data are robust, replicated in 4 trials, and the authors note that diminished avoidance is inherited in generations F1 and F2.

Strengths:

These results contrast with those reported by Gainey et al, who only observed intergenerational inheritance for a single generation. Although the authors' study does not explain why Gainey et el fail to reproduce the Murphy lab results, one possibility is that a difference in a media ingredient could be responsible.

Comments on revised version:

The responses to the reviewer comments appear reasonable for the most part.