DOI: 10.1101/2024.05.09.593334

Resource: (Thermo Fisher Scientific Cat# A-31573, RRID:AB_2536183)

Curator: @scibot

SciCrunch record: RRID:AB_2536183

---

What is this?

DOI: 10.1101/2024.05.09.593334

Resource: AB_2937041

Curator: @scibot

SciCrunch record: RRID:AB_2937041

---

What is this?

DOI: 10.1101/2024.05.09.593334

Resource: (Innovative Research Cat# AHB0042, RRID:AB_1502093)

Curator: @scibot

SciCrunch record: RRID:AB_1502093

---

What is this?

DOI: 10.1101/2024.05.09.593334

Resource: (Abcam Cat# ab51253, RRID:AB_869973)

Curator: @scibot

SciCrunch record: RRID:AB_869973

---

What is this?

DOI: 10.1101/2024.05.09.593334

Resource: (IMSR Cat# JAX_016123,RRID:IMSR_JAX:016123)

Curator: @scibot

SciCrunch record: RRID:IMSR_JAX:016123

---

What is this?

DOI: 10.1101/2024.05.09.593334

Resource: IMSR_JAX:010710

Curator: @scibot

SciCrunch record: RRID:IMSR_JAX:010710

---

What is this?

DOI: 10.1101/2024.05.09.593334

Resource: (IMSR Cat# JAX_000664,RRID:IMSR_JAX:000664)

Curator: @scibot

SciCrunch record: RRID:IMSR_JAX:000664

---

What is this?

DOI: 10.1101/2024.05.09.593334

Resource: (BD Biosciences Cat# 561908, RRID:AB_2869394)

Curator: @scibot

SciCrunch record: RRID:AB_2869394

---

What is this?

DOI: 10.1101/2024.05.09.593334

Resource: (Thermo Fisher Scientific Cat# MA1-940, RRID:AB_2136775)

Curator: @scibot

SciCrunch record: RRID:AB_2136775

---

What is this?

DOI: 10.1101/2024.05.09.593334

Resource: (Thermo Fisher Scientific Cat# MA1-164, RRID:AB_2536869)

Curator: @scibot

SciCrunch record: RRID:AB_2536869

---

What is this?

DOI: 10.1101/2024.05.09.593368

Resource: GraphPad Prism (RRID:SCR_002798)

Curator: @scibot

SciCrunch record: RRID:SCR_002798

---

What is this?

DOI: 10.1101/2024.05.09.593336

Resource: (Molecular Probes Cat# A-21202, RRID:AB_141607)

Curator: @scibot

SciCrunch record: RRID:AB_141607

---

What is this?

DOI: 10.1101/2024.05.06.592831

Resource: Human Islet Research Network (HIRN) (RRID:SCR_014393)

Curator: @scibot

SciCrunch record: RRID:SCR_014393

---

What is this?

DOI: 10.1101/2024.05.08.593169

Resource: RRID:Addgene_12259

Curator: @scibot

SciCrunch record: RRID:Addgene_12259

---

What is this?

DOI: 10.1101/2024.05.07.592305

Resource: Umap (RRID:SCR_018217)

Curator: @scibot

SciCrunch record: RRID:SCR_018217

---

What is this?

DOI: 10.1101/2024.05.07.592305

Resource: CellRank (RRID:SCR_022827)

Curator: @scibot

SciCrunch record: RRID:SCR_022827

---

What is this?

DOI: 10.1101/2024.05.07.592305

Resource: scanpy (RRID:SCR_018139)

Curator: @scibot

SciCrunch record: RRID:SCR_018139

---

What is this?

DOI: 10.1101/2024.05.07.592305

Resource: Velocyto (RRID:SCR_018167)

Curator: @scibot

SciCrunch record: RRID:SCR_018167

---

What is this?

DOI: 10.1101/2024.05.07.592305

Resource: Illumina NovaSeq 6000 Sequencing System (RRID:SCR_016387)

Curator: @scibot

SciCrunch record: RRID:SCR_016387

---

What is this?

DOI: 10.3390/brainsci14050484

Resource: (IMSR Cat# JAX_028861,RRID:IMSR_JAX:028861)

Curator: @scibot

SciCrunch record: RRID:IMSR_JAX:028861

---

What is this?

DOI: 10.1101/2024.05.06.592806

Resource: (BDSC Cat# 34771,RRID:BDSC_34771)

Curator: @scibot

SciCrunch record: RRID:BDSC_34771

---

What is this?

DOI: 10.1101/2024.05.06.592806

Resource: RRID:Addgene_172067

Curator: @scibot

SciCrunch record: RRID:Addgene_172067

---

What is this?

DOI: 10.1101/2024.05.06.592806

Resource: BDSC_93189

Curator: @scibot

SciCrunch record: RRID:BDSC_93189

---

What is this?

DOI: 10.1101/2024.05.06.592806

Resource: RRID:BDSC_93190

Curator: @scibot

SciCrunch record: RRID:BDSC_93190

---

What is this?

DOI: 10.1101/2024.05.06.592806

Resource: (BDSC Cat# 7473,RRID:BDSC_7473)

Curator: @scibot

SciCrunch record: RRID:BDSC_7473

---

What is this?

DOI: 10.1101/2024.05.06.592806

Resource: (BDSC Cat# 8765,RRID:BDSC_8765)

Curator: @scibot

SciCrunch record: RRID:BDSC_8765

---

What is this?

DOI: 10.1101/2024.05.06.592806

Resource: BDSC_3720

Curator: @scibot

SciCrunch record: RRID:BDSC_3720

---

What is this?

DOI: 10.1101/2024.05.06.592806

Resource: Addgene_172068

Curator: @scibot

SciCrunch record: RRID:Addgene_172068

---

What is this?

DOI: 10.1101/2024.05.06.592806

Resource: BDSC_44887

Curator: @scibot

SciCrunch record: RRID:BDSC_44887

---

What is this?

DOI: 10.1101/2024.05.06.592806

Resource: DGRC_1314

Curator: @scibot

SciCrunch record: RRID:DGRC_1314

---

What is this?

DOI: 10.1101/2024.05.06.592806

Resource: RRID:DGRC_1298

Curator: @scibot

SciCrunch record: RRID:DGRC_1298

---

What is this?

DOI: 10.1101/2024.05.07.592993

Resource: RRID:Addgene_101323

Curator: @scibot

SciCrunch record: RRID:Addgene_101323

---

What is this?

DOI: 10.1101/2024.05.07.592993

Resource: RRID:Addgene_12259

Curator: @scibot

SciCrunch record: RRID:Addgene_12259

---

What is this?

DOI: 10.1101/2024.05.07.592993

Resource: (LI-COR Biosciences Cat# 926-32213, RRID:AB_621848)

Curator: @scibot

SciCrunch record: RRID:AB_621848

---

What is this?

DOI: 10.1101/2024.05.07.592993

Resource: (LI-COR Biosciences Cat# 926-68072, RRID:AB_10953628)

Curator: @scibot

SciCrunch record: RRID:AB_10953628

---

What is this?

DOI: 10.1101/2024.05.07.592993

Resource: (Abcam Cat# ab8227, RRID:AB_2305186)

Curator: @scibot

SciCrunch record: RRID:AB_2305186

---

What is this?

DOI: 10.1101/2024.05.07.592993

Resource: (Cell Signaling Technology Cat# 3724, RRID:AB_1549585)

Curator: @scibot

SciCrunch record: RRID:AB_1549585

---

What is this?

DOI: 10.1101/2024.05.07.592909

Resource: RRID:Addgene_26161

Curator: @scibot

SciCrunch record: RRID:Addgene_26161

---

What is this?

DOI: 10.1101/2024.05.10.593481

Resource: (RGD Cat# 70508,RRID:RGD_70508)

Curator: @scibot

SciCrunch record: RRID:RGD_70508

---

What is this?

DOI: 10.1101/2024.05.07.593032

Resource: Addgene_174541

Curator: @scibot

SciCrunch record: RRID:Addgene_174541

---

What is this?

DOI: 10.3390/v16050758

Resource: RRID:Addgene_12260

Curator: @scibot

SciCrunch record: RRID:Addgene_12260

---

What is this?

DOI: 10.3390/v16050758

Resource: RRID:Addgene_99636

Curator: @scibot

SciCrunch record: RRID:Addgene_99636

---

What is this?

DOI: 10.3390/v16050758

Resource: Addgene_12529

Curator: @scibot

SciCrunch record: RRID:Addgene_12529

---

What is this?

DOI: 10.3390/v16050758

Resource: Cornell University BRC Imaging Core Facility (RRID:SCR_021741)

Curator: @scibot

SciCrunch record: RRID:SCR_021741

---

What is this?

DOI: 10.1101/2024.05.09.593449

Resource: SCR_025000

Curator: @scibot

SciCrunch record: RRID:SCR_025000

---

What is this?

DOI: 10.1101/2024.05.09.593444

Resource: RRID:Addgene_110770

Curator: @scibot

SciCrunch record: RRID:Addgene_110770

---

What is this?

DOI: 10.1101/2024.05.09.593444

Resource: RRID:Addgene_112866

Curator: @scibot

SciCrunch record: RRID:Addgene_112866

---

What is this?

DOI: 10.1101/2024.05.09.593444

Resource: RRID:Addgene_112864

Curator: @scibot

SciCrunch record: RRID:Addgene_112864

---

What is this?

DOI: 10.1101/2024.05.09.593444

Resource: RRID:Addgene_112863

Curator: @scibot

SciCrunch record: RRID:Addgene_112863

---

What is this?

DOI: 10.1101/2024.05.09.593444

Resource: RRID:Addgene_104964

Curator: @scibot

SciCrunch record: RRID:Addgene_104964

---

What is this?

DOI: 10.1101/2024.05.09.593444

Resource: Addgene_104054

Curator: @scibot

SciCrunch record: RRID:Addgene_104054

---

What is this?

DOI: 10.1101/2024.05.09.593444

Resource: RRID:Addgene_52925

Curator: @scibot

SciCrunch record: RRID:Addgene_52925

---

What is this?

DOI: 10.1101/2024.05.09.592848

Resource: (IMSR Cat# JAX_007909,RRID:IMSR_JAX:007909)

Curator: @scibot

SciCrunch record: RRID:IMSR_JAX:007909

---

What is this?

DOI: 10.1101/2024.05.09.592848

Resource: (IMSR Cat# JAX_016204,RRID:IMSR_JAX:016204)

Curator: @scibot

SciCrunch record: RRID:IMSR_JAX:016204

---

What is this?

DOI: 10.1101/2024.05.09.592848

Resource: (IMSR Cat# JAX_012569,RRID:IMSR_JAX:012569)

Curator: @scibot

SciCrunch record: RRID:IMSR_JAX:012569

---

What is this?

DOI: 10.1016/j.psychres.2024.115951

Resource: (Cell Signaling Technology Cat# 4060, RRID:AB_2315049)

Curator: @abever99

SciCrunch record: RRID:AB_2315049

---

What is this?

DOI: 10.1016/j.psychres.2024.115951

Resource: (Cell Signaling Technology Cat# 9272, RRID:AB_329827)

Curator: @abever99

SciCrunch record: RRID:AB_329827

---

What is this?

DOI: 10.1016/j.str.2024.04.016

Resource: AB_11213007

Curator: @abever99

SciCrunch record: RRID:AB_11213007

---

What is this?

DOI: 10.1101/2024.02.29.582855

Resource: (Millipore Cat# AB9610, RRID:AB_570666)

Curator: @AniH

SciCrunch record: RRID:AB_570666

---

What is this?

DOI: 10.1016/j.immuni.2024.04.019

Resource: (Thermo Fisher Scientific Cat# 50-0443-82, RRID:AB_10597449)

Curator: @abever99

SciCrunch record: RRID:AB_10597449

---

What is this?

DOI: 10.1016/j.celrep.2024.114205

Resource: (BD Biosciences Cat# 347580, RRID:AB_10015219)

Curator: @abever99

SciCrunch record: RRID:AB_10015219

---

What is this?

DOI: 10.1016/j.celrep.2024.114205

Resource: (Santa Cruz Biotechnology Cat# sc-376377, RRID:AB_10987841)

Curator: @abever99

SciCrunch record: RRID:AB_10987841

---

What is this?

DOI: 10.1016/j.celrep.2024.114205

Resource: (Proteintech Cat# 15651-1-AP, RRID:AB_2247477)

Curator: @abever99

SciCrunch record: RRID:AB_2247477

---

What is this?

DOI: 10.1101/2024.02.20.581241

Resource: WB-STRAIN:WBStrain00030220

Curator: @AniH

SciCrunch record: RRID:WB-STRAIN:WBStrain00030220

---

What is this?

DOI: 10.1101/2024.05.08.593219

Resource: (Abcam Cat# ab31940, RRID:AB_2200219)

Curator: @mzhang007

SciCrunch record: RRID:AB_2200219

---

What is this?

DOI: 10.1002/cne.25620

Resource: (DSHB Cat# GAD-6, RRID:AB_528264)

Curator: @AniH

SciCrunch record: RRID:AB_528264

---

What is this?

DOI: 10.1111/tra.12936

Resource: RRID:Addgene_39977

Curator: @AniH

SciCrunch record: RRID:Addgene_39977

---

What is this?

DOI: 10.1101/2024.05.08.593134

Resource: (MGI Cat# 5656552,RRID:MGI:5656552)

Curator: @mzhang007

SciCrunch record: RRID:MGI:5656552

---

What is this?

Les auteurs donnent leur point de vue à la suite de les enquêtes.

Conclusion des enquêtes. Qu'il s'agisse des observations ou des entretiens semi-directifs, il y a lieu de noter l'influence positive des technologies sur les personnes vivant en EHPAD. Ces technologies semblent les aider à mener une vie saine et productive.

Les critères pour participer à l'étude.

Les traits caractéristiques de la population étudiée.

Les effets du logiciel (l'outil) utilisé. L'accent est principalement mis sur l'estime de soi et le besoin d'être reconnu socialement.

La description de la méthode utilisée.

Ce paragraphe pourrait être considéré comme l'hypothèse 2 de la recherche.

La thèse (l'idée maitresse) de la recherche.

Le deuxième argument qui analyse l'apport des technologies sur la vie des personnes âgées en EHPAD.

Le premier argument analyse la situation des personnes âgées.

Ce cadre présente les arguments des auteurs.

Les modalités des enquêtes basées sur une méthode d'étude longitudinale, faite à partir de l'observation et d'entretiens semi-directifs.

La description de l'outil (logiciel) utilisé dans le cadre de la recherche.

L'hypothèse principale de la recherche

Nous avons ici le fil conducteur de l'étude, c'est-à-dire le plan de la recherche.

L'objectif de l'étude et les auteurs précisent en même temps la question du départ.

Le sous-titre de l'article nous donne déjà l'aperçu sur la problématique de la recherche.

Nous avons ici le cadre contextuel de l'étude et le constat général qui s'appuient sur la pensée de deux auteurs, Brangier et Pino (2001)

Proyecto "Anotación PFR", https://github.com/lmichan/PFR,

Tema/mesh/D000086382/COVID19,

TipoDePrueba/mesh/D002000/ForcedSpirometry,

TipoDePrueba/mesh/D001985/BronchialProvocationTests,

TipoDePrueba/mesh/D010993/PlethysmographyWholeBody,

TipoDePrueba/mesh/D011653/PulmonaryDiffusingCapacity,

TipoDePrueba/mesh/D000072277/MaximalRespiratoryPressures,

TipoDePrueba/mesh/D000403/AirwayResistance,

TipoDePrueba/mesh/D000089142/FractionalExhaledNitricOxideTesting,

TipoDePrueba/mesh/000070857/WalkTest,

EtapaPrueba/Operativa,

Enfermedad/mesh/D000086382/COVID19,

Enfermedad/mesh/D017563/InterstitialLungDisease,

Enfermedad/mesh/D009468/NeuromuscularDisease,

Enfermedad/mesh/D001249/Asthma,

Enfermedad/mesh/D003550/CysticFibrosis,

J.J. Short Associates, Inc for Typewriter Platens, Feed Rollers, Bail Rollers, Finger Rollers, and Power Rollers https://www.jjshort.com/typewriter-platen-repair.php

Proyecto "Anotación PFR", https://github.com/lmichan/PFR,

Tema/mesh/D012137/RespiratorySystem,

TipoDePrueba/mesh/D002000/ForcedSpirometry,

EtapaPrueba/Interpretacion,

PatronFuncional/Obstruccion,

PatronFuncional/PosibleRestriccion,

PatronFuncional/PosibleMixto,

PatronFuncional/Normal,

PatronFuncional/Broncodilatacion,

PatronFuncional/NoBroncodilatacion,

Enfermedad/mesh/D001249/Asthma,

Enfermedad/mesh/D029424/ChronicObstructivePulmonaryDisease,

Enfermedad/mesh/D009468/NeuromuscularDisease,

The opening paragraph's assertion that gender is not a fixed attribute but a dynamic and socially constructed concept is both insightful and essential for contemporary discourse. Emphasizing gender fluidity disrupts traditional binary notions, paving the way for a more inclusive understanding of identity. This perspective is crucial because it acknowledges that people's experiences of gender are diverse and evolving, shaped by social interactions rather than rigid biological determinants. Furthermore, the focus on intersectionality is invaluable, as it compels us to consider the complex, overlapping identities that individuals navigate.

这句话跟上一句话在一起，表示我们与自然之间的一种不想关的逻辑，自然资本被排除在资本体系之外的一种想法。

Could a local newsletterer do this?

One issue is running out of things to cover. Only so many eateries locally. County wise? Large metro?

Expand to bars, coffee etc?

Feature new menu items as year progresses?

Maybe accompanying website that has restaurant directory? Local eats etc. sponsorships and premium placements.

dentro para fora

fora para dentro

são peroxissomos

Love this idea in tandem with food newsletter above

• Cutsmart has 50 mM KoAc, 10 mM MgoAc
• Buffer 2.1 has 50 mM NaCl, 10 mM MgCl2
• Buffer 3.1 has 100 mM NaCl, 10 mM MgCl2

Did you mean reaction buffer? Because diluent is different from the reaction buffer. For HindIII this is diluent B

How is the diluent better than the reaction buffer?

Might be good to do on a cell lysate since there is unknown amount of DNA before direct ddPCR

desdobramento de parte de um tecido dentro de outra parte ou estrutura, criando uma dobradura que forma um bolso

dna

Procarionte = surgiu antes da carioteca (núcleo)

TODA CÉLULA possui membrana plasmática, citoplasma e material genético

Armazenamento de células em cultura - Esta técnica permite manter células e tecidos in vitro, conservando as suas propriedades fisiológicas, bioquímicas e genéticas.

para dar sustentação e forma para célula

all very wrong

Test

https://www.reddit.com/r/typewriters/comments/1csni4d/neat_find_on_clients_kmg/

Keyring pliers are used to remove the metal rings off of both circular and tombstone glass typewriter keys so that the legends can be replaced or even covered over with black paper circles for teaching or learning typing. They take some practice and skill to use, but speed up the replacement of legends significantly.

... autochtones, qui renforcent le dualisme en soulignant les différences entre occident et autochtones (2ème écueil),

sup.

t

il n'y a pas une loi omnibus, le terme omnibus désigne toute loi (ou plus souvent projet de loi) touchant plusieurs secteurs et thèmes non liés. Je dirais donc «la loi omnibus C-45»

une ? un mouvement de, pê ?

sugg : sans attribution ou bénéfice symbolique ou économique pour ces dernières.

(très optionnel) de retourner permet d'éviter trop d'occurrences du verbe être : L'approche de l'IPBES comporte l'avantage de permettre, à priori, une collaboration ... Cette collaboration est favorisée par le rejet des présupposés de supériorité épistémique de la science occidentale par rapport aux [scal].

au singulier, je crois

à priori, pê ? Bien que ce ne soit pas incohérent, il paraît un peu étrange au premier abord de parler d'avantage pratique théorique

«davantage épistémologiquement [...] pluraliste»

bénéficiera, puisque c'est l'avancement

This title makes it very easy to know what is going on. It lets the reader know you are now switching to the women's team

"Robust" This site shows a accessibility option which also shows how compatible it is through a wide variety of devices.

"Understandable" Having a separate menu where all languages can be shown is very helpful and makes the webpage understandable to a lot more people with this many options.

"Operable" Navigating using the keyboard to make it easier to get to the main page without needing a mouse.

"Perceivable" The text is made clear and the font has been changed for the reader so it can easily be read on a different background.

These titles for each category make it easy to move around the site knowing what you are looking at

"Robust" Available for region specific items, also built up to modern website standards as it has a good mobile, tablet and computer view of the website.

"Understandable" Very basic language being used in the titles, making it easy for everyone to understand while navigating the website.

"Operable" Can navigate bye searching using the keyboard without the use of a mouse. There are also drop down menu's which make it very accessible.

"Perceivable" assistive to users to skip content and get to where they want to get to.

Matthew seems to confirm a longheld belief i've had for a while (though, I think of it in differing ways). Mainly, that the world is ever changing, and that nothing is permanent (see permanent beta movement, as an example). If one wants to adapt to differing circumstances, one needs to learn.

Proyecto "Anotación PFR", https://github.com/lmichan/PFR,

Tema/mesh/D012137/RespiratorySystem,

TipoDePrueba/mesh/D002000/ForcedSpirometry,

EtapaPrueba/Estandar,

PatronFuncional/Obstruccion,

PatronFuncional/PosibleRestriccion,

PatronFuncional/PosibleMixto,

PatronFuncional/Normal,

PatronFuncional/Broncodilatacion,

PatronFuncional/NoBroncodilatacion,

This is very common in our current culture. Whether it be through social media, or video games, doxing can be a very dangerous and harmful act. As a user of the internet, everyone leaves significant digital footprints which can be used to reveal personal information, and by making this public, it allows anyone to harm this person.

A Twitter account is identifying participants from the white nationalist rally in Charlottesville by cross-referencing photos from the event with social media profiles. This public shaming has led to consequences for some of the marchers, such as job loss and social ostracism. The initiative raises questions about privacy, accountability, and the impact of online vigilantism.

One particularly striking detail from the article is its exploration of the historical context and evolution of doxing. Originally a tactic used within hacker communities to expose rival hackers, doxing has since evolved into a widespread tool for harassment and intimidation, often with devastating consequences for its victims. The article also delves into notable cases of doxing, illustrating the real-world impacts and the diverse motivations behind these attacks, from personal vendettas to political activism. This historical and case-study approach provides a well-rounded view of the phenomenon, complementing the chapter's discussion on the public forms of individual harassment and reinforcing the need for a multi-faceted approach to combating online harassment that includes legal, technical, and educational strategies.

The Wikipedia article on Gamergate (harassment campaign) details a misogynistic online harassment movement that targeted women in the video game industry, notably feminist media critic Anita Sarkeesian and game developers Zoë Quinn and Brianna Wu, primarily between 2014 and 2015. The campaign, propagated through the hashtag "#Gamergate," included severe harassment tactics like doxing, rape threats, and death threats. Gamergaters claimed to advocate for ethics in video game journalism and to protect the "gamer" identity against political correctness, though their actions and conspiracy theories, including false accusations of unethical relationships and press collusion, were widely dismissed as baseless. The movement highlighted a broader culture war over feminism, social criticism, and diversity in video games, and is noted for influencing discussions on online harassment and its prevention, as well as contributing to the rise of the alt-right.

From this article, i find author particularly focusing on the backlash following her online interactions with Courtney Stodden. This piece explores the broader implications of celebrity culture and internet accountability, illustrating how public figures are held responsible for their past actions in the digital age. Grady effectively captures the complexities of reputation management in the era of social media, where personal histories are perpetually accessible and subject to scrutiny.

I find this one discusses the phenomenon of 4chan raids, highlighting how this particular segment of the internet can influence broader digital and social environments. The focus on 4chan—a platform known for its anonymity and minimal moderation—underscores the challenges in managing online communities without stifling free expression. De Cristofaro's analysis is crucial for understanding the mechanisms through which online cultures can spill over into real-world actions, emphasizing the need for a balanced approach to internet governance.

I read this article, and it speaks about how powerful influence can be through media and communication. It would be hard to hold people accountable for inciting any acts of terrorism that happened because it could happen randomly.

I read "Study finds Reddit's controversial ban of its most toxic subreddits actually worked" from TechCrunch. This article talked over the impact banning of certain subreddits filled with hate speech. They found that the ban was pretty effective and not as many people migrated to different places to spew their hate speech. I think this is something that should be looked into further. How can we go about implementing things like this on other sites? I also wonder what consequences banning subreddits or other things will have? How will these people find other ways to show their hate - will they turn to violent actions instead of speech? Will they create their own site? I wonder if things like this already exist.

Doxing is a form of harassment where people’s personal information is made public so that they can receive further harassment from anyone to sees this information. It’s usually done on social media to those who have said or done something that has enraged the public which still doesn’t make it okay.

Doxing, or doxxing, involves publicly releasing someone's personal information without their consent, typically online, and can include data from both public and private sources. The practice is often associated with motives like online shaming, extortion, or hacktivism, and can result from both legal and illegal activities such as hacking or social engineering. Historically, the term "doxing" stems from hacker culture of the 1990s where "dropping dox" was a revenge tactic to expose and harass opponents. Modern examples of doxing have brought the term into mainstream awareness, especially highlighted by events like Gamergate, where doxing was used to target and harass individuals. Doxing raises significant concerns about privacy and security, illustrating the potential dangers of digital personal data exposure and misuse.

Doxxing is the act of finding and publicly releasing private information about someone, usually found through the internet. Information can include home address, work information, banking information, internet usage, and other personal information that could cause harm to the person or entity being affected. Sometimes, people will dox as a form of activism for different issues.

I remember watching the documentary on Meghan Markle. I had never felt so bad. She was constantly targeted for being a different race from her husband. The hate really got to the point where Meghan felt defeated but it never stopped her from exposing and standing firm. Her husband also seemed very supportive as well.

Recently, I have seen the actress from the Romeo and Juliet reboot be insanely harassed on Twitter, since she is Black. Pretty much all the harassment has been because of her looks and has been incredibly racist.

As I read Emiliano De Cristofaro's article on the influence of 4chan on internet culture and social behavior, I was struck by the crucial examination it offers into how unregulated spaces can foster toxic communities that impact wider social and political attitudes. While the insights provided are invaluable, I believe the article could have delved deeper into the mechanisms through which these communities exert influence beyond the digital realm, especially in shaping real-world events and policies.

In the article, the author interviews Kurt Braddock about how rhetorical strategies can lead to political violence. Kurt Braddock is an expert on communication techniques related to violence. He explains the concept of stochastic terror, where public figures make ambiguous statements that indirectly incite violence without explicit intention. This creates an environment of fear and unpredictability, as individuals influenced by such rhetoric may commit violent acts, thinking they are justified. Braddock suggests using "attitudinal inoculation" (educating people on recognizing and resisting manipulative messages) to combat stochastic terror.

Jacobson's story highlights the pervasive fear and anxiety caused by an online stalker, showing how digital harassment can persist over many years and significantly affect one's mental health and sense of security. This source underscores the urgent need for more effective protective measures and legal actions against cyberstalking to support victims who face such relentless harassment.

Proyecto "Anotación PFR", https://github.com/lmichan/PFR,

Tema/mesh/D012137/RespiratorySystem,

TipoDePrueba/mesh/D002000/ForcedSpirometry,

TipoDePrueba/mesh/D010993/PlethysmographyWholeBody,

TipoDePrueba/mesh/D011653/PulmonaryDiffusingCapacity,

EtapaPrueba/Interpretacion,

PatronFuncional/Obstruccion,

PatronFuncional/PosibleRestriccion,

PatronFuncional/PosibleMixto,

PatronFuncional/Normal,

PatronFuncional/Broncodilatacion,

PatronFuncional/NoBroncodilatacion,

PatronFuncional/RestriccionSimple,

PatronFuncional/RestriccionCompleja,

PatronFuncional/TrastornoMixto,

PatronFuncional/VolumenesNormales,

PatronFuncional/Hiperinflacion,

PatronFuncional/PulmonesGrandes,

PatronFuncional/AumentoFlujoSanguineo,

PatronFuncional/AnormalidadVascularPulmonar,

PatronFuncional/PerdidaVolumenLocalizada,

PatronFuncional/PerdidaAlveoloCapilar,

PatronFuncional/DifusionNormal,

Enfermedad/mesh/D001249/Asthma,

Enfermedad/mesh/D029424/ChronicObstructivePulmonaryDisease,

Enfermedad/mesh/D017563/InterstitialLungDisease,

Enfermedad/mesh/D009468/NeuromuscularDisease,

Enfermedad/mesh/D011649/PulmonaryAlveolarProteinosis,

Enfermedad/mesh/D013121/SpinalCurvatures,

Enfermedad/mesh/D012600/Scoliosis,

Enfermedad/mesh/D000081029/PulmonaryArterialHypertension,

Enfermedad/mesh/D006469/Hemoptysis,

Enfermedad/mesh/D011655/PulmonaryEmbolism,

Enfermedad/mesh/D011656/PulmonaryEmphysema,

Enfermedad/mesh/D001991/Bronchitis,

I have definitely have heard about intersectionality but never understood what it was. By looking at different factors, any one factor can affect different treatment, regardless of each individual factor. I think this is true because in the example given, even if there isn't a bias against women, a bias against black women still exists. I think this is very important because by understanding how just one factor can affect biases, we can look for solutions or at least be aware of this phenomenon.

To me, this approach is essential for understanding the complex layers of disadvantage that can affect individuals who belong to multiple marginalized groups simultaneously. The example of the resume filter underscores the importance of nuanced analyses in systems and policies to ensure fairness across all intersections of identity, avoiding oversimplified solutions that might inadvertently perpetuate discrimination.

To be honest, when reading online, we can feel some bias in comments about race, but I did not expect that the data would be so different. This shows that black women have been smeared by various negative and false rumors on the Internet all year round, and have been made assumptions by netizens with stereotypes. I think they have suffered from such painful misunderstandings for a long time, and they are treated so badly on the virtual network. Will the reality be even more tragic?

No, crowd harassment is never justified. Harassment, whether by an individual or a crowd, is harmful and abusive behavior that violates people's rights and dignity. It can cause significant emotional, psychological, and sometimes physical harm to the person or group being targeted.

The text highlights the troubling phenomenon of crowd harassment, which, amplified by social media, becomes a powerful and often devastating form of collective aggression. Reflecting on this, I find it deeply concerning how the anonymity and reach of digital platforms can transform individual acts of harassment into overwhelming waves of hostility. This not only magnifies the psychological impact on victims but also raises significant ethical questions about accountability. In my personal experience, witnessing how quickly a single negative post can escalate into widespread harassment underscores the urgent need for more robust mechanisms to combat such behavior online. It's imperative that we rethink our legal and social frameworks to address the unique challenges posed by crowd harassment in the digital age.

almost always it is not justified. most might see crowd harassment as cancel culture or worse. First a lot of individuals that have no internet presence or a small following “canceling” them is really unethical and can ruin lives. But for big celebrities or billionaires the cancellation that they deal with is not real. The person will still have all their money that they received from gaining a following and often for a lot of right wing grifters they become more popular when they perceived as being canceled.

The concept of crowd harassment, often manifested in the form of riots, mob violence, revolts, revolutions, and government persecution, is a complex and multifaceted phenomenon with deep historical roots. It involves collective behavior that can be both spontaneous and organized, driven by social, political, economic, or cultural motives.

Crowd harassment will never be justified. It makes no sense to bash individual people. Crowd harassment happens mostly on social media. Groups of people come together and target specific people whether that be in the comments or actual posts. It never makes sense why people feel the need to do such harmful actions. Most of the time people do not realize the mental impact it leaves on people.

No. I think it could never be justified unless there is a restriction on users' freedom of speech. This usually happens on social media because when people see one negative comment about a post, more people will join the group to put more negative comments on the post. I don't think there is a law that restrict people from doing that.

The exploration of individual harassment via social media vividly highlights the diverse and pernicious ways in which technology can be exploited to harm others. It strikes me how the anonymity and reach of social media platforms can amplify the impact of traditional forms of harassment, making them more pervasive and harder to escape. I am particularly alarmed by the concept of cyberstalking, as it illustrates a relentless invasion of privacy that can extend beyond the digital realm to the physical world, heightening the victim's sense of vulnerability. This chapter resonates with the broader discussion about digital ethics in our course, emphasizing the urgent need for robust legal frameworks and ethical guidelines to combat online harassment effectively. One question that arises for me is: What are the most effective strategies that social media platforms can implement to prevent such harassment while balancing the need for free expression?

Social media has made it easier for individual harassment to be done. For example, the suggested people to follow section. It suggests people that you may know or have any connection to. This makes harassment easier because if someone creates a new account, it could get suggested to them even if they have blocked them before.

This is something I've seen on Instagram where an account if either hacked and the person pretends to promote something impersonating as the person using their own account. I've also seen this where a fake account with similar information found publicly online impersonates them. In order to address impersonators, people post on their stories informing their followers they have been either hacked or the account is not theirs.

I had a friend that kept getting harassed by an anonymous account on Instagram. They knew that they knew the person in real life, because the harasser would reference things from my friend's life to intimidate them.

The chapter’s discussion on bullying and cyberstalking highlights how technology intensifies harassment, making it more pervasive and invasive. The relentless nature of cyberstalking, where perpetrators continuously create new accounts or track a person's physical location, is particularly alarming. This not only invades digital privacy but can also lead to severe emotional distress. It underscores the need for stricter regulations, improved reporting mechanisms on social media platforms, and increased awareness to help individuals protect themselves and seek justice effectively.

I believe cyberbullying is one of the greatest issue on social media. People on social media will judge posts and the user that create the post. I remember in a TV show I watched, it talked about how a girl got cyberbullying and eventually suicide after suffering in depression for a long time. This is not the only case, and there are thousands of cases similar to this one. Nowadays, people who want to be an influencer has to have the courage to face cyberbullying, and those who couldn't wouldn't have a good consequence.

This article is about reddit banning hateful subreddits and how that has made the site more pleasant. It is easier to spread hate if you are around other people who are also spreading hate. Dispersal of this hate subdues it and often makes the place (like reddit) become a better place. it probably also forces people off of reddit who cannot do the hating they once did.

I feel likee the comparison of the social contract to a medical scenario is particularly illustrative. Just as a patient might consent to a surgical procedure that restricts their physical autonomy temporarily in order to achieve a greater health outcome, members of a society consent to certain restrictions on their freedoms for the overall good and order of the community. This consent forms the basis of the legitimacy of the social constraints imposed, echoing the ethical underpinnings of both medical practice and societal governance.

Typo

Sobre [[Fenomenología del fin]] de [[Franco Berardi (Bifo)]].

De la transición tecnológica hacia el mundo digital y cómo eso modifica nuestra capacidad empática y de sociabilidad.

I feel like it is inevitable to feel differently depending on who the target is. Everyone has their own bias and it is impossible to avoid personal bias. There are definitely moments when harassment could possibly be justified if you are looking through different moral lenses. If someone is receiving harassment because they are a morally bad person or have set out and done bad actions, you could say the harassment is justified. If someone hurt an innocent person, or even a child, that harassment could be justified. We have an obligation to speak out against violent speech or actions, and one way people seem to do this is through harassment. There are definitely better ways to go about it and I hope we can stand up against violence is a more peaceful manner than harassment.

Although I do not want to admit it, I definitely feel differently when crowd harassment is done depending on the target's actions. The doxing of the KKK members, though not ethical, I feel is somewhat justified as it helps promote social harmony. Like Confucian ethics, doxing may raise ethical concerns, as it can lead to harm or social disruption, which may conflict with its principles. However, it may be justified if it is seen as a necessary action to protect the community, uphold justice, and reinforce societal norms against violence and racism. All in all, I understand the inclination to view crowd harassment in a different light due to the actions of the target, but I would never necessarily promote it. I think there are better avenues to create change.

This article is about a solar storm that hit Earth in 2024. It discusses the impact of the storm on technology, including power grids, GPS, and satellites.

The storm was the strongest in 20 years and caused disruptions worldwide. It is the first of its kind since 2003.

The Carrington Event of 1859 was the most powerful solar storm ever recorded. Solar storms are caused by the Sun releasing charged particles that can interfere with Earth's magnetic field.

malha energética

flutuações

rajadas de radiação

een te laag zuurstofgehalte van het bloed

Read and understood BS

Due Dates understood BS

Its 404 and a link that wont open

That is plain untrue

Yes, I think social media platforms should judge instances of harassment, even though I do not think there are any instances where harassment is warranted. First, I think platforms should review reports of harassment to tackle the issue initially. If someone makes the effort to report someone, it should definitely be investigated. Second, platforms should review if comments or actions are causing emotional distress upon someone else using the platform. This task is a bit more complex as actions are subjective and should be cautious and not engage in over-censorship.

wow. so if the grantType is authorization code flow type, this parseFromUrl function actually automatically does an entire extra step which is the POST to get the token. questionable API design, i mean you can't know that by looking at its name...

here's the key difference to implicit flow. the token and other info properties are return as the response of the POST request.

according to the next paragraph, at least in this example case of okta, we don't need manual access to the random secret generated by frontend code.

Assigning a disproportionate number of students from specific racial groups to segregated classrooms is harmful due to the numerous benefits of inclusive educational settings. These advantages encompass improved academic achievement, reduced behavioral issues, higher graduation rates, improved attendance, and better employment outcomes after graduation. Ensuring equitable access to inclusive education not only supports the academic and social development of students with disabilities but also promotes a more inclusive and equitable educational system overall. This underscores the importance of addressing disparities in special education placement to create more effective and supportive learning environments for all students.

This can be due to the many barriers that they face. They don't have many resources that can help them succeed in school. While others who are wealthy are able to hire tutors to help them throughout their education. Also students who are living in poverty also attend schools in which they don't have that many resources allocated towards students.

The disproportionate placement of students of color into special education settings exacerbates existing inequalities within schools. This practice can lead to serious negative outcomes, including racial segregation, stigmatization, and misrepresentation of racial groups within special education programs. Such outcomes not only affect individual students but also contribute to systemic issues of inequality and discrimination within the education system.

This statement highlights a concerning trend where low-income students are disproportionately labeled with subjective disability categories and segregated into separate classrooms. The reliance on subjective assessments can contribute to biased categorization and unequal access to mainstream educational settings. This underscores the need for equitable and unbiased practices in special education identification and placement to ensure that all students receive appropriate support and opportunities for inclusive education.

Author response:

This important manuscript uses circuit mapping, chemogenetics, and optogenetics to demonstrate a novel hippocampal lateral septal circuit that regulates social novelty behaviours and shows that downstream of the hippocampal septal circuit, septal projections to the ventral tegmental area are necessary for general novelty discrimination. The strength of the evidence supporting the claims is convincing but would be strengthened by the inclusion of additional functional assays. The work will be of interest to systems and behavioural neuroscientists who are interested in the brain mechanisms of social behaviours.

We thank the reviewers for their thoughtful and constructive feedback. We are excited that both reviewers thought that the manuscript was of “interest to specialists in the field and to the broad readership of the journal”, that the paper was “well-written and logically organized” and that the “study opens an avenue to study these circuits further to uncover the plasticity and synaptic mechanisms regulating social novelty preference.” Additionally, the reviewers wrote that the experiments were “well-designed” “with clever controls and conditions to provide compelling evidence for their conclusion.” The reviewers additionally provided constructive feedback which we address in our responses below.

Public Reviews:

Reviewer #1 (Public Review):

Summary:

The study investigated the neural circuits underlying social novelty preference in mice. Using viral circuit tracing, chemogenetics, and optogenetics in the vHPC, LS, and VTA, the authors found that vHPC to LS projections may contribute to the salience of social novelty investigations. In addition, the authors identify LS projections to the VTA involved in social novelty and familiar food responses. Finally, via viral tracing, they demonstrate that vHPC-LS neurons may establish direct monosynaptic connections with VTA dopaminergic neurons. The experiments are well-designed, and the conclusions are mostly very clear. The manuscript is well-written and logically organized, and the content will be of interest to specialists in the field and to the broad readership of the journal.

Strengths:

(1) The vHPC has been involved in social memory for novel and familiar conspecifics. Yet, how the vHPC conveys this information to drive motivation for novel social investigations remains unclear. The authors identified a pathway from the vHPC to the LS and eventually the VTA, that may be involved in this process.

(2) Mice became familiar with a novel conspecific by co-housing for 72h. This represents a familiarization session with a longer duration as compared to previous literature. Using this new protocol, the authors found robust social novelty preference when animals were given a choice between a novel and familiar conspecific.

(3) The effects of vHPC-LS inhibition are specific to novel social stimuli. The authors included novel food and novel object control experiments and those were not affected by neuronal manipulations.

(4) For optogenetic studies, the authors applied closed-loop photoinhibition only when the animals investigated either the novel conspecific or the familiar. This optogenetic approach allowed for the investigation of functional manipulations to selective novel or familiar stimuli approaches.

Weaknesses:

(1) The abstract and the overall manuscript pose that the authors identified a novel vHPC-LS-VTA pathway that is necessary for mice to preferentially investigate novel conspecifics. However, the authors assessed the functional manipulations of vHPC-LS and LS-VTA circuits independently and the sentence could be misleading. Therefore, a viral strategy specifically designed to target the vHPC-LS-VTA circuit combined with optogenetic/chemogenetic tools and behavior may be necessary for the statement of this conclusion.

The reviewer raises an important point. Although Figure 3 shows that vHPC (vCA1 and vCA3) is the source of the greatest number of monosynaptic inputs onto LS-VTA neurons, we did not perform any experiments that specifically manipulated vHPC neurons that project to LS-VTA neurons. While these experiments would be extremely interesting, they are technically challenging and beyond the scope of this study. We are happy to edit our manuscript to clarify this point.

(2) The authors combined males and females in their analysis, as neural circuit manipulation affected novelty discrimination ratios in both sexes. However, supplementary Figure 1 demonstrates the chemogentic inhibition of vHPC-LS circuit may cause stronger effects in male mice as compared to females.

The reviewer makes an interesting point. We can confirm that we found no significant differences in the effectiveness of our vHPC-LS inhibition between the males and females (2-factor ANOVA with sex (male/female) and drug condition (saline/CNO) as factors on the discrimination scores of hM4Di expressing animals: interaction p=0.2241, sex: p=0.1233, drug condition: p=0.0166). These data suggest that there are no significant sex differences in the effectiveness of inhibition of the vHPC-LS neurons. We will include these comparisons in the revised manuscript.

(3) In most experiments, the same animals were used for social novelty preference, for food or object novelty responses but washout periods between experiments are not mentioned in the methods section. In this line, the authors did not mention the time frame between the closed-loop optogenetic experiments that silenced the vHPC-LS only during familiar and then only novel social investigations. When using the same animals tested for social experiments in the same context there may be an effect of context-dependent social behaviors that could affect future outcomes.

We thank the reviewer for this important clarification. We apologize for not including these crucial details in our Methods section. For both the chemogenetic and optogenetic inhibition experiments, all conditions were separated by a minimum of 24 hours. In the chemogenetic inhibition experiments, saline and CNO conditions were counterbalanced between animals. Similarly, we counterbalanced the order of light ON vs light OFF conditions across animals during our optogenetic inhibition experiments. We will include these additional details in the revised manuscript.

(4) All the experiments were performed in a non-cell-type-specific manner. The viral strategies used targeted multiple neuronal subpopulations that could have divergent effects on social novelty preference. This constraint could be added in the discussion section.

We will expand our discussion section to address this essential point raised by the reviewer.

(5) The authors' assumptions were all based on experiments of necessity. The authors could use an experiment of sufficiency by targeting for instance the LS-VTA circuit and assess if animals reduce novel social investigations with LS-VTA photostimulation.

We agree with the reviewers that it would be interesting to determine if LS-VTA neurons are sufficient, in addition to being necessary, to drive social novelty. These will be interesting experiments to pursue in the future.

Reviewer #2 (Public Review):

Summary:

Rashid and colleagues demonstrate a novel hippocampal lateral septal circuit that is important for social recognition and drives the exploration of novel conspecifics. Their study spans from neural tracing to close-loop optogenetic experiments with clever controls and conditions to provide compelling evidence for their conclusion. They demonstrate that downstream of the hippocampal septal circuit, septal projections to the ventral tegmental area are necessary for general novelty discrimination. The study opens an avenue to study these circuits further to uncover the plasticity and synaptic mechanisms regulating social novelty preference.

Strengths:

Chemogenetic and optogenetic experiments have excellent behavioral controls. The synaptic tracing provides important information that informs the narrative of experiments presented and invites future studies to investigate the effects of septal input on dopaminergic activity.

Weaknesses:

There are unclear methodological important details for circuit manipulation experiments and analyses where multiple measures are needed but missing. Based on the legends, the chemogenetic experiment is done in a within-animal design. That is the same mouse receives SAL and CNO. However, the data is not presented in a within-animal manner such that we can distinguish if the behavior of the same animal changes with drug treatment. Similarly, the methods specify that the optogenetic manipulations were done in three different conditions, but the analyses do not report within-animal changes across conditions nor account for multiple measures within subjects.

Thank you for raising this important point. We are happy to include the repeated measure ANOVAs and paired t-tests in a revised version of the manuscript.

Finally, it is unclear if the order of drug treatment and conditions were counterbalanced across subjects.

As mentioned in the above response to Reviewer 1, for both the chemogenetic and optogenetic inhibition experiments, all conditions were separated by a minimum of 24 hours and we counterbalanced the order of chemogenetic (saline/CNO) and optogenetic (light ON/light OFF) experimental manipulations across animals. We will include these additional details in the revised manuscript.

I found this extremely interesting as I am a major theatre nerd. I love how this includes acting and even puppetry into the arts because it often is overlooked. However, this did spark a question in me: How can I as a teacher use art forms such as playwriting or poetry in my classroom to help engage my students? I want to become a math teacher and I would love to use some of these options in my own classroom!

This quote really moved me. I've always considered the arts to be such an incredible part of using creaitvity and imagination, but this helped me to understand that it is also essential! This just makes me think about the effects that art can have on our growing children and students and the benefits it can provide to our classrooms! In my own 3rd grade classes, our teacher took time out of her busy schedule to teach us how to watercolor, sketch, etc. This helped me to express myself and genuinely enjoy her class. I now can recognize how this not only helped me creatively, but also developmentally!

The Education Department had long been aware of evidence indicating that T.J.'s struggles were more severe than initially classified. This misclassification resulted in T.J. receiving insufficient services to address his needs. The failure to act on this evidence underscores systemic issues within the department regarding the assessment and support of students with special needs.

This is frustrating to read. This is a result of the lack of resources that he has not received through out his education. If he was actually given resources that could help him succeed he wouldn't be so far behind from the rest of his peers.

Note: This response was posted by the corresponding author to Review Commons. The content has not been altered except for formatting.

Learn more at Review Commons

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Reply to the reviewers

Description of the planned revisions

Reviewer #1 (Evidence, reproducibility and clarity (Required)):

• Again, in Figure 5, were FoxP3/CD4+ cells enumerated? Author Response: Fig 5 showed that the inflammatory score, and activation of CD4 and CD8 cells, were lower in the intestine of DSS-treated mice transplanted with Jag1Ndr/Ndr lymphocytes than in those transplanted with Jag1+/+ lymphocytes. However, in Figure 5 we had not quantified the number of FoxP3/CD4+ cells (Tregs). We agree that it would be interesting to know whether the dampened intestinal inflammation (in response to a classical inflammatory disease model (DSS-treatment)) is also mediated by excess Tregs. We will therefore now quantify Foxp3+ cells on the intestinal sections of experimental animals used for acquisition of data in Fig 5.

• *

Description of the revisions that have already been incorporated in the transferred manuscript.

Reviewer #1 (Evidence, reproducibility and clarity (Required)):

Reviewer 1 comment: This is an interesting study that examines defects in the Jag1ndr/ndr mouse model of Alagille syndrome. The novel aspects of this manuscript are the comparisons, at many levels, between the mouse model and ALG patient samples, including an examination of immune profiles. The conclusions that the Jag1ndr/ndr mouse model is an accurate representation of the human ALG syndrome appear valid. However the reported differences in immune profiles, particularly in the Jag1ndr/ndr mouse model are difficult to understand. The data presented indicate a reduction in CD4+ cells in the Jag1ndr/ndr mouse at day P3 in both liver and spleen. Additionally, the authors report differences between the the Jag1ndr/ndr mouse and controls at day P30 in the relative percentages of DN, DP and SP CD4 and CD8 cells in the thymus. When examining the peripheral lymphoid system, CD4+ numbers are the same in both the Jag1ndr/ndr animals and controls however CD8+ numbers are reduced and FoxP3/CD4+ cells are increased in both the spleen and the thymus. FoxP3/CD4+ T cells are usually assumed to be regulatory T cells that dampen the inflammatory responses of T cells. Therefore, the increase in this population in an animal model of what is assumed to be an inflammatory disease is confusing and confounding. The authors do not present a clear analysis of how they feel an increase of Tregs would lead to this disease. One possibility is that this population is not functioning as conventional Tregs and rather are promoting inflammation but this conclusion would require a functional analysis of this population of cells, at the very least in an in vitro analysis of T cell suppression. From an immunologist's point of view, their data are antithetical to what one would expect to find in an inflammatory disease. Perhaps this reviewer is missing an important point but if I am missing it, then other who read this manusgcript also may be confused.

Author Response: *We thank the reviewer for carefully assessing our work, and for noting which aspects of the immune analyses should be more thoroughly explained. We apologize for any confusion, which a clearer introduction will help to avoid. *

*Alagille syndrome is not thought of as an inflammatory disorder, it is a congenital disorder affecting bile duct development (Kohut et al 2021, Semin Liver Dis). During normal bile duct development, JAG1+ portal fibroblasts signal to NOTCH2+ hepatoblasts to instruct bile duct development. In the context of low JAG1 signaling, hepatoblasts either fail to adopt a cholangiocyte fate, or fail to undergo bile duct morphogenesis, resulting in bile duct paucity and cholestasis. This cholestasis should activate inflammatory processes leading to fibrosis, which is the subject of this study. *

• *

We agree with the reviewer that Tregs would be expected to suppress inflammation, and our data are consistent with Treg suppression of inflammation. We show, for the first time, that Tregs are enriched in Jag1Ndr/Ndr mice (Fig 4) and present evidence that they suppress inflammation (Fig 5) and fibrosis (Fig 6), which could explain the atypical fibrosis seen in patients with ALGS.

• *

*To clarify that ALGS is a genetic liver disease affecting bile duct formation, we: *

1. Modified and extended the following text in the Introduction (Page 2, lines 14-17): “ALGS is mainly caused by mutations in the Notch ligand JAGGED1 (JAG1, 94%) (Mašek & Andersson, 2017; Oda et al, 1997), affecting bile duct development and morphogenesis, resulting in bile duct paucity and cholestasis. Immune dysregulation has also been described (Tilib Shamoun et al, 2015), but how this might interact with liver disease in ALGS to affect fibrosis is not known.”
2. *Introduce the disease, the animal model, and the scientific question in a schematic in new Fig 1A. *
3. * Reviewer 1 comment: Minor points that should be addressed include: • The source cells used in the transfer experiments reported in Figure 5 is unclear. Are they using total spleen cells with T, B and myeloid cells or are they using purified T cells. And if it is the latter, have they assessed the ratio of CD4+ versus FoxP3/CD4+ cells in the transferred cells?

Author Response: *Total spleen cells including all lymphocytes were transplanted, as described in Materials and Methods. The constituent T-cell populations are characterized and shown in Fig 4F. To clarify this, we: *

1. *added the text “Adoptive transfer of lymphocytes” to the schematic in Fig 5A, FigS5A, and Fig 6A, and *
2. modified the opening paragraph related to results presented in Fig.5 and FigS5 in the following way (page 8, line 209): “To investigate Jag1Ndr/Ndr T cell function, we performed adoptive transfer of the splenic lymphocytes into Rag1-/- mice, which lack mature B- and T cell populations, but provide a host environment with normal Jag1 (Mombaerts et al, 1992).”
3. *

*To acknowledge that B-cells and innate lymphoid cells might contribute to the observed results, we include a following sentence in the Discussion: *

(page 12, lines 369-371) “Finally, our experimental setup does not exclude an additional contribution of other lymphocytes (B-cells or innate lymphoid cells) to the BDL-induced fibrosis, and selective testing of the individual subpopulations would be an intriguing follow up to this study.”

Reviewer 1 comment: In the DSS experiments in Figure 5, there does not appear to be a no DSS control. What does the architecture look like without DSS?

Author Response: The intestinal architecture and phenotype of mice transplanted with Jag1+/+ or Jag1Ndr/Ndr lymphocytes, not treated with DSS, are presented in Supplementary Figure 5. In the absence of DSS, Jag1+/+- or Jag1Ndr/Ndr -transplanted mice exhibit no overt differences in survival or weight gain/loss. The intestinal inflammatory score was not different in the two conditions and was *2.29 +/-0.44 and 2.03 +/-0.92 for Jag1+/+- or Jag1Ndr/Ndr -transplanted mice, respectively. *

To compare the results with and without DSS, we added the following text to the results section, when describing the DSS results (Page 9, lines 223-226):

“As expected, histological scoring of intestinal and colonic inflammation revealed elevated inflammation in Jag1+/+→Rag1-/- mice treated with DSS (Fig. 5C,D) compared to Jag1+/+→Rag1-/- mice not treated with DSS (Fig. S5). However, there was significantly less inflammation in Jag1Ndr/Ndr→Rag1-/- mice than in Jag1+/+→Rag1-/- mice (Fig. 5C,D)."

Reviewer 1 comment: The authors noted that splenomegaly was observed in the Jag1ndr/ndr mouse model. Again this is antithetical to what one would expect when one sees an increase in FoxP3/CD4+ T regs.

Author Response: *We thank the reviewer for pointing at a possible discrepancy, related to Fig1 in which we report the presence of splenomegaly. Although there can be multiple causes of splenomegaly, it is one of the hallmarks of portal hypertension (as also corroborated by Reviewer 2), tightly connected with liver fibrosis, present in patients with ALGS and we report it as such in the manuscript. To clarify this, we added the following text sections: *

1. Results (page 2, lines 37,38) “Liver fibrosis compresses blood vessels and reduces their blood flow, leading to portal hypertension, a serious consequence of liver disease which can manifest as splenomegaly.”
2. Discussion (page 13, line 394-401): “Splenomegaly has been described as a consequence of portal hypertension in ALGS (Kamath et al, 2020), but could also be attributed to immune-related pathology. Jag1Ndr/Ndr mice exhibit splenomegaly as early as P10, and is exacerbated at P30 ( 1E,F). Patients with other liver diseases display portal hypertension and cirrhosis, with both splenomegaly and hypersplenism associated with a high CD4+/CD8+ ratio, but a low Treg+/CD4+ ratio (Nomura et al, 2014). However, Jag1Ndr/Ndr mice present with splenomegaly but not hypersplenism. An overactive spleen (hypersplenism) would remove red blood cells which are instead enriched in Jag1Ndr/Ndr mice, and Tregs were enriched in Jag1Ndr/Ndr mice, not depleted as seen in cirrhosis/hypersplenism. These data are thus consistent with portal hypertension-induced splenomegaly rather than hypersplenism.*” *

Reviewer #1 (Significance (Required)):

Reviewer 1 comment: The strengths of this paper are the careful comparisons between the mouse model and the human ALG syndrome. These comparisons are valuable and worth publication.

Author Response: We thank the reviewer for these comments.

Reviewer 1 comment: Weaknesses are stated above. Needs a clearer explanation for their immune analysis.

Author Response: *We thank the reviewers for highlighting points requiring clarification and hope the proposed text changes and additional data presented in response to the comments of all three reviewers lead to a significant clarification of the immunological aspect of our study. *

Reviewer #2 (Evidence, reproducibility and clarity (Required)):

Reviewer 2 comment:

Summary: Masek and colleagues use multi-pronged studies on the Jag1[Ndr/Ndr] mouse model of Alagille syndrome (ALGS) combined with transcriptomic analysis on livers from patients with ALGS to elucidate the potential mechanisms regulating liver fibrosis in this disease. The authors first show that Jag1[Ndr/Ndr] animals develop pericellular and perisinusoidal fibrosis and exhibit evidence for portal hypertension, similar to patients with ALGS. Single-cell RNA-sequencing indicated more hepatoblasts and less hepatocytes, relatively speaking, in Jag1[Ndr/Ndr] P3 livers, which suggested hampering of hepatoblast differentiation to hepatocytes. Deconvolution of previously generated bulk RNA-seq data from Jag1[Ndr/Ndr] P10 livers and GESA on RNAseq data from livers of these mice and patients with ALGS confirmed the P3 scRNA-seq observations and indicated mild pro-inflammatory activation of immature hepatocytes in ALGS livers. GESA also suggested an inability of Jag1[Ndr/Ndr] livers to attract T cells upon cholestatic injury. Indeed, 25-color flow cytometry on liver and spleen from mutant and control mice indicated a defect in T cell response to cholestasis in this model. The authors then examined the effects of the Ndr mutation on T-cell development and function. They found that the Ndr/Ndr thymi were significantly smaller than control thymi. Moreover, Ndr/Ndr thymi showed an increase in CD4+ T-cells and Tregs at the expense of double-positive T-cells. The authors then performed lymphocyte transplantation studies and concluded that Ndr/Ndr T-cells fail to mount an adequate response to inflammation in a DSS model of ulcerative colitis. The authors tested the contribution of Ndr/Ndr immune cells to liver fibrosis in a model of experimentally induced cholestasis (bile duct ligation; BDL). Ndr/Ndr T-cells did not show any defects in migrating into the liver upon BDL. However, the periportal fibrosis observed in BDL model was reduced in animals receiving Ndr/Ndr immune cells compared to those receiving Jag1+/+ immune cells. This was accompanied by significantly less aSMA staining in these livers. Finally, reanalysis of bulk RNAseq data from liver samples from ALGS and other liver diseases suggested that the presence of FOXP3+ T-reg cells in the liver is associated with higher liver fibrosis in non-ALGS liver diseases but lower liver fibrosis in ALGS livers. The authors have used an impressive combination of single-cell RNA-sequencing, reanalysis of previous bulk RNA-sequencing data from their group and others, 25-color FACS analysis, and adoptive immune transfer experiments in this manuscript, and systematically provide quantification and statistical analysis for their data. Overall, this is an interesting and important study. Prior studies are referenced appropriately. The text and figures are clear and accurate. I don't think any additional experiments are essential. However, the issues listed under Major comments should be discussed and clarified in the manuscript, especially the first item.

Author Response: *We sincerely thank the reviewer for the comprehensive and insightful assessment of our manuscript. We are particularly gratified to note your acknowledgment of the thoroughness of our experimental approach and the clarity of our presentation. We are pleased that no further experiments would be required, and will address the points raised under Major comments which enhance our study's quality and accessibility. *

Reviewer 2 comment:

Major comments:

• Only a small fraction of the cells in scRNA-seq experiments have been assigned to hepatocytes/hepatoblast clusters, with the majority of these cells allocated to Hepato-Ery cluster. This suggests that many hepatocytes and potentially hepatoblasts have been lost during sample preparation. The authors should discuss this issue and its potential implications on the interpretation of the cell ratios and gene expression conclusions of scRNA-seq data. Author Response: We agree with the reviewer regarding this aspect of our study. We mentioned this limitation in the supplementary methods section: ”Liver parenchymal cells constituted ~6.5% of cells at E16.5, and ~7.5% of cells at P3 and included mesenchymal cells, endothelial cells, hepatoblasts and hepatocytes (Fig. S1D), this parenchymal proportion is lower than in vivo, but consistent with ex vivo liver digest (Guilliams et al, 2022).” We recognize it may be too inaccessible there, and we thus added the following text to the Discussion section of the manuscript: (Pages 11-12, lines 330-337) “A limitation of this study is the underrepresentation of the hepatoblast/cyte parenchymal cells in the scRNA-seq dataset (Fig. 2A-D), which constituted ~6.5% of analyzed cells at E16.5, and ~7.5% of cells at P3 (Fig. S1D). This parenchymal proportion is lower than in vivo, but is consistent with scRNA seq datasets obtained with ex vivo liver digest (Guilliams et al, 2022). One risk is that cell stress as a result of dissociation could result in further loss of injured Jag1Ndr/Ndr hepatocytes, impacting the interpretation of cell type abundance. Nuclear scRNAseq can overcome cell type-dependent dissociation sensitivity bias (Guilliams et al, 2022), and could provide further insights into Jag1Ndr/Ndr livers at the single cell level. Nonetheless, both bulk RNA seq deconvolution and histological analyses confirmed that patients and Jag1Ndr/Ndr mice exhibit hepatoblast enrichment and less differentiated hepatocytes.”

Reviewer 2 comment: The Jag1[Ndr/Ndr] strain is an excellent model for various aspects of ALGS phenotypes. However, when it comes to linking the effects of this mutation to the function of a specific cell type, it is worth considering that Jag1[Ndr/Ndr] might not recapitulate the effects of loss of one copy of JAG1 observed in most patients with ALGS. This is especially important given the sensitivity of various cellular and organ-level processes to the degree of Notch pathway activation. In the context of the present manuscript, it is possible that what the authors have observed in Jag1[Ndr/Ndr] lymphocytes does not mirror how a JAG1-heterozygous human lymphocyte behaves. This is not a major concern, but it is worth considering.

Author Response: We agree and thus added the following discussion paragraph (page 11, lines 315-321) “In patients with ALGS, who have a single mutation in either JAG1 or NOTCH2, the remnant healthy allele(s) could be expected to mediate signaling. However, some JAG1 mutations exhibit dominant negative effects (Ponio et al, 2007; Xiao et al, 2013; Guan et al, 2023), which could entail further repression of JAG1/NOTCH2 signaling. In this context, it is important to note that the Jag1Ndr/Ndr mice are homozygous for the missense mutation, but retain some JAG1 activity, and it is not clear to which degree this mimics JAG1 heterozygosity in humans. It would be of interest to test whether Jag1 potency affects hepatoblast differentiation or injury-induced reversion of hepatocytes in patients as a function of their genotype.”

Reviewer 2 comment: •The basis for the opposite type of correlation between COL1A1 expression and POXP3 level in ALGS versus non-ALGS liver disease is not clear.

Author Response: We thank the reviewer for pointing out the unclear interpretation of the patient data. In patients with ALGS, the extent of fibrosis is likely to be highly multifactorial, involving (as we show) hepatocyte immaturity, dampened inflammation, and immune system dysregulation (possibly involving more than T-cells). Since human patients ARE so heterogeneous, teasing apart the relative contribution of each is currently outside the scope of our study, but will be an important area of future research. Nonetheless we thought it was important and interesting to show these patterns in supplementary Fig 6, now extended with further data, and analyses, and described in the following manner:

• *

Results section: (page 10, lines 267-275) “Liver damage in non-ALGS liver disease (using liver injury marker LGALS3BP) (Yang et al, 2021), was positively correlated with recruitment of lymphocytes (including CD8A+,and FOXP3+ populations of T cells), as well as the extent of fibrosis (COL1A1 abundance) (Fig. S6G). However, in ALGS, the extent of liver damage, lymphocyte recruitment and fibrosis were unlinked (Fig. S6G). These data are in line with the observation that liver stiffness (a proxy for fibrosis) in ALGS is independent of biomarkers of liver disease (Leung et al, 2023). While Treg infiltration in ALGS was independent of liver damage, it exhibited a tendency towards a negative correlation with fibrosis (Fig. S6G), corroborating that elevated levels of Tregs may limit fibrosis in ALGS. Altogether, these data suggest that the liver and lymphocytes may be differentially affected in different patients with ALGS, a disorder that is well known for its heterogenous presentation.”

Minor comments:

• Page 2, last paragraph of Introduction, Page 12 last sentence, and Supplementary Methods: Please use "adoptive immune transfer" instead of "adaptive immune transfer". • Pages 3 and 4: Reference is made to Figures 3E-O, which appears to be Figure 2E-O. • Figure 3 legend: "Analysis in (E) is one-way ANOVA with Dunnett's multiple comparison test". Panel E compares two means, so ANOVA is not the appropriate statistical analysis for these data. Is this sentence related to panel D? • Page 9: Please correct misspelling: "response to intestinal insult (Fig. 5). W therefore". • The Science Translation Medicine references lack page number. Author Response: *We thank the reviewer deeply for taking the time to meticulously note and convey these errors, helping us to correct these. The suggested corrections have been implemented. Science Transl Med is an online journal and does not have page numbers – we have added an issue number to facilitate retrieval of these references. *

• *

Additionally, we noticed that the image of a consecutive liver section with CYP1A2 staining from Jag1Ndr/Ndr liver in Fig 2 L was accidentally flipped along the horizontal axis, which we have now corrected. We also changed the scRNAseq cell cluster naming from Hepatoblasts/cytes, Hepato_Ery, and Kupffer cells, Kuffer cells_Ery to Hepatoblasts/cytes I, and II, and Kupffer cells I and II, respectively, to match the Neutrophil progenitors I and II naming convention. Names were subsequently also changed in Fig S1 and methods.

**Referees cross-commenting**

To my knowledge, ALGS is not considered to be an inflammatory disorder. Furthermore, the splenomagaly observed in the mouse model could be due to portal hypertension rather than a primary immune disturbance. Having said that, I agree with the other reviewers that the manuscript will benefit from further discussion and clarification on the immune-related observations.

Author Response: We thank Reviewer 2 for indicating to Reviewer 1 that ALGS is not considered an inflammatory disorder, which we agree with. It was not our intention to convey this idea. To avoid confusion, we now:

1. *Added a schematic in Fig 1A. *
2. Modified and extended the following text in the Introduction: (Page 2, lines 14-17): “ALGS is mainly caused by mutations in the Notch ligand JAGGED1 (JAG1, 94%) (Mašek & Andersson, 2017; Oda et al, 1997), affecting bile duct development and morphogenesis, resulting in bile duct paucity and cholestasis. Immune dysregulation has also been described (Tilib Shamoun et al, 2015), but how this might interact with liver disease in ALGS to affect fibrosis is not known.” *Furthermore, we have addressed or will address all comments from reviewer 1 to clarify the immune-related observations. *

Reviewer #2 (Significance (Required)):

Despite severe cholestasis, ALGS patients do not show as much fibrosis as other cholestatic diseases, including biliary atresia (BA). A previous study had suggested that this phenomenon could be due to the difference in the nature of reactive hepatobiliary cells in ALGS compared to BA (Fabris et al, 2007). Moreover, a number of studies have suggested a role for Notch pathway activation in several cell types in the liver in the development of liver fibrosis (for example, Sawitza et al, Hepatology, 2009; Chen et al, Plos One, 2012; Duan et al, Hepatology, 2018; Yu et al, Science Translational Medicine, 2021). However, although a role for Notch signaling in T-cells is well established, it was not known whether impaired T-cell development/function contributes to reduced fibrosis in ALGS liver disease. Accordingly, the current manuscript provides novel insight into the mechanism of fibrosis in this disease. Moreover, the observation that Jag1-mutant T-cells do not confer as much protection as control T-cells to immunodeficient mice subjected to DSS-induced ulcerative colitis provides strong evidence for impaired T-cell immunity in this ALGS model and might help explain other aspects of ALGS phenotypes.

The manuscript will be of interest to broad audience (Notch signaling, cholestatic liver disease, mechanisms of liver fibrosis, T-cell development).

I have expertise in Notch signaling and in using animal models of human developmental disorders.

__Author Response: __We thank the reviewer for the balanced assessment of our manuscript in light of the current knowledge, and for highlighting its importance in the context of not only Notch and ALGS, but also other cholestatic and fibrotic liver diseases.

Reviewer #3 (Evidence, reproducibility and clarity (Required)):

The article entitled "Jag1 Insufficiency Disrupts Neonatal T Cell Differentiation and Impairs Hepatocyte Maturation, Leading to Altered Liver Fibrosis" by Mašek et al described the role of Notch ligand JAGGED1 (JAG1) in the T-cell differentiation contributing to liver fibrosis and immune system development in ALGS. This article is well written and has important preliminary findings that could establish Jag1 and its downstream signaling pathways as potential therapeutic targets to attenuate liver fibrosis.

Author Response: We thank the reviewer for recognizing our work and pointing out the therapeutical implications of our findings.

Reviewer 3 comment 1: Minor comments: In page 4, they mentioned that "the hepatoblast marker alpha fetoprotein (AFP) was 3.1-fold enriched (Fig. 3J,K), while the mature hepatocyte marker CYP1A2 protein was 1.7-fold less expressed (Fig. 3L-M)", the figure numbers should be changed to 2J, K, L-M etc.

Author Response:* We thank the reviewer for identifying these errors. The suggested corrections have been implemented. *

Reviewer 3 comment 2: In liver fibrosis the Th17 cells play crucial roles. Please show the level of IL17A mRNA level in the liver in the Jag1Ndr/Ndr mice compared to the Jag1+/+ mice.

Author Response: We thank the reviewer for the insightful comments. We indeed investigated the Th17 vs Treg immune response, however we detect neither Th17-expressed Il17, Il17a, Il17f, nor Il21 and Il22 mRNA in the bulk RNA data, suggesting their expression is either masked or they are not present in significant numbers within the liver tissue at P10, preventing us from drawing any conclusions about this cell population.

Reviewer ____3 comment 3: Also, please show the expression level of pro-inflammatory molecules, for example, TNFα, IL1β, MCP1 etc and the level of MMPs (especially MMP2, MMP8, MMP9) in the livers of the mice models used.

Author Response: *The expression of Il10, Il1b, Mcp1(Ccl2), was presented in the manuscript Fig. 2O, and we attach in the response to reviewers *

*a full list together with the expression levels of Mmp2/8/9, Tnfa, Ifng, Il17 receptor family and Tgfb1-3. Out of these, Mmp8 (0.9 Log2fold change = 1.9-fold), Ccl2 (2.2 Log2fold change = 4.7-fold), and Tl17rb (1.1 Log2fold change = 2.1-fold) were significantly upregulated, but do not indicate any specific leukocyte population’s response. This is in line with data in Fig S2E, demonstrating a dominance of myeloid over adaptive immune response in the GSEA of the immune KEGGs. *

*Since lymphocytes are underrepresented in the bulk transcriptomics, and individual genes might report activity of many different cell types, we chose to focus on the list of genes shown to be markers of activated hepatocytes, to avoid over interpretation of the RNA sequencing data. Instead, the immune analyses were based on flow cytometry data, which we expect should accurately report cell type abundance across organ systems. *

Reviewer 3 comment____ 4. Authors have shown significant alterations in the Treg population in their Jag1Ndr/Ndr mice of ALGS. Please also show the expression of IL10 and TGFβ in the liver and whether they are correlated with the level of Treg populations.

Author response:* IL10 and Tgfb mRNA levels in liver are shown in the heatmap in the response to reviewers, and were not significantly different between genotypes at P10. They were also not correlated with Foxp3 levels, as shown in the correlation matrices below (Pearson’s R values in top row, significance values in bottom row). *

Reviewer 3 comment 5. It would be interesting to know whether the IFNγ mRNA expression in the livers were altered in the Jag1Ndr/Ndr mice with altered populations of CD8 T cells.

Author Response: There was no significant difference in IFNγ mRNA expression levels between Jag1+/+ and Jag1Ndr/Ndr *livers at P10 (please see the heatmap in response to comment no.3, above). *

Reviewer #3 (Significance (Required)): Strength: This article is well written and has important preliminary findings that could establish Jag1 and its downstream signaling pathways as potential therapeutic targets to attenuate liver fibrosis.

Author Response: Thank you for these comments and pointing out the wider implications of our findings.

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Reviewer 3____ Limitations: This study lacked the detailed molecular pathways which could explain how the Jag1 altered the T-cell recruitment, development and hepatocyte maturation in the development of liver fibrosis in the ALGS model.

Author Response: We agree that this study does not focus on molecular pathways. The intention of this study was to identify which cell populations contribute to atypical neonatal fibrosis in ALGS. Because we expected this process to be multifactorial, Jag1Ndr/Ndr mice, carrying a systemic mutation, present both advantages (Jag1 abrogation in all cells --> ALGS-like organ interactions) and limitations (inability to identify contributions of individual cell types). However, by identifying maturing hepatocytes and Tregs as dysregulated, and demonstrating that Jag1Ndr/Ndr lymphocytes behave abnormally and suppress inflammation and fibrosis in Rag1-/- mice (with normal Jag1 expression), we establish a biological framework that can now be further investigated with conditional genetic tools and in vitro systems, to elucidate specific molecular pathways, that were beyond the scope of the current study.