첫 번째 Render (Mount) → React가 초기화 함수 실행 → loadFromStorage() 호출 → 100을 초기 State로 저장

두 번째 Render (Update) → 기존 State 사용 → loadFromStorage() 실행하지 않음

세 번째 Render (Update) → 기존 State 사용 → loadFromStorage() 실행하지 않음

첫 번째 Render → loadFromStorage() 실행 → useState(100)

두 번째 Render → loadFromStorage() 또 실행 → useState(100)

세 번째 Render → loadFromStorage() 또 실행 → useState(100)

React는 초기값 100을 첫 Mount에서만 사용하지만, JavaScript는 렌더링할 때마다 loadFromStorage()를 실행해 버려. 즉, 불필요한 계산이 반복되는 문제가 있어.

이전 State를 바탕으로 새 State를 계산할 때는 setCount(c => c + 1)을 사용하면 오래된 렌더링의 count 값에 의존하지 않아.

그 렌더링에서 받은 count는 바뀌지 않는다

React는 Hook 호출 순서로 저장 칸을 찾기 때문에, 매번 Render할 때 Hook이 같은 순서로 실행되어야 한다.

function Counter() {

function handleClick() {

const [count, setCount] = useState(0); // ❌

}

return <button onClick={handleClick}>Click</button>; }

These lines make me wonder if I have been a little unfair to Eliot. Throughout the poem, I have read his constant allusions as almost a way of blocking the door for others to understand, as though I need to know every language and every literary reference before I am allowed to understand what he is saying. However, the Upanishad behind “Datta. Dayadhvam. Damyata” offers another way to understand his borrowing, because the gods, humans, and demons all ask, “Teach us,” admitting that they still have something to learn. Even the gods do not know everything. This makes me consider whether Eliot’s movement between different traditions represents a thirst for knowledge, a search for something that the broken world of the poem cannot provide on its own. The commands to give, be compassionate, and exercise self-control also turn knowledge into something you have to practise, rather than something you possess to prove how intelligent you are. His references still make the poem difficult to enter, but perhaps I have mistaken some of his searching for showing off. He may be collecting these voices because he needs their guidance too, and the poem’s physical thirst becomes, in this reading, a thirst for understanding.

优化器状态、梯度、模型参数 ZeRO-2：每个人手里拿着完整的图纸（参数），负责拼凑属于自己的那一小块拼图（梯度）。拼好自己这块后，把不需要的边角料扔掉。

ZERO-3 计算前借全部，算完立刻扔，下次用再借

线性层的W,b，训练完成后就固定。而激活值是前向传播的输出结果，反向传播计算完后就释放。

The words tears and sadness in this sentence shows the extent of sorrow and pain in her life. This sentence also shows the burden of bills and life that was put on her mother shoulders after the father left.

According to psychology when people are going through very difficult time, they don't believe it. This sentence exactly shows this situation. In order to protect her, her mind had helped her to not believe what hell she was living in after her father left. This sentence shows that heaviness of her difficult time.

This beginning sentence gives reader at the very beginning the feeling that her happy family has gone through difficult times. Since the first sentence says that her family is a happy one, the second sentence says that their heart is broken from somewhere. This give the reader a sense that the family went through difficult days and overcome the difficult days.

Upgraded thesis statement: The family passed for two phases, one before the parents got divorced and after it, but it both you can see they had happy moments. The second had more sacrifices to do, but in the end they had a good life.

This part show us the sacrifice the mom had to do to give her children a future.

This sentence shows one of the most difficult causes that made the hard moment the family had to live. Struggling with that must be really difficult, because they were by themselves and no one else.

I think data constraints are essential for maintaining data accuracy. For example, limiting ages to 0–120 prevents unrealistic entries and makes datasets more reliable for statistical analysis.

I find floating-point precision issues interesting because even simple decimal calculations can produce unexpected results in Python. This shows why we need to be careful when comparing floating-point values in data analysis.

In vitro fertilization

but might play practical tole with important legislators, strategies are probably not mutually exclusive

Tie their expertise to other orgs

did this happen in history

Pretty manipulable

We would expect to see even less expansion because there aren't many politically homogenous electorates with a unifying attribute as broad as "female"

means that they needed to coalition build across race and class, or at least class in reality

force signalling

in reality if active constituent is making ideological decisions then there is probably a partisan split

Threat to major parties although of course now they are made to address sooner, except if third parties are more focused on another issue

As opposed to the previous model, we introduce the powerful tool of accountability

Existing voters may be ideological, progressive husbands say

Meaning the threat doesn't need to be imminent

Also if the interest group will benefit from the expansion, allows organizations beyond politics to dictate when the franchise expands

activists don't have to ally with a whole group just with an interest group

Might expand franchise even of women will vote republican

Suffrage may have bipartisan allies

doesn't solve capacity super majority paradox

Which implies there is not competition

Question the horizon of the status quo

Women = liberal, suffragettes make them less liberal

Hard to pass, uncertain payoff, no electoral accountability

Within party or by movements

Win the electorate preferences while not dealing with shifting median voter

Suffragettes create referendum support

Suggests you need a more dominant or popular party that foresees a downfall

So the current electorate preferences matters more than proposed enfranchised

Electoral pressure

movement pressure

Basically its a long process and stars have to align

differing strategic interests

has to pass multiple times

Tandem between elite preferences and movements, goals need to align even if motivations are different

The suffrage movements provide this certainty

New electorate is likely to be diverse but trying to shore up existing and divided voters

Ok so democrats and republicans

But social upheaval may have similar effects

not seamless

You might even be generous with their intentions, but they need power to do it

Because they might want to still have some wealth, not revolution

Don't share

This might be true, fukuyama bent to it, but there are material hiccups

Should non-citizens be allowed to vote?

Still creation of undercalss

Moral judgement

attract residents

expansion to poor whites coupled with excluding free black, cementing an underclass

also a comparative work

status quo protection

Gonna make a similair argument to the other book

Competition and mobilization

?

Second equation should be presented as IA

%

How does the process of reconstruction work

I heard that AI is developing too fast, to the point where we must but restrivtions on it for our own good

Knowing how to use AI ethically also contributes to it's development

For example, water usage in data centers

Highlights how big mainstream media, as well as AI is on a fundamental level

"No country...should be left behind" "having fair access to AI technologies" illustrates that AI is a powerful tool that can make a country more powerful than those without access.

The annual interest in only one year?

%

%

Erroneous nesting of equation structures.

How is that the answer?? 2x+5y+8z is not equal to 1 for (1,1,1)

first time I've seen (x, y, z).

My row-echelon matrix looks a little different, which is worth mentioning, but still find that the answer is (1,-1,2). I don't understand why your R1 shows fractions when the original set for R1 should be 1, -2, 3, 9 according to the first equation. BTW, I've never seen any solution sets in here for x, y, and z expressed in parenthesis.

I learned this in high school. Why is it in preCalculus?

rows 2 and 3 in the shown row-echelon form has numbers other than zero

missing answer: "Erroneous nesting of equation structures"

This text often introduces information as if it was new even though we've been working on it. It's irritating.

I understand now why Embodied Cognition was something challenging to define in lecture. It clearly lacks a definite structure and defining constraints to give it a definition.

Physics and spring energy as a far-flung factor is funny.

Is this to state that the world exists? That the world exists when it is not being percieved? I don't understand the pregiven world idea. Is the Egg position to reject that the world exists outside of our perception?

I see VTR as carrying more of a philosophical argument than a scientific one, they seem to be debating what amount of the world exists beyond how we percieve it.

In this argument it is important to agknowledge that these features of animals ie. 270 degrees of vision or electromagnetic spectrum detection have most likely evolved due to their newly gained survivability. Animals develop tools like this in order to survive. So perception would likely vary between creatures with different roles and functions in the environment.

These are not contrasting because the way in which Gibson theorized our cognition was in the end computational. By using the word "resonates" without definition in regards to how we perceive stimulus then saying that resonating is most likely a computational process, he inadvertently aligned himself with a standard cognitive science approach to perception.

Wouldn't bodies, motions, and interactions fall into the umbrella of standard cog sci visual perception. As in, we use our bodies to perceive the world around us and the motion of our body and the motion of the eye can all be explained with the nervous system and visual processing system?

Exactly what I said earlier. Happy we are clearing up what that could possibly mean.

I do think this is a very interesting way of thinking about sensory processing, but I don't fully understand what resonate means in this context other than being synonymous with process. Why use the word resonate if not to bring a specific mental image to mind?

It would be interesting to see researchers break away from what feels like a forcefully imposed binary, what makes embodied cognition and cognitive science entirely alternate to each other? Is there not some combined understanding of Cognition that shows how cognition extends to the environment while still appreciating the symbolic representation system of standard cog sci?

I do not follow the conclusion that Standard Cog Sci is not interpreting the interactions between a cognizer and their environment. Part of the foundation of Standard Cog Sci is studying the interaction between subject and environment, so how is there a lack of an examination of the interaction between the two?

• The hidden function of social meals and pauses:

  • Shared meals and drinks are not just about food; they provide built-in pauses that relieve participants from the constant pressure of having something profound to say.
  • For introverts and direct communicators, recognizing that conversation requires deliberate social buffers helps demystify why casual interactions feel demanding.

• Conversational warm-up as an emotional prerequisite:

  • When the author’s college freshman daughter called to talk about a problem, she insisted on talking about trivial matters first before gradually segueing into the uncomfortable topic.
  • Jumping straight to core problems can be overwhelming; people frequently need a prolonged, low-stakes warm-up before they feel safe enough to open up.

• Vulnerability, imprecision, and trust-building:

  • Human language is inherently imprecise, and articulating sensitive emotions exposes speakers to social risk, misunderstanding, or judgment.
  • Cultural rituals like small talk, idle chit-chat, and coffee breaks serve as protective mechanisms designed to establish mutual trust and lower emotional defenses.

• Why text-first social platforms break down:

  • Character limits and rapid formats (such as early Twitter) strip away conversational padding, forcing users to state conclusions without warm-up or shared rituals.
  • The lack of physical, ritualized buffers in online environments magnifies conflict, as digital platforms have not yet evolved organic social safety valves.

Hacker News Discussion

• Small talk as a modem handshake:

  • Multiple commenters analogize small talk to two modems negotiating a connection: a formal, zero-surprise protocol to test line conditions, compatibility, and receptiveness before transmitting high-bandwidth payload data.
  • Skipping the handshake risks jarring the other party or derailing communication before a stable emotional connection is formed.

• Assessing emotional safety and values:

  • Commenters emphasize that idle chatter is an exercise in emotional maturity, allowing speakers to gently map out the other person's mood, worldview, and potential sensitivities.
  • Being trustworthy is distinct from being perceived as trustworthy; superficial conversations give both sides time to sniff each other out and establish baseline safety.

• Evolutionary and cross-cultural rituals:

  • Participants cite evolutionary theories (such as Robin Dunbar's grooming hypothesis), arguing that speech primarily evolved to establish social bonds rather than purely transmit abstract facts.
  • Several contributors note that across diverse global business and diplomatic traditions, elaborate rituals around meals and drinking remain essential precursors to actual negotiation.

• Limitations in anonymous online spaces:

  • A key counterpoint raised is that small talk relies on the prospect of an ongoing relationship; in transient, anonymous online feeds with no persistent community, roundabout discussion often just leads to distraction rather than trust.
  • Commenters also pointed out technical trivia regarding the blog's domain, noting the discontinuation of third-level .name registrations by Verisign and debating web archiving longevity.

improvements in social science

• Frontier-level utility at a fraction of the price:
  • DeepSeek 4.1 Flash performs on par with top-tier proprietary models (like Claude Opus) in day-to-day development, coding sessions, and planning.
  • Despite lagging slightly behind leading frontier labs, distilled open models now handle identical workloads for significantly lower costs.
• Paradigm shift toward low-cost, unattended workflows:
  • Ultra-low inference costs (e.g., ~$10/month subscription or fractions of a cent per run) eliminate hesitation around running exploratory scripts, messy automated tests, or repetitive chores.
  • Development patterns shift from rationed token use to continuous, high-volume execution, reserving top-tier frontier models only for occasional secondary code reviews.
• KV-cache architectural breakthrough:
  • A ~437x reduction in KV cache size compared to V1 dramatically cuts GPU VRAM usage during long context windows.
  • Memory and inference efficiency not only makes day-long sessions cost under $1, but also significantly reduces power and water consumption compared to traditional large models.
• Disruption of the frontier business model:
  • Western frontier labs face immense R&D costs that force high consumer pricing, whereas fast-following open models operate like generic pharmaceuticals offering comparable utility at a ~90% discount.
  • Self-hosting remains economically impractical compared to cheap cloud inference, though future local deployments will benefit from these cache innovations.

Hacker News Discussion

• Heavily subsidized flat-rate subscriptions buffer frontier models:
  • Many developers do not feel the urge to switch because flat-rate plans (e.g., Claude Code, Cursor) deliver hundreds of dollars in API-equivalent tokens for ~$20/month.
  • Commenters point out that until proprietary providers stop subsidizing end-user access or raise subscription prices, open-weight price advantages remain less urgent for individual subscribers.
• Debate over DeepSeek's pricing model and margins:
  • Some argue open models benefit from an accounting double standard where original training R&D is ignored, while others cite architectural data (e.g., 50x KV cache reduction) showing that DeepSeek can run profitably at extremely low prices without artificial subsidies.
  • Previous price adjustments by DeepSeek were attributed to infrastructure capacity bottlenecks rather than the rollback of subsidized pricing.
• Model performance and task suitability:
  • Users find 4.1 Flash comparable to smaller frontier tiers like Haiku or Luna, but observe that models like Opus 5.5 still retain an edge on nuanced world knowledge and complex edge cases.
  • Several developers use DeepSeek as a primary coding driver while keeping top-tier models on hand for high-level guidance.
• Risks of platform lock-in and varying token consumption:
  • Commenters warn against over-reliance on centralized labs that control hidden system prompts, thinking tokens, and pricing tiers.
  • Discussions highlighted vast divides in consumption habits, noting that unoptimized workflows and brute-force context stuffing burn through token limits far faster than structured engineering workflows.

NYT Sep 27th 2026

Living in the Portland and Vancouver area, I estimated that a family of four would spend around $6,450 a month on housing, food, transportation, childcare, and other basic needs. That's about $77,400 a year, which is $55,187 more than the poverty line of $22,213 in 2010. It really shows how expensive it is for families just to get by.

The problem with how poverty is measured is that the system is outdated. It was created in the 1960s and mainly focused on food costs. Today, families have expenses like housing, healthcare, and transportation. Just because someone makes more than the poverty line doesn't mean they're financially stable.

Typo

Imagine throwing five darts at a dartboard: all five can land in a tight cluster (consistent), yet still be far from the bullseye (inaccurate).

I think Creon is saying that the people of Thebes deserve a king who can move forward instead of being connected to past problems and conflicts. When he says, “My name, thank God, is only Creon,” he emphasizes that he wants to be known for who he is, rather than for his family history.

I think this means that the person is expected to give a specific excuse for staying home. By saying they are sick and have not been out since yesterday, they make it sound like they need to rest and cannot go anywhere.

The passage shows how quickly the speaker puts the tobacco in their cheek, making the action seem ordinary and effortless.

As stated above, there are still gender stereotypes in job positions. Due to women being associated with communal traits in general, society believes they're best suited for nurturing roles despite individual traits. After all, those are nurturing jobs. Though it may seem positive that women are associated with warmth and care, that does not at all represent women as a whole. It is factual that women can be more agentic than nurturing, while men can be more nurturing than agentic. But because these descriptive stereotypes are generally recognized, people tend to believe anyway that women suit more communal jobs.

This section is an example of going against the western prescriptive stereotypes, in which women 'should' be the caretakers, whilst men 'should' be the providers. However in modern day, these stereotypes are falling out of fashion as decades of feminist protest and movements resulted in women being able to have more equal opportunities as men. Though it is still a stereotype recognized through most Americans that women are 'more nurturing', we at the same time have begun to normalize their positions in higher fields unlike in the past where it seemed 'pointless' and 'risky'.

너의 now State를 새 시간으로 업데이트하고 화면을 다시 계산해 줘. -> react에게 요청(rendering)

toLocaleTimeString()은 Date 객체를 시간 문자열로 바꾸는 메서드야.

new Date()는 현재 날짜와 시간을 나타내는 Date 객체를 만들어.

this.timerId = 5; // ✅ 일반 인스턴스 속성 변경 this.state.now = new Date(); // ❌ State 직접 수정 this.setState({ now: new Date() }); // ✅ State 업데이트

핵심은 this.를 붙였다고 전부 State가 되는 건 아니라는 거야!

• this.timerId → Clock 객체가 직접 가진 속성
• this.state → Clock 객체가 가진 State 객체
• this.state.now → State 객체 안에 있는 now 속성

한 줄 암기: state에 없어도 this.속성이름 = 값으로 인스턴스 속성을 새로 만들 수 있다. 단, React 화면에 영향을 주는 State는 setState()로 변경해야 한다.

document.title은 브라우저 탭에 표시되는 제목

setInterval()은 정해진 시간 간격으로 함수를 반복 실행하는 JavaScript 함수

setInterval(실행할함수, 시간간격);

단계 주요 흐름 Render Parent → Child Snapshot Child → Parent DOM Update 실제 변경 반영 DidUpdate Child → Parent

Mount = 내려갔다가 다시 올라오기! Parent render ↓ Child render ↓ Child mounted ↑ Parent mounted

Unmounting — 제거될 때

componentWillUnmount()

DOM Remove

Updating — State / Props가 변경될 때

shouldComponentUpdate()

render()

getSnapshotBeforeUpdate()

DOM Update

componentDidUpdate()

Mounting — 처음 생성될 때

constructor()

render()

DOM Update

componentDidMount()

React가 특정 Lifecycle 시점에 호출하는 메서드

메서드 역할 render() 현재 Props/State로 UI 계산

componentDidMount() 처음 DOM에 추가된 후 작업

componentDidUpdate() 업데이트가 반영된 후 작업

componentWillUnmount() DOM에서 제거되기 전 정리

Lifecycle = 컴포넌트가 생성 → 업데이트 → 제거되는 과정

StudentRow에서 Jane 클릭

onSelect({selectedId}) 호출

App.setState({{selectedId}}) 요청

React가 App.render() 재실행

Roster와 StudentRow 렌더링

selected 결과에 따라 > 표시 변경

<section>은 <h2>와 <ul>을 하나의 Root Element로 묶기 위해 사용한 거야.

반드시 <section>일 필요는 없으며 <div>나 Fragment로 대체할 수도 있어.

destructing -> 굳이 this.props.속성 이렇게 사용 안 해도 됨

const title = this.props.title;

const students = this.props.students;

const selectedId = this.props.selectedId;

const onSelect = this.props.onSelect;

함수 자체를 전달하는 거야

는 JavaScript 변수/속성의 값을 읽어 전달하니까 {}가 필요해.

여기서 {}는 JavaScript 객체를 만드는 중괄호야.

시험에서 가장 헷갈리는 중괄호 정리 코드 {} 의미 state = { … } State 객체 생성

{ id: 1, name: 'Joe' } 학생 객체 생성

(id) => { … } 함수 본문

this.setState({ selectedId: id }) 변경할 state 속성을 객체로 전달

students={this.state.students} JSX에서 JavaScript 값 사용

onSelect={this.handleSelectStudent} JSX에서 함수 객체 전달

function Greeting({ name })은 function Greeting(props)에서 props.name을 일일이 쓰지 않도록 name을 바로 꺼내는 축약 문법이야

React는 이 데이터를 Props로 전달해. props = { name: "Joe" }

비교 Class-based Functional

정의 방식 class Greeting extends Component function Greeting(props)

React 상속 필요 필요 없음

JSX 반환 render() 안에서 함수에서 직접

Props 접근 this.props.name props.name 사용 방법 <Greeting name="Joe" /> <Greeting name="Joe" />

여기서 jsx()는 React의 JSX 변환 과정에서 사용하는 함수야. 쉽게 다음 형태로 이해하면 돼.

jsx(태그이름, { 속성들, children: 자식내용 });

```function App() {

return (

<>
  <h1>Hello</h1>
  <p>Welcome</p>
</>

); }```

일반 JavaScript에서는 평소처럼 코드를 쓰고, JSX 내부에서 JavaScript 값을 끼워 넣을 때만 {}를 사용하면 돼!

코드 {} 의미 function App() { ... } 함수 본문을 감싸는 중괄호

<h1>{name}</h1> JSX 안에서 JS 표현식 사용

onClick={() => alert("Hi")} JSX 속성에 JS 함수 전달

틀린 코드 — 최상위 요소가 2개

function App() {

return (

<h1>Hello</h1>

<p>Welcome</p> ); }

JSX는 개발자가 편하게 작성하는 문법이고, Build Tool은 이를 브라우저에서 실행할 수 있는 JavaScript로 번역해 주는 거야.

Islam absorbs the "westernizing" idea almost ironically by quoting American facts against the Western ideologies.

This is mechanized by bourgeois culture, and though not the same, it bears a similarity to the communist revolution. As opposed to communism, muslim communities resorted to extremism, which gives muslims the reputation they have today.

This is similar to the Anderson reading. Many different cultures are blended and accounted for within education to make room for religious diversity, though they are often met with reproach. Hyphenating cultures is necessary for them to coexist.

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Lo importante si es que vota diferente, sino que tipo de agenda se instala ¿Qué secede con ello?

hola

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This is a test citation

Revisar la discretización de la ec. 3.5, compatible con las unidades de extracción

Remove space before line for screen-reader friendly experience

I highly recommend evaluating the current use of "equity" and "diversity" in the training material. The January 2025 DEIA -related Executive Orders prohibited agencies from conducting training on DEIA. You could instead discuss the advantages of including researchers with varying areas of expertise and experience as an aspect of "collaboration". You could also discuss making work easily accessed by everyone by incorporating open sharing practices.

看不懂

error ortográfico

week41 kah end 10.3 till 10.9

finantial institutions where dreaming for these to happen

Tangent brutal death of haitian president guillaume sam

https://en.wikipedia.org/wiki/Phthia

Mentioned in Plato's Crito

Achilles mentions arriving on the third day "in fertile Phthia" as does Socrates in Plato's Crito.

Relationship with Jesus' resurrection on the third day?

https://www.reddit.com/r/typewriters/comments/1o8vjc5/typewriter_type_action_chart_my_personal/

u/N0nob compares the weight versus feel of their typewriter collection:

Cross reference with https://boffosocko.com/2026/09/17/what-words-could-we-use-to-better-describe-the-soul-of-a-typewriter/

-gast (female dog, bitch) as a pejorative in Welsh

Arbo gast as "greedy bitch"

https://www.reddit.com/r/learnwelsh/comments/1wygupa/pejorative_terms_that_contain_gi/

비교 Class-based Functional

정의 방식 class Greeting extends Component function Greeting(props)

React 상속 필요 필요 없음

JSX 반환 render() 안에서 함수에서 직접

Props 접근 this.props.name props.name 사용 방법 <Greeting name="Joe" /> <Greeting name="Joe" />

틀린 코드 — 최상위 요소가 2개

function App() {

return (

```<h1>Hello</h1>```

```<p>Welcome</p>```

); }

값의 변경

console.log(this.state.selectedId); // null

this.setState({ selectedId: 2 });

console.log(this.state.selectedId); // ?

강의자료의 설명에 따르면 두 번째 console.log()도 null이야.

왜냐하면 setState()는 변경을 즉시 완료하는 게 아니라, React에 State 업데이트를 예약(schedule)하기 때문이야.

실행 흐름 (슬라이드 기준) 1. 현재 selectedId = null 2. setState({ selectedId: 2 }) 요청 3. 바로 다음 줄에서는 여전히 이전 값 null을 읽음 4. React가 State 변경을 처리 5. 새 State를 바탕으로 다시 Render

JavaScript 객체의 값은 바뀔 수 있지만, React는 이 대입만으로 화면을 다시 렌더링해야 한다는 사실을 알지 못해.

this.state.selectedId = 2;

class App extends Component {

componentDidMount() {

// 컴포넌트가 DOM에 추가된 후 실행
// Data fetch, timer 시작 등

}

render() {

// 화면에 보여줄 JSX 계산
```return <h1>Hello!</h1>;```

} }

반복 렌더링 유발 가능

React의 DOM 관리와 충돌할 수 있음

서버 요청이 반복될 수 있음

렌더마다 타이머 중복 생성 가능

setState() 반복 렌더링 유발 가능

StudentRow에서 Jane 클릭

onSelect({selectedId}) 호출

App.setState({{selectedId}}) 요청

React가 App.render() 재실행

Roster와 StudentRow 렌더링

selected 결과에 따라 > 표시 변경

<section>은 <h2>와 <ul>을 하나의 Root Element로 묶기 위해 사용한 거야.

반드시 <section>일 필요는 없으며 <div>나 Fragment로 대체할 수도 있어.

destructing -> 굳이 this.props.속성 이렇게 사용 안 해도 됨

const title = this.props.title;

const students = this.props.students;

const selectedId = this.props.selectedId;

const onSelect = this.props.onSelect;

function Greeting({ name })은 function Greeting(props)에서 props.name을 일일이 쓰지 않도록 name을 바로 꺼내는 축약 문법이야

https://www.facebook.com/groups/TypewriterCollectors/posts/10156067793359678/

David Born helped Richard Polt to update the list of typewriter repair shops. domarb@fastmail.cn

https://www.legacy.com/us/obituaries/toledoblade/name/donald-nichols-obituary?id=60525872

https://www.youtube.com/@donnichols2589/videos

Ok so clearly some tasks need models to act fast. However, im curious why we dont just have a super cheap subagent or some filtering method figure out if this is a time sensitive task or a reasoning task and delegate. For example a mother agent should be able to identify a time strict task and open a fast subagent to wait / execute it. I suppose these arent eval based considerations but certainly an implication to keep in mind for realworld harness deisng and IMO should be more clearly stated

https://www.theguardian.com/world/2026/mar/14/jurgen-habermas-german-philosopher-and-sociologist-dies-aged-96

self-assessment tax return

https://www.ebay.com/usr/1532xplane <br /> Darwin Raymond, Raymond's Office Machines, 927 Grand Ave., Glenwood Springs, CO 81601. (970) 945-5872.

eLife assessment

This study provides a useful inventory of genes that are up- or down-regulated during the early metamorphic development of male and female larvae and proposes that the microRNA cluster miR-277/34 is involved in the development of sexual differences during early metamorphosis of Drosophila melanogaster, although its precise role remains unclear. The strength of evidence, based on a combination of diverse methods including mRNA and small RNA sequencing, in silico analyses, in vitro assays, and loss-of-function experiments, is incomplete as it lacks a general model and an examination of the potential effects of the miR-277/34 mutations on phenotypes such as morphology or developmental time. This work will be of interest to developmental biologists interested in sexual dimorphism and in the interplay between hormones and microRNAs during development.

Reviewer #1 (Public Review):

Summary:

In this paper, Li and colleagues have found mircoRNAs that affect levels of metamorphosis-regulating genes that can also affect levels of sesquiterpenoids (juvenile hormone and related compounds) and ecdysteriods, which regulate the timing and stages of insects, respectively. They first compared the transcriptomes of Drosophila at the third larval instar and at the white pre-pupa stage. They found thousands of differences in gene transcript levels between males and females, and between the two different stages. Among those genes that were differentially regulated they saw that genes involved in insect hormone biosynthesis were disproportionately represented. Many of the differentially regulated genes were involved in the insect hormone biosynthesis pathway and ascorbate and alderete metabolism. MicroRNAs were also differentially expressed during metamorphosis and were separately identified. The authors then considered genes and whether the differentially expressed microRNAs might regulate transcripts known to be involved in sesquiterpenoid production. In silico analysis of microRNAs predicted a list of 17 microRNAs that can regulate transcripts of sesquiterpenoid biosynthesis genes. The authors then used an in vitro luciferase assay to validate the binding and downregulation of 10 of the microRNAs to genes involved with sesquiterpenoid production in S2 cells.

Li and colleagues then focus on two genes they found were bound by microRNAs that have established roles in metamorphosis. The microRNAs miR-34 and miR-277 bind transcripts of two protein-coding genes that regulate metamorphosis Kr-h1, which encodes a transcription factor that is a JH-inducible transcription factor, and Allatostatin C Receptor 1, (AstC-R1), a G-protein coupled receptor that regulates the corpora allatum, the gland that produces sesquiterpenoids. Using a LAMP assay, one of the microRNAs, miR-277 was shown to bind to both AstC-R1 and Kr-h1 in in vivo whole-animal extracts. There is no mention of binding between either protein-coding transcript and the miR-34 microRNA. Temporal expression of all four transcripts shows that their abundance is anti-correlated; stages of high miR-34 or miR-277 expression correlate with low AstC-R1 or Kr-h1 expression. Homozygous deletions of both mircroRNAs result in 23% lethality, five days after adult eclosion. The authors also generated specific mutants in miR-34 or miR-277 and find differences in the expression of AstC-R1 and Kr-h1 and sex-specific differences in both sesquiterpenoids and ecdysteroids in the knock-out lines. If there were phenotypes associated with the specific knock-outs, those were not mentioned. Next, the authors examined the transcriptomes of the miR-3277 and miR-34 mutants and found several other GO-terms enriched among the differentially expressed genes. However, the sesquiterpenoid pathway and ascorbate and alderete metabolism are not listed.

Strengths:

This is an interesting manuscript that could make an important contribution to our understanding of the roles of micro RNAs at metamorphosis, and potentially of how sex-specific differences arise during metamorphosis. Strengths of the paper include the functional validation of microRNA binding, in vitro and in vivo-, as well as the characterization of sesquiterpenoid and ecdysteroid titers. The authors have also used CRISPR to generate specific knock-outs of miR-34 and miR-277. The transcriptomes will be a resource for future work to mine for differences in gene expression during metamorphosis.

Weaknesses:

(1) Spatial Expression of miR-34 and miR-277. If miR-34 and miR-277 regulate AstC-R1 and Kr-h1, then they must be expressed in the same cells. Although the authors show that the microRNAs do bind to the transcripts of AstC-R1 and Kr-h1 in S2 cells, and miR-277 binds AstC-R1 and Kr-h1 in vivo whole-animal homogenates, we do not know if the microRNAs are ever in the cells where AstC-R1 or Kr-h1 are expressed. AstC-R1 is only expressed in a few cells in the brain, so it is not at all certain that it is co-expressed with either microRNA. The creation of enhancer lines or in situ hybridization in Drosophila is straightforward and would sort this out.

(2) Phenotypes. Although a double deletion was used and specific knock-outs of both miR-34 and miR-277 were generated, the analysis of the mutants is very superficial. For the homozygous deletion of both microRNAs miR-34 and miR-277, only a decrease in survivorship was observed a full six days after adult eclosion - after the end of metamorphosis. No phenotype for either miR-34KO or miR-277-KO was given. The authors cite the work of others who have found specific phenotypes after manipulation of sesquiterpenoids or ecdysteroids, like Riddiford and Ashburner, but do not use any of these many studies to help them characterize the phenotype. If the loss of miR-34 and miR-277 affects so many pathways (including MAPK signaling, TGF-beta signaling, FoxO signaling, and Wnt signaling), as well as global titers of metamorphic hormones, then there shouldn't there be something different in the development to discuss?

(3) I think the reliance on GO term enrichment is getting in the way of biology. For instance, I would not describe Kr-h1 as a sesquiterpenoid biosynthesis pathway gene. Yet the authors say they were motivated to examine microRNA regulation of Kr-h1 because they saw differences in levels of the sesquiterpenoid biosynthesis pathway between WL3 and WPP, a period which also saw differences in expression of some microRNAs. I understand that Kr-h1 expression is regulated by JH, a sesquiterpenoid, but it is not directly involved with JH production, so relying on GO term enrichment has made the decision to focus on Kr-h1 feel arbitrary.

(4) The transcriptomes of miR-34 and miR-277 should have revealed genes encoding members of the sesquiterpenoid biosynthesis pathway as well as AstC-R1 and Kr-h1, but neither was mentioned. The functional tests of miR-34 and miR-277 were performed because they were shown to affect the levels of expression of genes in the sesquiterpenoid biosynthesis pathway. Figure 2 shows a significant decrease in AstC-R1 and Kr-h1 transcripts after the loss of miR-34 and miR-277. However, the results do not mention either (Lines 250-264). Instead, there is a list of 10 different GO terms (like arginine and proline metabolism or fatty acid degradation) that were enriched in miR-34 and miR-277 transcriptomes. If any of those ten types have any relationship to Kr-h1, AstC-R1, or metamorphosis, that has not been explained.

(5) Not enough care was taken in describing the stages. The methods describe wandering larvae (WL3) and white pre-pupa (WPP) for the transcriptomes, but in the text, different terms are used, like "larva", "pupa" and "L3 larvae instars" "early pupae" "late L3". Also, it seems like the small RNA libraries for sequencing were taken from "L3 larvae", but the stage of the L3 larvae was not mentioned. Staging is important, especially during metamorphosis, since differences in expression are expected to exist between different stages of L3, between early vs late wandering, and between WPP and early pupal stages.

Reviewer #2 (Public Review):

Summary:

This study proposes that the microRNA cluster miR-277/34 controls the generation of sexual dimorphism in Drosophila melanogaster during metamorphosis by acting on specific hormonal and developmental gene pathways.

Strengths:

Using a combination of mRNA and small RNA sequencing together with genome-wide in silico and in vitro analyses the authors identified a microRNA cluster that may be involved in metamorphosis and the generation of sexual dimorphism in Drosophila melanogaster.

Weaknesses:

Biological validation of the identified sexually dimorphic genes and a detailed understanding of how the microRNA cluster miR-277/34 might be involved in the regulation of sesquiterpenoids are needed.

Major suggestions:

(1) If AstC-R1 and Kr-h1 are targets of the miR-277/34 cluster and cause their downregulation, it is not clear why there would also be a decrease in the levels of these genes in the miR-277/34 mutants. This would suggest that the mechanism is not straightforward and that further epistatic experiments should be carried out in order to clarify this issue.

(2) The changes in the expression levels of AstC-R1 in pupae of miR-277-KO and mir-34-KO flies must be accompanied by photos of the respective larvae and pupae, as well as an analysis of the larvae-pupa transition on the mutants by gender.

(3) Biological validation of the identified sexually dimorphic genes in vivo will be necessary for the support of this work.

Reviewer #3 (Public Review):

Summary:

The authors show convincingly the complexity of gene up- and down-regulation at the outset of metamorphosis and identify substantial differences between the two sexes, even at this early time in development. The complexity of microRNA expression and the difference between the sexes are also nicely laid out. The functional significance of these differences, though, is harder to establish. The authors have focused on the roles of two families of developmental hormones, the ecdysteroids, and the juvenile hormones. The emergence of sex-specific differentiation of organs during metamorphosis is clearly downstream of the action of ecdysteroids and/or JH, but there is no evidence that the presence or lack of these hormones has any effect on the sexual identity of organ systems - i.e., that manipulations of JH or ecdysteroid result in either the masculinization or feminization of individuals or their organs. The precedence for the linkage of these hormones to sex determination is the 2002, Belgacem & Martin study, which describes the effects of JH on fly locomotion. These authors show that the number of stop/start bouts is sexually dimorphic, and removal of JH in males shifts their frequency into the female range while giving treated males exogenous JH moves it back. While this is referred to as a "feminization" of male behavior, this quantitative shift in frequency is not as compelling as would be a qualitative shift -- for example, the removal of JH causing males to show egg-laying behavior (a result that has never been seen). Also, these effects are in a fully mature system, rather than at the early metamorphic time examined in the present paper. In driving and coordination metamorphosis, JH and ecdysteroids are intimately involved in sexual differentiation, but I know of no compelling evidence that they play a role in sex determination.

While the summary of the effects or removal of specific microRNAs on the components of the biosynthetic pathway for the JHs and ecdysteroids (Figure 2E ,F) is quite compelling, I am concerned about the effects of the removal of mir-277 and mir-34 on the levels of both the JHs and 20E. My concern centers around the data from the control group (w[1118] animals in Figure 2D). These data are the first report of a marked sex difference in the titer of either JH or ecdysteroid at the start of metamorphosis in Drosophila. As expected, males show a 10-20 increase in levels of JH III, JHB3, and 20E between the L3 stage and the white puparium, but, surprisingly, the levels of these hormones in female L3 larvae are equal to or greater than that seen at pupariation! These data for females run counter to over 50 years of work on the effects of ecdysteroids in Drosophila!

As far as I can gather from the paper, the L3 data were obtained using wandering larvae. This stage lasts for about 12 hours and ends with pupariation. Larvae from this period need to be used with caution for hormone studies. Levels of both JH and ecdysteroid are low as larvae leave the food but rapidly rise to their peak levels at the white puparium stage 12 hours later. To deal with the rapidly changing hormonal landscape through this period, the researchers have used physiological markers to track this progression. Initially, it was the sage of "puffing" of the giant salivary gland chromosomes, but, for bulk collection of staged larvae, larvae are fed on food containing a blue dye, and progression is tracked by the loss of blue coloring from the gut. I could not find if the authors had any criteria for selecting larvae during the wandering period. Male and female larvae grow to different sizes. Might this difference in growth be biased when larvae were selected during their wandering phase?

The other hormone-related issue is the expression of Kr-h1 during larval stages and metamorphosis (Figure 1G). Kr-h1 is the main target of JH and Kr-h1 expression is often used as a proxy for the JH titer. The authors report that peak Kr-h1 expression occurs in the L3 (when the JH titer should be lowest!) and that it drops at wandering. This pattern is counter to that reported in the literature (e.g., FlyBase, ModEncode).

Author response:

[Editors’ note: An Expression of Concern has been published for this article. The authors’ responses submitted during revision have been appended for further context]

eLife Assessment

This study provides a useful inventory of genes that are up- or down-regulated during the early metamorphic development of male and female larvae and proposes that the microRNA cluster miR-277/34 is involved in the development of sexual differences during early metamorphosis of Drosophila melanogaster, although its precise role remains unclear. The strength of evidence, based on a combination of diverse methods including mRNA and small RNA sequencing, in silico analyses, in vitro assays, and loss-of-function experiments, is incomplete as it lacks a general model and an examination of the potential effects of the miR-277/34 mutations on phenotypes such as morphology or developmental time. This work will be of interest to developmental biologists interested in sexual dimorphism and in the interplay between hormones and microRNAs during development.

We thank the editor and reviewers for the positive and constructive comments and suggestions. Point-to-point responses are listed below.

Reviewer #1:

Summary:

In this paper, Li and colleagues have found mircoRNAs that affect levels of metamorphosis-regulating genes that can also affect levels of sesquiterpenoids (juvenile hormone and related compounds) and ecdysteriods, which regulate the timing and stages of insects, respectively. They first compared the transcriptomes of Drosophila at the third larval instar and at the white pre-pupa stage. They found thousands of differences in gene transcript levels between males and females, and between the two different stages. Among those genes that were differentially regulated they saw that genes involved in insect hormone biosynthesis were disproportionately represented. Many of the differentially regulated genes were involved in the insect hormone biosynthesis pathway and ascorbate and alderete metabolism. MicroRNAs were also differentially expressed during metamorphosis and were separately identified. The authors then considered genes and whether the differentially expressed microRNAs might regulate transcripts known to be involved in sesquiterpenoid production. In silico analysis of microRNAs predicted a list of 17 microRNAs that can regulate transcripts of sesquiterpenoid biosynthesis genes. The authors then used an in vitro luciferase assay to validate the binding and downregulation of 10 of the microRNAs to genes involved with sesquiterpenoid production in S2 cells.

Li and colleagues then focus on two genes they found were bound by microRNAs that have established roles in metamorphosis. The microRNAs miR-34 and miR-277 bind transcripts of two protein-coding genes that regulate metamorphosis Kr-h1, which encodes a transcription factor that is a JH-inducible transcription factor, and Allatostatin C Receptor 1, (AstC-R1), a G-protein coupled receptor that regulates the corpora allatum, the gland that produces sesquiterpenoids. Using a LAMP assay, one of the microRNAs, miR-277 was shown to bind to both AstC-R1 and Kr-h1 in in vivo whole-animal extracts. There is no mention of binding between either protein-coding transcript and the miR-34 microRNA. Temporal expression of all four transcripts shows that their abundance is anti-correlated; stages of high miR-34 or miR-277 expression correlate with low AstC-R1 or Kr-h1 expression. Homozygous deletions of both mircroRNAs result in 23% lethality, five days after adult eclosion. The authors also generated specific mutants in miR-34 or miR-277 and find differences in the expression of AstC-R1 and Kr-h1 and sex-specific differences in both sesquiterpenoids and ecdysteroids in the knock-out lines. If there were phenotypes associated with the specific knock-outs, those were not mentioned. Next, the authors examined the transcriptomes of the miR-3277 and miR-34 mutants and found several other GO-terms enriched among the differentially expressed genes. However, the sesquiterpenoid pathway and ascorbate and alderete metabolism are not listed.

Strengths:

This is an interesting manuscript that could make an important contribution to our understanding of the roles of micro RNAs at metamorphosis, and potentially of how sex-specific differences arise during metamorphosis. Strengths of the paper include the functional validation of microRNA binding, in vitro and in vivo-, as well as the characterization of sesquiterpenoid and ecdysteroid titers. The authors have also used CRISPR to generate specific knock-outs of miR-34 and miR-277. The transcriptomes will be a resource for future work to mine for differences in gene expression during metamorphosis.

Weaknesses:

(1) Spatial Expression of miR-34 and miR-277. If miR-34 and miR-277 regulate AstC-R1 and Kr-h1, then they must be expressed in the same cells. Although the authors show that the microRNAs do bind to the transcripts of AstC-R1 and Kr-h1 in S2 cells, and miR-277 binds AstC-R1 and Kr-h1 in vivo whole-animal homogenates, we do not know if the microRNAs are ever in the cells where AstC-R1 or Kr-h1 are expressed. AstC-R1 is only expressed in a few cells in the brain, so it is not at all certain that it is co-expressed with either microRNA. The creation of enhancer lines or in situ hybridization in Drosophila is straightforward and would sort this out.

We thank the reviewer for the positive and constructive comments. We agree with the reviewer, and have pointed out the limitation in the main text as follows:

Lines 307-309 - “Further exploration of the spatial expression between miR-277, miR-34, Kr-h1 and AstC could bring new insights to the regulatory effects they have during metamorphosis”.

(2) Phenotypes. Although a double deletion was used and specific knock-outs of both miR-34 and miR-277 were generated, the analysis of the mutants is very superficial. For the homozygous deletion of both microRNAs miR-34 and miR-277, only a decrease in survivorship was observed a full six days after adult eclosion - after the end of metamorphosis. No phenotype for either miR-34KO or miR-277-KO was given. The authors cite the work of others who have found specific phenotypes after manipulation of sesquiterpenoids or ecdysteroids, like Riddiford and Ashburner, but do not use any of these many studies to help them characterize the phenotype. If the loss of miR-34 and miR-277 affects so many pathways (including MAPK signaling, TGF-beta signaling, FoxO signaling, and Wnt signaling), as well as global titers of metamorphic hormones, then there shouldn't there be something different in the development to discuss?

We thank for reviewer for pointing out the phenotypes. We did not detect morphological or developmental abnormalities during larval-pupal transitions in the different knockout mutants, and have now included the previously established research on miR-34 in the main text, shown as follows:

Lines 305-307 - “MiR-34 has been documented to maintain the neural proteostasis while suppressing age-associated decline (Liu et al. 2012; Kennerdell, Liu and Bonini 2018; Srinivasan, Tran and Bonini 2022).”

(3) I think the reliance on GO term enrichment is getting in the way of biology. For instance, I would not describe Kr-h1 as a sesquiterpenoid biosynthesis pathway gene. Yet the authors say they were motivated to examine microRNA regulation of Kr-h1 because they saw differences in levels of the sesquiterpenoid biosynthesis pathway between WL3 and WPP, a period which also saw differences in expression of some microRNAs. I understand that Kr-h1 expression is regulated by JH, a sesquiterpenoid, but it is not directly involved with JH production, so relying on GO term enrichment has made the decision to focus on Kr-h1 feel arbitrary.

We have stated out the rationale and rephrased that Kr-h1 as sesquiterpenoid responsive gene in relevant sentences in the Results, shown as follows:

Lines 135-145 - “Utilising an established cell culture dual-luciferase reporter assay (Hui et al 2013); we have validated 7 microRNAs (miR-8, miR-277, miR-958, miR-1000, miR-1002, miR-2283, miR-2280) that can downregulate the expression of Drosophila genes involved in JH synthesis, responsive, or signalling genes to varying degrees, including allatostatin C (AstC), allatostatin C receptor 1 (AstC-R1), allatostatin C receptor 2 (AstC-R2), Krüppel homolog 1 (Kr-h1), Chd64, juvenile hormone binding protein (JHBP), juvenile hormone diol kinase (JHDK) and juvenile hormone esterase (JHE) (Figure 1C). Among these genes, Kr-h1 or Kruppel homolog 1, is a JH-dependent transcription factor (responsive gene), and have been found to be regulated by another microRNA in cockroaches (Lozano et al 2015); while AstC-R1 or allatostatin C receptor 1, is a G-protein coupled receptor for neuropeptide allatostatin C which regulates juvenile hormone synthesis (Stay and Tobe 2007).”

(4) The transcriptomes of miR-34 and miR-277 should have revealed genes encoding members of the sesquiterpenoid biosynthesis pathway as well as AstC-R1 and Kr-h1, but neither was mentioned. The functional tests of miR-34 and miR-277 were performed because they were shown to affect the levels of expression of genes in the sesquiterpenoid biosynthesis pathway. Figure 2 shows a significant decrease in AstC-R1 and Kr-h1 transcripts after the loss of miR-34 and miR-277. However, the results do not mention either (Lines 250-264). Instead, there is a list of 10 different GO terms (like arginine and proline metabolism or fatty acid degradation) that were enriched in miR-34 and miR-277 transcriptomes. If any of those ten types have any relationship to Kr-h1, AstC-R1, or metamorphosis, that has not been explained.

We have now explicitly stated that Kr-h1 is significantly downregulated in the miR-277-KO transcriptomes consistent with our qPCR validation, as well as the transcriptomic analyses were used to understand genome-wide changes, shown as follows:

Lines 211-217 - “Among these differentially regulated genes, Kr-h1 was significantly downregulated in the miR-277-KO mutants (Supplementary Table S2), consistent with our qPCR validations (Figure 2B). On the other hand, AstC-R1 was not classified as differentially regulated based on the in silico analyses threshold of the whole-body RNA-seq data, and whether this was due to its restricted expression in a small subset of brain neurons would require further experiments to differentiate between different scenarios.”

Lines 255-259 - “It implies that miR-277 could have conserved regulatory roles of sesquiterpenoid and related pathways, and also divergent roles between sexes during metamorphosis with signaling pathways associated with cell proliferation, wing development, nutritional support, and endocrine regulation (Pan et al 2018; Jiang and Edgar 2009; Gibbens et al 2011; Toyama et al 2014).”

(5) Not enough care was taken in describing the stages. The methods describe wandering larvae (WL3) and white pre-pupa (WPP) for the transcriptomes, but in the text, different terms are used, like "larva", "pupa" and "L3 larvae instars" "early pupae" "late L3". Also, it seems like the small RNA libraries for sequencing were taken from "L3 larvae", but the stage of the L3 larvae was not mentioned. Staging is important, especially during metamorphosis, since differences in expression are expected to exist between different stages of L3, between early vs late wandering, and between WPP and early pupal stages.

We thank the reviewer for highlighting the importance of staging clarity, and have now revised both the Results and Methods to ensure all stages are defined explicitly and used consistently throughout the manuscript. In our dataset, all references to “L3 larvae” or “larva” correspond specifically to wandering third-instar larvae (WL3) which that have exited the food, display 1/3 to 1/4 gut contents, and show protruding anterior spiracles. All references to “pupa”, “pre-pupa”, or “early pupae” refer to white prepupae (WPP), defined as immobile individuals with fully everted spiracles and no movement when gently probed.

Recommendations:

(1) Lines 171-173. The genotype of the deletion line is given, but what is the genotype of the flies that were used? Were the balancers and chromosomes with mutations included in the analysis? I cannot tell from the description.

The miR-277-34-KO line was used and significant decrease in survivorship was observed in double knockout line. The information is now included as follows:

Lines 158-164 - “In the homozygous loss of function of miR-277/34 cluster fly (w[*]; Df(3R)mir-277-34-KO, TI{w[+mW.hs]=TI}mir-277-34-KO/TM3, P{w[+mC]=GAL4-twi.G}2.3, P{UAS-2xEGFP}AH2.3, Sb[1] Ser[1]; Chen et al 2014), we found that the heterozygous knockout individuals displayed low to almost no lethality, while the survivorship for the double knockout flies fell significantly with 77% of the double knockout flies dying within five days after eclosion (Figure 2A).”

(2) The introduction and discussion are too broad. I would expect some information about what is happening to ecdysone and juvenile hormone titers during metamorphosis. Additional detail should be given to the regulation of the corpora allatum, since the Ast-C1 gene and sesquiterpenoid levels were a major part of the results. I had hoped the discussion would help to synthesize the details within the results, particularly the differences between sexes. Instead, the discussion is very general, citing other papers that are related, but without tying the results within those papers to specific results in this paper.

We thank the reviewer. The following information is now included in the main text:

Lines 47-53 - “Multiple ecdysone titre peaks were observed during Drosophila development with each pulse corresponds to a developmental transition (Kannangara et al., 2023; Scanlan et al., 2023). For instance, there are one small and another big one triggering wandering in larvae and pupariation, respectively. For sesquiterpenoid juvenile hormone, its titre in corpora allatum changes in different developmental stages (Zhang et al., 2022) and generally decreases closer to the end of larval stages for pupariation (Dubrovsky 2005).”

(3) I found the text to be unclear, and important details were missing. There are many different cases where titers or gene expression went up or down, in males or females, but it is difficult to understand if these were expected or not, or what to make of all the differences. I also do not have a clear idea of what the hypothesis is. I had thought that the microRNAs were promoting the degradation of AstC-R1 or Kr-h1, so their loss should result in higher levels of AstC-R1 or Kr-h1 transcripts. However, Figures 2B and C show that there are lower levels of AstC-R1 and Kr-h1 transcripts in the loss of either miR-34 or miR-277. How does this fit with what is known of the progression of metamorphosis? A model built from the results to this point could help to organize the text.

The following information is now included in the main text, shown as follows:

- Lines 172-177 - “We then compared the expression of Kr-h1 and AstC-R1 of miR-277-KO, miR-34-KO, and between KO mutants and wild type flies using quantitative real-time PCR. The expression level of Kr-h1 was found to be significantly downregulated in both sexes of WL3 and WPP miR-277-KO mutants, coherent to the RNA-seq results; whereas a reduction was only noted in the male WL3 for miR-34-KO mutants but not in RNA-seq (Figure 2B; Supplementary Table S2).”

- Lines 178-186 - “On the other hand, expression level of AstC-R1 determined by real-time PCR was found to be upregulated during metamorphosis in wild-type (w1118) flies, but not for either miR-277-KO or miR-34-KO mutants (Figure 2C; Supplementary Figure 6). This is in agreement with the transcriptomic analyses where AstC-R1 was only up-regulated during male and female metamorphosis in the wild-type fly (Supplementary Table S2). In addition, we also found that the expression level of AstC-R1 in WPP of both miR-277-KO and mir-34-KO flies were significantly different to that of the w1118 flies (Figure 2C). Together, these data demonstrated that Kr-h1 and AstC-R are stage-specifically regulated by miR-277/34 in vivo.”

Reviewer #2:

Summary:

This study proposes that the microRNA cluster miR-277/34 controls the generation of sexual dimorphism in Drosophila melanogaster during metamorphosis by acting on specific hormonal and developmental gene pathways.

Strengths:

Using a combination of mRNA and small RNA sequencing together with genome-wide in silico and in vitro analyses the authors identified a microRNA cluster that may be involved in metamorphosis and the generation of sexual dimorphism in Drosophila melanogaster.

Weaknesses:

Biological validation of the identified sexually dimorphic genes and a detailed understanding of how the microRNA cluster miR-277/34 might be involved in the regulation of sesquiterpenoids are needed.

Major suggestions:

(1) If AstC-R1 and Kr-h1 are targets of the miR-277/34 cluster and cause their downregulation, it is not clear why there would also be a decrease in the levels of these genes in the miR-277/34 mutants. This would suggest that the mechanism is not straightforward and that further epistatic experiments should be carried out in order to clarify this issue.

We thank the reviewer for the positive and constructive comments. We agree that targets participate in endocrine feedback loops is dynamic, which could be the case for Kr-h1 and AstC-R1 during the larvae-pupae transition as they are components of the sesquiterpenoid hormone regulatory network. Epistatic analyses would be useful for future work and seem to be beyond the current scope of this study. The following information is now included in the main text:

Lines 184-187 - “Together, these data demonstrated that Kr-h1 and AstC-R are stage-specifically regulated by miR-277/34 in vivo. Further epistatic experiments would be needed to understand the dynamics between and the hormone actions.”

(2) The changes in the expression levels of AstC-R1 in pupae of miR-277-KO and mir-34-KO flies must be accompanied by photos of the respective larvae and pupae, as well as an analysis of the larvae-pupa transition on the mutants by gender.

We thank the reviewer for this suggestion. We have now added reference images illustrating the WL3 and WPP stages for w1118 in the Appendix as suggested (Author response image 1). We have analysed the larvae-pupae transition in both sexes of miR-277-KO and miR-34-KO mutants, directly comparing them with stage-matched w1118 controls, and we did not observe any detectable differences in larval morphology, pupal morphology, anterior spiracle eversion, gut-purge status, or timing of pupariation between the mutants and controls.

Author response image 1.

(3) Biological validation of the identified sexually dimorphic genes in vivo will be necessary for the support of this work.

We have now included additional quantitative RT-PCR validation of key insect hormone pathway genes in male and female WL3 and WPP stages of wild-type, miR-277-KO, and miR-34-KO flies (Figure 5). These additional analyses confirm sex-specific expression differences in both wild-type and mutant backgrounds, supporting dimorphic endocrine regulation. The main text are also revised and shown as follows:

Lines 276-287 - “To understand how different hormones such as juvenile hormone (Belgacem and Martin, 2002) involved during metamorphosis in both sexes, we selected and tested a selection of candidates from both sesquiterpenoid and ecdysteroid hormone pathways, and found JHAMT, Spok, CG40486, and CG10962 showed differential expression during transition from WL3 to WPP in mutant and/or wild-type fly lines in the transcriptome analyses (Supplementary Figure 16). To further examine these RNA-seq patterns, we performed RT-PCR on nine candidate genes from WL3 to WPP in WT and mutant fly lines. In the larval-pupal transition comparison, CG40486 was consistent down-regulated in WT males and females in both RNA-seq and RT-qPCR datasets. In the mutant-vs-WT comparison, CG9360 showed concordant downregulation in miR-277-KO WPP females. Other tested genes showed more condition-specific patterns matching between the transcriptomic and RT-qPCR analyses. Further validation and mechanistic studies will be required to define how these hormone-pathway changes contribute to the larval–pupal transition.”

Reviewer #3 (Public Review):

Summary:

The authors show convincingly the complexity of gene up- and down-regulation at the outset of metamorphosis and identify substantial differences between the two sexes, even at this early time in development. The complexity of microRNA expression and the difference between the sexes are also nicely laid out. The functional significance of these differences, though, is harder to establish. The authors have focused on the roles of two families of developmental hormones, the ecdysteroids, and the juvenile hormones. The emergence of sex-specific differentiation of organs during metamorphosis is clearly downstream of the action of ecdysteroids and/or JH, but there is no evidence that the presence or lack of these hormones has any effect on the sexual identity of organ systems - i.e., that manipulations of JH or ecdysteroid result in either the masculinization or feminization of individuals or their organs. The precedence for the linkage of these hormones to sex determination is the 2002, Belgacem & Martin study, which describes the effects of JH on fly locomotion. These authors show that the number of stop/start bouts is sexually dimorphic, and removal of JH in males shifts their frequency into the female range while giving treated males exogenous JH moves it back. While this is referred to as a "feminization" of male behavior, this quantitative shift in frequency is not as compelling as would be a qualitative shift -- for example, the removal of JH causing males to show egg-laying behavior (a result that has never been seen). Also, these effects are in a fully mature system, rather than at the early metamorphic time examined in the present paper. In driving and coordination metamorphosis, JH and ecdysteroids are intimately involved in sexual differentiation, but I know of no compelling evidence that they play a role in sex determination.

We thank the reviewer for the positive and constructive comments. We agree that our findings advanced on the sexual differences in gene regulation during metamorphosis rather than establishing direct evidence of juvenile hormone (JH) or ecdysteroids in influencing insect sex determination. In addition to rephrasing at various parts of the main text (see below), we have also carried out additional experiments in obtaining and analysing transcriptomes of late larvae and early pupae of both sexes in the wild-type of another species (Drosophila virilis), which showed similar KEGG pathways enriched. The revised parts are shown as follows:

Lines 289-303 – Results section

“Metamorphosis between different sexes in D. melanogaster and D. virilis

In D. melanogaster, the transition from WL3 to WPP shows increased carbohydrate/energy metabolism and ER protein processing, with decreased lipid metabolism, detoxification (cytochrome P450), and redox/cofactor pathways (folate, glutathione, retinol); while PM-LM exhibits broader negative enrichment in translational and RNA-processing machinery (ribosome, spliceosome, proteasome, RNA degradation, ubiquitin-mediated proteolysis) and in signaling modules (mTOR, MAPK, FoxO, Hippo), whereas PF-LF shifts are smaller and less coordinated by GSEA (Supplementary Figure S15). On the other hand in D. virilis, core features match that of D. melanogaster from WL3 to WPP, carbohydrate/energy metabolism and ER protein processing increase, while lipid, detoxification, and redox/cofactor pathways decrease. The sex bias is inverted relative to D. melanogaster as PM-LM shows limited up-regulation, whereas PF-LF displays broader, coordinated repression involving amino-acid metabolism (glycine/serine/threonine; cysteine/methionine), lysosome, ABC transporters, and longevity-related signaling, yielding the largest DEG set in females (Supplementary Figure S15). These data strongly suggested the sexual differences in gene regulation during metamorphosis in insects is not species-specific.”

Lines 354-361 – Methods section

“D. melanogaster w1118 and D. virilis were reared at 25 C, 45-50% relative humidity and 14:10 hour light-dark cycle. Sexes of wandering L3 larvae instar (WL3) were differentiated under the light dissecting microscope by the presence (male) or absence (female) of gonads (Supplementary Figure 1). Collections were restricted to larvae wandering out of food and ready for pupariation. WPP were defined as immobile puparia with everted spiracles and a uniform white cuticle that were no longer moving even with external stimuli. A total of three biological replicates (Whole-body of 1 individual was served as per biological replicate) per stage and sex were obtained for D. melanogaster and D. virilis.”

While the summary of the effects or removal of specific microRNAs on the components of the biosynthetic pathway for the JHs and ecdysteroids (Figure 2E ,F) is quite compelling, I am concerned about the effects of the removal of mir-277 and mir-34 on the levels of both the JHs and 20E. My concern centers around the data from the control group (w[1118] animals in Figure 2D). These data are the first report of a marked sex difference in the titer of either JH or ecdysteroid at the start of metamorphosis in Drosophila. As expected, males show a 10-20 increase in levels of JH III, JHB3, and 20E between the L3 stage and the white puparium, but, surprisingly, the levels of these hormones in female L3 larvae are equal to or greater than that seen at pupariation! These data for females run counter to over 50 years of work on the effects of ecdysteroids in Drosophila!

We thank the reviewer and agree that the endocrine profiles of WL3 females require careful interpretation, given the late-L3 to WPP transition represents a highly dynamic window in which ecdysteroid and sesquiterpenoid titres can fluctuate over short developmental intervals. The revised text is shown as follows:

Lines 322-326 – “Given the WL3 to WPP transition represents a highly dynamic window in which ecdysteroid and sesquiterpenoid titres can fluctuate over short developmental intervals, future experiments on narrower early-/mid-/late-wandering timepoints will be required to understand the dynamics of the microRNAs and hormones.”

As far as I can gather from the paper, the L3 data were obtained using wandering larvae. This stage lasts for about 12 hours and ends with pupariation. Larvae from this period need to be used with caution for hormone studies. Levels of both JH and ecdysteroid are low as larvae leave the food but rapidly rise to their peak levels at the white puparium stage 12 hours later. To deal with the rapidly changing hormonal landscape through this period, the researchers have used physiological markers to track this progression. Initially, it was the sage of "puffing" of the giant salivary gland chromosomes, but, for bulk collection of staged larvae, larvae are fed on food containing a blue dye, and progression is tracked by the loss of blue coloring from the gut. I could not find if the authors had any criteria for selecting larvae during the wandering period. Male and female larvae grow to different sizes. Might this difference in growth be biased when larvae were selected during their wandering phase?

We thank the reviewer and agree that WL3 represents a hormonally dynamic 12-hour window. To minimise developmental heterogeneity, WL3 larvae were selected for larvae that had completely exited the food and climbing upwards of the fly vials. To ensure tight developmental synchrony, pupating individuals from the same batch were collected 4 hours later, allowing us to match WL3 and WPP animals from a single, developmentally coordinated cohort. This paired-batch collection tried to minimise variation arising from differences in timing of wandering onset. As for the differences between the body sizes of male and female larvae, we minimise size-dependent bias, hormone titres measurement were measured to ng hormone per gram of tissue in our study, which corrects for sex-specific differences in mass. The information is now included in the main text as follows:

Lines 276-287 – “To understand how different hormones such as juvenile hormone (Belgacem and Martin, 2002) involved during metamorphosis in both sexes, we selected and tested a selection of candidates from both sesquiterpenoid and ecdysteroid hormone pathways, and found JHAMT, Spok, CG40486, and CG10962 showed differential expression during transition from WL3 to WPP in mutant and/or wild-type fly lines in the transcriptome analyses (Supplementary Figure 16). To further examine these RNA-seq patterns, we performed RT-PCR on nine candidate genes from WL3 to WPP in WT and mutant fly lines. In the larval-pupal transition comparison, CG40486 was consistent down-regulated in WT males and females in both RNA-seq and RT-qPCR datasets. In the mutant-vs-WT comparison, CG9360 showed concordant downregulation in miR-277-KO WPP females. Other tested genes showed more condition-specific patterns matching between the transcriptomic and RT-qPCR analyses. Further validation and mechanistic studies will be required to define how these hormone-pathway changes contribute to the larval–pupal transition”.

The other hormone-related issue is the expression of Kr-h1 during larval stages and metamorphosis (Figure 1G). Kr-h1 is the main target of JH and Kr-h1 expression is often used as a proxy for the JH titer. The authors report that peak Kr-h1 expression occurs in the L3 (when the JH titer should be lowest!) and that it drops at wandering. This pattern is counter to that reported in the literature (e.g., FlyBase, ModEncode).

We have now revised the main text as follows:

Lines 204-207 – “It should be noticed that the analyses of this study was based on RNA taken from whole-body rather than individual tissue, and further experiments will be required to dissect the dynamics of Kr-h1 and different hormones both spatially and temporally.”

Besides dealing with the issues above, it would be useful in the background to establish the difference between "sex determination" and "sexual differentiation". I was surprised that the authors could deal with this subject without ever mentioning the sex determination pathway - e.g., double sex, transformer, etc.

In this study, one major focus was on the differences between sex during metamorphosis rather than the sex determination pathway. We have included the reviewer’s suggestion by adding the following information in the main text, shown as follows:

Lines 73-75 - “In insects, various genes related to sex determination or showing sexual dimorphic features have been revealed, including doublesex and transformer (Shukla and Nagaraju, 2010; Deng et al, 2025).”

Line 61: at the time that the authors are considering, -- the transition from larva to pupa - the main controller of ecdysone production in the prothoracicotropic hormone, not ecdysis triggering hormone.

Thanks for the reviewer’s suggestions. The main text is now written as follows:

Lines 44-47 - “In general, these three hormones work in a cooperative manner, where ecdysteroid 20-hydroxyecdysone controls the timing of ecdysis or molting, prothoracicotropic hormone and the insulin-like peptides act as regulators of ecdysteroid synthesis, and sesquiterpenoid hormones such as juvenile hormone (JH) regulates metamorphosis based on their titre levels.”

Line 71: it would be useful to name the main microRNAs that have been implicated in regulating insect metamorphosis.

The main text is now written as follows:

Lines 60-63 - “Previous studies have identified a number of microRNAs contributing to the regulation of insect metamorphosis such as let-7, miR-125, and miR-2 family (e.g. Caygill and Johnston 2008; Sokol et al 2008; Lozano et al 2015).”

For Figures 2E and F as well as 3C (and a number of supplementary figures), it would be useful to provide some quantitative representation of the level of activation or suppression (as done in Figure 3B) rather than just Red or Green

The trend of expression change of genes in selected pathways and detailed quantitative information can now be found in the supplementary file.

hoort rectificatie niet bij schadevergoeding in andere vorm?

Test

Il me semble qu'il y a une contradiction entre ce paragraphe et le précédent. - le précédent nous dit que Warsh veut retourner aux "fondamentaux" de la politique monétaire qui prévalaient entre les années 80 et 2008, effectivement très discrétionnaire (et refermer la parenthèse ouverte par Bernanke) ; - celui-ci nous dit que il remet en cause une politique monétaire dont les fondements remontent aux années 80 et qui avaient mal préparé les BC aux périodes de crise auxquelles elles ont été confrontées depuis 2008.

Personnellement, j'ai le sentiment que les initiatives de Warsh visent davantage l'héritage de l'ère Bernanke (la politique de communication, que Bernanke a fait évoluer dès son arrivée en 2006, et l'instrumentalisation du bilan par la suite) que l'héritage Volker+Greenspan même s'il veut actualiser ce dernier.

Definition of VPNs, and how they work-Use for assignment. Eddleman