...

wow - awesome link to previous weeks' material...

ok...

self-acceptance!

So many good Frankl quotes!

honesty is the best policy (especially with oneself!) As it can prevent you from getting in over your head...

(this is related to the question I asked at the end of our last class -- do you envy people who lead an easy life? Where do they find meaning?)

Purpose in life!

positive relations!

Environmental mastery (tied to social settings and social skills in emotional intelligence...)

(mostly) the focus of the other article this week...

getting very philosophical here (but context is good)

all righty then...

telling us what is to follow...

hedonism - often associated with the pursuit of pleasure.

This article doesn't focus on it a lot (since it emphasizes the other main aspect of well-being), but I'd like to just highlight the connection between hedonism and happiness a little bit here by emphasizing the concept of the "hedonic treadmill".

The hedonic treadmill is a metaphor for the human tendency to pursue one pleasure after another. That's because the surge of happiness that's felt after a positive event is likely to return to a steady personal baseline over time.

So, it looks like this:

This is why I was so interested in the discussion we had at the end of the last class (which we'll continue this week) about money and happiness. Mo money, mo problems. We desire more riches, but we're never satisfied ...

Which is one of the reasons we need to look beyond hedonic well-being and explore eudaimonic well-being!

so, this is going to cover a lot of ground!

ok - we were already introduced to this distinction in the last article - we're going to get into it more here!

ok - we are familiar with this rhetorical contextextualizing... (it's the last half of our whole course)

oof

!

curious - given the next statement...

narrowing the focus...

ok

let's take the road less travelled!

probability of human behavior doesn't come from notions of free will as much as it comes from the variety of situations and possibilities that can't always be accounted for in strict categories

maybe Durkheim would differ

Did the actions of Protestants via 17th-England correlate with the ideal type of Protestant?

ideal type- the social construction of what something should be??

believes in necessity of generalizations and concepts but wary of over encompassing categories that try to explain literally everything. Additionally, wary of too narrow a focus of a case or a situation.

ooh! I really like this link to the previous week on perseverance and grit...

unless you're into stats and quantitative measurement, you can skip over these details ... but the social significance rather than the statistical significance of their findings is still trenchant!

like the lingo!

all right -- leading us into the next article!

generally accepted as entrepreneurs are assumed to get more meaning and purpose from their work than folk who work for someone/thing else other than themselves...

yep

• check it out in the quiz section this week...

wow! When listed in this way, it seems as though most workplaces or types of work might not encourage group productivity. How many of these apply to your work?

*

**

making a difference (for others...) or making a difference for oneself (being personally fulfilled)...

summing up perceptions of success

not as far as the marketplace is concerned...

hmmm

*

the testable proposition...

can you relate this to your own experiences?

remember what I said in my note about happiness and flow in the seminar prompt in week 7 (creative/cultural entrepreneurs)

sounds sensible enough (even if slightly utopian, moreso than ever in situations of precocity and neoliberal capitalist accumulation)

leading to the key insight...

!!

ooh! a key answer for one of the questions I asked as a seminar prompt in week 7 (about the propensity for people to feel intrinsically motivated to act/achieve goals....)

sad but true (and one of the reasons why entrepreneurial ventures are filled with such hope (because of all those times that one's mundane work life isn't...)

doesn't seem that controversial ... miserable people might be successful, but one doesn't expect it to be sustainable in the face of success (the misery...)

on the other hand, if you're happy (or satisfied), do you strive to achieve greater success?

this is quite the list -- kind of like success breeds more success, the happier you are, the more spillover there is likely to be in other related fields of achievement.

and we're going to focus on the latter... or at least the other reading does!

while entrepreneurial activities certainly relate to the well-being of one's physical body and one's relationships, we're going to focus on the psychological...

Feel free to ruminate on the others, though...

eudaemonia is quite distinct!

so, eudaemonia relates to self-improvement and self-actualization...

this should resonate with one of the questions I asked you to answer before the first class...

hedonism is focused on the pursuit of pleasure. What feels good...

whereas flow connects with a sense of intrinsic well-being, productivity relates to extrinsic forces too

focal points...

because there's a difference between being entrepreneurial and happy and being entrepreneurial to be happy (down the road...)

we're more likely to continue acting entrepreneurially if it satisfies us (if it gives us a sense of pleasure, or well-being) as a result of pursuing the dream, seeking and creating new opportunities... being bold and innovative and creative... (even though there are risks and great costs associated with these too)

*

already introduced 4 weeks ago...

where do you stand?

focusing on one's sense of subjective well-being (otherwise known as happiness).

Vítejte do světa Animatrixu, vizionářské fúze počítačové animace a japonské školy animace, od světově nejuznávanějších tvůrců animovanch filmů. Vychutnejte si snímek, který vznikl před filmem Matrix a dozvíte se o posledních městech lidské rasy, válce mezi stroji a lidmi a o závěrečném úpadku lidstva. Staňte se svědky posledního letu Osirise, který připravil půdu pro film Matrix Reloaded a videohru Enter The Matrix. Doplňte si své povědomí o Matrixu a získejte informace, které jinde nenajdete. Je čas zapojit se... (oficiální text distributora)

**

!!

that's quite the burn, for an academic article!

*

ok

how do you demonstrate this?

interesting!

this gets into persuasion & rhetoric. In essence, also "selling" but not selling things, but ideas, selling people on your version of reality...

!

connecting the 3 types of intelligence...

synergy! For success ...

you probably have experience with this ...

underwhelming. This really drives home the fact that this article is good mostly as a shell -- kind-of boring and potentially difficult to see (at least in an immediate fashion) how it can relate in a real sense to students' lives (beyond the idea of multiple intelligences). Still, the lens is really useful for helping us focus on social competence and tacit knowledge (practical intelligence) for moving us forward... and considering what might be necessary in order to achieve success in a given field...

the method...

classic job interview question...

link to independent spirit and counterfactual knowledge...

yowza! School can't teach tacit knowledge!

hah

(link to the growth mindset)

crucial insight

Their "street math" was not replicated in the classroom... They exhibited "street smarts" (not book smarts)

Have you ever been in a situation where your book smarts failed you in the "real" world? Contrarily, how often has "street" knowledge helped you in school?

crucial insight - we have access to multiple intelligences!

Please note - this is definitely related to the "theory of multiple intelligences" (which was the basis for the reality TV series "Canada's Smartest Person"):

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

Think about how often your ability to act in a "socially competent" manner came off as intelligent (a way of adapting to, shaping, and sometimes selecting your environment).

Knowing the right thing to say or how to act (cultural capital) comes off as "smart"

the moral of the story

** remember these 4 factors of social competence! (they will be very helpful next week!)

ok - getting right to the heart of it. Introduces the key concepts and emphasizes its 3 parts.

funny story - the punchline is at the end of the next paragraph!

what I was talking about at the end of last Thursday's class...

key concept, defined

link to divergent thinking...

link to opportunity discovery/creation

ok - definition by contextualization (and very similar to last week's material...)

completing the picture...

but practical intelligence guides the application of intellectual intelligence...

yep

VISUAL ANNOTATIONS

1. (3:55) Code Carbon helps to provide a visual for the emissions produced in a certain time period. Much like the nutrition information on the outside of a cereal box, though it is helpful to know how many calories or grams of sugar are contained in one serving, that information would be useless without knowing what a serving size is. Even more so if we are unaware of how many calories we should consume in a given time frame. This information allows us to make informed decisions about what we should consume.

2. (5:04) This visual of the photos that are generated when she inputs her own information provides a bit of shock factor to the audience. Not only are most of those images of someone who is not her, but they are quite illicit photos of someone who is not her. Taken out of context and unknown to be AI generated, photos such as these could be detrimental to someone's career or personal life.

REACTION ANOTAION:

I never paused to consider that AI models are not contained within the digital world and exist boldly in our physical world. I figured that, since AI is so easily available on a cell phone, it couldn't possibly require so much to sustain it.

RESTATEMENT ANNOTAIONS

1. (0:37) Though at times AI can be extremely helpful, as we often hear in the media, there is also a dark side to these functions. For instance, one chatbot suggested that the solution to one man's problems was to file for divorce. Another suggested a sure-to-please recipe for a large gathering, that contained chlorine gas among its list of ingredients.

2. (1:19) AI does not exist in solitude. Its affects have left the digital world and have begun to affect our physical world. AI models have been noted to contribute to climate change, use artist and authors work without their consent, and discriminate against entire groups of people.

3. (1:56) In regard to sustainability, the figurative cloud we imagine AI resides upon is not something only contained within the digital world. We often fail to acknowledge the large quantity of materials needed to build these models as well as the sizeable amount of energy required to sustain them.

Important concept: details the cycle of extracting natural (and finite) resources.

Style elements: speaks directly to the reader, repeats important phrases.

"Empty" land becomes a place to extract resources for profit.

It's almost like he's telling you to spend so much time "looking within" that you forget to look around and you become closed off to the world. He's advocating exceptionalism if you think about it.

According to Emerson The secret to a fulfilled life is to just do everything yourself no matter how daunting.

contradicts the message of keep to your self, do what you want, don't care about what others think

Getting a strong individualism vibe.

unpopular opinions cause estrangement.

Il est ici question des biais liés aux données collectées, aux biais des statistiques et du manque de neutralité de certaines études

Référence à notre réflexion plus haut concernant les biais préexistants, avant l'IA et Nouvelles Technologies

Un souci primordial à expliciter lorsqu'on parle de l'IA sur le marché du travail est celui de l'accéssiblité numérique, une accessiblité qui n'est pas encore étendue au monde entier et donc le sujet de l'IA dans le monde du travail ne concerne pour l'instant que les pays et les sociétés dit.e.s "développés" (pour la plupart en Occident) et donc opérer un shift majeur en faveur de technologies inacessibles pour beaucoup ne fera qu'alourdir le gap entre eux

Cela n'est pas inhérent à l'automatisation numérique en elle-même, il s'agit là d'un souci plus systémique -> l'IA et les Nouvelles Technologies ne font que refléter en prolongeant les problèmes qui existent déjà, ce ne sont que des outils

Même risque exposé que celui de l'article précédent.

Welcome back and by now you should understand the difference between stateless and stateful security protection.

In this lesson I want to talk about one security feature of AWS VPCs and a little bit more depth and that's network access control lists known as knuckles.

Now we do have a lot to cover so let's jump in and get started.

A network access control list we thought of as a traditional firewall available within AWS VPCs so let's look at a visual example.

A subnet within an AWS VPC which has two EC2 instances A and B.

The first thing to understand and this is core to how knuckles work within AWS is that they are associated with subnets.

Every subnet has an associated network ACL and this filters data as it crosses the boundary of that subnet.

In practice this means any data coming into the subnet is affected and data leaving the subnet is affected.

But and this is super important to remember connections between things within that subnet such as between instance A and instance B in this example are not affected by network ACLs.

Each network ACL contains a number of rules, two sets of rules to be precise.

We have inbound rules and outbound rules.

Now inbound rules only affect data entering the subnet and outbound rules affect data leaving the subnet.

Remember from the previous lesson this isn't always matching directly to request and response.

A request can be either inbound or outbound as can a response.

These inbound and outbound rules are focused only on the direction of traffic not whether it's request or response.

In fact and I'll cover this very soon knuckles are stateless which means they don't know if traffic is request or response.

It's all about direction.

Now rules match the destination IP or IP range, destination port or port range together with the protocol and they can explicitly allow or explicitly deny traffic.

Remember this one network ACLs offer both explicit allows and explicit denies.

Now rules are processed in order.

First a network ACL determines if the inbound or outbound rules apply.

Then it starts from the lowest rule number.

It evaluates traffic against each individual rule until it finds a match.

Then that traffic is either allowed or denied based on that rule and then processing stops.

Now this is critical to understand because it means that if you have a deny rule and an allow rule which match the same traffic but if the deny rule comes first then the allow rule might never be processed.

Lastly there's a catch all showed by the asterisk in the rule number and this is an implicit deny.

If nothing else matches then traffic will be denied.

So those are the basics.

Next let's move on to some more complex elements of network ACLs.

Now I just mentioned that network ACLs are stateless and this means that rules are required for both the request and the response part of every communication.

You need individual rules for those so one inbound and one outbound.

Take this example a multi-tiered application running in a VPC.

We've got a web server in the middle and an application server on the left.

On the right we have a user Bob using a laptop and he's accessing the website.

So he makes a connection using HTTPS which is TCP port 443 and this is the request as you know by now and this is also going to mean that a response is required using the ephemeral port range.

This ephemeral port is chosen at random from the available range decided by the operating system on Bob's laptop.

Now to allow for this initial communication if we're using network ACLs then we'll need to have one associated with the web subnet and it will need rules in the inbound and outbound sections of that network ACL.

Notice how on the inbound rule set we have rule number 110 which allows connections from anywhere and this is signified by 0.0.0.0 through this network ACL and this is allowed as long as it's using TCP port 443.

So this is what allows the request from Bob into the web server.

We also have on the outbound rule set rule number 120 and this allows outbound traffic to anywhere again 0.0.0.0 as long as the protocol is TCP using the port range of 1.0.2.4 to 65.5.3.5 and this is the ephemeral port range which I mentioned in the previous lesson.

Now this is not amazingly secure but with stateless firewalls this is the only way.

Now we also have the implicit denies and this is denoted by the rules with the star in the rule number and this means that anything which doesn't match rule 110 or 120 is denied.

Now it's also worth mentioning while I do have rule 110 and 120 number differently the rule numbers are unique on inbound and outbound so we could have the single rule number 110 on both rule sets and that would be okay.

It's just easier to illustrate this if I use unique rule numbers for each of the different rule sets.

Now let's move on and increase the complexity little.

So we have the same architecture we have Bob on the right, the web subnet in the middle and the application subnet on the left.

You know now that because network ACLs are stateless each communication requires one request rule and one response rule.

This becomes more complex when you have a multi-tiered architecture which operates across multiple subnets and let's step through this to illustrate why.

Let's say the pop initiates a connection to the web server we know about this already because I just covered it.

If we have a network ACL around the web subnet we'll need an inbound rule on the web network ACL.

There's also going to be response traffic so this is going to use the ephemeral port range and this is going to need an outbound rule on that same web network ACL so this should make sense so far.

But also the web server might need to communicate with the app server using some application TCP port.

Now this is actually crossing two subnet boundaries the web at subnet boundary and the application subnet boundary so it's going to need an outbound rule on the web at subnet knuckle and also an inbound rule on the application subnet knuckle.

Then we have the response for that as well from the app server through to the web server and this is going to be using ephemeral ports but this also crosses two subnet boundaries it leaves the application subnet which will need an outbound rule on that knuckle and it enters the web subnet which will also need an inbound rule on that network ACL and what if each of those servers need software updates it will get even more complex really quickly.

You always have to be aware of these rule pairs the application port request and the ephemeral response for every single communication in some cases you're going to have multi-tier architecture and this might mean the communications go through different subnets.

If you need software updates this will need more if you use network address translation or NAT you might need more rules still.

You'll need to worry about this if you use network ACLs within a vpc for traffic to a vpc or traffic from a vpc or traffic between subnets inside that vpc.

When a vpc is created it's created with a default network ACL and this contains inbound and outbound rule sets which have the default implicit deny but also a capsule allow and this means that the net effect is that all traffic is allowed so the default within a vpc is that knuckles have no effect they aren't used this is designed [Music] I need to be beginner friendly and reduce admin overhead.

AWS prefer using security groups which I'll be covering soon.

If you create your own custom network ACLs though that's a different story.

Custom knuckles are created for a specific vpc and initially they're associated with no subnets.

They only have one rule on both the inbound and outbound rule sets which is the default deny and the result is that if you associate this custom network ACL with any subnets all traffic will be denied so be careful with this it's radically different behavior than the default network ACL created with a vpc.

Now this point I just want to cover some finishing key points which you need to be aware of for any real-world usage and when you're answering exam questions.

So network access controlists remember they're known as knuckles they are stateless so they view request and response as different things so you need to add rules both for the request and for the response.

A knuckle only affects data which is crossing the subnet boundary so communications between instances in the same subnet is not affected by a network ACL on that subnet.

Now this can mean that if you do have data crossing between subnets then you need to make sure that each knuckle on both of those subnets has the appropriate inbound and outbound rules so you end up with a situation where one connection can in theory need two rules on each knuckle if that connection is crossing two different subnet boundaries.

Now knuckles are able to explicitly allow traffic and explicitly deny and the deny is important because as you'll see when I talk about security groups this is a capability that you need to network ACLs.

So network ACLs allow you to block specific IPs or specific IP ranges which are associated with bad actors so they're a really good security feature when you need to block any traffic attempting to exploit your systems.

Now network ACLs are not aware of any logical resources they only allow you to use IPs and cyber ranges ports and protocols you cannot reference logical resources within AWS and knuckles can also not be assigned two logical resources they're only assigned to subnets within VPCs within AWS.

Now knuckles are very often used together with security groups such as mentioned to add the capability to explicitly deny bad IPs or bad networks so generally you would use security groups to allow traffic and you use knuckles to deny traffic and I'll talk about exactly how this works in the next lesson.

Now each subnet within a VPC has one knuckle associated with it it's either going to be the default network ACL for that VPC or a custom one which you create and associate.

A single knuckle though can be associated with many different subnets so while a subnet can only have one network ACL one network ACL can be associated with many different subnets.

Now this point that is everything that I wanted to cover about network ACLs for this lesson so go ahead complete the video and when you're ready I'll look forward to you joining me in the next lesson.

此处提出了一个新方法

eLife Assessment

This important study provides proof of principle that C. elegans models can be used to accelerate the discovery of candidate treatments for human Mendelian diseases by detailed high-throughput phenotyping of strains harboring mutations in orthologs of human disease genes. The data are compelling and support an approach that enables the potential rapid repurposing of FDA-approved drugs to treat rare diseases for which there are currently no effective treatments. The authors should provide a clearer explanation of how the statistical analyses were performed, as well as a link to a GitHub repository to clarify how figures and tables in the manuscript were generated from the phenotypic data.

Reviewer #1 (Public review):

Summary:

As the scientific community identifies increasing numbers of genes and genetic variants that cause rare human diseases, a challenge in the field quickly identify pharmacological interventions to address known deficits. The authors point out that defining phenotypic outcomes required for drug screen assays is often a bottleneck, and emphasize how invertebrate models can be used for quick ID of compounds that may address genetic deficits. A major contribution of this work is to establish a framework for potential intervention drug screening based on quantitative imaging of morphology and mobility behavior, using methods that the authors show can define subtle phenotypes in a high proportion of disease gene knockout mutants. Overall, the work constitutes an elegant combination of previously developed high-volume imaging with highly detailed quantitative phenotyping (and some paring down to specific phenotypes) to establish proof of principle on how the combined applications can contribute to screens for compounds that may address specific genetic deficits, which can, in turn, suggest both mechanism and therapy.

In brief, the authors selected 25 genes for which loss of function is implicated in human neuro-muscular disease and engineered deletions in the corresponding C. elegans homologs. The authors then imaged morphological features and behaviors prior to, during, and after blue light stimuli, quantitating features, and clustering outcomes as they elegantly developed previously (PMID 35322206; 30171234; 30201839). In doing so, phenotypes in 23/25 tested mutants could be separated enough to distinguish WT from mutant and half of those with adequate robustness to permit high-throughput screens, an outcome that supports the utility of related general efforts to ID phenotypes in C. elegans disease orthologs. A detailed discussion of 4 ciliopathy gene defects, and NACLN-related channelopathy mutants reveals both expected and novel phenotypes, validating the basic approach to modeling vetted targets and underscoring that quantitative imaging approaches reiterate known biology.

The authors then screened a library of nearly 750 FDA-approved drugs for the capacity to shift the unc-80 NACLN channel-disrupted phenotype closer to the wild type. Top "mover" compounds shift outcome in the experimental outcome space; and also reveal how "side effects" can be evaluated to prioritize compounds that confer the fewest changes of other parameters away from the center.

Strengths:

Although the imaging and data analysis approaches have been reported and the screen is restricted in scope and intervention exposure, it is impressive, encouraging and important that the authors strongly combine tools to demonstrate how quantitative imaging phenotypes can be integrated with C. elegans genetics to accelerate the identification of potential modulators of disease (easily extendable to other goals). Generation of deletion alleles and documentation of their associated phenotypes (available in supplemental data) provide potentially useful reagents/data to the field. The capacity to identify "over-shooting" of compound applications with suggestions for scale back and to sort efficacious interventions to minimize other changes to behavioral and physical profiles is a strong contribution.

Weaknesses:

The work does not have major weaknesses, and in revision, the authors have expanded the discussion to potential utility and application in the field.

The authors have also taken into account minor modifications in writing.

Reviewer #2 (Public review):

Summary and strengths:

O'Brien et al. present a compelling strategy to both understand rare disease that could have a neuronal focus and discover drugs for repurposing that can affect rare disease phenotypes. Using C. elegans, they optimize the Brown lab worm tracker and Tierpsy analysis platform to look at movement behaviors of 25 knockout strains. These gene knockouts were chosen based on a process to identify human orthologs that could underlie rare diseases. I found the manuscript interesting and a powerful approach to make genotype-phenotype connections using C. elegans. Given the rate that rare Mendelian diseases are found and candidate genes suggested, human geneticists need to consider orthologous approaches to understand the disease and seek treatments on a rapid time scale. This approach is one such way. Overall, I have a few minor suggestions and some specific edits.

Weaknesses:

(1) Throughout the text on figures, labels are nearly impossible to read. I had to zoom into the PDF to determine what the figure was showing. Please make text in all figures a minimum of 10 point font. Similarly, Figure 2D point type is impossible to read. Points should be larger in all figures. Gene names should be in italics in all figures, following C. elegans convention.

(2) I have a strong bias against the second point in Figure 1A. Sequencing of trios, cohorts, or individuals NEVER identifies causal genes in the disease. This technique proposes a candidate gene. Future experiments (oftentimes in model organisms) are required to make those connections to causality. Please edit this figure and parts of the text.

(3) How were the high-confidence orthologs filtered from 767 to 543 (lines 128-131)? Also, the choice of the final list of 25 genes is not well justified. Please expand more about how these choices were made.

(4) Figures 3 and 4, why show all 8289 features? It might be easier to understand and read if only the 256 Tierpsy features were plotted in the heat maps.

(5) The unc-80 mutant screen is clever. In the feature space, it is likely better to focus on the 256 less-redundant Tierpsy features instead of just a number of features. It is unclear to me how many of these features are correlated and not providing more information. In other words, the "worsening" of less-redundant features is far more of a concern than "worsening" of 1000 correlated features.Reviewer #2 (Public review):

Summary and strengths:

O'Brien et al. present a compelling strategy to both understand rare disease that could have a neuronal focus and discover drugs for repurposing that can affect rare disease phenotypes. Using C. elegans, they optimize the Brown lab worm tracker and Tierpsy analysis platform to look at movement behaviors of 25 knockout strains. These gene knockouts were chosen based on a process to identify human orthologs that could underlie rare diseases. I found the manuscript interesting and a powerful approach to make genotype-phenotype connections using C. elegans. Given the rate that rare Mendelian diseases are found and candidate genes suggested, human geneticists need to consider orthologous approaches to understand the disease and seek treatments on a rapid time scale. This approach is one such way. Overall, I have a few minor suggestions and some specific edits.

Weaknesses:

(1) Throughout the text on figures, labels are nearly impossible to read. I had to zoom into the PDF to determine what the figure was showing. Please make text in all figures a minimum of 10 point font. Similarly, Figure 2D point type is impossible to read. Points should be larger in all figures. Gene names should be in italics in all figures, following C. elegans convention.

(2) I have a strong bias against the second point in Figure 1A. Sequencing of trios, cohorts, or individuals NEVER identifies causal genes in the disease. This technique proposes a candidate gene. Future experiments (oftentimes in model organisms) are required to make those connections to causality. Please edit this figure and parts of the text.

(3) How were the high-confidence orthologs filtered from 767 to 543 (lines 128-131)? Also, the choice of the final list of 25 genes is not well justified. Please expand more about how these choices were made.

(4) Figures 3 and 4, why show all 8289 features? It might be easier to understand and read if only the 256 Tierpsy features were plotted in the heat maps.

(5) The unc-80 mutant screen is clever. In the feature space, it is likely better to focus on the 256 less-redundant Tierpsy features instead of just a number of features. It is unclear to me how many of these features are correlated and not providing more information. In other words, the "worsening" of less-redundant features is far more of a concern than "worsening" of 1000 correlated features.

Reviewer #3 (Public review):

In this study, O'Brien et al. address the need for scalable and cost-effective approaches to finding lead compounds for the treatment of the growing number of Mendelian diseases. They used state-of-the-art phenotypic screening based on an established high-dimensional phenotypic analysis pipeline in the nematode C. elegans.

First, a panel of 25 C. elegans models was created by generating CRISPR/Cas9 knock-out lines for conserved human disease genes. These mutant strains underwent behavioral analysis using the group's published methodology. Clustering analysis revealed common features for genes likely operating in similar genetic pathways or biological functions. The study also presents results from a more focused examination of ciliopathy disease models.

Subsequently, the study focuses on the NALCN channel gene family, comparing the phenotypes of mutants of nca-1, unc-77, and unc-80. This initial characterization identifies three behavioral parameters that exhibit significant differences from the wild type and could serve as indicators for pharmacological modulation.

As a proof-of-concept, O'Brien et al. present a drug repurposing screen using an FDA-approved compound library, identifying two compounds capable of rescuing the behavioral phenotype in a model with UNC80 deficiency. The relatively short time and low cost associated with creating and phenotyping these strains suggest that high-throughput worm tracking could serve as a scalable approach for drug repurposing, addressing the multitude of Mendelian diseases. Interestingly, by measuring a wide range of behavioural parameters, this strategy also simultaneously reveals deleterious side effects of tested drugs that may confound the analysis.

Considering the wealth of data generated in this study regarding important human disease genes, it is regrettable that the data is not made accessible to researchers less versed in data analysis methods. This diminishes the study's utility. It would have a far greater impact if an accessible and user-friendly online interface were established to facilitate data querying and feature extraction for specific mutants. This would empower researchers to compare their findings with the extensive dataset created here.

Another technical limitation of the study is the use of single alleles. Large deletion alleles were generated by CRISPR/Cas9 gene editing. At first glance, this seems like a good idea because it limits the risk that background mutations, present in chemically-generated alleles, will affect behavioral parameters. However, these large deletions can also remove non-coding RNAs or other regulatory genetic elements, as found, for example, in introns. Therefore, it would be prudent to validate the behavioral effects by testing additional loss-of-function alleles produced through early stop codons or targeted deletion of key functional domains.

Author response:

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

Public Reviews:

Reviewer #1 (Public Review): 

Summary: 

As the scientific community identifies increasing numbers of genetic variants that cause rare human diseases, a challenge is how the field can most quickly identify pharmacological interventions to address known deficits. The authors point out that defining phenotypic outcomes required for drug screen assays is often challenging, and emphasize how invertebrate models can be used for quick ID of compounds that may address genetic deficits. A major contribution of this work is to establish a framework for potential intervention drug screening based on quantitative imaging of morphology and mobility behavior, using methods that the authors show can define subtle phenotypes in a high proportion of disease gene knockout mutants. 

Overall, the work constitutes an elegant combination of previously developed high-volume imaging with highly detailed quantitative phenotyping (and some paring down to specific phenotypes) to establish proof of principle on how the combined applications can contribute to screens for compounds that may address specific genetic deficits, which can suggest both mechanism and therapy. 

In brief, the authors selected 25 genes for which loss of function is implicated in human neuro-muscular disease and engineered deletions in the corresponding C. elegans homologs. The authors then imaged morphological features and behaviors prior to, during, and after blue light stimuli, quantitating features, and clustering outcomes as they elegantly developed previously (PMID 35322206; 30171234; 30201839). In doing so, phenotypes in 23/25 tested mutants could be separated enough to distinguish WT from mutant and half of those with adequate robustness to permit high-throughput screens, an outcome that supports the utility of general efforts to ID phenotypes in C. elegans disease orthologs using this approach. A detailed discussion of 4 ciliopathy gene defects, and NACLN-related channelopathy mutants reveals both expected and novel phenotypes, validating the basic approach to modeling vetted targets and underscoring that quantitative imaging approaches reiterate known biology. The authors then screened a library of nearly 750 FDA-approved drugs for the capacity to shift the unc-80 NACLN channel-disrupted phenotype closer to the wild type. Top "mover" compound move outcome in the experimental outcome space; and also reveal how "side effects" can be evaluated to prioritize compounds that confer the fewest changes of other parameters away from the center. 

Strengths: 

Although the imaging and data analysis approaches have been reported and the screen is limited in scope and intervention exposure, it is important that the authors strongly combine individual approach elements to demonstrate how quantitative imaging phenotypes can be integrated with C. elegans genetics to accelerate the identification of potential modulators of disease (easily extendable to other goals). Generation of deletion alleles and documentation of their associated phenotypes (available in supplemental data) provide potentially useful reagents/data to the field. The capacity to identify "over-shooting" of compound applications with suggestions for scale back and to sort efficacious interventions to minimize other changes to behavioral and physical profiles is a strong contribution. 

Weaknesses: 

The work does not have major weaknesses, although it may be possible to expand the discussion to increase utility in the field: 

(1) Increased discussion of the challenges and limitations of the approach may enhance successful adaptation application in the field. 

It is quite possible that morphological and behavioral phenotypes have nothing to do with disease mechanisms and rather reflect secondary outcomes, such that positive hits will address "off-target" consequences. 

This is possible and can only be determined with human data. We now discuss the possibility in the discussion.

The deletion approach is adequately justified in the text, but the authors may make the point somewhere that screening target outcomes might be enhanced by the inclusion of engineered alleles that match the human disease condition. Their work on sod-1 alleles (PMID 35322206) might be noted in this discussion. 

We agree and now mention this work in the discussion. We are currently working on a collection of strains with patient-specific mutations.

Drug testing here involved a strikingly brief exposure to a compound, which holds implications for how a given drug might engage in adult animals. The authors might comment more extensively on extended treatments that include earlier life or more extended targeting. The assumption is that administering different exposure periods and durations, but if the authors are aware as to whether there are challenges associated with more prolonged applications, larger scale etc. it would be useful to note them. 

More prolonged applications are definitely possible. We chose short treatments for this screen to model the potential for changing neural phenotypes once developmental effects of the mutation have already occurred. We now briefly discuss this choice and the potential of longer treatments in the discussion.

(2) More justification of the shift to only a few target parameters for judging compound effectiveness. 

- In the screen in Figure 4D and text around 313, 3 selected core features of the unc-80 mutant (fraction that blue-light pause, speed, and curvature) were used to avoid the high replicate requirements to identify subtle phenotypes. Although this strategy was successful as reported in Figure 5, the pared-down approach seems a bit at odds with the emphasis on the range of features that can be compared mutant/wt with the author's powerful image analysis. Adding details about the reduced statistical power upon multiple comparisons, with a concrete example calculated, might help interested scientists better assess how to apply this tool in experimental design. 

To empirically test the effect of including more features on the subsequent screen, we have repeated the analysis using increasing numbers of features. In a new supplementary figure we find increasing the number of features reduces our power to detect rescue. At 256 features, we would not be able to detect any compounds that rescued the disease model phenotype.

(3) More development of the side-effect concept. The side effects analysis is interesting and potentially powerful. Prioritization of an intervention because of minimal perturbation of other phenotypes might be better documented and discussed a bit further; how reliably does the metric of low side effects correlate with drug effectiveness? 

Ultimately this can only be determined with clinical trial data on multiple drugs, but there are currently no therapeutic options for UNC80 deficiency in humans. We have included some extra discussion of the side effect concept.

Reviewer #2 (Public Review): 

Summary and strengths: 

O'Brien et al. present a compelling strategy to both understand rare disease that could have a neuronal focus and discover drugs for repurposing that can affect rare disease phenotypes. Using C. elegans, they optimize the Brown lab worm tracker and Tierpsy analysis platform to look at the movement behaviors of 25 knockout strains. These gene knockouts were chosen based on a process to identify human orthologs that could underlie rare diseases. I found the manuscript interesting and a powerful approach to making genotype-phenotype connections using C. elegans. Given the rate at which rare Mendelian diseases are found and candidate genes suggested, human geneticists need to consider orthologous approaches to understand the disease and seek treatments on a rapid time scale. This approach is one such way. Overall, I have a few minor suggestions and some specific edits. 

Weaknesses: 

(1) Throughout the text on figures, labels are nearly impossible to read. I had to zoom into the PDF to determine what the figure was showing. Please make text in all figures a minimum of 10-point font. Similarly, the Figure 2D point type is impossible to read. Points should be larger in all figures. Gene names should be in italics in all figures, following C. elegans convention. 

We have updated all figures with larger labels and, where necessary, split figures to allow for better readability. We’ve also corrected italicisation.

(2) I have a strong bias against the second point in Figure 1A. Sequencing of trios, cohorts, or individuals NEVER identifies causal genes in the disease. This technique proposes a candidate gene. Future experiments (oftentimes in model organisms) are required to make those connections to causality. Please edit this figure and parts of the text. 

We have removed references to causation. We were thinking of cases where a known variant is found in a patient where causality has already been established rather than cases of new variant discovery.

(3) How were the high-confidence orthologs filtered from 767 to 543 (lines 128-131)? Also, the choice of the final list of 25 genes is not well justified. Please expand more about how these choices were made. 

We now explain the extra keyword filtering step. For the final filtering step, we simply examined the list and chose 25. There is therefore little justification to provide and we acknowledge these cannot be seen as representative of the larger set according to well-defined rules. The choice was based on which genes we thought would be interesting using their descriptions or our prior knowledge (“subjective interestingness” in the main text).

(4) Figures 3 and 4, why show all 8289 features? It might be easier to understand and read if only the 256 Tierpsy features were plotted in the heat maps. 

In this case, we included all features because they were all tested for differences between mutants and controls. By consistently using all features for each fingerprint we can be sure that the features that are different that we want to highlight in box plots can be referred to in the fingerprint.

(5) The unc-80 mutant screen is clever. In the feature space, it is likely better to focus on the 256 less-redundant Tierpsy features instead of just a number of features. It is unclear to me how many of these features are correlated and not providing more information. In other words, the "worsening" of less-redundant features is far more of a concern than the "worsening" of 1000 correlated features. 

This is a good point. We’ve redone the analysis using the Tierpsy 256 feature set and included this as a supplementary figure. We find that the same trend exists when looking at this reduced feature set.

Reviewer #3 (Public Review): 

In this study, O'Brien et al. address the need for scalable and cost-effective approaches to finding lead compounds for the treatment of the growing number of Mendelian diseases. They used state-of-the-art phenotypic screening based on an established high-dimensional phenotypic analysis pipeline in the nematode C. elegans. 

First, a panel of 25 C. elegans models was created by generating CRISPR/Cas9 knock-out lines for conserved human disease genes. These mutant strains underwent behavioral analysis using the group's published methodology. Clustering analysis revealed common features for genes likely operating in similar genetic pathways or biological functions. The study also presents results from a more focused examination of ciliopathy disease models. 

Subsequently, the study focuses on the NALCN channel gene family, comparing the phenotypes of mutants of nca-1, unc-77, and unc-80. This initial characterization identifies three behavioral parameters that exhibit significant differences from the wild type and could serve as indicators for pharmacological modulation. 

As a proof-of-concept, O'Brien et al. present a drug repurposing screen using an FDA-approved compound library, identifying two compounds capable of rescuing the behavioral phenotype in a model with UNC80 deficiency. The relatively short time and low cost associated with creating and phenotyping these strains suggest that high-throughput worm tracking could serve as a scalable approach for drug repurposing, addressing the multitude of Mendelian diseases. Interestingly, by measuring a wide range of behavioural parameters, this strategy also simultaneously reveals deleterious side effects of tested drugs that may confound the analysis. 

Considering the wealth of data generated in this study regarding important human disease genes, it is regrettable that the data is not actually made accessible. This diminishes the study's utility. It would have a far greater impact if an accessible and user-friendly online interface were established to facilitate data querying and feature extraction for specific mutants. This would empower researchers to compare their findings with the extensive dataset created here. Otherwise, one is left with a very limited set of exploitable data. 

We have now made the feature data available on Zenodo (https://doi.org/10.5281/zenodo.12684118) as a matrix of feature summaries and individual skeleton timeseries data (the feature matrix makes it more straightforward to extract the data from particular mutants for reanalysis). We have also created a static html version of the heatmap in Figure 2 containing the entire behavioural feature set extracted by Tierpsy. This can be opened in a browser and zoomed for detailed inspection. Mousing over the heatmap shows the names of features at each position making it easier to arrive at intuitive conclusions like ‘strain A is slow’ or ‘strain B is more curved’.

Another technical limitation of the study is the use of single alleles. Large deletion alleles were generated by CRISPR/Cas9 gene editing. At first glance, this seems like a good idea because it limits the risk that background mutations, present in chemically-generated alleles, will affect behavioral parameters. However, these large deletions can also remove non-coding RNAs or other regulatory genetic elements, as found, for example, in introns. Therefore, it would be prudent to validate the behavioral effects by testing additional loss-of-function alleles produced through early stop codons or targeted deletion of key functional domains. 

We have added a note in the main text on limitations of deletion alleles. We like the idea of making multiple alleles in future studies, especially in cases where a project is focussed on just one or a few genes.

Recommendations for the authors

Reviewer #1 (Recommendations For The Authors): 

Note that none of the above suggestions or the one immediately below are considered mandatory. 

One additional minor point: The dual implication of mevalonate perturbations for NACLM deficiencies is striking. At the same time, the mevalonate pathway is critical for embryo viability among other things, which prompts questions about how reproductive physiology is integrated in this screen approach. It appears that sterilization protocols are not used to prepare screen target animals, but it would be useful to know if there were a signature associated with drug-induced sterility that might help identify one potential common non-interesting outcome of compound treatments in general. In this work, the screen treatment is only 4 hours, which is probably too short to compromise reproduction, but as noted above, it is likely users would intend to expose test subjects for much longer than 4-hour periods. 

This is an interesting point. In its current form our screen doesn’t assess reproductive physiology. This is something that we will consider in ongoing projects.

Figures 

Figure 1D might be omitted or moved to supplement. 

We have removed 1D and moved figure 1E as a standalone table (Table 1) to improve readability.

Figure 2D "key" is hard to make out size differences for prestim, bluelight, and poststim -more distinctive symbols should be used. 

We have increased the size of the symbols so that the key is easier to read.

Line 412 unc-25 should be in italics 

Corrected

Reviewer #2 (Recommendations For The Authors): 

Specific edits: 

All of the errors below have been corrected.

Line 47, "loss of function" should be hyphenated because it is a compound adjective that modifies mutations. 

Line 50, "genetically-tractable" should not be hyphenated because it is not a compound adjective. It is an adverb-adjective pair. Line 102 has the same grammatical issue. 

Line 85, "rare genetic diseases" do not "affect nervous system function". The disease might have deficits in this function, but the disease does not do anything to function. 

Line 86, it should be mutations not mutants. Mutations are changes to DNA. Mutants are individuals with mutations. 

Throughout, wild-type should be hyphenated when it is used as a compound adjective. 

Figure 4, asterisks is spelled incorrectly. 

Reviewer #3 (Recommendations For The Authors): 

- As stated in the public review, the utility of the study is limited by the lack of access to the complete dataset. The wealth of data produced by the study is one of its major outputs. 

We have made the data publicly available on Zenodo. We appreciate the request.

- Describe the exact break-points of the different alleles, because it was not readily feasible to derive them from the gene fact sheets provided in the supplementary materials. 

We have now provided the start position and total length of deletion for each gene in the gene fact sheets.

- Figure 1C: what does "Genetic homology"/"sequence identity" refer to? How were these values calculated? 

UNC-49 is clearly not 95% identical to vertebrate GABAR subunits at the protein level. 

We have changed the axis label to “BLAST % Sequence Identity” to clarify that these values are calculated from BLAST sequence alignments on WormBase and the Alliance Genom Resources webpages.

- Figure 1E : The data presented in Figure 1E appears somewhat unreliable. For example, a cursory check showed: 

(1) Wrong human ortholog: unc-49 is a Gaba receptor, not a Glycine receptor as indicated in the second column. 

(2) Wrong disease association: dys-1 is not associated with Bardet-Biedl syndrome; overall the data indicated in the table does not seem to fully match the HPO database. 

(3) Inconsistent disease association: why don't the avr-14 and glc-2 (and even unc-49) profiles overlap/coincide given that they present overlapping sets of human orthologs. 

Thank you for catching this! We have corrected gene names which were mistakenly pasted. We have also made this a standalone table (Table 1) for improved readability.

- Error in legend to figure 4I : "with ciliopathies and N2" > ciliopathies should be "NALCN disease". 

- Error at line 301: "Figures 2E-H" should be "Figures 4E-H". 

Corrected.

departure from tuck and yang

Important to realize that if referring to a model with auxiliary view, focus on the top and front views to get accurate dimensions. Auxiliary is not true to its sizing

Welcome back and in this video I want to cover the differences between stateful and stateless firewalls.

And to do that I need to refresh your knowledge of how TCP and IP function.

So let's just jump in and get started.

In the networking fundamentals videos I talk about how TCP and IP worked together.

You might already know this if you have networking experience in the real world, but when you make a connection using TCP, what's actually happening is that each side is sending IP packets to each other.

These IP packets have a source and destination IP and are carried across local networks and the public internet.

Now TCP is a layer 4 protocol which runs on top of IP.

It adds error correction together with the idea of ports, so HTTP runs on TCP port 80 and HTTPS runs on TCP port 443 and so on.

So keep that in mind as we continue talking about the state of connections.

So let's say that we have a user here on the left Bob and he's connecting to the Categoram application running on a server on the right.

What most people imagine in this scenario is a single connection between Bob's laptop and the server.

So Bob's connecting to TCP port 443 on the server and in doing so he gets information back, in this case many different categories.

Now you know that below the surface at layer 3 this single connection is handled by exchanging packets between the source and the destination.

Conceptually though you can imagine that each connection, in this case it's an outgoing connection from Bob's laptop to the server.

Each one of these is actually made up of two different parts.

First we've got the request part where the client requests some information from a server, in this case from Categors, and then we have the response part where that data is returned to the client.

Now these are both parts of the same interaction between the client and server, but strictly speaking you can think of these as two different components.

What actually happens as part of this connection setup is this.

First the client picks a temporary port and this is known as an ephemeral port.

Now typically this port has a value between 1024 and 65535, but this range is dependent on the operating system which Bob's laptop is using.

Then once this ephemeral port is chosen the client initiates a connection to the server using a well-known port number.

Now a well-known port number is a port number which is typically associated with one specific popular application or protocol.

In this case TCP port 443 is HTTPS.

So this is the request part of the connection, it's a stream of data to the server.

You're asking for something, some cat pictures or a web page.

Next the server responds back with the actual data.

The server connects back to the source IP of the request part, in this case Bob's laptop, and it connects to the source port of the request part, which is the ephemeral port which Bob's laptop has chosen.

This part is known as the response.

So the request is from Bob's laptop using an ephemeral port to a server using a well-known port.

The response is from the server on that well-known port to Bob's laptop on the ephemeral port.

Now it's these values which uniquely identify a single connection.

So that's a source port and source IP.

And a destination IP and a destination port.

Now hope that this makes sense so far, if not then you need to repeat this first part of the video again because this is really important to understand.

If it does make sense then let's carry on.

Now let's look at this example in a little bit more detail.

This is the same connection that we looked at on the previous screen.

We have Bob's laptop on the left and the Catering Server on the right.

Obviously the left is the client and the right is the server.

I also introduced the correct terms on the previous screen so request and response.

So the first part is the client talking to the server asking for something and that's the request.

And the second part is the server responding and that's the response.

But what I want to get you used to is that the directionality depends on your perspective and let me explain what I mean.

So in this case the client initiates the request and I've added the IP addresses on here for both the client and the server.

So what this means is the packets will be sent from the client to the server and these will be flowing from left to right.

These packets are going to have a source IP address of 119.18.36.73, which is the IP address of the client.

So Bob's laptop and they will have a destination IP of 1.3.3.7, which is the IP address of the server.

Now the source port will be a temporary or ephemeral port chosen by the client and the destination port will be a well-known port.

In this case we're using HTTPS so TCP port 443.

Now if I challenge you to take a quick guess, would you say that this request is outbound or inbound?

If you had to pick, if you had to define a firewall rule right now, would you pick inbound or outbound?

Well this is actually a trick question because it's both.

From the client perspective this request is an outbound connection.

So if you're adding a firewall rule on the client you would be looking to allow or deny an outbound connection.

From the server perspective though it's an inbound connection so you have to think about perspective when you're working with firewalls.

But then we have the response part from the server through to the client.

This will also be a collection of packets moving from right to left.

This time the source IP on those packets will be 1.3.3.7, which is the IP address of the server.

The destination IP will be 119.18.36.73, which is the IP address of the client.

So Bob's laptop.

The source port will be TCP port 443, which is the well-known port of HTTPS and the destination port will be the ephemeral port chosen originally by the client.

Now again I want you to think about the directionality of this component of the communication.

Is it outbound or inbound?

Well again it depends on perspective.

The server sees it as an outbound connection from the server to the client and the client sees it as an inbound connection from the server to itself.

Now this is really important because there are two things to think about when dealing with firewall rules.

The first is that each connection between a client and a server has two components, the request and the response.

So the request is from a client to a server and the response is from a server to a client.

The response is always the inverse direction to the request.

But the direction of the request isn't always outbound and isn't always inbound.

It depends on what that data is together with your perspective.

And that's what I want to talk about a bit more on the next screen.

Let's look at this more complex example.

We still have Bob and his laptop on the CaterGram server, but now we have a software update server on the bottom left.

Now the CaterGram server is inside a subnet which is protected by a firewall.

And specifically this is a stateless firewall.

A stateless firewall means that it doesn't understand the state of connections.

What this means is that it sees the request connection from Bob's laptop to CaterGram and the response of from CaterGram to Bob's laptop as two individual parts.

You need to think about allowing or denying them as two parts.

You need two rules.

In this case one inbound rule which is the request and one outbound rule for the response.

This is obviously more management overhead.

Two rules needed for each thing.

Each thing which you as a human see as one connection.

But it gets slightly more confusing than that.

For connections to the CaterGram server, so for example when Bob's laptop is making a request, then that request is inbound to the CaterGram server.

The response logically enough is outbound, sending data back to Bob's laptop, which is possible to have the inverse.

Consider the situation where the CaterGram server is performing software updates.

Well in this situation the request will be from the CaterGram server to the software update server, so outbound, and the response will be from the software update server to the CaterGram server, so this is inbound.

So when you're thinking about this, start with the request.

Is the request coming to you or going to somewhere else?

The response will always be in the reverse direction.

So this situation also requires two firewall rules.

One outbound for the request and one inbound for the response.

Now there are two really important points I want to make about stateless firewalls.

First, for any servers where they accept connections and where they initiate connections, and this is common with web servers which need to accept connections from clients, but where they also need to do software updates.

In this situation you'll have to deal with two rules for each of these, and they will need to be the inverse of each other.

So get used to thinking that outbound rules can be both the request and the response, and inbound rules can also be the request and the response.

It's initially confusing, but just remember, start by determining the direction of the request, and then always keep in mind that with stateless firewalls you're going to need an inverse rule for the response.

Now the second important thing is that the request component is always going to be to a well-known port.

If you're managing the firewall for the category application, you'll need to allow connections to TCP port 443.

The response though is always from the server to a client, but this always uses a random ephemeral port, because the firewall is stateless, it has no way of knowing which specific port is used for the response, so you'll often have to allow the full range of ephemeral ports to any destination.

This makes security engineers uneasy, which is why stateless firewalls which I'll be talking about next are much better.

Just focus on these two key elements, that every connection has a request and a response, and together with those keep in mind the fact that they can both be in either direction, so a request can be inbound or outbound, and a response will always be the inverse to the directionality of the request.

Also you'll keep in mind that any rules that you create for the response will need to often allow the full range of ephemeral ports.

That's not a problem with stateless firewalls which I want to cover next.

So we're going to use the same architecture, we've got Bob's laptop on the top left, the category server on the middle right, and the software update server on the bottom left.

A stateless firewall is intelligent enough to identify the response for a given request, since the ports and IPs are the same, it can link one to the other, and this means that for a specific request to category from Bob's laptop to the server, the firewall automatically knows which data is the response, and the same is true for software updates, for a given connection to a software update server, the request, the firewall is smart enough to be able to see the response or the return data from the software update server back to the category server, and this means that with a stateful firewall, you'll generally only have to allow the request or not, and the response will be allowed or not automatically.

This significantly reduces the admin overhead and the chance for mistakes, because you just have to think in terms of the directionality and the IPs and ports of the request, and it handles everything else.

In addition, you don't need to allow the full ephemeral port range, because the firewall can identify which port is being used, and implicitly allow it based on it being the response to a request that you allow.

Okay, so that's how stateless and stateful firewalls work, and now it's been a little bit abstract, but this has been intentional, because I want you to understand how they work, and sexually, before I go into more detail with regards to how AWS implements both of these different security firewall standards.

Now at this point, I've finished with the abstract description, so go ahead and finish this video, and when you're ready, I'll look forward to you joining me in the next.

I thought this was interesting because in Octavia Butler's Parable of the Sower there was this sense of "denial and desiring." I interpreted this in a way that Lauren desired to fit in with the world around her but also despised that she had to change herself to fit in. This was very distinct to me in Lauren's world because when she had to leave her homeland, one of the first things she did was cut her hair to make herself resemble a man. The reason for this is that she knows in society she will be more feared if she presents herself as a man. We see throughout the novel how she suffers with her own femininity because of it. Where she finds being a woman sad a disadvantage. It's interesting to me how Frazier was able to point out this complex idea that both Butler and Mutu portray in their works.

Would you be willing to provide the code you used for this project as a GitHub repo or a gist?

It would be interesting to see a side-by-side comparison to PDB and ViralZone. Where does BFDV have more coverage? Where does ViralZone or PDB have more depth (more sequences per represented sequence in BFDV)

As commented above, it would be interesting to know more details on how you did this splitting

not sure what to make of this statement...

*

hmmm

marketization is subsumed into a creative solution (another aspect of problem-solving!)

for creative entrepreneurs, the creative tends to come first!

yep

very interesting How do you relate to this?

huh

ok - this statement is, on the other hand, exactly like Peter's. creativity = problem solving. A curious definition. But it also opens the door to allow everybody in ... (independent of entrepreneurial labels...)

!

we all exist as (relative) experts within some domain/field or other, acting creatively within it (with others) -- not just folk in the creative industry!

(it's just that they have this specific focus...)

quite a distinct outlook from Peter's point of view! Which do you find more persuasive?

the arbitrariness of it all! Purposeful entrepreneurial identity nowhere near as significant as the serendipity of retroactively putting the pieces together...

interesting

getting into the different case studies

...

So, can we speak of all entrepreneurs as creative entrepreneurs, not just those employed in creative fields (much like the debate about "cultural" entrepreneurs)?

this is fascinating to me.

surprise and randomness are not factors to be expunged but in fact are integral to the discovery process

what about the "creative" entrepreneur -- who might use knowledge, but even moreso, perhaps intuition... and inspiration. Perhaps knowledge is not so much in the head but in the gut, a feeling, an emotional need that must be met, a chord that must be struck...

I'm digging this too...

it reminds me of the statement that genius is nothing more nor less than doing well what anyone can do badly.

an interesting concept...

but it makes me wonder if this somehow constrains the potential of individuals to CREATIVELY seek and create opportunities where others couldn't see them... hmmm

*

a different factor for creative entrepreneurs!

the conundrum...

interesting

nice overview

!

*

creativity has to come from somewhere (people interact with and are influenced by their environment!)

*

it's a broad field! -- note the use of the term "autotelic" -- see my "journal prompts" for further explanation of this!

Yep!

all of this seems self-evident...

the only constant is change...

yep - this is basically how the other article concluded. This one provides actual narratives...

ok - that seems to fit with the overall arc of this course material for sure!

ok...

Just like (traditional) discourses surrounding entrepreneurship!

focusing specifically upon the 2nd type of cultural entrepreneur (as identified by the other article, which was really just trying to outline the importance of the concept and understanding the implications of how we use the terms...)

justifying the narrative-based approach

interesting - kind of the antithesis of the opening argument of this course...

I use the term "connective tissue" all the time!

ooh! -- on the road to turning creativity into a commodity...

what's your opinion of this? Does the existence of a tribe change this?

hah - relates to one of my seminar prompts... Funny enough, though, there's the contrary perspective that some world famous artists (like Damien Hirst in the world of contemporary art or Drake (!) a musical artist - pictured below, together for an article titled "Damien Hirst Simply Cannot Stop Painting Things for Drake") are commercially successful AND artistic innovators...

...

View 2: people engaged in the "arts" do it ... providing cultural good and services

ok - so not anyone can change people's beliefs ... but the best examples of cultural entrepreneurs are those who change society's belief structures...

View 1: everyone engages in culture!

I like this lingo. T'is true - the artistic innovation sphere might hardly ever be highlighted in "conventional" entrepreneurship...

I like this!

culture (and business) being revolutionized alongside each other. But seriously, Henry Ford changed culture even if that's not what we predominantly associate with him

the expertise of the cultural entrepreneur goes beyond possessing cultural capital - but affecting how we perceive cultural capital, how we negotiate its contours ... interesting...

I like this distinction, though, as it clarifies that everyone can be a cultural entrepreneur, as long as you're working "creatively"

interesting - cultural entrepreneurs as those who can change the beliefs of others...

the commercial aspect is never (entirely) absent...

this is an interesting definition (also circular in nature) -- the cultural entrepreneur will exhibit all the best qualities of someone undertaking a career in cultural sectors...

so we've come full circle - a cultural entrepreneur CAN be someone working in "industry" who has a profound impact on the wider culture (not just someone working in the "cultural industry/ies")

much more basic... a cultural entrepreneur produces a cultural good...

hah

not a circular argument at all!

interesting - a values (morality) based argument for entrepreneurship!

interesting - this might also serve as an inclusive definition of entrepreneur (without the "cultural" modifier). But people in the arts definitely highlight the cultural (and maybe the social) value of their creations, while also recognizing the economic nature of their practices...

yep!

seems like something of a theme in a lot of the weeks we've looked at so far!

interesting ... what makes us "cultured" or "cultivated"?

creativity --> creation --> culture --> cultivation --> GROWTH

ok ... an industrious person doesn't necessarily work in an industry, but labours to create ... and cultural industries create all manner of material (that may "elevate" their citizens...)

ok - so let's explore these different directions in greater depth...