Check this out! It is interesting and a good place to start the Guide activity in class today...

Yes! I could see doing a study about the value of contract grading for resolving issues with misuse of AI...

personal evidence from a real source/informant (ethos)

male bias towards AI use...(not what I've found among teachers...) What do we think about male AI voices vs. female?

Yes! This is an important distinction

I wonder if prompted by their teachers or on their own initiative/exploration... probably the latter..

the most important voice, since educators should be here to teach...many in higher ed are here to research (and supposed to teach, but don't very well...)

You have to know your audience before interacting with a group of people

Cultural expectations can differ depending on the context and audience that you are presenting your information to

for - definition - environmental steering - interlevel communications via environmental steering - interlevel control - interlevel communications

this is good sentence and explains that although prehistoric groups were usually small and isolated. they occasionally gathered with other groups. these gatherings likely happened during seasonal festivals. such events were important for sharing information and cultural knowledge strengthening social connections, and helping younger members interact beyond their small community.

Grammar in discourse focuses on structure and clarity, much like consistent layouts, typography, and design systems help users understand interfaces easily. Logic deals with reasoning and flow, which mirrors how UX designers build intuitive navigation and logical information hierarchies that guide users smoothly from one step to the next. Finally, rhetoric aims to persuade and appeal to emotion, similar to how persuasive visuals, tone, and storytelling in UX encourage user engagement or action. In this way, the principles of classical discourse continue to shape how we communicate meaning effectively through modern digital design.

UX Designers are very similar to rhetoricians as they are both working on getting a message/persuading their audience/users towards a certain goal. UX Designers must think about what might interest/appeal to a userto get them to click a certain area or link.

eLife Assessment

This manuscript presents an in-depth analysis of gene expression across multiple brown algal species with differing life histories, providing convincing evidence for the conservation of life cycle-specific gene expression. While largely descriptive, the study is an important step forward in understanding the core cellular processes that differ between life cycle phases, and its findings will be of broad interest to developmental and evolutionary biologists.

Reviewer #2 (Public review):

Summary:

The manuscript by Ratchinski et al presents a comprehensive analysis of developmental and life history gene expression patterns in brown algal species. The manuscript shows that the degree of generation bias or generation-specific gene expression correlates with the degree of dimorphism. It also reports conservation of life cycle features within generations and marked changes in gene expression patterns in Ectocarpus in the transition between gamete and early sporophyte. The manuscript also reports considerable conservation of gene expression modules between two representative species, particularly in genes associated with conserved functional characteristics.

Strengths:

The manuscript represents a considerable "tour de force" dataset and analytical effort. While the data presented is largely descriptive, it is likely to provide a very useful resource for studies of brown algal development and for comparative studies with other developmental and life cycle systems.

Comments on revisions

The authors have provided in their response (point 1) a good clarification for their rationale in excluding fucoid algae from the study, based on the diploid nature of the fucoid life cycle. Similarly, they have noted (point 2) that "the relationship between changes in gene expression during very early sporophyte development and during alternation of life cycle generations could be investigated further using a highlydimorphic kelp model system such as Saccharina latissima." For the benefit of the reader who may not be too familiar with the different life cycles in brown algae, I would recommend that these clarifications are included in the Discussion.

Otherwise the authors have addressed my previous comments adequately.

for - like - Michael Levins - Richard Sutton - youtube interview

Summary - interesting talk on learning - reminds me of Michael Levin's work - the priority is on goal directed activity

I think mixed-race people don't have boundaries to have a definition/to exist such as an identity group, but there's a social truth to it.

Both can be true. The common ground can be shared, but the experience, the identity is personal, individual.

There's also a white perspective on that.

I assume by passing the embeddings through the embeddings parameter.

The default method.

“Gene track” refers to the ‘GENCODE V48’ track, that should be located at the top of your screen. This is the track influenced by your GENCODE V48 selection down at the “genes and gene predictions” settings.

A lot of students get very overwhelmed with the tracks and don't know what to do

This paragraph can be confusing for some students. Here is a revision that potentially is easier to follow:

When studying a disease-associated SNP found in a GWAS, researchers look at the surrounding genomic region to identify possible disease related genes. This involves examining nearby genes and regulatory elements to see if they could play a role in the disease or trait. A typical GWAS only tests a subset of all SNPs in the genome, so the SNP that shows the strongest association is usually not the actual cause. Instead, it lies very close to the true causal variant. Because SNPs that are near each other on the genome tend to be inherited together, the associated SNP acts as a marker for the real causal variant.

That was the most interesting part, and the one less “common”.

But, and maybe it's philosophical or even a metaphysical thought, can things be essentially true without having a social influence? Like even math is based on theories, we say 1+1=2 because it fits, but it's a theory. So real definitions are also based on verbal agreements. (It's more a questioning about the definition of these definitions, I get the difference and how it applies to game study.)

// transcultural perspective?

“how we perceive things” // when we create somethings, those things are representative of our biases, we like it or not.

But at this time, sex between men took place everywhere else because the model of the bourgeois family had not become hegemonic yet.

Becoming a monk becomes a way to explore feminine and androgynous gender expression which wouldn't be possible anywhere else in the society

it wasn’t common anymore to see two men sharing a bed while during Middle-Ages laborers who were working at the same place could easily share a bed and have sex.

Between 1536 and 1541, by which Henry VIII disbanded all Catholic monasteries, priories, convents, and friaries in England, Wales, and Ireland; => seized their wealth; disposed of their assets; destroyed buildings and relics; dispersed or destroyed libraries; and provided for their former personnel and functions. => At the time of the Reformation, monasteries were some of the most egregious examples of catholic materialistic excess to be found. During the counter-reformation, monastic orders were then some of the most reactionary enemies of Protestantism. This led to a general antipathy towards the institution as a whole.

Appears as a stigma reversal in the 19th Century (while homosexuality was confused with the sin of sodomy).

Members of the Society of Jesus make profession of "perpetual poverty, chastity, and obedience" and "promise a special obedience to the sovereign pontiff in regard to the missions." A Jesuit is expected to be totally available and obedient to his superiors, accepting orders to go anywhere in the world, even if required to live in extreme conditions (wikipedia) => a huge engagement, especially in an english context where Catholics are a minority among the believers

eLife Assessment

In this preregistered study, Kunkel and colleagues set out to compare the magnitude and duration of placebo versus nocebo effects in healthy volunteers, and also to examine the different factors contributing to these effects. The authors follow a rigorous methodology in a within-subjects design, taking into consideration standard conventions for manipulation of expectations, and using an appropriate sham condition. They present compelling evidence of long-lasting placebo and nocebo effects, with nocebo responses demonstrating consistently greater strength. These valuable results have the potential for a great impact in the field of experimental and clinical pain.

Reviewer #1 (Public review):

Summary:

The study aimed to: (1) assess the magnitude of placebo and nocebo effects immediately after induction through verbal instructions and conditioning, (2) examine the persistence of these effects one week later, and (3) identify predictors of sustained placebo and nocebo responses over time.

Strengths:

An innovation was to use sham TENS stimulation as the expectation manipulation. This expectation manipulation was reinforced not only by the change in pain stimulus intensity, but also by delivery of non-painful electrical stimulation, labelled as TENS stimulation.

Questionnaire-based treatment expectation ratings were collected before conditioning and after conditioning, and after the test session, which provided an explicit measure of participant's expectations about the manipulation.

The finding that placebo and nocebo effects are influenced by recent experience provides a novel insight into a potential moderator of individual placebo effects.

Weaknesses:

There are a limited number of trials per test condition (10) which means that the trajectory of responses to the manipulation may not be explored, which would be an interesting future study.

The differences between the nocebo and control condition in pain ratings during conditioning could be explained by differing physiological effects of the different stimulus intensities, so it is difficult to make any claims about the expectation effects here. A a randomisation error meant that 25 participants received an unbalanced number 448 of trials per condition (i.e., 10 x VAS 40, 14 x VAS 60, 12 x VAS 80), although the authors accounted for this during analysis so it is not of major concern.

This manuscript presents a study on expectation manipulation to induce placebo and nocebo effects in healthy participants. The study follows standard placebo experiment conventions with use of TENS stimulation as the placebo manipulation. The authors were able to achieve their aims. A key finding is that placebo and nocebo effects were predicted by recent experience, which is a novel contribution to the literature. The findings provide insights into the differences between placebo and nocebo effects and the potential moderators of these effects.

Comments on revisions:

I am satisfied with the author's revisions to the manuscript and have no further comments.

Reviewer #2 (Public review):

Summary:

Kunkel et al aim to answer a fundamental question: Do placebo and nocebo effects differ in magnitude or longevity? To address this question, they used a powerful within-participants design, with a very large sample size (n=104), in which they compared placebo and nocebo effects - within the same individuals - across verbal expectations, conditioning, testing phase, and a 1-week follow-up. With elegant analyses, they establish that different mechanisms underlie the learning of placebo vs nocebo effects, with the latter being acquired faster and extinguished slower. This is an important finding for both the basic understanding of learning mechanisms in humans and for potential clinical applications to improve human health.

Strengths:

Beyond the above - the paper is well-written and very clear. It lays out nicely the need for the current investigation and what implications it holds. The design is elegant, and the analyses are rich, thoughtful, and interesting. The sample size is large which is highly appreciated, considering the longitudinal, in-lab study design. The question is super important and well-investigated, and the entire manuscript is very thoughtful with analyses closely examining the underlying mechanisms of placebo versus nocebo effects.

Comments on revisions:

The authors have addressed all of my concerns and comments - one point for them to verify is that indeed analyses that have not been preregistered will be flagged as such. The provided pre-registration link doesn't specify much about the analysis plans and specific tests used.

Author response:

The following is the authors’ response to the original reviews

Public Reviews:

Reviewer #1 (Public review):

Summary:

This manuscript presents a study on expectation manipulation to induce placebo and nocebo effects in healthy participants. The study follows standard placebo experiment conventions with the use of TENS stimulation as the placebo manipulation. The authors were able to achieve their aims. A key finding is that placebo and nocebo effects were predicted by recent experience, which is a novel contribution to the literature. The findings provide insights into the differences between placebo and nocebo effects and the potential moderators of these effects.

Specifically, the study aimed to:

(1) assess the magnitude of placebo and nocebo effects immediately after induction through verbal instructions and conditioning

(2) examine the persistence of these effects one week later, and

(3) identify predictors of sustained placebo and nocebo responses over time.

Strengths:

An innovation was to use sham TENS stimulation as the expectation manipulation. This expectation manipulation was reinforced not only by the change in pain stimulus intensity, but also by delivery of non-painful electrical stimulation, labelled as TENS stimulation.

Questionnaire-based treatment expectation ratings were collected before conditioning and after conditioning, and after the test session, which provided an explicit measure of participants' expectations about the manipulation.

The finding that placebo and nocebo effects are influenced by recent experience provides a novel insight into a potential moderator of individual placebo effects.

We thank the reviewer for their thorough evaluation of our manuscript and for highlighting the novelty and originality of our study.

Weaknesses:

There are a limited number of trials per test condition (10), which means that the trajectory of responses to the manipulation may not be adequately explored.

We appreciate the reviewer’s comment regarding the number of trials in the test phase. The trial number was chosen to ensure comparability with previous studies addressing similar research questions with similar designs (e.g. Colloca et al., 2010). Our primary objective was to directly compare placebo and nocebo effects within a within-subject design and to examine their persistence one week after the first test session. While we did not specifically aim to investigate the trajectory of responses within a single testing session, we fully agree that a comprehensive analysis of the trajectories of expectation effects on pain would be a valuable extension of our work. We have now acknowledged this limitation and future direction in the revised manuscript.

The paragraph reads as follows: “It is important to note that our study was designed in alignment with previous studies addressing similar questions (e.g., Colloca et al., 2010). Our primary aim was to directly compare placebo and nocebo effects in a within-subject design and assess their persistence of these effects one week following the first test session. One limitation of our approach is the relatively short duration of each session, which may have limited our ability to examine the trajectory of responses within a single session. Future studies could address this limitation by increasing the number of trials for a more comprehensive analysis.”

On day 8, one stimulus per stimulation intensity (i.e., VAS 40, 60, and 80) was applied before the start of the test session to re-familiarise participants with the thermal stimulation. There is a potential risk of revealing the manipulation to participants during the re-familiarization process, as they were not previously briefed to expect the painful stimulus intensity to vary without the application of sham TENS stimulation.

We thank the reviewer for the opportunity to clarify this point. Participants were informed at the beginning of the experiment that we would use different stimulation intensities to re-familiarize them with the stimuli before the second test session. We are therefore confident that participants perceived this step as part of a recalibration rather than associating it with the experimental manipulation. We have added this information to the revised version of the manuscript.

The paragraph now reads as follows: “On day 8, one stimulus per stimulation intensity (i.e., VAS 40, 60 and 80) was applied before the start of the test session to re-familiarise participants with the thermal stimulation. Note that participants were informed that these pre-test stimuli were part of the recalibration and refamiliarization procedure conducted prior to the second test session.”

The differences between the nocebo and control conditions in pain ratings during conditioning could be explained by the differing physiological effects of the different stimulus intensities, so it is difficult to make any claims about expectation effects here.

We appreciate the reviewer’s comment and agree that, despite the careful calibration of the three pain stimuli, we cannot entirely rule out the possibility that temporal dynamics during the conditioning session were influenced by differential physiological effects of the varying stimulus intensities (e.g., intensity-dependent habituation or sensitization). We have addressed this in the revision of the manuscript, but we would like to emphasize that the stronger nocebo effects during the test phase are statistically controlled for any differences in the conditioning session.

The paragraph now reads: “This asymmetry is noteworthy in and of itself because it occurred despite the equidistant stimulus calibration relative to the control condition prior to conditioning. It may be the result of different physiological effects of the stimuli over time or amplified learning in the nocebo condition, consistent with its heightened biological relevance, but it could also be a stronger effect of the verbal instructions in this condition.”

A randomisation error meant that 25 participants received an unbalanced number of 448 trials per condition (i.e., 10 x VAS 40, 14 x VAS 60, 12 x VAS 80).

We agree that this is indeed unfortunate. However, we would like to point out that all analyses reported in the manuscript have been controlled for the VAS ratings in the conditioning session, i.e., potential effects of the conditioned placebo and nocebo stimuli. Moreover, we have now conducted additional analyses, presented here in our response to the reviewers, to demonstrate that this imbalance did not systematically bias the results. Importantly, the key findings observed during the test phase remain robust despite this issue.

Specifically, when excluding these 25 participants from the analyses, the reported stronger nocebo compared to placebo effects in the test session on day 1 remain unchanged. Likewise, the comparison of placebo and nocebo effects between days 1 and 8 shows the same pattern when excluding the participants in question. The only exception is the interaction between effect (placebo vs nocebo) x session (day 1 vs day 8), which changed from a borderline significant result (p = .049) to insignificant (p = .24). However, post hoc tests continued to show the same pattern as originally reported: a significant reduction in the nocebo effect from day 1 to day 8 and no significant change in the placebo effect.

Reviewer #2 (Public review):

Summary:

Kunkel et al aim to answer a fundamental question: Do placebo and nocebo effects differ in magnitude or longevity? To address this question, they used a powerful within-participants design, with a very large sample size (n=104), in which they compared placebo and nocebo effects - within the same individuals - across verbal expectations, conditioning, testing phase, and a 1-week follow-up. With elegant analyses, they establish that different mechanisms underlie the learning of placebo vs nocebo effects, with the latter being acquired faster and extinguished slower. This is an important finding for both the basic understanding of learning mechanisms in humans and for potential clinical applications to improve human health.

Strengths:

Beyond the above - the paper is well-written and very clear. It lays out nicely the need for the current investigation and what implications it holds. The design is elegant, and the analyses are rich, thoughtful, and interesting. The sample size is large which is highly appreciated, considering the longitudinal, in-lab study design. The question is super important and well-investigated, and the entire manuscript is very thoughtful with analyses closely examining the underlying mechanisms of placebo versus nocebo effects.

We thank the reviewer for their positive evaluation of our manuscript and for acknowledging the methodological rigor and the significant implications for clinical applications and the broader research field.

Weaknesses:

There were two highly addressable weaknesses in my opinion:

(1) I could not find the preregistration - this is crucial to verify what analyses the authors have committed to prior to writing the manuscript. Please provide a link leading directly to the preregistration - searching for the specified number in the suggested website yielded no results.

We thank the reviewer for pointing this out. We included a link to the preregistration in the revised manuscript. This study was pre-registered with the German Clinical Trial Register (registration number: DRKS00029228; https://drks.de/search/de/trial/DRKS00029228).

(2) There is a recurring issue which is easy to address: because the Methods are located after the Results, many of the constructs used, analyses conducted, and even the main placebo and nocebo inductions are unclear, making it hard to appreciate the results in full. I recommend finding a way to detail at the beginning of the results section how placebo and nocebo effects have been induced. While my background means I am familiar with these methods, other readers will lack that knowledge. Even a short paragraph or a figure (like Figure 4) could help clarify the results substantially. For example, a significant portion of the results is devoted to the conditioning part of the experiment, while it is unknown which part was involved (e.g., were temperatures lowered/increased in all trials or only in the beginning).

We thank the reviewer for their helpful comment and agree that the Results section requires additional information that would typically be provided by the Methods section if it directly followed the Introduction. In response, we have moved the former Figure 4 from the Methods section to the beginning of the Results section as a new Figure 1, to improve clarity. Further, we have revised the Methods section to explicitly state that all trials during the conditioning phase were manipulated in the same way.

Recommendations for the Authors:

Reviewer #1 (Recommendations for the authors):

(1) Given that the authors are claiming (correctly) that there is only limited work comparing placebo/nocebo effects, there are some papers missing from their citations:

Nocebo responses are stronger than placebo responses after subliminal pain conditioning - - Jensen, K., Kirsch, I., Odmalm, S., Kaptchuk, T. J. & Ingvar, M. Classical conditioning of analgesic and hyperalgesic pain responses without conscious awareness. Proc. Natl. Acad. Sci. USA 112, 7863-7 (2015)

We thank the reviewer and have now included this relevant publication into the introduction of the revised manuscript.

Hird, E.J., Charalambous, C., El-Deredy, W. et al. Boundary effects of expectation in human pain perception. Sci Rep 9, 9443 (2019). https://doi.org/10.1038/s41598-019-45811-x

We thank the reviewer for suggesting this relevant publication. We have now included it into the discussion of the revised manuscript by adding the following paragraph:

“Recent work using a predictive coding framework further suggests that nocebo effects may be less susceptible to prediction error than placebo effects (Hird et al., 2019), which could contribute to their greater persistence and strength in our study.”

(2) The trial-by-trial pain ratings could have been usefully modelled with a computational model, such as a Bayesian model (this is especially pertinent given the reference to Bayesian processing in the discussion). A multilevel model could also be used to increase the power of the analysis. This is a tentative suggestion, as I appreciate it would require a significant investment of time and work - alternatively, the authors could acknowledge it in the Discussion as a useful future avenue for investigation, if this is preferred.

We thank the reviewer for this thoughtful suggestion. While we agree that computational modelling approaches could provide valuable insights into individual learning, our study was not designed with this in mind and the relatively small number of trials per condition and the absence of trial-by-trial expectancy ratings limit the applicability of such models. We have therefore chosen not to pursue such analysis but highlight it in the discussion as a promising direction for future research.

“Notably, the most recent experience was the most predictive in all three analyses; for instance, the placebo effect on day 8 was predicted by the placebo effect on day 1, not by the initial conditioning. This finding supports the Bayesian inference framework, where recent experiences are weighted more heavily in the process of model updating because they are more likely to reflect the current state of the environment, providing the most relevant and immediate information needed to guide future actions and predictions24. Interestingly, while a change in pain predicted subsequent nocebo effects, it seemed less influential than for placebo effects. This aligns with findings that longer conditioning enhanced placebo effects, while it did not affect nocebo responses10 and the conclusion that nocebo instruction may be sufficient to trigger nocebo responses. Using Bayesian modeling, future studies could identify individual differences in the development of placebo and nocebo effects by integrating prior experiences and sensory inputs, providing a probabilistic framework for understanding the underlying mechanisms.”

(3) The paper is missing any justification of sample size, i.e. power analysis - please include this.

We apologize for the missing information on our a priori power analysis. As there is a lack of prior studies investigating within-subjects comparisons of placebo and nocebo effects that could inform precise effect size estimates for our research question, we based our calculation on the ability detect small effects. Specifically, the study was powered to detect effect sizes in the range of d = 0.2 - 0.25 with α = .05 and power = .9, yielding a required sample size of N = 83-129. We have now added this information to the methods section of the revised manuscript.

(4) "On day 8, one stimulus per stimulation intensity (i.e., VAS 40, 60 and 80) was applied before the start of the test session to re-familiarise participants with the thermal stimulation."

What were the instructions about this? Was it before the electrode was applied? This runs the risk of unblinding participants, as they only expect to feel changes in stimulus intensity due to the TENS stimulation.

We thank the reviewer for pointing out the potential risk of unblinding participants due to the re-familiarization process prior to the second test session. We would like to clarify that we followed specific procedures to prevent participants from associating this process with the experimental manipulation. The re-familiarisation with the thermal stimuli was conducted after the electrode had been applied and re-tested to ensure that both stimulus modalities were re-introduced in a consistent and neutral context. Participants were explicitly informed that both procedures were standard checks prior to the actual test session (“We will check both once again before we begin the actual measurement.”). For the thermal stimuli, we informed participants that they would experience three different intensities to allow the skin to acclimate (e.g., “...we will test the heat stimuli in 3 trials with different temperatures, allowing your skin to acclimate to the stimuli. …”), without implying any connection to the experimental conditions.

Importantly, this re-familiarization procedure mirrored what participants had already experienced during the initial calibration session on day 1. We therefore assume that participants interpreted as a routine technical step rather than part of the experimental manipulation. We have now clarified this procedure in the methods section of the revised manuscript.

(5) "For a comparison of pain intensity ratings between time-points, an ANOVA with the within-subject factors Condition (placebo, nocebo, control) and Session (day 1, day 8) was carried out. For the comparison of placebo and nocebo effects between the two test days, an ANOVA with the with-subject factors Effect (placebo effect, nocebo effect) and Session (day 1, day 8) was used."

It seems that one ANOVA is looking at raw pain scores and one is looking at difference scores, but this is a bit confusing - please rephrase/clarify this, and explain why it is useful to include both.

We thank the reviewer for highlighting this point. Our primary analyses focus on placebo and nocebo effects, which we define as the difference in pain intensity ratings between the control and the placebo condition (placebo effect) and the nocebo and the control condition (nocebo effect), respectively.

To examine whether condition effects were present at each time-point, we first conducted two separate repeated measures ANOVAs - one for day 1 and one for day 8 - with the within-subject factor CONDITION (placebo, nocebo, control).

To compare the magnitude and persistence of placebo and nocebo effects over time, we then calculated the above-mentioned difference scores and submitted these to a second ANOVA with within-subject factors EFFECT (placebo vs. nocebo effect) and SESSION (day 1 vs. day 8). We have now clarified this approach on page 19 of the revised manuscript. To avoid confusion, the Condition x Session ANOVA has been removed from the manuscript.

(6) Please can the authors provide a figure illustrating trial-by-trial ratings during test trials as well as during conditioning trials?

In response to the reviewer’s point, we now provide the trial-by-trial ratings of the test phases on days 1 and 8 as an additional figure in the Supplement (Figure S1) and would like to clarify that trial-by-trial pain intensity ratings of the conditioning phase are displayed in Figure 2C of the manuscript,

(7) "Separate multiple linear regression analyses were performed to examine the influence of expectations (GEEE ratings) and experienced effects (VAS ratings) on subsequent placebo and nocebo effects. For day 1, the placebo effect was entered as the dependent variable and the following variables as potential predictors: (i) expected improvement with placebo before conditioning, (ii) placebo effect during conditioning and (iii) the expected improvement with placebo before the test session at day 1"

The term "placebo effect during conditioning" is a bit confusing - I believe this is just the effect of varying stimulus intensities - please could the authors be more explicit on the terminology they use to describe this? NB changes in pain rating during the conditioning trials do not count as a placebo/nocebo effect, as most of the change in rating will reflect differences in stimulation intensity.

We agree with the reviewer that the cited paragraph refers to the actual application of lower or higher pain stimuli during the conditioning session, rather than genuinely induced placebo or nocebo effect. We thank the reviewer for this helpful observation and have revised the terminology, accordingly, now referring to these as “pain relief during conditioning” and “pain worsening during conditioning”.

(8) Supplementary materials: "The three temperature levels were perceived as significantly different (VAS ratings; placebo condition: M= 32.90, SD= 16.17; nocebo condition: M= 56.62, SD= 17.09; control condition: M= 80.84, SD= 12.18"

This suggests that the VAS rating for the control condition was higher than for the nocebo condition. Please could the authors clarify/correct this?

We thank the reviewer for spotting this error. The values for the control and the nocebo condition had accidentally been swapped. This has now been corrected in the manuscript: control condition: M= 56.62, SD= 17.09; nocebo condition: M= 80.84, SD= 12.18.

(9) "To predict placebo responses a week later (VAScontrol - VASplacebo at day 8), the same independent variables were entered as for day 1 but with the following additional variables (i) the placebo effect at day 1 and (ii) the expected improvement with placebo before the test session at day 8."

Here it would be much clearer to say 'pain ratings during test trials at day 1".

We agree with the reviewer and have revised the manuscript as suggested.

(10) For completeness, please present the pain intensity ratings during conditioning as well as calibration/test trials in the figure.

Please see our answer to comment (6).

(11) In Figure 1a, it looks like some participants had rated the control condition as zero by day 8. If so, it's inappropriate to include these participants in the analysis if they are not responding to the stimulus. Were these the participants who were excluded due to pain insensitivity?

On day 8, the lowest pain intensity ratings observed were VAS 3 in the placebo condition and VAS 2 in the control condition, both from the same participant. All other participants reported minimum values of VAS 11 or higher (all on a scale from 0-100). Thus, no participant provided a pain rating of VAS 0, and all ratings indicated some level of pain perception in response to the stimulus. We did not define an exclusion criterion based on day 8 pain ratings in our preregistration, and we did not observe any technical issues with the stimulation procedure. To avoid post-hoc exclusions and maintain consistency with our preregistered analysis plan, we therefore decided to include all participants in the analysis.

(12) "Comparison of day 1 and day 8. A direct comparison of placebo and nocebo effects on day 1 and day 8 pain intensity ratings showed a main effect of Effect with a stronger nocebo effect (F(1,97)= 53.93, 131 p< .001, η2= .36) but no main effect of Day (F(1,97)= 2.94, p= .089, η2 = .029). The significant Effect x Session interaction indicated that the placebo effect and the nocebo effect developed differently over time (F(1,97)= 3.98, p= .049, η2 = .039)"

This is confusing as it talks about a main effect of "day" and then interaction with "session" - are they two different models? The authors need to clarify.

We thank the reviewer for pointing this out. In our analysis, “Session” is the correct term for the experimental factor, which has two factor levels, “day 1” and “day 8”. This has now been corrected in the revised manuscript.

Reviewer #2 (Recommendations for the authors):

(1) More information on how "size of the effect" in Figures 1b and 2b was calculated is needed; this can be in the legend. If these are differences between control and each condition, then they were reversed for one condition (nocebo?), which is ok - but this should be clearly explained.

We agree with the reviewer and have now revised the figure legends to improve clarity. The legends now read:

1b: “Figure 1. Pain intensity ratings and placebo and nocebo effects during calibration and test sessions. (A) Mean pain intensity ratings in the placebo, nocebo and control condition during calibration, and during the test sessions at day 1 and day 8. (B) Placebo effect (control condition - placebo condition, i.e., positive value of difference) and nocebo effect (nocebo condition - control condition, i.e., positive value of difference) on day 1 and day 8. Error bars indicate the standard error of the mean, circles indicate mean ratings of individual participants. *: p < .001, : p < .01, n.s.: non-significant.”

2b: “Figure 2. Mean and trial-by-trial pain intensity ratings, placebo and nocebo effects during conditioning. (A) Mean pain intensity ratings of the placebo, nocebo and control condition during conditioning. (B) Placebo effect (control condition - placebo condition, i.e., positive value of difference) and nocebo effect (nocebo condition - control condition, i.e., positive value of difference) during conditioning. (C) Trial-by-trial pain intensity ratings (with confidence intervals) during conditioning. Error bars indicate the standard error of the mean, circles indicate mean ratings of individual participants. ***: p < .001.”

(2) In the methods, I was missing a clear understanding of how many trials there were in the conditioning phase, and then how many in the other testing phases. Also, how long did the experiment last in total?

We apologize that the exact number of trials in the testing phases was not clear in the original manuscript. We now indicate on page 18 of the revised manuscript that we used 10 trials per condition in the test sessions. We have also added information on the duration of each test day (i.e., three hours on day 1 and one hour on day 8) on page 15.

(3) In expectancy ratings, line 186 - are improvement and worsening expectations different from expected pain relief? It is implied that these are two different constructs - it would be helpful to clarify that.

We agree that this is indeed confusing and would like to clarify that both refer to the same construct. We used the Generic rating scale for previous treatment experiences, treatment expectations, and treatment effects (GEEE questionnaire, Rief et al. 2021) that discriminates between expected symptom improvement, expected symptom worsening, and expected side effects due to a treatment. We now use the terms “expected pain relief” and “expected pain worsening” throughout the whole manuscript.

(4) In the last section of the Results, somatosensory amplification comes out of nowhere - and could be better introduced (see point 2 above).

We agree with the reviewer that introducing the concept of somatosensory amplification and its potential link to placebo/nocebo effects only in the Methods is unhelpful, given that this section appears at the end of the manuscript. We therefore now introduce the relevant publication (Doering et al., 2015) before reporting our findings on this concept.

(5) In line 169, if the authors want to specify what portion of the variance was explained by expectancy, they could conduct a hierarchical regression, where they first look at R2 without the expectancy entered, and only then enter it to obtain the R2 change.

We fully agree that hierarchical regression can be a useful approach for isolating the contribution of variables. However, in our case, expectancy was assessed at different time points (e.g., before conditioning and before the test session on day 1), and there was no principled rationale for determining the order in which these different expectancy-related variables should be entered into a hierarchical model.

That said, in response to the reviewer’s suggestion, we have now conducted hierarchical regression analyses in which all expectancy-related variables were entered together as a single block (see below). These analyses largely confirmed the findings reported so far and are provided here in the response to the reviewers below. Given the exploratory nature of this grouping and the lack of an a priori hierarchy, we feel that the standard multiple regression models remain the most appropriate for addressing our research question because it allows us to evaluate the total contribution of expectancy-related predictors while also examining the individual contribution of each variable within the block. We would therefore prefer to retain these as the primary analyses in the manuscript.

Results of the hierarchical regression analyses:

Day 1 - Placebo response: In step 1, we entered the difference in pain intensity ratings between the control and the placebo condition during conditioning as a predictor. In step 2, we added the two variables reflecting expectations (i.e., expected improvement with placebo (i) before conditioning and (ii) before the test session on day 1). This allowed us to assess whether expectation-related variables explained additional variance beyond the effect of conditioning.

The overall regression model at step 1 was significant, F(1, 102) = 13.42, p < .001, explaining 11.6% of the variance in the dependent variable (R² = .116). Adding the expectancy-related predictors in step 2 did not lead to a significant increase in explained variance, ΔR² = .007, F(2, 100) = 0.384, p = .682. Thus, the conditioning response significantly predicted placebo-related pain reduction on day 1, but additional information on expectations did not account for further variance.

Day 1 - Nocebo response: The equivalent analysis was run for the nocebo response on day 1. In step 1, the pain intensity difference between the nocebo and the control condition was entered as a predictor before adding the two expectancy ratings (i.e., expected worsening with nocebo (i) before conditioning and (ii) before the test session on day 1).

In step 1, the regression model was not statistically significant, F(1, 102) = 2.63, p = .108, and explained only 2.5% of the variance in nocebo response (R² = .025). Adding the expectation-related predictors in Step 2 slightly increased the explained variance by ΔR² = .027, but this change was also non-significant, F(2, 100) = 1.41, p = .250. The overall variance explained by the full model remained low (R² = .052). These results suggest that neither conditioning nor expectation-related variables reliably predicted nocebo-related pain increases on day 1.

Day 8 - Placebo response: For the prediction of the placebo effect on day 8, the following variables reflecting perceived effects were entered as predictors in step 1: the difference in pain intensity ratings between the control and the placebo condition (i) during conditioning and (ii) on day 1. In step 2, the variables reflecting expectations were added: the expected improvement with placebo (i) before conditioning, (ii) before the test session on day 1 and (iii) before the test session on day 8.

In step 1, the model was statistically significant, F(3, 95) = 14.86, p < .001, explaining 23.8% of the variance in the placebo response (R² = .238, Adjusted R² = .222). In step 2, the addition of the expectation-related predictors resulted in a non-significant improvement in model fit, ΔR² = .051, F(3, 92) = 2.21, p = .092. The overall variance explained by the full model increased modestly to 29.0%.

Day 8 - Nocebo response: For the equivalent analyses of nocebo responses on day 8, the following variables were included in step 1: the difference in pain intensity ratings between the nocebo and the control condition (i) during conditioning and (ii) on day 1. In step 2, we entered the variables reflecting nocebo expectations including expected worsening with nocebo (i) before conditioning, (ii) before the test session on day 1 and (iii) before the test session on day 8. In step 1, the model significantly predicted the day 8 nocebo response, F(3, 95) = 6.04, p = .003, accounting for 11.3% of the variance (R² = .113, Adjusted R² = .094). However, the addition of expectation-related predictors in Step 2 resulted in only a negligible and non-significant improvement, ΔR² = .006, F(3, 92) = 0.215, p = .886. The full model explained just 11.9% of the variance (R² = .119).

Typos:

(6) Abstract - 104 heathy xxx (word missing).

(7) Line 61 - reduce or decrease - I think you meant increase.

Thank you, we have now corrected both sentences.

References

Colloca L, Petrovic P, Wager TD, Ingvar M, Benedetti F. How the number of learning trials affects placebo and nocebo responses. Pain. 2010

Doering BK, Nestoriuc Y, Barsky AJ, Glaesmer H, Brähler E, Rief W. Is somatosensory amplification a risk factor for an increased report of side effects? Reference data from the German general population. J Psychosom Res. 2015

eLife Assessment

This work describes a highly complex automated algorithm for analyzing vascular imaging data from two-photon microscopy. This tool has the potential to be extremely valuable to the field and to fill gaps in knowledge of hemodynamic activity across a regional network. The solid biological application provides a demonstration of their pipeline's capabilities and suggests intriguing hypotheses around prolonged vascular tone changes, but will need to be followed up by further experiments to be conclusively demonstrated.

Reviewer #1 (Public Review):

Summary:

In this manuscript, the authors describe a new pipeline to measure changes in vasculature diameter upon opt-genetic stimulation of neurons.

The work is interesting and the topic is quite relevant to better understand the hemodynamic response on the graph/network level.

Strengths:

The manuscript provides a pipeline that allows for the detection of changes in the vessel diameter as well as simultaneously allowing for the location of the neurons driven by stimulation.

The resulting data could provide interesting insights into the graph-level mechanisms of regulating activity-dependent blood flow.

The interesting findings include that vessel radius changes depend on depth from the cortical surface and that dilations on average happen closer to the activated neurons.

Reviewer #2 (Public Review):

Summary:

The authors develop a highly detailed pipeline to analyze hemodynamic signals from in vivo two-photon fluorescence microscopy. This includes motion correction, segmentation of the vascular network, diameter measurements across time, mapping neuronal position relative to the vascular network, and analyzing vascular network properties (interactions between different vascular segments). For the segmentation, the authors use a Convolution Neural Network to identify vessel (or neural) and background pixels and train it using ground truth images based on semi-automated mapping followed by human correction/annotation. Considerable processing was done on the segmented images to improve accuracy, extract vessel center lines, and compute frame-by-frame diameters. The model was tested with artificial diameter increases and Gaussian noise and proved robust to these manipulations.

Network-level properties include Assortativity - a measure of how similar a vessel's response is to nearby vessels - and Efficiency - the ease of flow through the network (essentially, the combined resistance of a path based on diameter and vessel length between two points).

Strengths:

This is a very powerful tool for cerebral vascular biologists as many of these tasks are labor intensive, prone to subjectivity, and often not performed due to the complexity of collecting and managing volumes of vascular signals. Modelling is not my specialty so I cannot speak too specifically, but the model appears to be well-designed and robust to perturbations. It has many clever features for processing the data.

The authors rightly point out that there is a real lack in the field of knowledge of vascular network activity at single-vessel resolution. Network anatomy has been studied, but hemodynamics are typically studied either with coarse resolution or in only one or a few vessels at a time. This pipeline has the potential to change that.

[Editors' note: this work has been through three rounds of revisions, and most recently the authors have added caveats to the discussion. This version of the paper has been assessed by the editors and the weaknesses identified previously remain with earlier versions of the work.]

Author response:

The following is the authors’ response to the previous reviews

Reviewer #1 (Public review):

Summary:

In the manuscript the authors describe a new pipeline to measure changes in vasculature diameter upon optogenetic stimulation of neurons. The work is useful to better understand the hemodynamic response on a network /graph level.

Strengths:

The manuscript provides a pipeline that allows to detect changes in the vessel diameter as well as simultaneously allows to locate the neurons driven by stimulation.

The resulting data could provide interesting insights into the graph level mechanisms of regulating activity dependent blood flow.

Weaknesses:

(1) The manuscript contains (new) wrong statements and (still) wrong mathematical formulas.

The symbols in these formulas have been updated to disambiguate them, and the accompanying statements have been adjusted for clarity.

(2) The manuscript does not compare results to existing pipelines for vasculature segmentation (opensource or commercial). Comparing performance of the pipeline to a random forest classifier (illastik) on images that are not preprocessed (i.e. corrected for background etc.) seems not a particularly useful comparison.

We’ve now included comparisons to Imaris (a commercial) for segmentation and VesselVio (open-source) for graph extraction software.

For the ilastik comparison, the images were preprocessed prior to ilastik segmentation, specifically by doing intensity normalization.

Example segmentations utilizing Imaris have now been included. Imaris leaves gaps and discontinuities in the segmentation masks, as shown in Supplementary Figure 10. The Imaris segmentation masks also tend to be more circular in cross-section despite irregularities on the surface of the vessels observable in the raw data and identified in manual segmentation. This approach also requires days to months to generate per image stack.

A comparison to VesselVio has now also been generated, and results are visualized in Supplementary Figure 11. VesselVio generates individual graphs for each time point, resulting in potential discrepancies in the structure of the graphs from different time points. Furthermore, Vesselvio uses distance transformation to estimate the vascular radius, which renders the vessel radius estimates highly susceptible to variation in the user selected methodology used to obtain segmentation results; while our approach uses intensity gradient-based boundary detection from centerlines in the image instead mitigating this bias. We have added the following paragraph to the Discussion section on the comparisons with the two methods:

“Comparison with commercial and open-source vascular analysis pipelines

To compare our results with those achievable on these data with other pipelines for segmentation and graph network extraction, we compared segmentation results qualitatively with Imaris version 9.2.1 (Bitplane) and vascular graph extraction with VesselVio [1]. For the Imaris comparison, three small volumes were annotated by hand to label vessels. Example slices of the segmentation results are shown in Supplementary Figure 10. Imaris tended to either over- or under-segment vessels, disregard fine details of the vascular boundaries, and produce jagged edges in the vascular segmentation masks. In addition to these issues with segmentation mask quality, manual segmentation of a single volume took days for a rater to annotate. To compare to VesselVio, binary segmentation masks (one before and one after photostimulation) generated with our deep learning models were loaded into VesselVio for graph extraction, as VesselVio does not have its own method for generating segmentation masks. This also facilitates a direct comparison of the benefits of our graph extraction pipeline to VesselVio. Visualizations of the two graphs are shown in Supplementary Figure 11. Vesselvio produced many hairs at both time points, and the total number of segments varied considerably between the two sequential stacks: while the baseline scan resulted in 546 vessel segments, the second scan had 642 vessel segments. These discrepancies are difficult to resolve in post-processing and preclude a direct comparison of individual vessel segments across time. As the segmentation masks we used in graph extraction derive from the union of multiple time points, we could better trace the vasculature and identify more connections in our extracted graph. Furthermore, VesselVio relies on the distance transform of the user supplied segmentation mask to estimate vascular radii; consequently, these estimates are highly susceptible to variations in the input segmentation masks.We repeatedly saw slight variations between boundary placements of all of the models we utilized (ilastik, UNet, and UNETR) and those produced by raters. Our pipeline mitigates this segmentation method bias by using intensity gradient-based boundary detection from centerlines in the image (as opposed to using the distance transform of the segmentation mask, as in VesselVio).”

(3) The manuscript does not clearly visualize performance of the segmentation pipeline (e.g. via 2d sections, highlighting also errors etc.). Thus, it is unclear how good the pipeline is, under what conditions it fails or what kind of errors to expect.

On reviewer’s comment, 2D slices have been added in the Supplementary Figure 4.

(4) The pipeline is not fully open-source due to use of matlab. Also, the pipeline code was not made available during review contrary to the authors claims (the provided link did not lead to a repository). Thus, the utility of the pipeline was difficult to judge.

All code has been uploaded to Github and is available at the following location: https://github.com/AICONSlab/novas3d

The Matlab code for skeletonization is better at preserving centerline integrity during the pruning of hairs from centerlines than the currently available open-source methods.

- Generalizability: The authors addressed the point of generalizability by applying the pipeline to other data sets. This demonstrates that their pipeline can be applied to other data sets and makes it more useful.  However, from the visualizations it's unclear to see the performance of the pipeline, where the pipelines fails etc. The 3d visualizations are not particularly helpful in this respect . In addition, the dice measure seems quite low, indicating roughly 20-40% of voxels do not overlap between inferred and ground truth. I did not notice this high discrepancy earlier. A thorough discussion of the errors appearing in the segmentation pipeline would be necessary in my view to better assess the quality of the pipeline.

2D slices from the additional datasets have been added in the Supplementary Figure 13 to aid in visualizing the models’ ability to generalize to other datasets.

The dice range we report on (0.7-0.8) is good when compared to those (0.56-86) of 3D segmentations of large datasets in microscopy [2], [3], [4], [5], [6]. Furthermore, we had two additional raters segment three images from the original training set. We found that the raters had a mean inter class correlation  of 0.73 [7]. Our model outperformed this Dice score on unseen data: Dice scores from our generalizability tests on C57 mice and Fischer rats on par or higher than this baseline.

Reviewer #2 (Public review):

The authors have addressed most of my concerns sufficiently. There are still a few serious concerns I have. Primarily, the temporal resolution of the technique still makes me dubious about nearly all of the biological results. It is good that the authors have added some vessel diameter time courses generated by their model. But I still maintain that data sampling every 42 seconds - or even 21 seconds - is problematic. First, the evidence for long vascular responses is lacking. The authors cite several papers:

Alarcon-Martinez et al. 2020 show and explicitly state that their responses (stimulus-evoked) returned to baseline within 30 seconds. The responses to ischemia are long lasting but this is irrelevant to the current study using activated local neurons to drive vessel signals.

Mester et al. 2019 show responses that all seem to return to baseline by around 50 seconds post-stimulus.

In Mester et al. 2019, diffuse stimulations with blue light showed a return to baseline around 50 seconds post-stimulus (cf. Figure 1E,2C,2D). However, focal stimulations where the stimulation light is raster scanned over a small region focused in the field of view show longer-lasting responses (cf. Figure 4) that have not returned to baseline by 70 seconds post-stimulus [8]. Alarcon-Martinez et al. do report that their responses return baseline within 30 seconds; however, their physiological stimulation may lead to different neuronal and vessel response kinetics than those elicited by the optogenetic stimulations as in current work.

O'Herron et al. 2022 and Hartmann et al. 2021 use opsins expressed in vessel walls (not neurons as in the current study) and directly constrict vessels with light. So this is unrelated to neuronal activity-induced vascular signals in the current study.

We agree that optogenetic activation of vessel-associated cells is distinct from optogenetic activation of neurons, but we do expect the effects of such perturbations on the vasculature to have some commonalities.

There are other papers including Vazquez et al 2014 (PMID: 23761666) and Uhlirova et al 2016 (PMID: 27244241) and many others showing optogenetically-evoked neural activity drives vascular responses that return back to baseline within 30 seconds. The stimulation time and the cell types labeled may be different across these studies which can make a difference. But vascular responses lasting 300 seconds or more after a stimulus of a few seconds are just not common in the literature and so are very suspect - likely at least in part due to the limitations of the algorithm.

The photostimulation in Vazquez et al. 2014 used diffuse photostimulation with a fiberoptic probe similar to Mester et al. 2019 as opposed to raster scanning focal stimulation we used in this study and in the study by Mester et al. 2019  where we observed the focal photostimulation to elicited longer than a minute vascular responses. Uhlirova et al. 2016 used photostimulation powers between 0.7 and 2.8 mW, likely lower than our 4.3 mW/mm² photostimulation. Further, even with focal photostimulation, we do see light intensity dependence of the duration of the vascular responses. Indeed, in Supplementary Figure 2, 1.1 mW/mm² photostimulation leads to briefer dilations/constrictions than does 4.3 mW/mm²; the 1.1 mW/mm² responses are in line, duration wise, with those in Uhlirova et al. 2016.

Critically, as per Supplementary Figure 2, the analysis of the experimental recordings acquired at 3-second temporal resolution did likewise show responses in many vessels lasting for tens of seconds and even hundreds of seconds in some vessels.

Another major issue is that the time courses provided show that the same vessel constricts at certain points and dilates later. So where in the time course the data is sampled will have a major effect on the direction and amplitude of the vascular response. In fact, I could not find how the "response" window is calculated. Is it from the first volume collected after the stimulation - or an average of some number of volumes? But clearly down-sampling the provided data to 42 or even 21 second sampling will lead to problems. If the major benefit to the field is the full volume over large regions that the model can capture and describe, there needs to be a better way to capture the vessel diameter in a meaningful way.

In the main experiment (i.e. excluding the additional experiments presented in the Supplementary Figure 2 that were collected over a limited FOV at 3s per stack), we have collected one stack every 42 seconds. The first slice of the volume starts following the photostimulation, and the last slice finishes at 42 seconds. Each slice takes ~0.44 seconds to acquire. The data analysis pipeline (as demonstrated by the Supplementary Figure 2) is not in any way limited to data acquired at this temporal resolution and - provided reasonable signal-to-noise ratio (cf. Figure 5) - is applicable, as is, to data acquired at much higher sampling rates.

It still seems possible that if responses are bi-phasic, then depth dependencies of constrictors vs dilators may just be due to where in the response the data are being captured - maybe the constriction phase is captured in deeper planes of the volume and the dilation phase more superficially. This may also explain why nearly a third of vessels are not consistent across trials - if the direction the volume was acquired is different across trials, different phases of the response might be captured.

Alternatively, like neuronal responses to physiological stimuli, the vascular responses elicited by increases in neuronal activity may themselves be variable in both space and time.

I still have concerns about other aspects of the responses but these are less strong. Particularly, these bi-phasic responses are not something typically seen and I still maintain that constrictions are not common. The authors are right that some papers do show constriction. Leaving out the direct optogenetic constriction of vessels (O'Herron 2022 & Hartmann 2021), the Alarcon-Martinez et al. 2020 paper and others such as Gonzales et al 2020 (PMID: 33051294) show different capillary branches dilating and constricting. However, these are typically found either with spontaneous fluctuations or due to highly localized application of vasoactive compounds. I am not familiar with data showing activation of a large region of tissue - as in the current study - coupled with vessel constrictions in the same region. But as the authors point out, typically only a few vessels at a time are monitored so it is possible - even if this reviewer thinks it unlikely - that this effect is real and just hasn't been seen.

Uhlirova et al. 2016 (PMID: 27244241) observed biphasic responses in the same vessel with optogenetic stimulation in anesthetized and unanesthetized animals (cf Fig 1b and Fig 2, and section “OG stimulation of INs reproduces the biphasic arteriolar response”). Devor et al. (2007) and Lindvere et al. (2013) also reported on constrictions and dilations being elicited by sensory stimuli.

I also have concerns about the spatial resolution of the data. It looks like the data in Figure 7 and Supplementary Figure 7 have a resolution of about 1 micron/pixel. It isn't stated so I may be wrong. But detecting changes of less than 1 micron, especially given the noise of an in vivo prep (brain movement and so on), might just be noise in the model. This could also explain constrictions as just spurious outputs in the model's diameter estimation. The high variability in adjacent vessel segments seen in Figure 6C could also be explained the same way, since these also seem biologically and even physically unlikely.

Thank you for your comment. To address this important issue, we performed an additional validation experiment where we placed a special order of fluorescent beads with a known diameter of 7.32 ± 0.27um, imaged them following our imaging protocol, and subsequently used our pipeline to estimate their diameter. Our analysis converged on the manufacturer-specified diameters, estimating the diameter to be 7.34 ± 0.32. The manuscript has been updated to detail this experiment, as below:

Methods section insert

“Second, our boundary detection algorithm was used to estimate the diameters of fluorescent beads of a known radius imaged under similar acquisition parameters. Polystyrene microspheres labelled with Flash Red (Bangs Laboratories, inc, CAT# FSFR007) with a nominal diameter of 7.32um and a specified range of 7.32 ± 0.27um as determined by the manufacturer using a Coulter counter were imaged on the same multiphoton fluorescence microscope set-up used in the experiment (identical light path, resonant scanner, objective, detector, excitation wavelength and nominal lateral and axial resolutions, with 5x averaging). The images of the beads had a higher SNR than our images of the vasculature, so Gaussian noise was added to the images to degrade the SNR to the same level of that of the blood vessels. The images of the beads were segmented with a threshold, centroids calculated for individual spheres, and planes with a random normal vector extracted from each bead and used to estimate the diameter of the beads. The same smoothing and PSF deconvolution steps were applied in this task. We then reported the mean and standard deviation of the distribution of the diameter estimates. A variety of planes were used to estimate the diameters.”

Results Section Insert

“Our boundary detection algorithm successfully estimated the radius of precisely specified fluorescent beads. The bead images had a signal-to-noise ratio of 6.79 ± 0.16 (about 35% higher than our in vivo images): to match their SNR to that of in vivo vessel data, following deconvolution, we added Gaussian noise with a standard deviation of 85 SU to the images, bringing the SNR down to 5.05 ± 0.15. The data processing pipeline was kept unaltered except for the bead segmentation, performed via image thresholding instead of our deep learning model (trained on vessel data). The bead boundary was computed following the same algorithm used on vessel data: i.e., by the average of the minimum intensity gradients computed along 36 radial spokes emanating from the centreline vertex in the orthogonal plane. To demonstrate an averaging-induced decrease in the uncertainty of the bead radius estimates on a scale that is finer than the nominal resolution of the imaging configuration, we tested four averaging levels in 289 beads. Three of these averaging levels were lower than that used on the vessels, and one matched that used on the vessels (36 spokes per orthogonal plane and a minimum of 10 orthogonal planes per vessel). As the amount of averaging increased, the uncertainty on the diameter of the beads decreased, and our estimate of the bead's diameter converged upon the manufacturer's Coulter counter-based specifications (7.32 ± 0.27um), as tabulated below in Table 1.”

Bibliography

(1) J. R. Bumgarner and R. J. Nelson, “Open-source analysis and visualization of segmented vasculature datasets with VesselVio,” Cell Rep. Methods, vol. 2, no. 4, Apr. 2022, doi: 10.1016/j.crmeth.2022.100189.

(2) G. Tetteh et al., “DeepVesselNet: Vessel Segmentation, Centerline Prediction, and Bifurcation Detection in 3-D Angiographic Volumes,” Front. Neurosci., vol. 14, Dec. 2020, doi: 10.3389/fnins.2020.592352.

(3) N. Holroyd, Z. Li, C. Walsh, E. Brown, R. Shipley, and S. Walker-Samuel, “tUbe net: a generalisable deep learning tool for 3D vessel segmentation,” Jul. 24, 2023, bioRxiv. doi: 10.1101/2023.07.24.550334.

(4) W. Tahir et al., “Anatomical Modeling of Brain Vasculature in Two-Photon Microscopy by Generalizable Deep Learning,” BME Front., vol. 2020, p. 8620932, Dec. 2020, doi: 10.34133/2020/8620932.

(5) R. Damseh, P. Delafontaine-Martel, P. Pouliot, F. Cheriet, and F. Lesage, “Laplacian Flow Dynamics on Geometric Graphs for Anatomical Modeling of Cerebrovascular Networks,” ArXiv191210003 Cs Eess Q-Bio, Dec. 2019, Accessed: Dec. 09, 2020. (Online). Available: http://arxiv.org/abs/1912.10003

(6) T. Jerman, F. Pernuš, B. Likar, and Ž. Špiclin, “Enhancement of Vascular Structures in 3D and 2D Angiographic Images,” IEEE Trans. Med. Imaging, vol. 35, no. 9, pp. 2107–2118, Sep. 2016, doi: 10.1109/TMI.2016.2550102.

(7) T. B. Smith and N. Smith, “Agreement and reliability statistics for shapes,” PLOS ONE, vol. 13, no. 8, p. e0202087, Aug. 2018, doi: 10.1371/journal.pone.0202087.

(8) J. R. Mester et al., “In vivo neurovascular response to focused photoactivation of Channelrhodopsin-2,” NeuroImage, vol. 192, pp. 135–144, May 2019, doi: 10.1016/j.neuroimage.2019.01.036.

heavy use fo alliteration -> there are fleeeting moments of meaning through the alliteration

Wasteland -> modernity as a vision of Hell

stoccato-like metre -> marching, seperate/sharp

biblivala -> Echoes Eliot's life and where he is on his religious journey

Objective correlative -> images of death, deserted land and barren ideologies repressent Eliot's emotions about modernity and the crisis of sspirituality

Grotesque imagery -> humanity is in the state of decay

• alliteration -> facades again (link to Rhapsody, Preludes and Prufrock

repeated to emphasise the distance in society's spirituality -->> this modern life has made things harder.

positive cannotations biblical -> possible link to Dante and Beatrice: his muse --> in 'Divine comedy' her eyes are a powerful symbol, representing divide grace and the pathway to God.

synecdoche: disembodied -> window to the soul (links to Rhapsody, Prufrock and Preludes) --> repeated idea of eyes and what they can and cannot see

ffirst person -> moves away from the collective experience to the personal but only of 'I' --> almost 3rd person ---> seperation from self

repetition: stripping humanity away -> nightmarish private and collective identity is lost

Hell: biblical allusion -> death of spirituality (metonymy) (link to prufrock)

'hollow men' and 'stuffed men empty/unfulfilled

collective: the speaker is one of them -> link to spirituality individuality is reduced

Everything is breaking down. What is left for humanity?

knowledge and culture is being lost in the new society

Eliot had not yet converted to Anglicanism and still had an issue with religious purpose.

Modernity: where does religion belong? Science and industry led too less spiritualaity in society.

The presence of God is deliberately redacted (抹除) throughout -> lament (哀悼) of humanity?

unlike Guy Fawke's Night (Bonfire Night) with all of the fireworks.

repetition -> meaninglessness of life --> Apocalyptic (濒临崩溃、失去信仰与意义的) modern world

repeated religious imageery

Repeetition of 'Falls the Shadow'

Repeetition of 'Falls the Shadow'

Repeetition of 'Falls the Shadow' * possible link to war

refer to previous note on all the 'between's

refer to previous note on all the 'between's

refer to all previous notes on the 'between's

Gaps -> every part of life seems unfulfilled --> sexual desire unfulfulled

refer to previous note on all the 'between's

refer to previous note on all the 'between's

biblical allusion: Lord's prayer -> corrupted by humanity

Psalm 23 - shadow of death -> motif -->humanity shrouded in darkness

allusion to the nursery rhyme 'Here we go arounnd the mullery bush' -> showing desolation (荒凉) and emptiness in a more barren landscape/ world

bathetic moment（情感突降的反高潮) -> hope doesnt seem to last in the poem --> Eliot 将原本象征希望与循环的童谣改写为空洞的仪式，使短暂的希望立刻坠入荒凉与虚无之中。

the only think that will save humanity -> we are all thhe 'empty men'

biblical allusion to the birth of christ -> follow star to a better life linked to spirituality

truncated and clumsy language

Natural absolute

Illuminate

Don't reach

Transcendence

Impassable permeable

Cut two perpenducular

And they are not atoms

All relative

Complete

Limits of knowledge

Goedel

Debunk

Homoiconicity

Cancel

Prove completeness

Whole > parts

Analitical details

Nice

Real is reasonable

only The reasonable can be realized

elmélet-e?

e suffix possessive

and at the same time question

is that so

Objective correlative -> images of death, deserted land and barren ideologies repressent Eliot's emotions about modernity and the crisis of spirituality

repetition of eyes thorough out the poem

first person -> moves away from the collective experience to the personal but only of 'I' --> almost 3rd person ---> seperation from self

bathetic moment（情感突降的反高潮) -> hope doesnt seem to last in the poem --> Eliot 将原本象征希望与循环的童谣改写为空洞的仪式，使短暂的希望立刻坠入荒凉与虚无之中。

• alliteration -> facades again (link to Rhapsody, Preludes and Prufrock

synecdoche: disembodied -> window to the soul (links to Rhapsody, Prufrock and Preludes) --> repeated idea of eyes and what they can and cannot see

Hell: biblical allusion -> death of spirituality (metonymy) (link to prufrock)

Eliot had not yet converted to Anglicanism and still had an issue with religious purpose.

Modernity: where does religion belong? Science and industry led too less spiritualaity in society.

The presence of God is deliberately redacted (抹除) throughout -> lament (哀悼) of humanity?

Everything is breaking down. What is left for humanity?

knowledge and culture is being lost in the new society

unlike Guy Fawke's Night (Bonfire Night) with all of the fireworks.

repetition -> meaninglessness of life --> Apocalyptic (濒临崩溃、失去信仰与意义的) modern world

repeated religious imageery

Repeetition of 'Falls the Shadow'

Repeetition of 'Falls the Shadow'

Repeetition of 'Falls the Shadow' * possible link to war

refer to all previous notes on the 'between's

Gaps -> every part of life seems unfulfilled --> sexual desire unfulfulled

refer to previous note on all the 'between's

refer to previous note on all the 'between's

refer to previous note on all the 'between's

refer to previous note on all the 'between's

biblical allusion: Lord's prayer -> corrupted by humanity

Psalm 23 - shadow of death -> motif -->humanity shrouded in darkness

allusion to the nursery rhyme 'Here we go arounnd the mullery bush' -> showing desolation (荒凉) and emptiness in a more barren landscape/ world

the only think that will save humanity -> we are all thhe 'empty men'

biblical allusion to the birth of christ -> follow star to a better life linked to spirituality

truncated and clumsy language

Wasteland -> modernity as a vision of Hell

stoccato-like metre -> marching, seperate/sharp

biblivala -> Echoes Eliot's life and where he is on his religious journey

Grotesque imagery -> humanity is in the state of decay

heacy use fo alliteration -> there are fleeeting moments of meaning through the alliteration

repeated to emphasise the distance in society's spirituality -->> this modern life has made things harder.

positive cannotations biblical -> possible link to Dante and Beatrice: his muse --> in 'Divine comedy' her eyes are a powerful symbol, representing divide grace and the pathway to God.

repetition: stripping humanity away -> nightmarish private and collective identity is lost

'hollow men' and 'stuffed men empty/unfulfilled

collective: the speaker is one of them -> link to spirituality individuality is reduced

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What is this?

L'Échec comme Moteur : Analyse et Perspectives

Résumé

L'analyse des contextes fournis révèle une vision contre-intuitive et multifacette de l'échec, le présentant non pas comme une finalité mais comme un processus fondamental, voire essentiel, au développement humain, artistique et social.

Loin d'être une simple absence de succès, l'échec est dépeint comme une ressource active : il est le carburant de la créativité pour les artistes et les clowns, un catalyseur de transformation personnelle profonde pour l'individu confronté à l'effondrement de ses rêves, et une méthodologie d'innovation pour des projets communautaires.

Les thèmes centraux qui émergent sont la capacité de l'échec à humaniser en brisant l'illusion de la perfection, son pouvoir de générer du lien social par le rire et l'empathie, et sa fonction libératrice qui, bien que douloureuse, peut ouvrir la voie à une existence plus authentique.

Il est également souligné que la capacité à surmonter un échec majeur est un "luxe" qui dépend d'un soutien social et structurel, mettant en lumière une inégalité face à la possibilité même de "chuter".

1. L'Échec comme Ressource Créative et Artistique

Le document met en évidence comment plusieurs disciplines artistiques intègrent l'échec non seulement comme un risque inhérent mais comme une composante centrale de leur processus créatif et de leur message.

L'Art du Clown : L'Échec Humanisant

Pour les clowns du cirque Ronkali, l'échec est la matière première de leur art. Ils le décrivent comme un élément qui les "alimente" et le considèrent comme de "l'or". Cette approche repose sur plusieurs principes clés :

• Humanisation : L'échec est perçu comme ce qui rend l'être humain, en opposition à une quête de perfection jugée "inhumaine".

En se "cassant la figure", le clown rappelle au public que l'erreur fait partie de la condition humaine.

• Thérapie et Lien Social : L'échec mis en scène a une fonction quasi thérapeutique pour le public et les artistes.

Le rire qu'il provoque n'est pas moqueur mais un "nous ri" fondamental, créant une communauté et un sentiment de partage.

Le clown invite le public à participer, créant "une atmosphère où l'échec n'est pas quelque chose de tragique".

• Improvisation et Transformation : Un échec technique, comme une fausse note de trompette, n'est pas une fin en soi. Il devient une opportunité de jeu, transformé en "blague" ou en "message". La solution est de "saisir ce diamant et le polir" pour en faire un effet comique.

L'Artiste et la "Méditation sur l'Échec"

L'artiste américaine Cassidi Toner a fait de l'échec le cœur de sa pratique artistique, qu'elle qualifie de "méditation sur l'échec".

• Figure Totémique : Elle s'inspire de la figure tragicomique de Vil Coyote, qui échoue sans cesse à attraper Bip Bip.

Citant Mark Twain, elle décrit le coyote comme une "allégorie vivante du désir", toujours affamé mais jamais rassasié, incarnant cet échec constant.

• Valorisation du Ratage : Le titre de son exposition, "besides the point" (hors sujet), est une "invitation au ratage joyeux".

Elle considère que si l'on accepte le ratage non comme un échec mais comme un "potentiel créatif", la réussite prend une tout autre signification.

• Le Paradoxe de la Maîtrise : Son succès à transformer l'échec en art l'a conduite à un paradoxe : "j'ai tellement récupéré l'échec que même si j'essaie d'échouer [...] je ne peux pas".

Un véritable ratage de sa part serait interprété par le public comme une démarche intentionnelle.

2. L'Échec comme Crise Personnelle et Voie de Reconstruction

Le témoignage de l'auteur Alexander Crutfelt illustre la dimension dévastatrice de l'échec personnel et social, mais aussi son potentiel de reconstruction et de lucidité.

Le Récit de l'Effondrement

Le projet de rénovation d'une ferme familiale se transforme en cauchemar, menant à une cascade de faillites :

• Échec Matériel : Le toit de la maison s'effondre, le projet de rénovation est interdit, et la propriété finit par être une "maison hantée" qui "détruit une famille".

• Échec Financier : Il accumule une "dette à six chiffres" et doit déclarer une "faillite personnelle".

• Échec Familial : Le couple se sépare, et il décrit le moment où il a dû annoncer son départ à ses enfants comme la "pire horreur de [sa] vie" et le "moment culminant de l'échec".

La Dimension Sociale et la Libération par la Parole

L'échec de Crutfelt met en lumière la pression sociale et le pouvoir de la vulnérabilité.

• Le Poids du Regard Social : Il ressent l'échec "aux yeux de la société" axée sur la performance, qui le marginalise.

Il prend conscience que, contrairement à ce qu'il pensait, il n'est pas indifférent à "ce que les autres pensaient de [lui]". Il critique l'illusion de perfection véhiculée par les réseaux sociaux comme Instagram.

• La Vertu du Partage : En écrivant sur son expérience et en en parlant publiquement, il découvre qu'il n'est pas seul. Son récit incite d'autres personnes à partager leurs propres échecs, créant un sentiment de connexion et de validation mutuelle.

La Gratitude Paradoxale et le "Luxe d'Échouer"

Malgré la douleur, l'échec a été une expérience profondément transformatrice pour Crutfelt.

• Libération du "Moi Antérieur" : Il se dit "reconnaissant à la maison de s'être écroulée", car "sans cet échec, je ne serai jamais sorti de mon ancien moi". Cela lui a permis de vivre dans le présent et de trouver le bonheur dans une vie plus simple.

• La Conscience du Privilège : Il reconnaît qu'échouer et se relever est un "immense luxe".

Il a bénéficié du soutien de sa famille, d'un thérapeute et de l'État-providence, ce qui lui a permis "d'atterrir sur un doux édredon".

Il oppose sa situation à celle des "personnes qui n'ont aucune chance de se retrouver dans la situation d'un possible échec parce qu'elles ne peuvent tout simplement pas tomber plus bas".

3. L'Échec comme Méthodologie d'Innovation et de Cohésion Sociale

Le projet du "Bosk" à Leeuwarden, mené par un duo de designers, illustre comment l'acceptation de l'échec et de l'imperfection peut devenir une méthode de travail productive et un vecteur de lien social.

Un Projet Ancré dans un Échec Historique

Le projet prend racine dans l'histoire de la ville, symbolisée par la tour Oldehove, une cathédrale dont la construction a "magnifiquement échoué" il y a 500 ans en s'enfonçant dans le sol.

Le "Bosk" se propose de "finir la cathédrale" non pas architecturalement, mais comme un "projet social" temporaire de 100 jours.

L'Acceptation de l'Imperfection dans le Processus

Les designers critiquent la culture néerlandaise qui, par excès de planification, cherche à "se préserver de l'échec".

Leur approche consiste à tester les projets publiquement, comme une "maquette grandeur nature", et à s'adapter aux imprévus.

Problème / Échec Rencontré

Solution / Adaptation

Résultat

Les voiles de bateau frison ne passent pas le test de sécurité incendie.

Utilisation de toile de vieux ballons à air chaud.

Le rendu est "plus coloré" et "convient bien mieux au projet".

Les fragments de ballon ne pendent pas en triangles géométriques parfaits comme prévu.

Ils décident de "continuer et on verra ce qui va se passer".

L'imperfection est acceptée comme une part inévitable de la réalisation.

Ils affirment : "quand on essaie de traduire [un plan] en une image réelle, on échoue forcément".

Le Succès par le Lien Social

Le véritable succès du projet ne réside pas dans la perfection de la structure, mais dans son impact humain.

La maison communautaire est "vraiment devenue la maison des gens d'ici".

Le plus beau retour qu'ils aient reçu est que des gens "se sont fait de nouveaux amis".

La valeur du projet, qui sera démantelé, "restera dans le cœur et dans l'âme des gens".

chief seatlle will not mourn or dwell on the deaths nor take revenge upon the whites for doing it, as he considers that they are somewhat in blame as well

Their tribe used to be a large number and occupied land like waves of a wind ruffled sea covering the shell paved floor. But that time is long gone which is now a mournful memory.

It is generous of the white men to allow them to live peacefully after buying their land, he says. He considers that the offer may be good because since they are so less in number, they don't need such a big country.

Contrast between the two people; Chief Seattle compares how the whites are many and the red indians are only a remaining dew like scattering trees

The big chief of washington send greetings of friendship and goodwill only as a formality; the red indians know that they don't actually need their friendship

The nature for e.g the sky which shed tears for red indians for centuries may change even though it appears changeless and eternal

This called to me because it is something that I have experienced in the past and have only recently discovered that it was wrong.

This stood out to me because response to errors is one of the biggest ways that I help myself keep a growth mindset.

13:30 Ricardo Leppe ist der deutsche John Taylor Gatto:<br /> Weapons of Mass Instruction: A Schoolteacher's Journey Through the Dark World of Compulsory Schooling.

Wednesdays : Synthèse et Analyse Approfondie

Résumé Exécutif

Ce document présente une analyse détaillée du jeu vidéo Wednesdays, coédité et coproduit par Arte, sorti le 26 mars 2025 sur PC (Steam et Itch.io).

Conçu par l'auteur et directeur créatif Pierre et l'illustratrice Exaeva, ce jeu narratif aborde les thématiques complexes et sensibles de l'inceste, de la pédocriminalité et des violences intrafamiliales.

Malgré la dureté des sujets, le jeu adopte un ton qualifié de "lumineux et bienveillant".

D'une durée moyenne de deux heures, Wednesdays se distingue par une direction artistique unique, inspirée de la bande dessinée indépendante, où les personnages victimes sont représentés avec des têtes cubiques.

Un pilier central du projet est son accessibilité, pensée à la fois pour les non-joueurs et les personnes en situation de handicap, avec un travail approfondi sur la lisibilité des couleurs et des mécaniques de jeu simplifiées.

Le développement, mené par une petite équipe de sept personnes travaillant à distance, a été marqué par des choix créatifs forts, notamment la création de l'espace de décompression "Orcopark" et une conception sonore immersive qui pallie l'absence de doublage.

Wednesdays se positionne comme une œuvre cherchant à libérer la parole et à utiliser le médium du jeu vidéo comme un outil de prise de conscience et d'écoute.

I. Présentation du Jeu "Wednesdays"

A. Concept et Thématiques Abordées

Wednesdays est un jeu vidéo narratif qui plonge le joueur dans les souvenirs fragmentés de Timothé, un personnage victime d'inceste.

Le but est de reconstituer son histoire en explorant différentes scènes de sa vie, de l'enfance à l'âge adulte. Le jeu traite frontalement de sujets difficiles comme la pédocriminalité et les violences intrafamiliales.

Malgré la gravité de ces thèmes, la démarche des créateurs est de proposer une expérience "lumineuse et bienveillante".

L'approche narrative et visuelle évite toute représentation graphique de la violence, privilégiant la suggestion, la pédagogie et l'émotion.

Des avertissements de contenu (trigger warnings) sont intégrés directement dans le jeu pour permettre aux joueurs de se préserver.

B. Équipe de Développement et Édition

Le jeu est le fruit d'une collaboration entre plusieurs talents de la scène indépendante, sous l'égide d'Arte qui coproduit et coédite des jeux vidéo depuis plus de dix ans.

Membre

Rôle

Contributions Notables

Pierre

Auteur et Directeur Créatif

Conception du projet, écriture du scénario et des dialogues.

Exaeva

Illustratrice

Création de toute la direction artistique, des personnages et des décors.

Virginia

Sound Designer

Conception de l'univers sonore, incluant les gimmicks sonores des personnages.

Florent Morin (The Pixel Hunt)

Éditeur

Accompagnement du projet, gestion administrative, conseils créatifs.

Chris

Programmeur

Développement technique, lui-même concerné par le sujet du jeu.

Nico Novac

Artiste Pixel Art

Création des visuels pour la section "Orcopark".

Dianne

Programmeuse (renfort)

Aide à la programmation sur des aspects spécifiques du jeu.

L'équipe principale de sept personnes a travaillé majoritairement à distance via Discord, sans réunions formelles, démontrant une grande autonomie de chaque membre.

C. Données Clés

Caractéristique

Détail

Date de sortie

26 mars 2025

Plateformes

PC (via Steam et Itch.io)

Durée de jeu moyenne

Environ 2 heures à 2 heures 30

Genre

Jeu narratif, Bande dessinée interactive

II. Direction Artistique et Conception Visuelle

A. Un Style "Bande Dessinée Interactive"

La direction artistique de Wednesdays est l'un de ses aspects les plus marquants. Elle s'inspire fortement de la bande dessinée indépendante franco-belge et américaine, avec des références citées comme Frédéric Peeters, Craig Thompson et Tillie Walden.

Le processus de création est traditionnel et méticuleux :

1. Dessin sur papier : Exaeva réalise tous les dessins des décors et des personnages sur papier, son support de prédilection. Les personnages sont dessinés sur des calques en papier "layout", un peu transparent, utilisé en animation traditionnelle.

2. Numérisation : Tous les éléments graphiques sont ensuite scannés.

3. Colorisation numérique : Les couleurs sont ajoutées digitalement, en respectant une technique de bichromie, qui consiste à utiliser principalement deux teintes dominantes par image pour créer des ambiances colorées et lumineuses spécifiques.

Cette approche donne au jeu une texture unique, avec un aspect crayonné très personnel qui va à contre-courant des productions 3D ultra-réalistes.

B. Le Symbolisme des "Têtes Cubiques"

Un choix visuel central du jeu est la représentation des personnages victimes d'inceste avec des têtes cubiques. Cette idée, présente dès la genèse du projet, a plusieurs fonctions :

• Visibilité de l'invisible : Elle rend les victimes, souvent invisibles dans la société, immédiatement identifiables pour le joueur.

• Faciliter la projection : En s'appuyant sur les théories de Scott McCloud (L'Art invisible), un visage moins détaillé et réaliste permet au joueur de se projeter plus facilement dans le personnage.

• Défi artistique : Contrairement à l'idée initiale que cela simplifierait le travail, l'absence d'expressions faciales a représenté un défi majeur. Toute l'émotion des personnages doit être transmise par la corporalité, les postures et la gestuelle, ce qui a demandé un travail d'animation et de dessin très poussé.

C. Processus Créatif et Influences

La collaboration entre Pierre et Exaeva a été fondamentale. Pierre arrivait avec des idées de scènes, parfois sous forme de placeholders (visuels de substitution) très simples, et Exaeva les transformait en scènes complètes.

De nombreuses décisions de mise en scène ont été prises lors de sessions de travail à Bruxelles, autour d'un verre. Le jeu alterne entre les scènes dessinées par Exaeva et l'univers en pixel art d'Orcopark, créant un contraste visuel fort.

III. Conception Sonore et Narrative

A. Sound Design sans Voice Acting

Le jeu ne contient pas de dialogues parlés (voice acting), un choix justifié par le budget mais aussi par une volonté artistique.

La sound designer Virginia a créé un univers sonore immersif basé sur des sons réalistes et des "gimmicks" sonores pour chaque personnage, tous liés à l'univers du papier et de l'écriture :

• Timothé : Bruit de machine à écrire.

• Les enfants : Bruits de Crayola ou de feutres.

• Joël (le père) : Son de stylo-plume.

• Fatia (l'institutrice) : Bruit de craie sur un tableau.

Cette approche permet non seulement d'identifier auditivement qui parle, mais renforce aussi l'idée que l'histoire est en train de s'écrire ou de se reconstituer.

B. La Libération de la Parole par le Gameplay

La structure narrative et les mécaniques de jeu sont conçues pour servir le thème principal : la difficulté et les étapes de la libération de la parole.

• Souvenirs fragmentés : Le joueur peut choisir les souvenirs dans un ordre non linéaire, reflétant le processus non chronologique de la mémoire traumatique.

• Mécaniques de dialogue : Dans certaines scènes, comme celle de la voiture avec le personnage de Yeram, le gameplay joue avec les bulles de dialogue.

Le joueur sélectionne une option, mais le personnage peine à la formuler, la phrase change ou est remplacée par des points de suspension.

Cela représente la lutte interne pour verbaliser le trauma. Pierre note que près de 4% des bulles de dialogue du jeu sont des silences ("..."), soulignant l'importance de ce qui n'est pas dit.

IV. L'Accessibilité : Un Pilier du Projet

L'accessibilité a été une priorité dès le début du développement. L'objectif était double :

1. Rendre le jeu jouable par des non-joueurs : Avec des contrôles simples et une interface claire.

2. Inclure les personnes en situation de handicap.

Pour y parvenir, l'équipe a collaboré avec Game Accessibility Hub, une société spécialisée. Des tests ont été menés avec des joueurs ayant différents handicaps.

Un exemple marquant est celui d'un testeur achromate (qui ne voit aucune couleur).

Il a trouvé le jeu parfaitement lisible et a même ressenti une différence dans la seule scène conçue en noir et blanc pur, validant ainsi l'efficacité des contrastes et de la direction artistique.

Le travail sur les palettes de couleurs a été systématiquement testé à l'aide d'outils simulant différentes formes de daltonisme. Arte a soutenu cette démarche en allouant un budget supplémentaire dédié à l'accessibilité.

V. Genèse et Coulisses de la Production

A. D'un "One-Man Show" au Jeu Vidéo

L'idée de Wednesdays est née de l'inspiration de Pierre après avoir vu L'Imposture, un spectacle de marionnettes de Lucie Arnodin.

Fasciné par la capacité du spectacle à traiter de sujets graves avec légèreté et une narration éclatée, il a d'abord tenté d'écrire un one-man show sur le sujet.

Après un retour mitigé d'un ami proche, il a abandonné cette idée pour se tourner vers un médium qu'il maîtrisait : le jeu vidéo, tout en conservant le ton et l'approche narrative du projet initial.

B. Orcopark : L'Espace de Décompression

L'interface de sélection des chapitres a connu une évolution significative. Le concept initial était un bureau sur lequel le joueur cliquait sur différents objets pour lancer les souvenirs. Jugée "un peu chiante" par Arte, cette idée a été remplacée par Orcopark, un parc d'attractions rétro en pixel art.

Orcopark sert de hub central mais aussi d'espace "safe" pour le joueur.

Entre des scènes émotionnellement intenses, il peut prendre le temps de se détendre, de ramasser des débris, de cliquer sur des éléments interactifs et de décorer son parc.

Cet espace a été développé plus que prévu initialement, à l'encouragement d'Arte, pour renforcer son rôle de sas de décompression.

C. Anecdotes de Développement

• Moustache le chat : Le chat Moustache a été ajouté dans une scène finale à la demande de Nil, le fils de Pierre.

• Joël, l'alter ego vieilli : Le design du personnage du père, Joël, est basé sur une version vieillie de l'auteur, Pierre.

• Figurine en argile : L'objet mystère de l'émission était une statuette en argile réalisée par la grand-mère de Pierre, qui a aussi servi de base pour une marionnette dans un autre de ses projets de jeu sur Game Boy Camera.

VI. Réflexions sur l'Impact et la Réception

A. Le Jeu Vidéo comme Média d'Écoute

Les créateurs soulignent la position particulière de Wednesdays, un "OVNI" qui se situe à l'intersection du jeu vidéo et de l'œuvre culturelle. Cette position hybride pose des défis de réception :

• Les journalistes spécialisés jeu vidéo peuvent être déroutés par un jeu qui ne correspond pas aux critères d'évaluation habituels (gameplay, durée de vie, etc.).

• Les journalistes culturels généralistes peuvent être réticents en raison d'un mépris ou d'une méconnaissance du médium.

Malgré cela, le jeu a reçu une bonne couverture en France et a trouvé son public.

B. Un Outil pour la Prise de Conscience

Le retour le plus gratifiant pour l'équipe vient des joueurs.

De nombreux témoignages font état de l'impact positif du jeu, y compris de la part de personnes victimes qui se sont senties comprises ou qui ont eu des prises de conscience sur leur propre vécu en jouant.

Le jeu semble ainsi atteindre son objectif : non seulement libérer la parole de son personnage, mais aussi potentiellement celle de ses joueurs, et sensibiliser l'entourage aux réalités de l'inceste.

Short Story

so not so much about a single Superintelligent AI, as society gradually drowning in AI enshittification. it may not be existential to society but it still really sucks

Like the camel paper to protect against prompt injection

So the capability reliability gap implies that if you’re willing to accept self driving cars mowing through droves of people until they can reliably not crush people on the street under their wheels, then we’d likely end up with self driving cards pretty quickly, but for the most part we don’t see this as acceptable

This is essentially what uber and various gig economy projects do - they externalise the otherwise minimise the negative externalities in favour of more iterations

It’s interesting because it shows how deeply the blind man influenced her inner life. Even years later, she still felt the need to share her emotions and experiences with him through poetry.

It surprises me because I never imagined that touching someone’s face could be such an intimate and meaningful act, especially between two people who were not romantically involved.

This The ideology that men have to be extra [assertive, competitive and tough, Is heavily embedded into black culture I would say a lot of it comes from past times to wear a black man. Survival depended on his strength..

These benefits of being a male in society may seem pretty insignificant, but they are very important. A huge one that sold out to me was being able to take public transportation like subway Bart, even Uber without the fear of somehow someway, being violated or harassed or course into doing anything unwanted.

It shows how big the language gap is between families of different income levels. I think it helps explain why some kids start school already ahead in vocabulary and communication skills—it’s not about ability, but about early exposure and environment.

I think this is a really important approach prioritizing the perspective and opinion of humans inputs. I also think you can receive more insight and information through this approach, considering that you're not just considering the viewpoint of people affected or people who share their input as they face implications on these designs, but also bringing them along the process gives more than understanding, but perspective and opportunity to make change, to adhere to other individuals needs.

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As I learned in my previous writing classes, it is critical to rely on verified information, and I wrote research papers that contain cited information from scholarly articles. I was not allowed to use Wikipedia because many articles do not include any proper citations.

I would bring in the consultant because it would be less costly than publishing falsified information and damaging the company’s reputation. Companies that have poor reputations will suffer from financial problems and could go bankrupt.

These are the factors of fair use, a doctrine that allows for otherwise prohibited use of copyrighted material. It is important to determine if your use of copyrighted material falls under all these factors. If any of these factors are violated, then you could be sued or charged for copyright infringement.

When I took a statistics course, I learned that data sampling can be influenced by the race, proximity to a location, etc. of those who are surveyed. I believe this is unethical because readers look for unbiased information in scholarly articles.

The image of a “strange mad flower” growing or “dripping in the dark” contrasts beauty with madness and decay

Why does the speaker say “take me home”? Are they asking to escape suffering or death? And who is “he” a god, a king, or someone else the speaker fears?

This poem personifies death as dirty and unstoppable “filth” that destroys the broken. Reckon means to judge or measure (Merriam-Webster). The final line lists victims, showing death’s human cost.

In school, students are usually taught the teacher's method of learning. How they draw it up is how most students process the information during the class. Allowing students to learn their own ways encourages growth while keeping the material exciting and engaging.

Replication is key, especially in science. It allows people to effectively prove their ideas and/ or claims to themselves or others and keep it grounded and valuable.

I believe that being open-minded is extremely important as it allows you to gain information, even though it can be false. This can allow for a better exchange of ideas and claims with proper criticism.

It’s not surprise to me personally that male dominated places or countries have higher counts of sexual harassment towards women and rape against women. However, I wonder in women dominated the countries and places is there as high of a sexual assault problem against men? Are men also more likely to be sexually assaulted, harassing or raped in women dominated countries or societies?

The ideology that women are asking for an Unwanted action, movement or attack is very harmful. And unfortunately, Has been or for some cases still currently is a way of thinking for many.

There’s clearly an astronomical extent of sexual assault and rape happening towards women. As a woman myself I find it very understandable that the majority of cases go on reported

It surprised me that her husband, who is a doctor, doesn’t believe she is sick at all. I would think a doctor should trust his patient’s feelings, especially when the patient is his wife.

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Definition of patriarchy, Defined from medical feminism.

Also the term of intersexuality which was specifically created for black women to face sexism and racism. Which encourage the idea that because they were black and a woman that was the only identity that could be reorganized. there was no in between. they cannot be black and woman. They had to be a black woman.

The idea that women cannot do certain things because of our biological factors and gender. Usually Promoting the idea of inferiority to the opposite sex/gender.

Essentially equal opportunities to work equal opportunities to advancement equals in political work, life, and social environments.

So Someone who grew up in a Community that discussed gender identity and fluidity in gender would be be more open to not sticking to the preset, gender identities?

Definition of femininity and masculinity. Essentially explained the differences in how society expects women or girls to behave and present themselves, as well as masculinity, refers to the expectations society has on boys and men

Definition of gender explained, gender as a social concept. In sociology one I got a nice overview of how a Gender, social concepts, or gender roles have been grouped and linked to individuals.

Definition of sex. Sex is strictly biological factors that explains the differences between male and female. strictly focused on biology and chromosomes.

For expatriated Americans seeking work or partnerships in southern Africa, including my own job search in Namibia and South Africa, U.S.–South Africa tensions could shape economic prospects and trade stability.

The potential sentence could significantly weaken one of South Africa’s most visible opposition voices.

The incident behind the conviction underscores Malema’s use of provocative, militant imagery in his politics

His party frames the conviction as part of a broader struggle against Western interference

Malema portrays his prosecution as politically motivated and influenced by U.S. pressure

The use of this footage fueled international outrage and linked Malema’s rhetoric to racial violence narratives

Trump’s direct criticism of Malema shows how U.S. politics can spill into African domestic affairs

the story’s focus is Malema’s latest conviction, which deepens political tensions between South Africa and the U.S

This is were, as future health promotion practicians, can start to change the messaging around health and to stop looking for a quick fix to fool the masses that 'change' is being implemented. The 'little by little' approach, instead of an 'all or nothing' mindset needs to be pushed.

Asking today's society to give up some of their 'stuff' to give it to others is a very hard sell. This type of societal change will never occur in the short term, with people's individual sense on entitlement. This will only occur when the next generation of leadership come into office, who have the courage to undertake this.

I think it point out that inequality in schools isn’t just about individual effort, it’s built into the structure itself. “Tracking,” or sorting students into different academic levels, often reflects race and class more than actual ability. For example, wealthier or white students might be placed in honors classes while students of color are steered toward lower tracks, limiting their access to advanced opportunities.

I think this is how students like Chantelle can fall behind, not because they lack ability, but because they lack guidance and support. Without someone to explain how course choices affect college eligibility, she unknowingly limited her future options. In real life, this happens often when first-generation students don’t realize how crucial early course decisions are, for example, missing Algebra I in ninth grade can block the path to advanced math and science later.