eLife assessment

This useful study reports an unexpected phenotype of atrophy of the male reproductive system and infertility upon combined knockout in adult mice of the genes encoding the two kinases CDK8 and CDK19. While the morphological evidence and single-cell transcriptomic data are solid, the proposed mechanism remains unconvincing as there is little evidence for causality, and some controls are missing. This work will be of interest to reproductive biologists, developmental biologists, and andrologists.

Reviewer #1 (Public Review):

Summary:

In this paper, Bruter and colleagues report the effects of inducible deletion of the genes encoding the two paralogous kinases of the Mediator complex in adult mice. The physiological roles of these two kinases, CDK8 and CDK19, are currently rather poorly understood; although conserved in all eukaryotes, and among the most highly conserved kinases in vertebrates, individual knockouts of genes encoding CDK8 homologues in different species have revealed generally rather mild and specific effects, in contrast to Mediator itself. Here, the authors provide evidence that neither CDK8 nor CDK19 are required for adult homeostasis but they are functionally redundant for maintenance of reproductive tissue morphology and fertility in males.

Strengths:

The morphological data on the atrophy of the male reproductive system and the arrest of spermatocyte meiosis are solid and are reinforced by single-cell transcriptomics data, which is a challenging technique to implement in vivo. The main findings are important and will be of interest to scientists in the fields of transcription and developmental biology.

Weaknesses:

There are several major weaknesses.

The first is that data on the general health of mice with single and double knockouts is not shown, nor is there any data on effects in any other tissues. This gives the impression that the only phenotype is in the male reproductive system, which would be misleading if there were phenotypes in other tissues that are not reported. Furthermore, data for the genitourinary system in single knockouts are very sparse; data are described for fertility in Figure 1H, ploidy, and cell number in Figures 2B and C, plasma testosterone and luteinizing hormone levels in Figures 5C and 5D, and morphology of testis and prostate tissue for single Cdk8 knockout in Supplementary Figure 1C (although in this case the images do not appear very comparable between control and CDK8 KO, thus perhaps wider fields should be shown), but, for example, there is no analysis of different meiotic stages or of gene expression in single knockouts. It is worth mentioning that single knockouts seem to show a corresponding upregulation of the level of the paralogue kinase, indicating that any lack of phenotypes might be due to feedback compensation, which would be an interesting finding if confirmed; this has not been mentioned.

The second major weakness is that the correlation between double knockout and reduced expression of genes involved in steroid hormone biosynthesis is portrayed as a causal mechanism for the phenotypes observed. While this is a possibility, there are no experiments performed to provide evidence that this is the case. Furthermore, there is no evidence showing that CDK8 and/or CDK19 are directly responsible for the transcription of the genes concerned.

Finally, the authors propose that the phenotypes are independent of the kinase activity of CDK8 or CDK19 because treatment of mice for a month with an inhibitor does not recapitulate the effects of the knockout, and nor does expression of two steroidogenic genes change in cultured Leydig cells upon treatment with an inhibitor. However, there are no controls for effective target inhibition shown.

Reviewer #2 (Public Review):

Summary:

The authors tried to test the hypothesis that Cdk8 and Cdk19 stabilize the cytoplasmic CcNC protein, the partner protein of the Mediator complex including CDK8/19 and Mediator protein via a kinase-independent function by generating induced double knockout of Cdk8/19. However, the evidence presented suffers from a lack of focus and rigor and does not support their claims.

Strengths:

This is the first comprehensive report on the effect of a double knockout of CDK8 and CDK19 in mice on male fertility, hormones, and single-cell testicular cellular expression. The inducible knockout mice led to male sterility with severe spermatogenic defects, and the authors attempted to use this animal model to test the kinase-independent function of CDK8/19, previously reported for humans. Single-cell RNA-seq of knockout testis presented a high resolution of molecular defects of all the major cell types in the testes of the inducible double knockout mice. The authors also have several interesting findings such as reentry into cell cycles by Sertoli cells, and loss of Testosterone in induced dko that could be investigated further.

Weaknesses:

The claim of reproductive defects in the induced double knockout of CDK8/19 resulted from the loss of CCNC via a kinase-independent mechanism is interesting but was not supported by the data presented. While the construction and analysis of the systemic induced knockout model of Cdk8 in Cdk19KO mice is not trivial, the analysis and data are weakened by the systemic effect of Cdk8 loss, making it difficult to separate the systemic effect from the local testis effect.

The analysis of male sterile phenotype is also inadequate with poor image quality, especially testis HE sections. The male reproductive tract picture is also small and difficult to evaluate. The mice crossing scheme is unusual as you have three mice to cross to produce genotypes, while we could understand that it is possible to produce pups of desired genotypes with different mating schemes, such a vague crossing scheme is not desirable and of poor genetics practice. Also using TAM-treated wild type as control is ok, but a better control will be TAM-treated ERT2-cre; CDK8f/f or TAM-treated ERT2 Cre CDK19/19 KO, so as to minimize the impact from the well-recognized effect of TAM.

While the authors proposed that the inducible loss of CDK8 in the CDK19 knockout background is responsible for spermatogenic defects, it was not clear in which cells CDK8/19 genes are interested and which cell types might have a major role in spermatogenesis. The authors also put forward the evidence that reduction/loss of Testosterone might be the main cause of spermatogenic defects, which is consistent with the expression change in genes involved in steroigenesis pathway in Leydig cells of inducible double knockout. However it is not clear how the loss of Testosterone contributed to the loss of CcnC protein.

The authors should clarify or present the data on where CDK8 and CDK19 as well as CcnC are expressed so as to help the readers understand which tissues both CDK might be functioning in and cause the loss of CcnC. It should be easier to test the hypothesis of CDK8/19 stabilizing CcnC protein using double knock-out primary cells, instead of the whole testis.

Since CDK8KO and CDK19KO both have significantly reduced fertility in comparison with wildtype, it might be important to measure the sperm quantity and motility among CDK8 KO, CDK19KO, and induced DKO to evaluate spermatogenesis based on their sperm production.

Some data for the inducible knockout efficiency of Cdk8 were presented in Supplemental Figure 1, but there is no legend for the supplemental figures, it was not clear which band represented the deletion band, and which tissues were examined. Tail or testis? It seems that two months after the injection of Tam, all the Cdk8 were completely deleted, indicating extremely efficient deletion of Tam induction by two months post administration. Were the complete deletion of Cdk8 happening even earlier? An examination of time points of induced loss would be useful and instructional as to when is the best time to examine phenotypes.

The authors found that Sertoli cells re-entered the cell cycle in the inducible double knockout but stopped short of careful characterization other than increased expression of cell cycle genes.

Overall this work suffered from a lack of focus and rigor in the analysis and lack of sufficient evidence to support their main conclusions.

Minor:

Dko should be appropriately named iDKO (induced dKO).

"suppress spermatogenesis and male fertility" in the title does not fit the evidence presented.

"DKO males, had an understized and dedifferentiated reproductive system?" what is the evidence for "undifferentiated"?

We performed necropsy ? not the right wording here.

Colchicine-lke apoptotic bodies ? what does this mean? Not clear.

Images throughout the manuscript suffer from poor resolution and are often blurry and hard to evaluate.

To pinpoint the meiotic stage defect of iDKO, it is better to use the meiotic chromosome spread approach.

eLife assessment

This manuscript presents an important analysis of the role that the nucleosome acidic patch plays in SWR1-catalyzed histone exchange. This manuscript contains convincing data which significantly expands our understanding of the complex process of H2A.Z deposition by SWR1 and therefore would be of interest to a broad readership.

Reviewer #1 (Public Review):

Summary:

The authors presented here a novel 3D fibroblast culture and transdifferentiation approach for potential meat production with GelMA hydrogel.

Strengths:

(1) Reduced serum concentration for 3D chicken fibroblast culture and transdifferentiation is optimized.<br /> (2) Efficient myogenic transdifferentiation and lipogenesis as well as controlled fat deposition are achieved in the 3D GelMA.

Reviewer #2 (Public Review):

The manuscript by Ma et al. tries to develop a protocol for cell-based meat production using chicken fibroblasts as three-dimensional (3D) muscle tissues with fat accumulation. The authors used genetically modified fibroblasts, which can be forced to differentiate into muscle cells, and formulated 3D tissues with these cells and a biphasic material (hydrogel). The degrees of muscle differentiation and lipid deposition in culture were determined by immunohistochemical, biochemical, and molecular biological evaluations. Notably, the protocol successfully achieved the process of myogenic and lipogenic stimulation in the 3D tissues.

As addressed after the initial review process, the manuscript is clearly written with well-supportive figures. The study design is reasonable with adequate analysis. In the revised manuscript, the authors further discussed the ideas in terms of the approach using genetic modification for cell-based meat production. However, more careful considerations may still be helpful when actually using the technology for cultivated meat production.

eLife assessment

The authors present an important theoretical framework that describes the interplay between liquid-liquid phase separation and protein aggregation within a mean-field model. This work will be of high interest to the biophysics and molecular biology communities, as it will understand and analyse assembly within biomolecular condensates in cells or in-vitro. Major strengths of this convincing work are the consideration of aggregates with various dimensionality and the possibility for protein gelation. A relative weakness is the lack of intuitive interpretation of some of the results and the work could be more accessible to non-experts.

Reviewer #1 (Public Review):

Summary:

The authors present a mean-field model that describes the interplay between (protein) aggregation and phase separation. Different classes of interaction complexity and aggregate dimensionality are considered, both in calculations concerning (equilibrium) phase behavior and kinetics of assembly formation.

Strengths:

The present work is, although purely theoretical, of high interest to understanding biological processes that occur as a result of a coupling between protein aggregation and phase separation. Of course, such processes are abundant, in the living cell as well as in in-vitro experiments. I appreciate the consideration of aggregates with various dimensionality, as well as the categorization into different "interaction classes", together with the mentioning of experimental observations from biology. The model is convincing and underlines the complexity associated with the distribution of proteins across phases and aggregates in the living cell.

Weaknesses:

There are a few minor weaknesses.

Reviewer #3 (Public Review):

Summary:

The authors combine classical theories of phase separation and self-assembly to establish a framework for explaining the coupling between the two phenomena in the context of protein assemblies and condensates. By starting from a mean-field free energy for monomers and assemblies immersed in solvent and imposing conditions of equilibrium, the authors derive phase diagrams indicating how assemblies partition into different condensed phases as temperature and the total volume fraction of proteins are varied. They find that phase separation can promote assembly within the protein-rich phase, providing a potential mechanism for spatial control of assembly. They extend their theory to account for the possibility of gelation. They also create a theory for the kinetics of self-assembly within phase separated systems, predicting how assembly size distributions change with time within the different phases as well as how the volumes of the different phases change with time.

Strengths:

The theoretical framework that the authors present is an interesting marriage of classic theories of phase separation and self-assembly. Its simplicity should make it a powerful general tool for understanding the thermodynamics of assembly coupled to phase separation, and it should provide a useful framework for analyzing experiments on assembly within biomolecular condensates.

The key advance over previous work is that the authors now account for how self-assembly can change the boundaries of the phase diagram.

A second interesting point is the explicit theoretical consideration for the possibility that gelation (i.e. self-assembly into a macroscopic aggregate) could account for widely observed solidification of condensates. While this concept has been broadly discussed, to date I have yet to see a rigorous theoretical analysis of the possibility.

The kinetic theory in sections 5 and 6 is also interesting as it extends on previous work by considering the kinetics of phase separation as well as those of self-assembly.

Weaknesses:

A key point the authors make about their theory is that it allows, as opposed to previous research, to study non-dilute limits. It is true that they consider gelation when the 3D assemblies become macroscopic. However, dilute solution theory assumptions seem to be embedded in many aspects of their theory, and it is not always clear where else the non-dilute limits are considered. Is it in the inter-species interaction \chi_{ij}? Why then do they never explore cases for which \chi_{ij} is nonzero in their analysis?

The connection between this theory and biological systems is described in the introduction but lost along the main text. It would be very helpful to point out, for instance, that the presence of phase separation might induce aggregation of proteins. This point is described formally at the end of Section 3, but a more qualitative connection to biological systems would be very useful here.

Building on the previous point, it would be helpful to give an intuitive sense of where the equations derived in the Appendices and presented in the main text come from and to spell out clear physical interpretations of the results. For example, it would be helpful to point out that Eq. 4 is a form of the law of mass action, familiar from introductory chemistry.

It would be useful to better explain how the current work extends on existing previous work from these authors as well as others. Along these lines, closely related work by W. Jacobs and B. Rogers [O. Hedge et al. 2023, https://arxiv.org/abs/2301.06134; T. Li et al. 2023, https://arxiv.org/abs/2306.13198] should be cited in the introduction.

The results discussed in the first paragraph of Section 3 on assembly size distributions in a homogeneous system are well-known from classic theories of self-assembly. This should be acknowledged and appropriate references should be added; see for instance Rev. Mod. Phys. 93, 025008 and Statistical Thermodynamics Of Surfaces, Interfaces, And Membranes by Sam Safran.

Equation 14 for the kinetic of volume fractions is given with a reference to Bauermann et al 2022, but it should be accompanied by a better intuitive interpretation of its terms in the main text. In particular, how should one understand the third term in this equation? Why does the change in volume impact the change of volume fraction in this way?

The discussion in the last paragraph of Section 6 should be clarified. How can the total amount of protein in both phases decrease? This would necessarily violate either mass or volume conservation. Also, the discussion of why the volume is non-monotonic in time is not clear.

difference of data samples and simulation samples

eLife assessment

This important study describes how PhoP regulates cyclic-AMP production in the human pathogen Mycobacterium tuberculosis. The authors provide convincing evidence that PhoP acts as a repressor of the cyclic-AMP-specific phosphodiesterase, Rv0805, which can degrade cyclic-AMP. The revised manuscript has addressed all outstanding comments and the work will be of interest to bacteriologists.

Reviewer #1 (Public Review):

Summary:

This paper provides a straightforward mechanism of how mycobacterial cAMP level is increased under stressful conditions and shows that the increase is important for the survival of the bacterium in animal hosts. The cAMP level is increased by decreasing the expression of an enzyme that degrades cAMP.

Strengths:

The paper shows that under different stresses the response regulator PhoP represses a phosphodiesterase (PDE) that degrades cAMP specifically. Identification of PhoP as a regulator of cAMP is significant progress in understanding Mtb pathogenesis, as an increase in cAMP apparently increases bacterial survival upon infection. On the practical side, reduction of cAMP by increasing PDE can be a means to attenuate the growth of the bacilli. The results have wider implications since PhoP is implicated in controlling diverse mycobacterial stress responses and many bacterial pathogens modulate host cell cAMP levels. The results here are straightforward, internally consistent, and of both theoretical and applied interests. The results also open considerable future work, especially how increases in cAMP level help to increase survival of the pathogen.

Weaknesses:

It is not clear whether PhoP-PDE Rv0805 is the only pathway to regulate cAMP level under stress.

Comments on revised submission:

The authors have addressed my comments adequately, actually except for all but one. I have only one comment to do with the last line of the abstract. First, "genetic manipulation" usually means changing DNA. In Mtb pathogenesis I hope there is no DNA modification or change in the bacterial DNA. Also, the authors did not really inactivate the whole PhoP- rv0805-cAMP pathway. It would be best if the last line is made more fact based: Thus, inactivation of PhoP decreases cAMP level, thereby stress tolerance and intracellular survival of the bacillus.

Reviewer #2 (Public Review):

Summary:

In the manuscript, the authors have presented new mechanistic details to show how intracellular cAMP levels are maintained and linked to the phosphodiesterase enzyme which in turn is controlled by PhoP. Later, they showed the physiological relevance linked to altered cAMP concentrations.

Strengths:

Well-thought-out experiments. The authors carefully planned the experiments well to uncover the molecular aspects of it diligently.

Weaknesses:

None. The authors have meticulously responded to all my queries and concerns through multiple rounds of review.

Genau genommen gründete Snell schon 1944 (Anregungen von Ernst Kapp aufgreifend) das „Archiv für griechische Lexikographie“ (vgl. dazu Lohse, Gerhard, Bruno Snell (1896–1986). Geisteswissenschaft und politische Erfahrung im 20. Jahrhundert, Göttingen 2023, 122–124), das 1950 zum Thesaurus linguae Graecae wurde, aus dem der Index Hippocraticus (1989 abgeschlossen) und das Lexikon des frühgriechischen Epos (2010 abgeschlossen) entstanden.

eLife assessment

This valuable work presents the latest version of CTFFIND, which is the most popular software for determination of the contrast transfer function (CTF) in cryo-electron microscopy. CTFFIND5 estimates and considers acquisition geometry and sample thickness, which leads to improved CTF determination. The paper describes convincing evidence that CTFFIND5 finds better CTF parameters than previous methods, in particular for tilted samples (e.g. for cryo-electron tomography) or where thickness is an issue (e.g. cellular samples, or electron microscopy at low voltages).

Reviewer #1 (Public Review):

This work presents CTFFIND5, a new version of the software for determination of the Contrast Transfer Function (CTF) that models the distortions introduced by the microscope in cryoEM images. CTFFIND5 can take acquisition geometry and sample thickness into consideration to improve CTF estimation.

To estimate tilt (tilt angle and tilt axis), the input image is split into tiles and correlation coefficients are computed between their power spectra and a local CTF model that includes the defocus variation according to a tilted plane. As a final step, by applying a rescaling factor to the power spectra of the tiles, an average tilt-corrected power spectrum is obtained and used for diagnostic purposes and to estimate the goodness of fit. This global procedure and the rescaling factor resemble those used in Bsoft, Warp, etc, with determination of the tilt parameters being a feature specific of CTFFIND5 (and formerly CTFTILT). The performance of the algorithm is evaluated with tilted 2D crystals and tilt-series, demonstrating accurate tilt estimation in some cases and some limitations in others. Further analysis of CTF determination with tilt-series, particularly showing whether there is accurate or stable estimation at high tilts, might be helpful to show the robustness of CTFFIND5 in cryoET.

CTFFIND5 represents the first CTF determination tool that considers the thickness-related modulation envelope of the CTF firstly described by McMullan et al. (2015) and experimentally confirmed by Tichelaar et al. (2020). To this end, CTFFIND5 uses a new CTF model that takes the sample thickness into account. CTFFIND5 thus provides more accurate CTF estimation and, furthermore, gives an estimation of the sample thickness, which may be a valuable resource to judge the potential for high resolution. To evaluate the accuracy of thickness estimation in CTFFIND5, the authors use the Lambert-Beer law on energy-filtered data and also tomographic data, thus demonstrating that the estimates are reasonable for images with exposure around 30 e/A2. While consideration of sample thickness in CTF determination sounds ideally suited for cryoET, practical application under the standard acquisition protocols in cryoET (exposure of 3-5 e/A2 per image) is still limited. In this regard, the authors are honest in the conclusions and clearly identify the areas where thickness-aware CTF determination will be valuable at present: e.g. in situ single particle analysis and in vitro single particle cryoEM of purified samples at low voltages.

In conclusion, the manuscript introduces novel methods inside CTFFIND5 that improve CTF estimation, namely acquisition geometry and sample thickness. The evaluation demonstrates the performance of the new tool, with fairly accurate estimates of tilt axis, tilt angle and sample thickness and improved CTF estimation. The manuscript critically defines the current range of application of the new methods in cryoEM.

Reviewer #2 (Public Review):

Summary:

This paper describes the latest version of the most popular program for CTF estimation for cryo-EM images: CTFFIND5. New features in CTFFIND5 are the estimation of tilt geometry, including for samples, like FIB-milled lamellae, that are pre-tilted along a different axis than the tilt axis of the tomographic experiment, plus the estimation of sample thickness from the expanded CTF model described by McMullan et al (2015). The results convincingly show the added value of the program for thicker and tilted images, such as are common in modern cryo-ET experiments. The program will therefore have a considerable impact on the field.

I have only minor suggestions for improvement below:

Abstract: "[CTF estimation] has been one of the key aspects of the resolution revolution"-> This is a bit over the top. Not much changed in the actual algorithms for CTF estimation during the resolution revolution.<br /> L34: "These parameters" -> Cs is typically given, only defocus (and if relevant phase shift) are estimated.<br /> L110-116: The text is ambiguous: are rotations defined clockwise or counter-clockwise? It would be good to explicitly state what subsequent rotations, in which directions and around which axes this transformation matrix (and the input/output angles in CTFFIND5) correspond to.<br /> L129-130: As a suggestion: it would be relatively easy, and possibly beneficial to the user, to implement a high-resolution limit that varies with the accumulated dose on the sample. One example of this exists in the tomography pipeline of RELION-5.<br /> Substituting Eq (7) into Eq (6) yields ksi=pi, which cannot be true. If t is the sample thickness, then how can this be a function of the frequency g of the first node of the CTF function? The former is a feature of the sample, the latter is a parameter of the optical system. This needs correction.

Reviewer #3 (Public Review):

In this manuscript, the authors detail improvements in the core CTFFIND (CTFFIND5 as implemented in cisTEM) algorithm that better estimates CTF parameters from titled micrographs and those that exhibit signal attenuation due to ice thickness. These improvements typically yield more accurate CTF values that better represent the data. Although some of the improvements result in slower calculations per micrograph, these can be easily overcome through parallelization.

There are some concerns outlined below that would benefit from further evaluation by the authors.

For the examples shown in Figure 3b, given the small differences in estimated defocus1 and 2, what type of improvements would be expected in the reconstructed tomograms? Do such improvements in estimates manifest in better tilt-series reconstruction?

Similarly, the data shown in Figure 3C shows minimal improvements in the CTF resolution estimate (e.g., 4.3 versus 4.2 Å), but exhibited several hundred Å difference in defocus values. How do such differences impact downstream processing? Is such a difference overcame by per-particle (local) CTF refinements (like the authors mention in the discussion, see below)?

At which point does the thickness of the specimen preclude the ice thickness modulation to be included for "accurate" estimate? 500Å? 1000Å? 2000Å? Based on the data shown in Figure 3B, as high as 969 Å thick specimens benefit moderately (4.6 versus 3.4 Å fit estimate), but perhaps not significantly, from the ice thickness estimation. Considering the increased computational time for ice thickness estimation, such an estimate of when to incorporate for single-particle workflows would be beneficial.

It would seem that this statement could be evaluated herein: "the analysis of images of purified samples recorded at lower acceleration voltages, e.g., 100 keV (McMullan et al., 2023), may also benefit since thickness-dependent CTF modulations will appear at lower resolution with longer electron wavelengths". There are numerous examples of 300kV, 200kV, and 100kV EMPIAR datasets to be compared and recommendations would be welcomed.

Although logical, this statement is not supported by the data presented in this manuscript: "The improvements of CTFFIND5 will provide better starting values for this refinement, yielding better overall CTF estimation and recovery of high-resolution information during 3D reconstruction."

Moreso, the lack of single-particle data evaluation does present a concern. Naively, these improvements would benefit all cryoEM data, regardless of modality.

eLife assessment

This work is of fundamental significance and has a compelling level of evidence for a new population that protects against obesity-induced hypothalamic inflammation. This topic will attract attention from a broad base of readers, from hypothalamic neuroscientists to immunologists with an interest in metabolism.

Reviewer #1 (Public Review):

Summary:

The present work from Velloso and collaborators investigated the transcription profiles of resident and recruited hypothalamic microglia. They found sex-dependent differences between males and females and identified the protective role of chemokine receptor CXCR3 against diet-induced obesity.

Strengths:

(1) Novelty<br /> (2) Relevance, since this work provides evidence about a subset of recruited microglia that has a protective effect against DIO. This provides a new concept in hypothalamic inflammation and obesity.

Weaknesses:

(1) Lack of mechanistic insight into the sex-dependent effects.<br /> (2) Analysis of indirect calorimetry data requires more depth.<br /> (3) A deeper analysis of hypothalamic inflammation and ER stress pathways would strengthen the manuscript.

Reviewer #2 (Public Review):

Summary:

This study by Mendes et al provides novel key insights into the role of chemotaxis and immune cell recruitment into the hypothalamus in the development of diet-induced obesity. Specifically, the authors reveal that although transcriptional changes in hypothalamic resident microglia following exposure to high-fat feeding are minor, there are compelling transcriptomic differences between resident microglia and microglia recruited to the hypothalamus, and these are sexually dimorphic. Using independent loss-of-function studies, the authors also demonstrate an important role of CXCR3 and hypothalamic CXCL10 in the hypothalamic recruitment of CCR2+ positive cells on metabolism following exposure to high-fat diet-feeding in mice. This manuscript puts forth conceptually novel evidence that inhibition of chemotaxis-mediated immune cell recruitment accelerates body weight gain in high-fat diet-feeding, suggesting that a subset of microglia that express CXCR3 may confer protective, anti-obesogenic effects.

Strengths:

The work is exciting and relevant given the prevalence of obesity and the consequences of inflammation in the brain on perturbations of energy metabolism and ensuant metabolic diseases. Hypothalamic inflammation is associated with disrupted energy balance, and activated microglia within the hypothalamus resulting from excessive caloric intake and saturated fatty acids are often thought to be mediators of impairment of hypothalamic regulation of metabolism. The present work reports a novel notion in which immune cells recruited into the hypothalamus that express chemokine receptor CXCR3 may have a protective role against diet-induced obesity. In vivo studies reported herein demonstrate that inhibition of CXCR3 exacerbates high-fat diet-induced body weight gain, increases circulating triglycerides and fasting glucose levels, worsens glucose tolerance, and increases the expression of orexigenic neuropeptides, at least in female mice.

This work provides a highly interesting and needed overview of preclinical and clinical brain inflammation, which is relevant to readers with an interest in metabolism and immunometabolism in the context of obesity.

Using flow cytometry, cell sorting, and transcriptomics including RNA-sequencing, the manuscript provides novel insights into transcriptional landscapes of resident and recruited microglia in the hypothalamus. Importantly, sex differences are investigated.

Overall, the manuscript is perceived to be highly interesting, relevant, and timely. The discussion is thoughtful, well-articulated, and a pleasure to read and felt to be of interest to a broad audience.

Weaknesses:

There were no major weaknesses perceived. Some comments for potential textual additions to the results/discussion are listed in recommendations to authors.

eLife assessment

This fundamental study advances our understanding of the cell specific treatment of cone photoreceptor degeneration by Txnip. The evidence supporting the conclusions is compelling with rigorous genetic manipulation of Txnip mutations. The work will be of broad interest to vision researchers, cell biologists and biochemists.

Reviewer #1 (Public Review):

Summary:

This is a follow-up study to the authors' previous eLife report about the roles of an alpha-arrestin called protein thioredoxin interacting protein (Txnip) in cone photoreceptors and in the retinal pigment epithelium. The findings are important because they provide new information about the mechanism of glucose and lactate transport to cone photoreceptors and because they may become the basis for therapies for retinal degenerative diseases.

Strengths:

Overall, the study is carefully done and, although the analysis is fairly comprehensive with many different versions of the protein analyzed, it is clearly enough described to follow. Figure 4 greatly facilitated my ability to follow, understand and interpret the study. The authors have appropriately addressed a few concerns about statistical significance and the relationship between their findings and previous studies of the possible roles of Txnip on GLUT1 expression and localization on the surfaces of RPE cells.

Reviewer #2 (Public Review):

The hard work of the authors is much appreciated. With overexpression of a-arrestin Txnip in RPE, cones and the combined respectively, the authors show a potential gene agnostic treatment that can be applied to retinitis pigmentosa. Furthermore, since Txnip is related to multiple intracellular signaling pathway, this study is of value for research in the mechanism of secondary cone dystrophy as well.

Strengths

- The follow-up study builds on innovative ground by exploring the impact of TxnipC247S and its combination with HSP90AB1 knockdown on cone survival, offering novel therapeutic pathways.<br /> - Testing of different Txnip deletion mutants provides a nuanced understanding of its functional domains, contributing valuable insights into the mechanism of action in RP treatment.<br /> - The findings regarding GLUT1 clearance and the differential effects of Txnip mutants on cone and RPE cells lay the groundwork for targeted gene therapy in RP.

Comments on revised version:

The researchers answered our questions and included additional discussion in the manuscript.

1 +

1 δειγμα

3 ερωτ

3 ερωτ

3 ερωτ

3 ερωτ

3 ερωτ

4

Connant is the physicist (cosmic ray specialist) and on night watch with the creature.

Những câu chuyện mang tính cảnh giác hoặc đạo đức, trong đó trẻ ngoan được khen thưởng và trẻ hư bị trừng phạt thích đáng, nhìn chung ít được quan tâm đến hình ảnh minh họa hơn so với truyện cổ tích. Nhiều cuốn sách về tôn giáo được viết dưới ảnh hưởng của những người theo đạo Tin lành Anh giáo, và chúng đã được xuất bản với số lượng lớn trong suốt thế kỷ 18 và đầu thế kỷ 19 bởi các công ty như Newbery và Marshall. Sự gia tăng nhanh chóng các ấn bản của những cuốn sách như Những bài hát thiêng liêng của Isaac Watts (1715; xem mèo. số 213) chứng tỏ sự phổ biến lâu dài của những tác phẩm biến các bài học tôn giáo thành một hình thức thú vị hơn. Trong số các tác giả nữ viết truyện sùng đạo hoặc truyện đạo đức đáng chú ý nhất ở Anh có Trimmer, Anna Laetitia Barbauld (1743–1825) và Mary Martha Sherwood.

Những câu chuyện cổ tích, cũng như những câu chuyện phiêu lưu nổi tiếng như Robinson Crusoe của Daniel Defoe (1719; cat. nos. 115, 121), thường gây ra nhiều lời chỉ trích trong thế kỷ 18 và đầu thế kỷ 19. Sarah Trimmer (1741–1810), một tác giả nổi tiếng về các cuốn sách dạy đạo đức, đã lên án “những sinh vật tưởng tượng dành cho trẻ em” trong bài phê bình Truyện cổ tích của Mẹ Bunch năm 1773.9 Thật vậy, mặc dù nhiều ấn bản sách giáo khoa của Perrault đã được xuất bản trong suốt thế kỷ 18, chúng nhìn chung bị lu mờ bởi những cuốn sách mang tính mô phạm hơn đề cập đến các vấn đề đạo đức hoặc tôn giáo. Mãi đến thế kỷ 19, truyện cổ tích mới thống trị thị trường sách thiếu nhi.

Hai thể loại quan trọng nhất của văn học thiếu nhi thế kỷ 18 là truyện cổ tích và truyện đạo đức. Truyện cổ tích được truyền từ thế hệ này sang thế hệ khác thông qua truyền miệng, lần đầu tiên được sưu tầm và in tại triều đình Pháp của Louis XIV bởi các nhà văn như Nữ bá tước d'Aulnoy, Madame de Villeneuve và Madame Le Prince de Beaumont. . Cuốn sách Histoires ou contes du temps passé năm 1697 của Charles Perrault (Những câu chuyện về cách đây đã lâu; xem cat. nos. 144, 146–48, 150, 192) chứa các phiên bản viết đầu tiên của "Cô bé lọ lem", "Người đẹp ngủ trong rừng", "Cô bé quàng khăn đỏ", “Blue Beard,” “Hop o' My Thumb,” và “Puss in Boots.”8 Phiên bản của Perrault về những câu chuyện này đã thống trị văn học thiếu nhi Anh và Mỹ kể từ thế kỷ thứ mười tám. Mặt trước của ấn bản gốc của ông (hình 11) là hình một bà già kể chuyện cho một nhóm trẻ em, với dòng chữ Contes de ma mère l'oye ("Truyện kể về mẹ ngỗng", một cách diễn đạt dân gian của Pháp gần tương đương với "chuyện xưa". chuyện của vợ”). Đây là lần xuất hiện đầu tiên của nhân vật mà sau này gắn liền với những bài đồng dao dành cho trẻ mẫu giáo khi hãng Newbery gắn tên vào một tuyển tập được xuất bản với tựa đề Giai điệu của Mẹ Ngỗng; hoặc, Sonnets for the Cradle (1781; cat. no. 154).

Thái độ mới đối với trẻ em và nền giáo dục của chúng bắt đầu phát triển vào cuối thế kỷ 17, khi nhiều nhà giáo dục kêu gọi quan tâm nhiều hơn đến những nhu cầu đặc biệt của trẻ và khi khái niệm về niềm vui học tập ngày càng được chấp nhận rộng rãi hơn.3 Biểu hiện rõ ràng nhất của sự phát triển ý tưởng này là các bài viết của triết gia John Locke (1632–1704) và Jean-Jacques Rousseau (1712–78). Năm 1693, Locke viết trong Một số suy nghĩ liên quan đến giáo dục rằng “trẻ em nên được đối xử như những sinh vật có lý trí…. Chúng không được bị cản trở để trở thành trẻ con, cũng như không được chơi và làm như trẻ con, mà là bị bệnh.” Rousseau coi tuổi thơ như một đứa trẻ. một trạng thái trong sáng và tự nhiên - khác biệt với trạng thái trưởng thành - và tin rằng mục tiêu trọng tâm của giáo dục phải là bảo tồn bản chất nguyên thủy của trẻ. Ông cũng tin rằng điều cần thiết là giáo viên phải nhìn sự việc như trẻ em nhìn.5 Các tác phẩm của Locke và Rousseau đã ảnh hưởng đến các nhà giáo dục Anh, và ý tưởng của họ cuối cùng đã dẫn tới một cách tiếp cận giáo dục nhân đạo hơn, trong đó niềm vui được coi là một phương tiện hỗ trợ cho việc học.

Một trong những thể loại tiểu thuyết lâu đời nhất, truyện ngụ ngôn (xem cat. nos. 15–32) ban đầu được đọc bằng tiếng Latinh trong lớp học hơn là để giải trí ở nhà. Những câu chuyện được cho là của Aesop (được cho là một người kể chuyện Hy Lạp ở thế kỷ thứ sáu trước Công nguyên nhưng gần như chắc chắn là một nhân vật huyền thoại) nằm trong số những câu chuyện được xuất bản và minh họa thường xuyên nhất (xem hình 5, 6). Truyện ngụ ngôn của Aesop được xuất bản trong bản dịch tiếng Anh đầu tiên của William Caxton (khoảng 1422–91) vào năm 1484. Nó nhanh chóng trở thành một trong những cuốn sách có tranh minh họa phổ biến nhất dành cho trẻ em, mặc dù trong nhiều ấn bản đầu tiên có rất ít nỗ lực chuyển thể các câu chuyện để tạo nên một tác phẩm hay. chúng dễ dàng hơn cho trẻ em hiểu và liên hệ với

Sách bảng chữ cái minh họa một trong những cách sử dụng hình ảnh sớm nhất trong sách hướng dẫn dành cho trẻ em (xem hình 2; mèo. số 33–77). Từ thế kỷ 16 cho đến tận thế kỷ 18, trẻ em học bảng chữ cái bằng cách nghiên cứu sách sừng (xem cat. nos. 33, 38, 39, 50), những mái chèo bằng gỗ có ghi các bảng chữ cái thường được kết hợp với các tác phẩm tôn giáo như Kinh Lạy Cha. Ngoài truyền thống sách kèn đã phát triển thêm battledore bằng hình ảnh (xem cat. nos. 41, 45, 47, 54, 56, 58, 59), một mảnh bìa cứng gấp lại với bảng chữ cái minh họa, được đặt tên theo một trò chơi truyền thống trong đó sách sừng dùng làm mái chèo. Battledore tồn tại cho đến giữa thế kỷ 19. Vào đầu thế kỷ 19, các loại trò chơi có hình ảnh minh họa khác đã được phát triển để dạy ABC cũng như toán, ngữ pháp và khoa học.

Para la actualidad la tecnología sigue evolucionando cada vez más y con ellos los programas educativos que se encuentran disponibles para cada usuario a menos de que sea una aplicación pagada, pero por lo general la mayoría de ellas están a nuestra disposición al momento que deseemos.

Can you add these as icons below this section so they stand out?

Like a logo icon bar where you list them side by side with a matching icon.

Can you possibly add a private podcast seeing that your students are busy mums? Just a thought. ;-)

Can you be more specific here around how many recipes and what type of recipes, as well as the benefits of having these?

This one also doesn't seem to apply

Delete this one too

This doesn't really apply because they get access to the replay, right? I'd just delete this and change it to something like... how long do I get access to the class for?

Change this to something like 'How the first aid skills you'll uncover have helped others'

I'd also delete this to keep it short and sharp

Change this to 'the xx-min medicinal first aid class video'

Shorten this to 'Get instant access to...'

delete this and change to 'Imagine solving those inevitable ouchies with the wisdom...'

And remove the code because it should link to the discounted checkout cart, right?

Cross out the $47 original price here

I'd delete the term 'the class' to keep it super concise

I would change the wording of the Deadline Funnel bar to 'Special One-Time Offer: SAVE $20 RIGHT NOW'

This comment is not properly reflected in Figure 2.6.1. Specifically, Figure 2.6.1 makes it appear that all n=3 electrons, whether s, p, or d, will shield at 0.35. While the table calls out that l less than the probe electron value (ie l=2 for 3d electron) receives a shielding constant of 1 (ie 3s and 3p electrons) even though the n value is the same for 3s, 3p, and 3d electrons.

Las investigaciones realizadas sobre el nivel de efectividad de la gamificación aumentaron de manera significativa, ya que muchos maestros empezaron a adoptar enfoques tecnológicos y mediante esta gamificación se logró identificar que los niños aumentaban el grado de motivación y rendimiento académico.

Imagery. Details of what is found around them, makes the reader feel like they are right there.

Personification. Stoves aren't actually capable of cooking themselves.

Genre: science fiction, dealt a lot with the idea of technology taking over. This story feels as if it was in the Postmodernist movement

Explaining what is going to be told in this collection of short stories. The stories that are on the illustrated man are the stories the narrator is going to see.

imagery, making the reader feel as if they are present in this part of the story.

Personification. Pictures don't actually move.

Overall I think this is my best source. They only reported actual facts and kept opinions out but still talked about the conspiracies but more importantly, why they started.

See this is an actual good quote. It isn't from a random internet sluth bored at home, but an actual royal spokesperson explaining things from their perspective.

Great fact to add. Seems like they are actually just reporting the facts.

This was a big part of the conspiracies.

Well of course if she was undergoing abdomen surgery I wouldn't want to be awake either. Weird thing to site.

Yes I am glad this was pointed out. When King Charles was sick he was still photographed all the time in public but Kate was not. I think that is what sparked the she was killed or kidnapped rumors.

Gossipy is a strong word choice. Could be pushing a narrative of people are too bored and nosey.

This wording was used in another article... weird choice and I do not know what it means.

Using old photos could be confusing for those who do not read the dates.

From what I know about Forbes is that they are a legitimate news source? Interested to see how this play out.

Los programas y herramientas tecnológicas han facilitado la forma de enseñar a los estudiante, por que hoy en día existe un sin número de actividades que se puede desarrollar de manera mas lúdica y creativa, de esta manera también logramos captar la concentración y atención de los estudiantes de forma mas sencilla y eficaz.

After a little more research, this article is not under the opinions page, which is just interesting to note based on the tones and wording used.

Again with these random quotes. This quote does not make me any more convinced because they asked a random Oklahoma lady.

Have not heard this yet. I think this article has some different information that the others.

These are some new theories not mentioned in the other articles.

A lot of big words used already two paragraphs in. I think they are trying to establish some kind of legitimacy?

What on Earth is this opening line.

Como se evidencia hoy en día, los avances tecnológicos son de gran ayuda en el ámbito educativo ya que son implementados en cada aula de clases para generar un mejor ambiente de aprendizaje, de igual manera cada docente busca mejorar su proceso de enseñanza-aprendizaje por medio de estos programas de innovación.

For a gossip site they do show a lot of evidence as to why they think what they think. Somewhat convincing.

The palace is a great source of evidence here.

No one ever mentioned that the palace said she would be out until Easter or else I think a lot of people would not have believed the conspiracies.

Most people know cosmo is not a "news" source in a sense, more just a gossip site, kind of like a better TMZ. But for those who don't know, this line could be misleading.

I know my topic is before people found out she has cancer and the truth, but this article looks back at the conspiracies which I think could be interesting.

I could see why the author may have took offense to the term "that" being used to describe the population of students at the school. It is dismissive, and almost others the population of students at the school, by labeling them as different from the "normal" or "typical" student bodies at other more affluent high schools. There is a largely negative connotation towards these students categorized as low-income in an urban classroom setting.

I'm more than halfway thought this article and this is the first mention of William. The title of the article is about what he says and thinks about all of this. Should have come up sooner.

This is the only article to say what actually happened and not in an accusatory tone. I was curious if the news sites just took it down because there was speculation so I am glad they cleared this up.

I get what Kate was trying to do but it does not take an expert to notice bad photoshop. Plus not putting her husband in the picture was a bad choice because it made more rumors.

I do like that this article is starting with the facts in the beginning and then listing the rumors.

A lot of the other article says jan 17th...

The journalist is saying we do these things and make up these elaborate stories because we are bored. Another factor of a conspiracy theory.

Making light of the situation and stating that these are not facts at all which is good because there are stupid people on the internet that would actually believe this.

Playing into the conspiracies by using the word ominous.

They are pointing out exactly what makes a conspiracy theory. I like that.

Niche audience, but this is exactly who reads Vogue though. They understand their audience. Also, makes it kinda known its an opinion piece right off the bat. Respectable.

This passage shed light on an interesting idea. Even though both of Alexander's parents felt that there may be racial prejudice in the school, Alexander's mom, Ms. Williams argued that Mr. Williams should not voice these concerns around Alexander. I could see how even if there was a concern about prejudice, Ms. Williams wanted Alexander to have an open mindset. She nonetheless kept a commitment to vigilance, as did Mr. Alexander.

The wording of this whole article seems harsh and like they want to prove a point...

This is one of the only article to say this. This is why the rumors started. Also, the royal family has updated the public every second of everyone else's health issues in the family, so I will admit since there were no updates that I believed in a conspiracy or two as well.

I remember people being way more suspicious after this video because she looked so different than before.

The whole article this Intelligencer magazine is so mean and critical of The Sun. Wonder what the relation is there.

Abbink, Jon. “Dam Controversies: Contested Governance and Developmental Discourse on the Ethiopian Omo River Dam.” Social Anthropology/Anthropologie Sociale 20, no. 2 (2012): 125–144.

Again a grasp for straws. I don't even see this one.

Bloom in March is such a grasp for straws because they clearly did not have ten things but wanted an article. Makes people look at the image and say "oh they are right" when it literally means nothing.

Why did they chose this man to quote? Anyone could look at the picture and say that. Also, why a professor of computer sciences? Why one at Cal Berkeley? Just a weird quote.

Yanking being the wording here is very harsh, making the news outlets that published the photos seem embarrassed or like they are covering something up.

Well not exactly... according to the post from Kate shown in the article, she did not clearly say this was photoshoped. She said she does experiment with editing sometimes but just says happy mother's day. Bad wording.

Abbink, Jon. “Dam Controversies: Contested Governance and Developmental Discourse on the Ethiopian Omo River Dam.” Social Anthropology/Anthropologie Sociale 20, no. 2 (2012): 125–144.

Leauthaud, Crystele, Stéphanie Duvail, Olivier Hamerlynck, Jean-Luc Paul, Hubert Cochet, Judith Nyunja, Jean Albergel, and Olivier Grünberger. “Floods and Livelihoods: The Impact of Changing Water Resources on Wetland Agro-Ecological Production Systems in the Tana River Delta, Kenya.” Global Environmental Change 23, no. 1 (February 1, 2013): 252–63. doi:10.1016/j.gloenvcha.2012.09.003.

"Water Harvesting has Added Benefit for Kenya - Less Flooding." AllAfrica.Com, Jan 03, 2020. https://ezproxy.lafayette.edu/login?url=https://www.proquest.com/wire-feeds/water-harvesting-has-added-benefit-kenya-less/docview/2334201005/se-2.

"Water Harvesting has Added Benefit for Kenya - Less Flooding." AllAfrica.Com, Jan 03, 2020. https://ezproxy.lafayette.edu/login?url=https://www.proquest.com/wire-feeds/water-harvesting-has-added-benefit-kenya-less/docview/2334201005/se-2.

"Water Harvesting has Added Benefit for Kenya - Less Flooding." AllAfrica.Com, Jan 03, 2020. https://ezproxy.lafayette.edu/login?url=https://www.proquest.com/wire-feeds/water-harvesting-has-added-benefit-kenya-less/docview/2334201005/se-2.

“Lead and Copper Rule.” EPA. Accessed April 9, 2024. https://www.epa.gov/dwreginfo/lead-and-copper-rule.

Katner, Adrienne, Kelsey J. Pieper, Yanna Lambrinidou, Komal Brown, Chih-Yang Hu, Howard W. Mielke, and Marc A. Edwards. “Weaknesses in Federal Drinking Water Regulations and Public Health Policies That Impede Lead Poisoning Prevention and Environmental Justice.” Environmental Justice 9, no. 4 (August 2016): 109–17. https://doi.org/10.1089/env.2016.0012.

Lubrano, Jonathon. “Water, Lead, and Environmental Justice: Easing the Flint Water Crisis With a Public Water Contamination Liability Fund.” William and Mary Environmental Law and Policy Review 42, no. 1 (January 1, 2017): 331. https://scholarship.law.wm.edu/cgi/viewcontent.cgi?article=1697&context=wmelpr.

Clark, Karen. “The Value of Water: The Flint Water Crisis as a Devaluation of Natural Resources, Not a Matter of Racial Justice.” Environmental Justice 9, no. 4 (August 2016): 99–102. https://doi.org/10.1089/env.2016.0007.

Rodgers, Dennis, and Bruce O’Neill. “Infrastructural Violence: Introduction to the Special Issue.” Ethnography 13, no. 4 (October 23, 2012): 401–12. https://doi.org/10.1177/1466138111435738.

London, J.K.; Fencl, A.L.; Watterson, S.; Choueiri, Y.; Seaton, P.; Jarin, J.; Dawson, M.; Aranda, A.; King, A.; Nguyen, P.; Pannu, C.; Firestone, L. and Bailey, C. “Disadvantaged unincorporated communities and the struggle for water justice in California.” Water Alternatives 14(2) (2021): 536. https://www.water-alternatives.org/index.php/alldoc/articles/vol14/v14issue2/626-a14-2-4/file

London, J.K.; Fencl, A.L.; Watterson, S.; Choueiri, Y.; Seaton, P.; Jarin, J.; Dawson, M.; Aranda, A.; King, A.; Nguyen, P.; Pannu, C.; Firestone, L. and Bailey, C. “Disadvantaged unincorporated communities and the struggle for water justice in California.” Water Alternatives 14(2) (2021): 534. https://www.water-alternatives.org/index.php/alldoc/articles/vol14/v14issue2/626-a14-2-4/file

London, J.K.; Fencl, A.L.; Watterson, S.; Choueiri, Y.; Seaton, P.; Jarin, J.; Dawson, M.; Aranda, A.; King, A.; Nguyen, P.; Pannu, C.; Firestone, L. and Bailey, C. “Disadvantaged unincorporated communities and the struggle for water justice in California.” Water Alternatives 14(2) (2021): 529. https://www.water-alternatives.org/index.php/alldoc/articles/vol14/v14issue2/626-a14-2-4/file

The United States Congress, Environmental Protection Agency, Safe Drinking Water Act, 104th Congress, Washington D.C., 1974 https://www.epa.gov/sdwa

Henry, Lois, “Contamination a looming possibility for groundwater bank to be built using public funds”, San Joaquin Valley Water (February 24, 2022). https://sjvwater.org/contamination-a-looming-possibility-for-groundwater-bank-to-be-built-using-public-funds/

“Actions for Cleaner Water., United States Environmental Protection Agency (May 17, 2023). https://www.epa.gov/sanjoaquinvalley/actions-cleaner-water.

“Actions for Cleaner Water., United States Environmental Protection Agency (May 17, 2023). https://www.epa.gov/sanjoaquinvalley/actions-cleaner-water.

The United States Congress, Environmental Protection Agency, Safe Drinking Water Act, 104th Congress, Washington D.C., 1974 https://www.epa.gov/sdwa

“Human Right to Water”, California Department of Water Resources, https://water.ca.gov/Programs/All-Programs/Human-Right-to-Water#:~:text=California%20was%20the%20first%20state,%2C%20cooking%2C%20and%20sanitary%20purposes.

Angel Santiago Fernandez-Bou, José M. Rodríguez-Flores, Alexander Guzman, J. Pablo Ortiz-Partida, Leticia M. Classen-Rodriguez, Pedro A. Sánchez-Pérez, Jorge Valero-Fandiño, Chantelise Pells, Humberto Flores-Landeros, Samuel Sandoval-Solís, Gregory W. Characklis, Thomas C. Harmon, Michael McCullough, Josué Medellín-Azuara. “Water, environment, and socioeconomic justice in California: A multi-benefit cropland repurposing framework” Science of The Total Environment, Volume 858, Part 3 (February 1, 2023), 340 and 343. https://doi.org/10.1016/j.scitotenv.2022.159963

“New Drinking Water Report: Communities of Color More Likely to Suffer Drinking Water Violations For Years” National Resources Defence Council, September 24, 2019 https://www.nrdc.org/press-releases/new-drinking-water-report-communities-color-more-likely-suffer-drinking-water

Huang, Ganlin, and Jonathan K. London. "Cumulative Environmental Vulnerability and Environmental Justice in California’s San Joaquin Valley" International Journal of Environmental Research and Public Health9, no. 5: 1600 (March 19, 2012). https://doi.org/10.3390/ijerph9051593

Huang, Ganlin, and Jonathan K. London. "Cumulative Environmental Vulnerability and Environmental Justice in California’s San Joaquin Valley" International Journal of Environmental Research and Public Health9, no. 5: 1600 (March 19, 2012). https://doi.org/10.3390/ijerph9051593

Boyden, Hollynd, Mayela Gillan, Javier Molina, Ashok Gadgil, and Winston Tseng. "Community Perceptions of Arsenic Contaminated Drinking Water and Preferences for Risk Communication in California’s San Joaquin Valley" International Journal of Environmental Research and Public Health 20, no. 1: 813 (January 1, 2023). https://doi.org/10.3390/ijerph20010813

Balazs, Caroline L., Rachel Morello-Frosch, Alan Hubbard, and Isha Ray. “Just Water?: Social Disparities and Drinking Water Quality in California’s San Joaquin Valley”, (University of California, Berkeley, 2011): 32. https://www.proquest.com/docview/1526000633?pq-origsite=gscholar&fromopenview=true&sourcetype=Dissertations%20&%20Theses

Cole, Luke, and Susan Senger Bowyer. “Pesticides and the Poor in California.” Race, Poverty & the Environment 2, no. 1 (1991): 17. http://www.jstor.org/stable/41555051.

“Farmworkers in California: A Brief Introduction”, California Research Bureau, October 2013, 1. https://latinocaucus.legislature.ca.gov/sites/latinocaucus.legislature.ca.gov/files/CRB%20Report%20on%20Farmworkers%20in%20CA%20S-13-017.pdf

London, J.K.; Fencl, A.L.; Watterson, S.; Choueiri, Y.; Seaton, P.; Jarin, J.; Dawson, M.; Aranda, A.; King, A.; Nguyen, P.; Pannu, C.; Firestone, L. and Bailey, C. “Disadvantaged unincorporated communities and the struggle for water justice in California.” Water Alternatives 14(2) (2021): 533. https://www.water-alternatives.org/index.php/alldoc/articles/vol14/v14issue2/626-a14-2-4/file

“New Drinking Water Report: Communities of Color More Likely to Suffer Drinking Water Violations For Years” National Resources Defence Council, September 24, 2019 https://www.nrdc.org/press-releases/new-drinking-water-report-communities-color-more-likely-suffer-drinking-water

U.S. Census Bureau, “Population by Race (San Joaquin County)”, San Joaquin Council of Governments, 2020. https://www.sjcog.org/236/Demographics

Matthew 6:34 “Take therefore no thought for the morrow: for the morrow shall take thought for the things of itself. Sufficient unto the day is the evil thereof.”

I did not previously think about how age impacts the way you experience poverty, but I can see how this may be true. When we are little kids, we are still unaware of a lot of the different facets of identity that set us apart, and are more likely to be open minded to more things-- we are still very easily impressionable. As we become older though, and cliques form, there is a way clearer understanding of what may be deemed as cool or desirable for teenagers and what is embarrassing or something to be ashamed of.

To me this passage ties back to flawed pedagogical structures in k-12 schools; so often the academics and even fun seeming extracurricular activities are tailored to a white middle class (nuclear families). They do not seem to be inclusive or take into consideration that certain children may not have or live with fathers, and that a Father's Day activity might not feel right to them.

Blake, Sarah B., “Spatial Relationships among Dairy Farms, Drinking Water Quality, and Maternal-Child Health Outcomes in the San Joaquin Valley.” Public Health Nursing, 31 (2014): 492. https://doi.org/10.1111/phn.12166

“Water & Health in the Valley: Nitrate Concentration of Drinking Water and the Health of San Joaquin Valley Residents”, California State Water Resources Control Board, 5408 and 5410. https://www.waterboards.ca.gov/public_notices/petitions/water_quality/docs/a2239/overview/Documents/AR-Docs%20(296).pdf

Gunier, Robert B., Martha E. Harnly, Peggy Reynolds, Andrew Hertz, and Julie Von Behren. “Agricultural Pesticide Use in California: Pesticide Prioritization, Use Densities, and Population Distributions for a Childhood Cancer Study.” Environmental Health Perspectives 109, no. 10 (2001): 1071–78. https://doi.org/10.2307/3454963.

Boyden, Hollynd, Mayela Gillan, Javier Molina, Ashok Gadgil, and Winston Tseng. "Community Perceptions of Arsenic Contaminated Drinking Water and Preferences for Risk Communication in California’s San Joaquin Valley" International Journal of Environmental Research and Public Health 20, no. 1: 813 (January 1, 2023). https://doi.org/10.3390/ijerph20010813

Meadows, Robin, “Living in California’s San Joaquin Valley May Harm Your Health”, The New Humanitarian, July 5, 2017. https://deeply.thenewhumanitarian.org/water/articles/2017/07/05/living-in-californias-san-joaquin-valley-may-harm-your-health

Meadows, Robin, “Living in California’s San Joaquin Valley May Harm Your Health”, The New Humanitarian, July 5, 2017. https://deeply.thenewhumanitarian.org/water/articles/2017/07/05/living-in-californias-san-joaquin-valley-may-harm-your-health

Cole, Luke, and Susan Senger Bowyer. “Pesticides and the Poor in California.” Race, Poverty & the Environment 2, no. 1 (1991): 16. http://www.jstor.org/stable/41555051.

“Water & Health in the Valley: Nitrate Concentration of Drinking Water and the Health of San Joaquin Valley Residents”, California State Water Resources Control Board, 5409. https://www.waterboards.ca.gov/public_notices/petitions/water_quality/docs/a2239/overview/Documents/AR-Docs%20(296).pdf

“Water & Health in the Valley: Nitrate Concentration of Drinking Water and the Health of San Joaquin Valley Residents”, California State Water Resources Control Board, 5410. https://www.waterboards.ca.gov/public_notices/petitions/water_quality/docs/a2239/overview/Documents/AR-Docs%20(296).pdf

Diekman, Kristine, “Sandra Garcia, Fighting for Clean Water”, Run Dry, last modified 2017, http://www.kristinediekman.net/run-dry-1

I thought this was a very interesting point! I can absolutely see the way that certain school cultures can shape a student's understanding of their capabilities or influence them to think they fit into a certain mold. I think there is definitely room for reform in these types of pedagogical structures and biases. Empowering students in underprivileged schools could be great way to help them realize their full potential and not feel as stuck in a box of label as being low SES.

the landlord laughs as because they are foreigners they do not understand the lands local currency, implying they are not to pay because they do not have money from the kingdom. This highlights the hospitability of the innkeeper and suggestive of the cultural norms within the society

highlights the value and principles of individual freedom, but also the king warns them of the challenges they will face if they attempt to leave venice.

the landlord laughs as because they are foreigners they do not understand the lands local currency, implying they are not to pay because they do not have money from the kingdom. This highlights the hospitability of the innkeeper and suggestive of the cultural norms within the society

抗议的结果

I think that it is important to have a team working together to ensure the best practices for the student. It take a village and everyone needs to working to ensure the best support.

This is very important and not just because it is in bold print. This statement helps to create a positive learning community where all learners feel valued. This leads to an enjoyment of learning, as well as, a culturally rich classroom which helps to promote diversity and inclusion.

This finding makes a lot of sense to me-- over 1/3 of families living in poverty experienced six or more risk factors. Thus, with risk factors happening, poverty a lot more likely to occur ( this made me think back to situational poverty, and how one risky or unpredictable situation can land an individual in poverty so quickly). I think all of the risk factors-- emotional and social challenges, acute and chronic stressors, cognitive lags, and health and safety issues-- can become escalated and have the potential to result in a family being in poverty.

I remember when everyone used to hate Rebecca Black because of this song. However, I have recently seen that her other music is extremely critically acclaimed and has one of the "best pop albums" of the 2000s. How do we alter social media to not allow something to impact an artists' impact and artistry?

This demonstrates that the government did not follow the constitution, because if they had, they would have known that this implied that slavery was unconstitutional.

Important because it describes what the federal government did enact.

South Carolina seceded from the United States.

In other words, if an African American flees to another state where they may have greater freedom, if captured, they will be returned to their masters in the state from which they escaped.

Yes

Real

Heck yeah

Ok

Rodgers, Dennis, and Bruce O’Neill. “Infrastructural Violence: Introduction to the Special Issue.” Ethnography 13, no. 4 (October 23, 2012): 401–12. https://doi.org/10.1177/1466138111435738., 402.

Rodgers, Dennis, and Bruce O’Neill. “Infrastructural Violence: Introduction to the Special Issue.”, 402

Fearnside, Philip M. “The Roles and Movements of Actors in the Deforestation of Brazilian Amazonia.”, 23.

Fearnside, Philip M. “The Roles and Movements of Actors in the Deforestation of Brazilian Amazonia.” Ecology and Society 13, no. 1 (2008).

Florêncio dos Reis, Deusiano, Ayala Eduardo Salazar, Mayana Mendes Dias Machado, Sheyla Regina Marques Couceiro, and Paula Benevides de Morais. “Measurement of the Ecological Integrity of CerradoStreams Using Biological Metrics and the Index ofHabitat Integrity.”, 1.

Nixon, Rob. “Slow Violence and the Environmentalism of the Poor.” ,2.

Nixon, Rob. “Slow Violence and the Environmentalism of the Poor.” Princeton University, August 26, 2021. https://english.princeton.edu/research/slow-violence-and-environmentalism-poor., 2

From what I noticed, the majority of old memes were this kind of stuff where people could tease at others who posted themselves doing stuff. Have the standards for our humor changed? Today although making fun of people is common online, it isn't as direct as it used to be.

DOI: 10.1016/j.celrep.2021.108889

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

Curator: @Naa003

SciCrunch record: RRID:AB_141607

What is this?

DOI: 10.1016/j.celrep.2021.108883

Resource: (Sigma-Aldrich Cat# HPA016740, RRID:AB_1855153)

Curator: @Naa003

SciCrunch record: RRID:AB_1855153

What is this?

DOI: 10.1093/g3journal/jkab183

Resource: RRID:DGGR_130448

Curator: @abever99

SciCrunch record: RRID:DGGR_130448

What is this?

DOI: 10.1093/g3journal/jkab183

Resource: (BDSC Cat# 25709,RRID:BDSC_25709)

Curator: @abever99

SciCrunch record: RRID:BDSC_25709

What is this?

DOI: 10.1093/g3journal/jkab183

Resource: RRID:DGGR_106629

Curator: @abever99

SciCrunch record: RRID:DGGR_106629

What is this?

DOI: 10.1093/g3journal/jkab183

Resource: RRID:DGGR_107955

Curator: @abever99

SciCrunch record: RRID:DGGR_107955

What is this?

DOI: 10.1093/g3journal/jkab183

Resource: RRID:DGGR_105669

Curator: @abever99

SciCrunch record: RRID:DGGR_105669

What is this?

DOI: 10.1093/g3journal/jkab183

Resource: RRID:DGGR_109643

Curator: @abever99

SciCrunch record: RRID:DGGR_109643

What is this?

DOI: 10.1093/g3journal/jkab183

Resource: RRID:BDSC_54592

Curator: @abever99

SciCrunch record: RRID:BDSC_54592

What is this?

DOI: 10.1093/g3journal/jkab183

Resource: (BDSC Cat# 54594,RRID:BDSC_54594)

Curator: @abever99

SciCrunch record: RRID:BDSC_54594

What is this?

DOI: 10.25387/g3.13245053

Resource: (BDSC Cat# 55814,RRID:BDSC_55814)

Curator: @abever99

SciCrunch record: RRID:BDSC_55814

What is this?

DOI: 10.25387/g3.13245053

Resource: (BDSC Cat# 24749,RRID:BDSC_24749)

Curator: @abever99

SciCrunch record: RRID:BDSC_24749

What is this?

DOI: 10.1016/j.xcrm.2024.101523

Resource: (KCLB Cat# 30080, RRID:CVCL_0026)

Curator: @abever99

SciCrunch record: RRID:CVCL_0026

What is this?

DOI: 10.1016/j.xcrm.2024.101523

Resource: (NCBI_Iran Cat# C558, RRID:CVCL_0152)

Curator: @abever99

SciCrunch record: RRID:CVCL_0152

What is this?

DOI: 10.1016/j.xcrm.2024.101523

Resource: (ICLC Cat# HTL97008, RRID:CVCL_1644)

Curator: @abever99

SciCrunch record: RRID:CVCL_1644

What is this?

DOI: 10.1016/j.xcrm.2024.101523

Resource: (ICLC Cat# HTL01016, RRID:CVCL_0397)

Curator: @abever99

SciCrunch record: RRID:CVCL_0397

What is this?

DOI: 10.1016/j.xcrm.2024.101523

Resource: (ATCC Cat# CCL-251, RRID:CVCL_1581)

Curator: @abever99

SciCrunch record: RRID:CVCL_1581

What is this?

DOI: 10.1016/j.xcrm.2024.101523

Resource: (KCLB Cat# 0000C1, RRID:CVCL_1708)

Curator: @abever99

SciCrunch record: RRID:CVCL_1708

What is this?

DOI: 10.1016/j.xcrm.2024.101523

Resource: (BCRC Cat# 60343, RRID:CVCL_0547)

Curator: @abever99

SciCrunch record: RRID:CVCL_0547

What is this?

DOI: 10.1016/j.xcrm.2024.101523

Resource: (CLS Cat# 300457/p707_LS-513, RRID:CVCL_1386)

Curator: @abever99

SciCrunch record: RRID:CVCL_1386

What is this?

DOI: 10.1016/j.xcrm.2024.101523

Resource: (KCLB Cat# 22158, RRID:CVCL_1382)

Curator: @abever99

SciCrunch record: RRID:CVCL_1382

What is this?

DOI: 10.1016/j.xcrm.2024.101523

Resource: (ATCC Cat# CCL-237, RRID:CVCL_0632)

Curator: @abever99

SciCrunch record: RRID:CVCL_0632

What is this?

DOI: 10.1016/j.xcrm.2024.101523

Resource: (JCRB Cat# JCRB1022, RRID:CVCL_3088)

Curator: @abever99

SciCrunch record: RRID:CVCL_3088

What is this?

DOI: 10.1016/j.xcrm.2024.101523

Resource: (RRID:CVCL_0220)

Curator: @abever99

SciCrunch record: RRID:CVCL_0220

What is this?