Effects of Red Sorghum-Derived Deoxyanthocyanidins and Their O-ß-D-Glucosides on E-Cadherin Promoter Activity in PC-3 Prostate Cancer Cells.

Although much less common than anthocyanins, 3-Deoxyanthocyanidins (3-DAs) and their glucosides can be found in cereals such as red sorghum. It is speculated that their bioavailability is higher than that of anthocyanins. Thus far, little is known regarding the therapeutic effects of 3-DAs and their O-ß-D-glucosides on cancer, including prostate cancer. Thus, we evaluated their potential to decrease cell viability, to modulate the activity of transcription factors such as NFκB, CREB, and SOX, and to regulate the expression of the gene CDH1, encoding E-Cadherin. We found that 4',7-dihydroxyflavylium chloride (P7) and the natural apigeninidin can reduce cell viability, whereas 4',7-dihydroxyflavylium chloride (P7) and 4'-hydroxy-7-O-ß-D-glucopyranosyloxyflavylium chloride (P3) increase the activities of NFkB, CREB, and SOX transcription factors, leading to the upregulation of CDH1 promoter activity in PC-3 prostate cancer cells. Thus, these compounds may contribute to the inhibition of the epithelial-to-mesenchymal transition in cancer cells and prevent the metastatic activity of more aggressive forms of androgen-resistant prostate cancer.

Regulatory mechanisms of SoxD transcription factors and their influences on male fertility.

Members of the SRY-related box (SOX) subfamily D (SoxD) of transcription factors are well conserved among vertebrate species and play important roles in different stages of male reproductive development. In mammals, the SoxD subfamily contains three members: SOX5, SOX6 and SOX13. Here, we describe their implications in testicular development and spermatogenesis, contributing to fertility. We also cover the mechanisms of action of SoxD transcription factors in gene regulation throughout male development. The specificity of activation of target genes by SoxD members depends, in part, on their post-translational modifications and interactions with other partners. Sperm production in adult males requires the coordination in the regulation of gene expression by different members of the SoxD subfamily of transcription factors in the testis. Specifically, the regulation of genes promoting adequate spermatogenesis by SoxD members is discussed in comparison between species.

Classical cadherins in the testis: how are they regulated?

Cadherins (CDH) are crucial intercellular adhesion molecules, contributing to morphogenesis and creating tissue barriers by regulating cells' movement, clustering and differentiation. In the testis, classical cadherins such as CDH1, CDH2 and CDH3 are critical to gonadogenesis by promoting the migration and the subsequent clustering of primordial germ cells with somatic cells. While CDH2 is present in both Sertoli and germ cells in rodents, CDH1 is primarily detected in undifferentiated spermatogonia. As for CDH3, its expression is mainly found in germ and pre-Sertoli cells in developing gonads until the establishment of the blood-testis barrier (BTB). This barrier is made of Sertoli cells forming intercellular junctional complexes. The restructuring of the BTB allows the movement of early spermatocytes toward the apical compartment as they differentiate during a process called spermatogenesis. CDH2 is among many junctional proteins participating in this process and is regulated by several pathways. While cytokines promote the disassembly of the BTB by enhancing junctional protein endocytosis for degradation, testosterone facilitates the assembly of the BTB by increasing the recycling of endocytosed junctional proteins. Mitogen-activated protein kinases (MAPKs) are also mediators of the BTB kinetics in many chemically induced damages in the testis. In addition to regulating Sertoli cell functions, follicle stimulating hormone can also regulate the expression of CDH2. In this review, we discuss the current knowledge on regulatory mechanisms of cadherin localisation and expression in the testis.

RAS-independent ERK activation by constitutively active KSR3 in non-chordate metazoa.

During early development of the sea urchin embryo, activation of ERK signalling in mesodermal precursors is not triggered by extracellular RTK ligands but by a cell-autonomous, RAS-independent mechanism that was not understood. We discovered that in these cells, ERK signalling is activated through the transcriptional activation of a gene encoding a protein related to Kinase Suppressor of Ras, that we named KSR3. KSR3 belongs to a family of catalytically inactive allosteric activators of RAF. Phylogenetic analysis revealed that genes encoding kinase defective KSR3 proteins are present in most non-chordate metazoa but have been lost in flies and nematodes. We show that the structure of KSR3 factors resembles that of several oncogenic human RAF mutants and that KSR3 from echinoderms, cnidarians and hemichordates activate ERK signalling independently of RAS when overexpressed in cultured cells. Finally, we used the sequence of KSR3 factors to identify activating mutations of human B-RAF. These findings reveal key functions for this family of factors as activators of RAF in RAS-independent ERK signalling in invertebrates. They have implications on the evolution of the ERK signalling pathway and suggest a mechanism for its co-option in the course of evolution.

The transcription factors Creb1 and CEBPB regulate Sox9 promoter activity in TM4 Sertoli cells.

In Sertoli cells, the Sox9 gene is essential for testicular development and normal spermatogenesis. SOX9 is critical for postnatal Sertoli cells differentiation and proliferation in the testis. However, the molecular mechanisms that specifically regulate its expression are not entirely understood. Sox9 expression is regulated by CREB1 and CEBPB in other biological contexts such as during chondrogenesis and in rat thyroid follicular cells. We hypothesized that Sox9 promoter activity is regulated by CREB1 and CEBPB in Sertoli cells. Our results show that Sox9 expression is dependent on the activation of these transcription factors by the cAMP/PKA signaling pathway in TM4 Sertoli cells. Chromatin immunoprecipitation and promoter/reporter luciferase assays with 5' promoter deletions and site-directed mutagenesis demonstrated that CREB1 is being recruited to a DNA regulatory element at -141 bp of the Sox9 promoter region. Such regulation is dependent on the cAMP/PKA signaling pathway, resulting in phosphorylation of CREB1. Activation of Sox9 expression by CEBPB may involve its recruitment to the proximal promoter region by protein-protein interaction with CREB1. Thus, we have shown that the Sox9 promoter is being regulated by the transcription factors CREB1 and CEBPB in TM4 Sertoli cells and involve their recruitment to the proximal promoter region.

A Proof-of-Concept Evaluation of the 1616 Story-Based Positive Youth Development Program.

The 1616 Program is a newly developed and evidence-informed story-based positive youth development (PYD) program for young ice hockey players (10-12 years of age) in North America. The program uses elite ice hockey players as role models-through story-telling-to serve as inspirational figures to engage youth athletes and important social agents (i.e., parents, coaches) with evidence-informed PYD concepts. The objective of this study was to use a Proof-of-Concept evaluation to assess whether the 1616 Program 'worked' in enhancing PYD outcomes and to determine if the concepts were engaging and enjoyable for youth, their parents, and coaches. The 5 week Proof-of-Concept evaluation was conducted with 11 ice hockey teams (n = 160 youths, 93 parents, and 11 coaches), encompassing both qualitative (e.g., focus groups) and quantitative (e.g., retrospective pretest-posttest questionnaires) processes and outcome assessments. Results showed that the program was well received by participants and positively impacted the intended outcomes. Overall, the data presented in this Proof-of-Concept evaluation was deemed to support the development and implementation of the full-scale 1616 Program for a more comprehensive evaluation.

Regulation of Cdh2 by the AP-1 family transcription factor Junb in TM4 Sertoli cells.

Cadherins are transmembrane proteins that mediate cell-to-cell adhesion and various cellular processes. In Sertoli cells of the testis, Cdh2 contributes to the development of the testis and the formation of the blood-testis barrier, being essential for germ cells' protection. Analyses of chromatin accessibility and epigenetic marks in adult mouse testis have shown that the region from -800 to +900 bp respective to Cdh2 transcription start site (TSS) is likely the active regulatory region of this gene. In addition, the JASPAR 2022 matrix has predicted an AP-1 binding element at about -600 bp. Transcription factors of the activator protein 1 (AP-1) family have been implicated in the regulation of the expression of genes encoding cell-to-cell interaction proteins such as Gja1, Nectin2 and Cdh3. To test the potential regulation of Cdh2 by members of the AP-1 family, siRNAs were transfected into TM4 Sertoli cells. The knockdown of Junb led to a decrease in Cdh2 expression. ChIP-qPCR and luciferase reporter assays with site-directed mutagenesis confirmed the recruitment of Junb to several AP-1 regulatory elements in the proximal region of the Cdh2 promoter in TM4 cells. Further investigation with luciferase reporter assays showed that other AP-1 members can also activate the Cdh2 promoter albeit to a lesser extent than Junb. Taken together, these data suggest that in TM4 Sertoli cells, Junb is responsible for the regulation of Cdh2 expression which requires its recruitment to the proximal region of the Cdh2 promoter.

The who, what, and how of teamwork research in medical operating rooms: A scoping review.

Despite the importance of teamwork in the operating room (OR), teamwork can often be conflated with teamwork components (e.g., communication, cooperation). We reviewed the existing literature pertaining to OR teamwork to understand which teamwork components have been assessed. Following PRISMA guidelines for scoping reviews, 4,233 peer-reviewed studies were identified using MEDLINE and Embase. Eighty-seven studies were included for synthesis and analysis. Using the episodic model of teamwork as an organizing framework, studies were grouped into the following teamwork categories: (a) transition processes (e.g., goal specification), (b) action processes (e.g., coordination), (c) interpersonal processes (e.g., conflict management), (d) emergent states (e.g., psychological safety), or (e) omnibus topics (a combination of higher-order teamwork processes). Results demonstrated that action processes were most frequently explored, followed by transition processes, omnibus topics, emergent states, and interpersonal processes. Although all studies were framed as investigations of teamwork, it is important to highlight that most explored only one or a few constructs under the overarching umbrella of teamwork. We advocate for enhanced specificity with descriptions of OR teamwork, reporting practices pertaining to interprofessional demographics and outcomes, and increased diversity in study design and surgery type to advance understanding of teamwork and its implications.

A systematic review examining associations between physical activity, sedentary behaviour, and sleep duration with quality of life in older adults aged 65 years and above.

This systematic review examined the associations between movement behaviours (i.e., physical activity, sedentary behaviour, and sleep duration) and quality of life (QOL) in adults ≥65 years of age. Four databases were searched in June 2021. Studies were eligible for inclusion if published within the last 20 years, peer-reviewed, examined apparently healthy older adults, and analysed ≥2 movement behaviours together. QOL was represented by the World Health Organization Quality of Life measure which conceptualizes QOL by distinct domains. Study results were categorized and presented by domain. Risk of bias was completed for all included studies using methods described in the Cochrane Handbook. Thirty-one studies with 307 292 participants were included that examined QOL outcomes across seven domains: superdomain (composite measures), perceived physical health, mental and psychological states, level of independence, social relationships, environment, and general health. Findings indicated that moderate-to-vigorous intensity physical activity was favourably associated with QOL. Time re-allocation studies that showed moving time into physical activity from sedentary behaviour were associated with favourable QOL changes. The evidence regarding sedentary behaviour and sleep duration was inconsistent. The quality of evidence was very low for all domains. In conclusion, there is consistent evidence that physical activity improves QOL in adults ≥65 years of age. International Prospective Register of Ongoing Systematic Reviews (PROSPERO) registration No.: CRD42021260566.

Moderators of the coach leadership and athlete motivation relationship.

The purpose of this study was to determine whether the relationship between coach leadership and athlete motivation was moderated by age, gender, competition level, and seasons spent with a coach. This study involved data from two previous studies that explored this relationship yet provides a novel perspective through the lens of important moderators. Three-hundred and three athletes (Mage = 17.6 years; SD = 3.20; 49.7% women and 50.3% men) responded to questionnaires pertaining to their coaches' leadership behaviours and their own sport motivation. Multiple regression analyses using moderators were conducted. Age, competition level, and seasons spent with the coach significantly moderated the relationships of interest. Coach transformational leadership predicted intrinsic and extrinsic motivation to a greater extent when athletes were younger than 20.8 and 18.2 years of age, respectively. Further, coach transactional leadership predicted intrinsic and extrinsic motivation to a greater extent when athletes had trained for more than two seasons with their coach. Results emphasize the need to consider athlete characteristics from both research and practitioner perspectives. Herein, we advocate for increased awareness amongst key sport stakeholders on the influence that a coach can have on younger athletes' motivation and the importance of developing coach-athlete relationships over time.

The transcription factors Junb and Fosl2 cooperate to regulate Cdh3 expression in 15P-1 Sertoli cells.

In Sertoli cells of the testis, cadherins (Cdh) are important cell-to-cell interaction proteins and contribute to the formation of the blood-testis barrier being essential for germ cells' protection. P-cadherin or Cdh3 is only expressed in Sertoli cells from embryonic to prepubertal development. Interestingly, the expression profile of Cdh3 correlates with that of activating protein-1 (AP-1) transcription factors during Sertoli cells development. To assess their potential implications in the regulation of Cdh3, different AP-1 transcription factors were overexpressed in 15P-1 Sertoli cells. We found that the overexpressions of Junb and Fosl2 activated Cdh3 promoter. ChIP-qPCR assay and luciferase reporter assay with 5' promoter deletions and site-directed mutagenesis confirmed the recruitment of Junb and Fosl2 to an AP-1 regulatory element at -47 bp in the proximal region of Cdh3 promoter in 15P-1 cells. These findings were further supported by histone modification markers and chromatin accessibility surrounding Cdh3 promoter in mouse testis. Moreover, the knockdowns of Junb and/or Fosl2 by siRNA decreased Cdh3 protein levels. Taken together, these data suggest that in 15P-1 Sertoli cells, the AP-1 family members Junb and Fosl2 are responsible for the regulation of Cdh3 expression, which requires the recruitment of both factors to the proximal region of the Cdh3 promoter.

Sinapic and ferulic acid phenethyl esters increase the expression of steroidogenic genes in MA-10 tumor Leydig cells.

During aging, the production of androgens by the testis Leydig cells gradually decreases. Phenolic compounds can improve testosterone biosynthesis and delay the onset of hypogonadal symptoms in males. In this study, sinapic acid phenethyl ester was evaluated for its ability to regulate gene expression and steroid production in Leydig cells. Specifically, the effects of this ester on the transcriptome of MA-10 Leydig cells were investigated by RNA-Seq. To better establish a structure-function relationship of the hydroxy-methoxyphenyl moiety of sinapic and phenethyl ester, its influences on gene expression were compared to those of its ferulic acid phenethyl ester analogue. According to the transcriptomic analysis, most genes encoding enzymes related to cholesterol biosynthesis are increased in response to sinapic acid phenethyl ester treatment of MA-10 Leydig cells. Interestingly, treatments with 10 µM of ferulic acid phenethyl ester increased cAMP-dependent Star promoter activation, gene expression and protein levels. In addition, treatments of MA-10 Leydig cells with 10 µM of sinapic or ferulic acid phenethyl ester resulted in increased progesterone production. Thus, our results indicate that sinapic and ferulic acid phenethyl esters can improve cholesterol and steroid biosynthesis in testicular Leydig cells.

Involvement of calmodulin-dependent protein kinase I in the regulation of the expression of connexin 43 in MA-10 tumor Leydig cells.

Connexin 43 (Cx43, also known as Gja1) is the most abundant testicular gap junction protein. It has a crucial role in the support of spermatogenesis by Sertoli cells in the seminiferous tubules as well as in androgen synthesis by Leydig cells. The multifunctional family of Ca2+/calmodulin-dependent protein kinases (CaMK) is composed of CaMK I, II, and IV and each can serve as a mediator of nuclear Ca2+ signals. These kinases can control gene expression by phosphorylation of key regulatory sites on transcription factors. Among these, AP-1 members cFos and cJun are interesting candidates that seem to cooperate with CaMKs to regulate Cx43 expression in Leydig cells. In this study, the Cx43 promoter region important for CaMK-dependent activation is characterized using co-transfection of plasmid reporter-constructs with different plasmids coding for CaMKs and/or AP-1 members in MA-10 Leydig cells. Here we report that the activation of Cx43 expression by cFos and cJun is increased by CaMKI. Furthermore, results from chromatin immunoprecipitation suggest that the recruitment of AP-1 family members to the proximal region of the Cx43 promoter may involve another uncharacterized AP-1 DNA regulatory element and/or protein-protein interactions with other partners. Thus, our data provide new insights into the molecular regulatory mechanisms that control mouse Cx43 transcription in testicular Leydig cells.

Basic Leucine Zipper Transcription Factors as Important Regulators of Leydig Cells' Functions.

Transcription factors members of the basic leucine zipper (bZIP) class play important roles in the regulation of genes and functions in testicular Leydig cells. Many of these factors, such as cAMP responsive element binding protein 1 (CREB1) and CCAAT enhancer binding protein beta (CEBPB), are regulated by the cAMP/protein kinase A (PKA) pathway, the main signaling pathway activated following the activation of the luteinizing hormone/choriogonadotropin membrane receptor LHCGR by the - hormone LH. Others, such as X-box binding protein 1 (XBP1) and members of the cAMP responsive element binding protein 3 (CREB3)-like superfamily, are implicated in the endoplasmic reticulum stress by regulating the unfolded protein response. In this review, the influences of bZIP transcription factors, including CREB1, CEBPB and activator protein 1 (AP-1) family members, on the regulation of genes important for cell proliferation, steroidogenesis and Leydig cell communication will be covered. In addition, unresolved questions regarding the mechanisms of actions of bZIP members in gene regulation will be identified.

Efficacy of plasma exchange in patients with anti-MDA5 rapidly progressive interstitial lung disease.

BACKGROUND: Rapidly progressive interstitial lung disease (RP-ILD) is a frequent and severe manifestation of anti-MDA5 dermatomyositis (MDA5-DM) associated with poor outcome. The optimal treatment regimen for MDA5-DM RP-ILD is yet to be determined. Specifically, the value of adding plasma exchange (PLEX) to corticosteroids and immunosuppressants remains unclear. We aimed to evaluate the effect of PLEX on the outcome of patients with MDA5-DM RP-ILD. METHODS: This French nationwide multicentre retrospective study included all MDA5-DM RP-ILD patients from 2012 to 2021 admitted to 18 centres. The primary endpoint was one-year transplant-free survival. RESULTS: 51 patients with MDA5-DM RP-ILD (female 67%; mean age at disease onset: 51 ± 11.6 years) were included. Thirty-two (63%) patients required mechanical ventilation and twenty-five (49%) received PLEX. One-year mortality or lung transplant occurred in 63% cases after a median follow-up of 77 [38-264] days. The Cox proportional hazards multivariable model only retained mechanical ventilation but not PLEX (p = 0.7) as independent predictor of the primary endpoint. One-year transplant-free survival rates in PLEX + vs. PLEX-were 20% vs. 54% (p = 0.01), respectively. The Kaplan-Meier estimated probabilities of one-year transplant-free survival was statistically higher in PLEX-compared to PLEX + patients (p = 0.05). PLEX + compared to PLEX-patients more frequently received mechanical ventilation and immunosuppressants suggesting PLEX + patients had a more severe disease. CONCLUSION: MDA5-DM RP-ILD is associated with poor rate of one-year transplant-free survival. The use of PLEX was not associated with a better outcome albeit they were mainly given to more severe patients. While our study reports the largest series of MDA5-DM RP-ILD given PLEX, these results needs to be interpreted with caution owing the numerous selection, indication and interpretation bias. Further studies are needed to evaluate their efficacy in this setting.

Non-Markovian Feedback Control and Acausality: An Experimental Study.

Causality is an important assumption underlying nonequilibrium generalizations of the second law of thermodynamics known as fluctuation relations. We here experimentally study the nonequilibrium statistical properties of the work and of the entropy production for an optically trapped, underdamped nanoparticle continuously subjected to a time-delayed feedback control. Whereas the non-Markovian feedback depends on the past position of the particle for a forward trajectory, it depends on its future position for a time-reversed path, and is therefore acausal. In the steady-state regime, we show that the corresponding fluctuation relations in the long-time limit exhibit a clear signature of this acausality, even though the time-reversed dynamics is not physically realizable.

Transcriptomic analysis of MA-10 tumor Leydig cells treated with adipose derived hormones adiponectin and resistin.

Obesity contributes to a decrease in testosterone production in men. Indeed, adipose tissue produces several hormones, including adiponectin and resistin, and these may influence the activity of signaling pathways responsible for regulating the expression of genes related to steroidogenesis. In this study, we wanted to identify which genes are directly regulated by these hormones using the MA-10 tumor Leydig cell model. To do this, we treated these cells with adiponectin or resistin, followed by RNA extraction and RNA-Seq transcriptome analysis. Interestingly, genes upregulated by the globular form of adiponectin (gACRP30) were associated to steroid hormones biosynthesis, whereas resistin had no effect on the transcriptome of MA-10 Leydig cells. Moreover, the expression of the Star gene, encoding the steroidogenic acute regulatory protein, was increased in response to treatments with 0.5 mM 8Br-cAMP. Such stimulation was further increased by adiponectin, resulting in increased progesterone production. However, resistin had no effect on steroid production from MA-10 tumor Leydig cells under the treatment conditions investigated. Thus, our data suggest that a direct regulation of steroidogenic genes' expressions in Leydig cells by adipose derived hormones involves cooperation between the cAMP/PKA pathway and adiponectin, but not resistin, to activate Star expression and improve progesterone synthesis.

"Beyond the Rink": A Multilevel Analysis of Social Identity Behaviors Captured Using the Electronically Activated Recorder.

This study used ecological sampling methods to examine associations between youth athletes' experiences receiving and engaging in behaviors indicative of in-group ties, cognitive centrality, and in-group affect (i.e., social identity) during a 3-day competitive ice hockey tournament. Forty-five youth (Mage = 12.39 years; SDage = 1.14 years; 94% male) from nine teams wore an electronically activated recorder that captured brief (50-s) audio observations throughout the tournament. Participants also completed daily diary questionnaires for each day of competition. Multilevel structural equation modeling demonstrated that athletes were more likely to engage in behaviors indicative of in-group affect and cognitive centrality on days when they received as higher-than-average frequency of behaviors indicative of cognitive centrality from teammates, coaches, and parents. The findings suggest that when team members interact in ways that demonstrate they are thinking about their team, they influence fellow members to behave in ways that promote a sense of "us."

The AP-1 family of transcription factors are important regulators of gene expression within Leydig cells.

PURPOSE: Members of the AP-1 family of transcription factors are immediate early genes being modulated by different extracellular signals. The aim of this review is to highlight the important roles of AP-1 members in transcriptional regulation of genes important for testicular Leydig cell function and male testosterone production. METHODS: A search of the relevant literature was performed in Google Scholar and NCBI Pubmed for AP-1 members and Leydig cells. Additional information was accessed from references of relevant articles. Only primary data from original peer-reviewed articles was considered for this review. RESULTS: Different signaling pathways important for Leydig cells' functions are involved in the regulation of the activity of AP-1 members. These transcription factors participate in the regulation of genes related to different biological processes important for Leydig cells. CONCLUSIONS: We conclude that members of the AP-1 family of transcription factors play critical roles in the regulation of Leydig cell proliferation, steroidogenesis, and cell-to-cell communication.

KLF10 integrates circadian timing and sugar signaling to coordinate hepatic metabolism.

The mammalian circadian timing system and metabolism are highly interconnected, and disruption of this coupling is associated with negative health outcomes. Krüppel-like factors (KLFs) are transcription factors that govern metabolic homeostasis in various organs. Many KLFs show a circadian expression in the liver. Here, we show that the loss of the clock-controlled KLF10 in hepatocytes results in extensive reprogramming of the mouse liver circadian transcriptome, which in turn alters the temporal coordination of pathways associated with energy metabolism. We also show that glucose and fructose induce Klf10, which helps mitigate glucose intolerance and hepatic steatosis in mice challenged with a sugar beverage. Functional genomics further reveal that KLF10 target genes are primarily involved in central carbon metabolism. Together, these findings show that in the liver KLF10 integrates circadian timing and sugar metabolism-related signaling, and serves as a transcriptional brake that protects against the deleterious effects of increased sugar consumption.