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ImportanceFew studies are available on informed consent (IC) among detained persons, even with ethics being a critical aspect of prison research. In IC research, audiovisual material seems to improve understanding and satisfaction compared with conventional paper-based material, but findings remain unclear.ObjectiveTo compare audiovisual and paper-based materials for 1-time general IC for research in prisons.Design, Setting, and ParticipantsThis cross-sectional randomized clinical trial was conducted in 2 corrections facilities in Switzerland (an adult prison and a juvenile detention center). The study was conducted from December 14, 2019, to December 2, 2020, in the adult prison and from January 15, 2020, to September 9, 2021, in the juvenile detention center. In the adult prison, study participation was offered to detained persons visiting the medical unit (response rate, 84.7%). In the juvenile detention center, all newly incarcerated adolescents were invited to participate (response rate, 98.0%).InterventionsParticipants were randomized to receive paper-based conventional material or to watch a 4-minute video. Materials included the same legal information, as required by the Swiss Federal Act on Research Involving Human Beings.Main Outcomes and MeasuresThe main outcome was acceptance to sign the IC form. Secondary outcomes included understanding, evaluation, and time to read or watch the IC material.ResultsThe study included 190 adults (mean [SD] age, 35.0 [11.8] years; 190 [100%] male) and 100 adolescents (mean [SD] age, 16.0 [1.1] years; 83 [83.0%] male). In the adult prison, no significant differences were found between groups in acceptance to sign the IC form (77 [81.1%] for paper-based material and 81 [85.3%] for audiovisual material; P = .39) and to evaluate it (mean [SD] correct responses, 5.09 [1.13] for paper-based material and 5.01 [1.07] for audiovisual material; P = .81). Understanding was significantly higher in the audiovisual material group (mean [SD] correct responses, 5.09 [1.84]) compared with the paper-based material group (mean [SD] correct responses, 4.61 [1.70]; P = .04). In the juvenile detention center, individuals in the audiovisual material group were more likely to sign the IC form (44 [89.8%]) than the paper-based material group (35 [68.6%], P = .006). No significant difference was found between groups for understanding and evaluation. Adults took a mean (SD) of 5 (2) minutes to read the paper material, and adolescents took 7 (3) minutes.Conclusions and RelevanceGiven the small benefit of audiovisual material, these findings suggest that giving detained adults and prison health care staff a choice regarding IC material is best. For adolescents, audiovisual material should be provided. Future studies should focus on increasing understanding of the IC process.Trial RegistrationClinicalTrials.gov Identifier: NCT05505058

Voltage-gated sodium channels are pore-forming transmembrane proteins that selectively allow sodium ions to flow across the plasma membrane according to the electro-chemical gradient thus mediating the rising phase of action potentials in excitable cells and playing key roles in physiological processes such as neurotransmission, skeletal muscle contraction, heart rhythm, and pain sensation. Genetic variations in the nine human genes encoding these channels are known to cause a large range of diseases affecting the nervous and cardiac systems. Understanding the molecular effect of genetic variations is critical for elucidating the pathologic mechanisms of known variations and in predicting the effect of newly discovered ones. To this end, we have created a Web-based tool, the Ion Channels Variants Portal, which compiles all variants characterized functionally in the human sodium channel genes. This portal describes 672 variants each associated with at least one molecular or clinical phenotypic impact, for a total of 4,658 observations extracted from 264 different research articles. These data were captured as structured annotations using standardized vocabularies and ontologies, such as the Gene Ontology and the Ion Channel ElectroPhysiology Ontology. All these data are available to the scientific community via neXtProt at https://www.nextprot.org/portals/navmut.

Ovarian cancer is the fifth leading cause of cancer-related death in the world. Some ovarian cancer patients present large amount of ascites at the time of diagnosis which may play an active role in tumor development. In earlier studies, we demonstrated that the acellular fraction of ascites can induce apoptosis of ovarian cancer cells. The current study identifies pigment epithelium derived factor (PEDF) as the molecule responsible for the apoptotic effect of ascites and evaluates the Sleeping Beauty transposon (SBT) system as a new tool for PEDF gene therapy against ovarian cancer. We utilize gel filtration, mass spectrometry, affinity column, cell viability assay, tumor development on chick chorioallantoic membrane and molecular biology techniques for these purposes. PEDF was thus identified as the agent responsible for the effects of ascites on ovarian cancer cell viability and tumor growth. Interestingly, the PEDF expression is decreased in ovarian cancer cells compared to healthy ovarian cells. However, the level of PEDF is higher in ascites than in serum of ovarian cancer patients suggesting that cells present in the tumor environment are able to secrete PEDF. We then used the SBT system to stably induce PEDF expression in ovarian cancer cells. The overexpression of PEDF significantly reduced the tumor growth derived from these cells. In conclusion, the results presented here establish that PEDF is a therapeutic target and that PEDF from ascites or SBT could be utilized as a therapeutic strategy for the treatment of ovarian cancer.

neXtProt (http://www.nextprot.org/) is a new human protein-centric knowledge platform. Developed at the Swiss Institute of Bioinformatics (SIB), it aims to help researchers answer questions relevant to human proteins. To achieve this goal, neXtProt is built on a corpus containing both curated knowledge originating from the UniProtKB/Swiss-Prot knowledgebase and carefully selected and filtered high-throughput data pertinent to human proteins. This article presents an overview of the database and the data integration process. We also lay out the key future directions of neXtProt that we consider the necessary steps to make neXtProt the one-stop-shop for all research projects focusing on human proteins.

Type 1 diabetes develops when most insulin-producing β cells of the pancreas are killed by an autoimmune attack. The in vivo conditions modulating the sensitivity and resistance of β cells to this attack remain largely obscure. Here, we show that connexin 36 (Cx36), a trans-membrane protein that forms gap junctions between β cells in the pancreatic islets, protects mouse β cells against both cytotoxic drugs and cytokines that prevail in the islet environment at the onset of type 1 diabetes. We documented that this protection was at least partially dependent on intercellular communication, which Cx36 and other types of connexin channels establish within pancreatic islets. We further found that proinflammatory cytokines decreased expression of Cx36 and that experimental reduction or augmentation of Cx36 levels increased or decreased β cell apoptosis, respectively. Thus, we conclude that Cx36 is central to β cell protection from toxic insults.

OBJECTIVE Pancreatic β-cells exposed to proinflammatory cytokines display alterations in gene expression resulting in defective insulin secretion and apoptosis. MicroRNAs are small noncoding RNAs emerging as key regulators of gene expression. Here, we evaluated the contribution of microRNAs to cytokine-mediated β-cell cytotoxicity. RESEARCH DESIGN AND METHODS We used global microarray profiling and real-time PCR analysis to detect changes in microRNA expression in β-cells exposed to cytokines and in islets of pre-diabetic NOD mice. We assessed the involvement of the microRNAs affected in cytokine-mediated β-cell failure by modifying their expression in insulin-secreting MIN6 cells. RESULTS We found that IL-1β and TNF-α induce the expression of miR-21, miR-34a, and miR-146a both in MIN6 cells and human pancreatic islets. We further show an increase of these microRNAs in islets of NOD mice during development of pre-diabetic insulitis. Blocking miR-21, miR-34a, or miR-146a function using antisense molecules did not restore insulin-promoter activity but prevented the reduction in glucose-induced insulin secretion observed upon IL-1β exposure. Moreover, anti–miR-34a and anti–miR-146a treatment protected MIN6 cells from cytokine-triggered cell death. CONCLUSIONS Our data identify miR-21, miR-34a, and miR-146a as novel players in β-cell failure elicited in vitro and in vivo by proinflammatory cytokines, notably during the development of peri-insulitis that precedes overt diabetes in NOD mice.

Some twenty-five years ago it was reported for the first time that indoleamine 2,3-dioxygenase (IDO) activity was particularly important in rodent epididymis protein extracts. Several years ago, it was also shown that the epididymal expression of IDO was constitutive and not driven by the interferon-gamma, inflammatory cytokine, as is the case in most of the other tissues tested. Since then, the epididymal expression of IDO has not been investigated further. Recently, we have reported a more detailed study showing that indeed the mouse epididymis expresses both IDO transcript and protein at a high level while this is not the case for the other tryptophan-catabolizing enzyme, TDO. On the basis of the different functions that have been assigned to IDO we discuss here the putative roles of IDO expression as well as tryptophan metabolism on the physiology of the mammalian epididymis.

Les gametes mâles elabores dans le testicule ou se deroule la spermatogenese poursuivent leur maturation hors de la gonade dans les conduits genitaux. L’epididyme est au centre de cette maturation post-testiculaire des spermatozoides responsable de l’acquisition de leur aptitude a se mouvoir ainsi qu’a reconnaitre et a penetrer un ovocyte, evenements essentiels pour la fecondation. Durant leur transit dans le canal epididymaire les gametes evoluent dans un environnement luminal en perpetuel changement via le jeu complexe des activites de secretion et de reabsorption de l’epithelium epididymaire. Ces activites de secretion regulees dans l’espace et dans le temps ce qui fait de cet epithelium epididymaire un tissu d’une grande complexite soumis a un reseau elabore de regulations croisees. Outre les regulations classiques de type endocrine et paracrine, la region proximale de l’organe est soumise a des controles lumicrines par des facteurs d’origine testiculaire qui ajoutent a la complexite des reseaux de regulations mis en jeu. Le travail realise ci-dessous se veut etre un etat des lieux exhaustif des acteurs dont la litterature a montre qu’ils exercaient une modulation de l’activite epididymaire.

Previous reports have suggested that indoleamine 2,3-dioxygenase (IDO) activity is particularly important in mouse epididymis tissue. We show here, using reverse transcription/polymerase chain reaction assays, Northern assays, Western blotting experiments, and immunohistochemistry that IDO is indeed highly expressed in mouse epididymis, and that IDO mRNA distribution and protein location are precisely regionalized within the organ and within sub-territories of the proximal part of the epididymal duct, the so-called caput epididymidis. Within the caput epididymidis, both the principal and the apical cells have been shown to express IDO. On the contrary, tryptophan dioxygenase (TDO), a sister enzyme of IDO, is weakly and uniformly expressed in mouse epididymis and, in contrast to IDO, is also expressed in testis. In the epididymis, TDO protein expression has been found in a totally different cell type in the smooth muscle layer surrounding the epididymal tubules. Finally, IDO is not secreted into the epididymal lumen, whereas the testis-expressed TDO is present on the head of spermatozoa retrieved from the cauda epididymidis. On the basis of the various functions that have been associated with IDO/TDO, we discuss the putative impacts of IDO/TDO expression on the physiology of mammalian epididymis and spermatozoa.

Les recepteurs nucleaires des oxysterols LXR (Liver X receptor) α et LXRs sont des facteurs de transcription appartenant a la superfamille des recepteurs nucleaires. Ils sont actives par une serie particuliere d’oxysterols. Des antagonistes naturels ont ete egalement identifies comme les acides gras poly-insatures ou certains sulfates de cholesterol plasmatiques. L’etude des souris deficientes en LXRs a permis de les associer a la regulation de nombreux metabolismes (cholesterol, acides gras, glucose, steroides). Les LXRs et leur partenaire RXR (recepteur de l’acide retinoique 9-cis) sont exprimes dans le tractus genital mâle et les testicules, et leurs ligands y sont a des concentrations physiologiquement actives. Dans ces organes, l’homeostasie du cholesterol doit etre strictement regulee car 1) le cholesterol est un precurseur indispensable pour la synthese des steroides testiculaires; 2) pendant la maturation epididymaire, la membrane plasmique des spermatozoides subit des changements de composition notamment la diminution de cholesterol et de lecithines. L’analyse des souris deficientes en recepteurs LXR α et LXRs a mis en evidence une destructuration de la couche epitheliale du segment 2 de la tete de l’epididyme, ainsi qu’une fragilite des spermatozoides recueillis. Au total, les analyses de physiologie integrative et moleculaire mettent en evidence le role des recepteurs nucleaires LXRs dans la physiologie de la reproduction chez le mâle.

Using various molecular approaches, including reverse transcription–polymerase chain reaction (RT–PCR), rapid amplification of cDNA ends–PCR, sequencing, northern and western blotting, we found that the mouse GPX5 gene gives rise to at least three different transcripts that are not expressed at the same levels in the mouse epididymis. In addition to the major GPX5 transcript, we show that minor GPX5 transcripts exist, arising either from precocious termination of transcription or an alternative splicing event within intron 4 of the 5 exon-encoding GPX5 single copy gene. Furthermore, we demonstrate that variants of the GPX5 protein that are correlated with the shorter GPX5 transcripts can be detected in caput epididymidis protein extracts and that the various GPX5 isoforms are subject to differential post-transcriptional maturation processes in the mouse epididymis that essentially involve the addition of O-glycosyl extensions. Using a sensitive poly-A+ mRNA tissue blot, as well as RT–PCR and northern assays, we further show that in addition to being expressed in the epididymis, the GPX5 gene is also expressed, albeit at lower levels, in other tissues of the male genital tract, including the testis and prostate. Finally, we present evidence suggesting that the GPX5 gene is expressed in a temporally regulated manner during mouse embryonic development.

The emergence of pancreatic islets has necessitated the development of a signalling system for the intra- and inter-islet coordination of beta cells. With evolution, this system has evolved into a complex regulatory network of partially cross-talking pathways, whereby individual cells sense the state of activity of their neighbours and, accordingly, regulate their own level of functioning. A consistent feature of this network in vertebrates is the expression of connexin (Cx)-36-made cell-to-cell channels, which cluster at gap junction domains of the cell membrane, and which adjacent beta cells use to share cytoplasmic ions and small metabolites within individual islets. This chapter reviews what is known about Cx36, and the mechanism whereby this beta-cell connexin significantly regulates insulin secretion. It further outlines other less established functions of the protein and evaluates its potential relevance for the development of novel therapeutic approaches to diabetes.

In this study we looked at the epididymides and spermatozoa of mice knocked-out for nuclear oxysterol receptors (LXR). We have shown that LXR-deficient mice exhibited upon ageing a severe disruption of their caput epididymides associated with abnormal accumulation of neutral lipids. The epididymis defaults were correlated with sperm head fragility and infertility. In agreement with the observed caput defect in transgenic animals in which both LXRα and LXRβ isoforms were disrupted, we have shown here that both receptors are expressed in caput and cauda epididymides regions. LXRβ was predominantly expressed throughout the mouse epididymis while the expression of LXRα was weaker. In addition, the expression of selected genes that can be considered as markers of adult epididymis function was monitored via Northern blots in the different single and double LXR-deficient backgrounds. Altogether, the data presented here suggest that LXR receptors are important actors in epididymis function.

International audience; We report here on the characterization of tissue-culture cell lines derived from primary cultures of the mouse caput epididymidis epithelium. The cell lines were spontaneously immortalized without the use of transforming oncogenes. In defined conditions, our epididymal cells adopted various morphological features that resembles that of the in vivo epididymis epithelium such as a polarized organization and the presence of junctional structures at their apical/lateral membranes as revealed by electron microscopy analyses. Flow cytometry analysis revealed that we were dealing with homogenous cell populations that had reached a near-tetraploid state. RT-PCR assays were used in order to show that several genes that can be considered as markers of in vivo caput epididymidis epithelium activity were expressed in our cell lines confirming that these cells were indeed in a differentiated state close to their endogenous state.

It is well documented that a dietary deficiency in magnesium can induce oxidative stress and an inflammatory response in animal models. In our study, we have investigated these responses in the mouse epididymis after mice had been fed a magnesium-deficient diet for a 2-week duration. The extracellular and intracellular concentrations of magnesium where shown to be depleted on this diet. This was followed, however, only in the liver of the Mg-deficient animals, by an increase in both alpha 2-macroglobulin (alpha-2m), an acute phase marker, and interleukin-6 transcripts suggesting that an inflammatory response had been initiated. These changes were correlated with a decrease in circulating neutrophils. To address the question of whether or not peroxidation was induced in mouse epididymis following hypomagnesia, we have monitored the level of endogenous peroxidation, their ability to respond to induced peroxidation as well as the expression and activity of the enzymatic glutathione peroxidase (GPX) antioxidant family. To evaluate if the epididymis had evolved specific protections against peroxidation, other organs such as the liver and the kidney were monitored in parallel. We detected no evidence for increased peroxidation in any of the mouse organs tested. However, GPX activity was found to be significantly lower in the liver and the kidney of Mg-deficient animals while it was unchanged in the epididymides of the same animals during the deficiency. Histological analysis of the epididymis showed no major difference in the overall cytological aspect of the organ. Segment 2 of the caput, however presented a significant increase in the number of apically located cells or blebbing cells. Immunohistochemical analysis proved that these cells were epididymal apical cells and not infiltrated leukocytes. These observations suggested that the mouse caput epididymidis segment 2 specifically responded to Mg deficiency via the apical cells. Finally, a comparative analysis of stress response genes was conducted in control and magnesium-deficient caput epididymidis samples. It brought forward some genes that might be involved in the peculiar response of the caput epithelium following hypomagnesia.

Introduction Le diabete de type 1 resulte d'une destruction auto-immune de la majorite des cellules beta pancreatiques. Nous avons teste si les connexines (Cxs), des proteines transmembranaires permettant le couplage intercellulaire via les jonctions gap, etaient impliquees dans la modulation de l'apoptose des cellules beta exposees a divers toxines in vivo et in vitro mimant l'environnement cellulaire en debut de diabete de type 1. Materiels et methodes Des souris RIP-Cx36 (qui surexpriment la Cx36 native), RIP-Cx43 et RIP-Cx32 (qui expriment les Cxs 43 ou 32 en plus de la Cx36) et KO-Cx36 (invalidees pour la Cx36) ont ete injectees avec 200mg/kg de streptozotocine (STZ) ou 70mg/kg d'alloxane (AX). Des ilots isoles de ces souris ont ete exposes in vitro a 4,4mm de STZ ou a un cocktail de 0,25ng/ml d'IL-1 beta + 0,1μg/ml d'IFN gamma + 9,1ng/ml de TNF alpha. Le couplage intercellulaire a ete quantifie par micro-injection de jaune de lucifer ou de bromure d'ethidium. Resultats L'injection de STZ ou d'AX a des souris controles presentant un niveau de couplage normal a immediatement resulte en une hyperglycemie. Les souris presentant un couplage intercellulaire augmente (les souris heterozygotes ou homozygotes des lignees RIP-Cx36, RIP-Cx32 et RIP-Cx43) sont restees normoglycemiques. Les souris presentant un couplage diminue ou aboli (les souris heterozygotes ou homozygotes de la lignee KO-Cx36) sont devenues plus diabetiques que les controles. L'exposition in vitro d'ilots isoles des souris RIP-Cx36 et KO-Cx36 a la STZ ou aux cytokines a montre que l'apoptose des cellules beta etait inversement proportionnelle aux niveaux de Cx36 et de couplage. Conclusion La resistance de cellules beta a la STZ, l'AX ou aux cytokines diabetogenes est proportionnelle au couplage intercellulaire confere par les connexines. Des strategies visant a augmenter pharmacologiquement le couplage intercellulaire pourraient representer une nouvelle option therapeutique.