The germ cell-specific RNA binding protein RBM46 is essential for spermatogonial differentiation in mice.

Control over gene expression is exerted, in multiple stages of spermatogenesis, at the post-transcriptional level by RNA binding proteins (RBPs). We identify here an essential role in mammalian spermatogenesis and male fertility for 'RNA binding protein 46' (RBM46). A highly evolutionarily conserved gene, Rbm46 is also essential for fertility in both flies and fish. We found Rbm46 expression was restricted to the mouse germline, detectable in males in the cytoplasm of premeiotic spermatogonia and meiotic spermatocytes. To define its requirement for spermatogenesis, we generated Rbm46 knockout (KO, Rbm46-/-) mice; although male Rbm46-/- mice were viable and appeared grossly normal, they were infertile. Testes from adult Rbm46-/- mice were small, with seminiferous tubules containing only Sertoli cells and few undifferentiated spermatogonia. Using genome-wide unbiased high throughput assays RNA-seq and 'enhanced crosslinking immunoprecipitation' coupled with RNA-seq (eCLIP-seq), we discovered RBM46 could bind, via a U-rich conserved consensus sequence, to a cohort of mRNAs encoding proteins required for completion of differentiation and subsequent meiotic initiation. In summary, our studies support an essential role for RBM46 in regulating target mRNAs during spermatogonia differentiation prior to the commitment to meiosis in mice.

Cleft lip and cleft palate in Esrp1 knockout mice is associated with alterations in epithelial-mesenchymal crosstalk.

Cleft lip is one of the most common human birth defects. However, there remain a limited number of mouse models of cleft lip that can be leveraged to characterize the genes and mechanisms that cause this disorder. Crosstalk between epithelial and mesenchymal cells underlies formation of the face and palate, but the basic molecular events mediating this crosstalk remain poorly understood. We previously demonstrated that mice lacking the epithelial-specific splicing factor Esrp1 have fully penetrant bilateral cleft lip and palate. In this study, we further investigated the mechanisms leading to cleft lip as well as cleft palate in both existing and new Esrp1 mutant mouse models. These studies included a detailed transcriptomic analysis of changes in ectoderm and mesenchyme in Esrp1-/- embryos during face formation. We identified altered expression of genes previously implicated in cleft lip and/or palate, including components of multiple signaling pathways. These findings provide the foundation for detailed investigations using Esrp1 mutant disease models to examine gene regulatory networks and pathways that are essential for normal face and palate development - the disruption of which leads to orofacial clefting in human patients.

Converting adults with sickle cell disease from full agonist opioids to buprenorphine: A reliable method with safety and early evidence of reduced acute care utilization.

Buprenorphine, a novel opioid with complex pharmacology, is effective for treating pain and is qualitatively safer than high-dose full agonist opioid therapy; but transitioning to buprenorphine can be technically complex and carries some risk of precipitated withdrawal. We report our clinic's experience converting 36 patients with sickle cell disease (SCD) from full agonist opioids to buprenorphine using a method developed in the past 10 years. Thirty of these patients were induced using a standard outpatient protocol and six were induced during medical admissions. Typically, patients were on high-dose chronic opioid therapy (COT) with inadequate response, and often with very high acute care utilization. Unlike prior case series, the method of induction, dosing, and management of withdrawal are detailed, as are post-induction adverse events. There were seven adverse events in the first 3 days following standard induction, and two of which were judged to be definitely related to the induction but none with any lasting sequelae. At 6 months follow-up, five participants had discontinued buprenorphine (16.67%), and overall acute care visits dropped from a mean of 10.50 (SD 11.35) in the 6 months pre-induction to 2.89 (SD 3.40) in the 6 months post-induction. In an appropriately interdisciplinary care setting, buprenorphine shows promise as a safe alternative to COT with early evidence of benefit for high-utilizing patients with SCD.

The global Protein-RNA interaction map of ESRP1 defines a post-transcriptional program that is essential for epithelial cell function.

The epithelial splicing regulatory proteins, ESRP1 and ESRP2, are essential for mammalian development through the regulation of a global program of alternative splicing of genes involved in the maintenance of epithelial cell function. To further inform our understanding of the molecular functions of ESRP1, we performed enhanced crosslinking immunoprecipitation coupled with high-throughput sequencing (eCLIP) in epithelial cells of mouse epidermis. The genome-wide binding sites of ESRP1 were integrated with RNA-Seq analysis of alterations in splicing and total gene expression that result from epidermal ablation of Esrp1 and Esrp2. These studies demonstrated that ESRP1 functions in splicing regulation occur primarily through direct binding in a position-dependent manner to promote either exon inclusion or skipping. In addition, we also identified widespread binding of ESRP1 in 3' and 5' untranslated regions (UTRs) of genes involved in epithelial cell function, suggesting that its post-transcriptional functions extend beyond splicing regulation.

Prolonged and variable copulation durations in a promiscuous insect species: No evidence of reproductive benefits for females.

Within promiscuous mating systems, copulation often functions as more than a means of fertilization, and copulation durations can vary widely. Copulating for prolonged durations can enhance both female and male reproductive success, but can also result in costs: females of some insect species experience increased fecundity and fertility through male-provided nutrition during prolonged copulations, but also decreased longevity due to male-driven mechanisms. Here, for a common, promiscuous insect species (the squash bug, Anasa tristis), we first describe the range of copulation duration, which spans from 2 min to over 23 h. To investigate whether female A. tristis benefit from prolonged copulation, we next manipulated copulation duration and female diet, and we documented the resulting fecundity, fertility, and longevity of each female. We found no evidence that prolonged copulation durations affect female reproductive success. Females produced fertile eggs after a single 30 min copulation, and they subsequently produced fertile eggs for an additional 4 weeks. Our findings suggest that females do not benefit from prolonged copulations, that sperm transfer occurs very early during copulations, and that females can store sperm for long durations. Consequently, we suggest that female harassment avoidance and male mate-guarding may explain prolonged copulations in this species.

Use of a Central Catheter Maintenance Bundle in Long-Term Acute Care Hospitals.

OBJECTIVE: Evidence-based guidelines have resulted in decreases in bloodstream infections associated with central catheters (CLABSIs) in hospital intensive care units. However, relatively little is known about CLABSI incidence and prevention in long-term acute care hospitals (LTACHs). METHODS: A central catheter maintenance bundle was implemented in 30 LTACHs, and compliance with the bundle was tracked for 6 months. CLABSI rates were monitored for 14 months before and 14 months after the bundle was implemented. RESULTS: The pooled mean CLABSI rate (No. of infections per 1000 days with a central catheter) was 1.28 before the bundle and 0.96 after the bundle (repeated measures general linear model; F1,58 = 6.973; P = .01; partial η(2) = .11). From 14 months before to 14 months after the bundle was implemented, the mean number of CLABSIs per LTACH decreased by 4.5 (95% CI, 1.85-7.15). Time series modeling showed a significant decrease in the mean hospital CLABSI rate after the bundle was implemented (-0.511 CLABSI/1000 catheter days, SE = 0.050), indicating an immediate effect of the bundle. The mean hospital CLABSI rate was decreasing slightly before the bundle was implemented and continued to decrease at a reduced rate after the bundle was implemented. CONCLUSION: The bundle resulted in a significant and sustained reduction in CLABSI rates in 30 LTACHs for 14 months. These results encourage the development and implementation of similar bundles as effective strategies for infection reduction in LTACHs.