Saunderssantos0913

Moreover, two effective siRNAs targeting Mrfem-1 were found and their effectiveness verified in vitro. These results on Mrfem-1 will help us to better understand the fem family and provide a new resource for subsequent investigations of siRNA-mediated regulation on ovarian development in M. rosenbergii.The 50 kDa N-terminal product of the cellular transcription factor E4F1 (p50E4F1) mediates E1A289R trans-activation of the adenovirus E4 gene, and suppresses E1A-mediated transformation by sensitizing cells to cell death. This report shows that while both E1A289R and E1A243R stimulate p50E4F1 DNA binding activity, E1A289R trans-activation, as measured using GAL-p50E4F1 fusion proteins, involves a p50E4F1 transcription regulatory (TR) region that must be promoter-bound and is dependent upon E1A CR3, CR1 and N-terminal domains. Trans-activation is promoter-specific, as GAL-p50E4F1 did not stimulate commonly used artificial promoters and was strongly repressive when competing against GAL-VP16. p50E4F1 and E1A289R stably associate in vivo using the p50E4F1 TR region and E1A CR3, although their association in vitro is indirect and paradoxically disrupted by MAP kinase phosphorylation of E1A289R, which stimulates E4 trans-activation in vivo. Multiple cellular proteins, including TBP, bind the p50E4F1 TR region in vitro. The mechanistic implications for p50E4F1 function are discussed.Background Congenital hypogonadotropic hypogonadism (CHH) is a rare genetically heterogeneous disorder. We aimed to determine the prevalence and pathogenesis of NECL2 (Nectin-like molecule 2) variants in a cohort of female patients with CHH. Methods We sequenced and determined the prevalence of NECL2 variants in 68 female patients with CHH and 243 healthy controls collected from an academic medical center. Further cellular and animal studies were performed to verify the pathogenicity of the mutations. Necl2 knockout female mice were generated, and their puberty development was observed. Results A novel NECL2 variant (c.1052_1060del, p.Thr351_Thr353del) was detected in 4 of 68 (5.9%) patients with CHH. Its prevalence was significantly higher in CHH patients than in healthy controls (0%). At the cellular level, the necl2 variant leads to a decrease in gonadotropin-releasing hormone. In animal models, we found that the Necl2 protein was expressed in the hypothalamus, especially in the ventromedial hypothalamic nucleus of mice. Necl2 knockout female mice showed delayed puberty and an irregular estrous cycle, consistent with CHH patient phenotypes. Conclusions Our findings predict that NECL2 may be a new candidate gene for CHH and that the NECL2 protein plays a critical role in the progression of puberty development.Continuous Manufacturing (CM) of pharmaceutical drug products is a new approach within the pharmaceutical industry. In the presented paper, a GMP continuous wet granulation line for production of solid dosage forms was investigated. The line was composed of the subsequent continuous unit operations feeding - twin-screw wet-granulation - fluid-bed drying - sieving and tableting. The formulation of a commercial entity was selected for this study. Several critical process parameters were evaluated in order to probe the process and to characterize the impact on quality attributes. Seven critical process parameters have been selected after a risk analysis API and excipient mass flows of the two feeders, liquid feed rate and rotation speed of the extruder and rotation speed, temperature and airflow of the dryer. Eight quality attributes were controlled in real time by Process Analytical Technologies (PAT) API content after blender, after dryer, in tablet press feed frame and of tablet, LOD after dryer and PSD afterntinuous manufacturing line and increase the knowledge of this innovative production line and the products that it makes.CLEC12B is a C-type lectin-like receptor expressed on myeloid cells. In this study, we have characterized the porcine homologue of CLEC12B (poCLEC12B). To this end, we have generated constructs encoding a c-myc tagged version of the whole receptor, or its ectodomain fused to the Fc portion of human IgG1, from a cDNA clone obtained from an alveolar macrophage library, and raised monoclonal antibodies (mAb) against this molecule. Using these mAbs, poCLEC12B was found to be expressed on alveolar macrophages and, at lower levels, on blood conventional type 1 dendritic cells (cDC1) and plasmacytoid DCs. No binding was detected on monocytes, monocyte-derived macrophages or monocyte-derived DCs. Engagement of CLEC12B on alveolar macrophages with mAbs had no apparent effect on cytokine production (TNF-α, IL-8) induced by LPS. These results provide the basis for future investigations aimed to assess the role of poCLEC12B in different microbial infections and to evaluate its potential in vaccination strategies targeting DCs.Background The utility of cerebrospinal fluid drainage (CSFD) for prevention of spinal cord ischemia (SCI) after thoracic endovascular aortic repair (TEVAR) remains unclear. We previously published our institutional algorithm restricting preoperative CSFD to patients deemed high risk for SCI. see more Since that publication, our algorithm has evolved with preoperative CSFD avoided in all patients undergoing isolated descending +/- arch TEVAR. This study evaluates the updated algorithm in a contemporary cohort. Methods Patients who underwent TEVAR for descending aortic +/- arch pathology between 2/2012 - 9/2018 at a single center were identified from an institutional aortic surgery database. The algorithm includes left subclavian artery (LSA) revascularization in cases of coverage with no preservation of antegrade flow, permissive hypertension, and use of evoked potential monitoring. The primary endpoints were SCI or postoperative CSFD. Results N=225 patients underwent descending +/- arch TEVAR during the study interval. 2 patients (0.9%) had CSFD prior to TEVAR in violation of the algorithm and were excluded from the study cohort. 81% had endograft coverage below T6. The LSA was fully covered in 100 patients (47%), all of whom underwent LSA revascularization. Following the updated algorithm, the incidence of temporary or permanent SCI was 0%. No patient required postoperative CSFD. Conclusions A restrictive lumbar CSFD algorithm including permissive hypertension and LSA revascularization in the setting of descending +/- arch TEVAR appears safe with a 0% incidence of SCI in 223 consecutive patients treated over a 6.5-year interval. We recommend consideration of further prospective study to evaluate this algorithm.Despite recent progress in the understanding of cardiac ion channel function and its role in inherited forms of ventricular arrhythmias, the molecular basis of cardiac conduction disorders often remains unresolved. We aimed to elucidate the genetic background of familial atrioventricular block (AVB) using a whole exome sequencing (WES) approach. In monozygotic twins with a third-degree AVB and in another, unrelated family with first-degree AVB, we identified a heterozygous nonsense mutation in the POPDC2 gene causing a premature stop at position 188 (POPDC2W188⁎), deleting parts of its cAMP binding-domain. Popeye-domain containing (POPDC) proteins are predominantly expressed in the skeletal muscle and the heart, with particularly high expression of POPDC2 in the sinoatrial node of the mouse. We now show by quantitative PCR experiments that in the human heart the POPDC-modulated two-pore domain potassium (K2P) channel TREK-1 is preferentially expressed in the atrioventricular node. Co-expression studies in Xenopus oocytes revealed that POPDC2W188⁎ causes a loss-of-function with impaired TREK-1 modulation. Consistent with the high expression level of POPDC2 in the murine sinoatrial node, POPDC2W188⁎ knock-in mice displayed stress-induced sinus bradycardia and pauses, a phenotype that was previously also reported for POPDC2 and TREK-1 knock-out mice. We propose that the POPDC2W188⁎ loss-of-function mutation contributes to AVB pathogenesis by an aberrant modulation of TREK-1, highlighting that POPDC2 represents a novel arrhythmia gene for cardiac conduction disorders.Based on extensive studies on gonadotropin-releasing hormone (GnRH) it was assumed that GnRH is the only hypothalamic neurohormone regulating gonadotropin release in vertebrates. In 2000, however, Tsutsui's group discovered gonadotropin-inhibitory hormone (GnIH), a novel hypothalamic neuropeptide that inhibits gonadotropin release, in quail. Subsequent studies by Tsutsui's group demonstrated that GnIH is conserved among vertebrates, acting as a new key neurohormone regulating reproduction. GnIH inhibits gonadotropin synthesis and release through actions on gonadotropes and GnRH neurons via GnIH receptor, GPR147. Thus, GnRH is not the sole hypothalamic neurohormone controlling vertebrate reproduction. The following studies by Tsutsui's group have further demonstrated that GnIH has several important functions in addition to the control of reproduction. Accordingly, GnIH has drastically changed our understanding about reproductive neuroendocrinology. This review summarizes the discovery of GnIH, progress in GnIH research on reproductive physiology and behavior and perspective of GnIH research on neuroendocrine regulation of reproduction.Background The data on acute kidney injury (AKI) in patients without chronic kidney disease (CKD) after transcatheter aortic valve replacement (TAVR) are limited. The study sought to compare the incidence of AKI and its impact on 5-year mortality after TAVR and surgical aortic valve replacement (SAVR) in patients without CKD. Methods This registry included data from 6463 consecutive patients who underwent TAVR or SAVR. CKD was defined as estimated glomerular filtration rate less then 60 mL/min/1.73 m2. AKI was defined according to the Kidney Disease Improving Global Outcomes criteria. For sensitivity analysis, propensity-score matching between TAVR and SAVR was performed. Results The study included 4555 consecutive patients (TAVR, n = 1215 and SAVR, n = 3340) without CKD. Propensity-score matching identified 542 pairs. Patients who underwent TAVR had a significantly lower incidence of AKI in comparison to those who underwent SAVR (unmatched 4.7% vs 16.4%, P less then 0.001, multivariable analysis odds ratio, 0.29, 95% confidence interval [CI], 0.20-0.41; matched 5.9% vs 19.0%, P less then 0.001). Patients with AKI had significantly increased 5-year mortality compared with those without AKI (unmatched 36.0% vs 19.1%, log-rank P less then 0.001; matched 36.3% vs 24.0%, log-rank P less then 0.001). The adjusted hazard ratios for 5-year mortality were 1.58 (95% CI, 1.20-2.08) for AKI grade 1, 3.27 (95% CI, 2.09-5.06) for grade 2, and 4.82 (95% CI, 2.93-8.04) for grade 3. Conclusions TAVR in patients without CKD was associated with a significantly less frequent incidence of AKI compared with SAVR. AKI significantly increased the risk of 5-year mortality after either TAVR or SAVR, and increasing severity of AKI was incrementally associated with 5-year mortality.Prolonged cardiac hypertrophy, a pathological compensatory response of the heart, finally leads to heart failure. Numerous studies have illustrated the vital roles of non-coding RNAs (ncRNAs) in cardiac hypertrophy. Here, we probed into the role and probable mechanism of microRNA-30e-5p (miR-30e-5p) in Angiotensin II (Ang-II)-stimulated hypertrophic cardiomyocytes. link2 Intriguingly, the expression of hypertrophic markers, cell surface area and protein/DNA ratio were all reduced in Ang-II-induced hypertrophic cardiomyocytes when miR-30e-5p expression was augmented. link3 Then, ADAM9 was screened out as the target of miR-30e-5p and ADAM9 overexpression rescued the effect of miR-30e-5p upregulation in Ang-II-treated cardiomyocytes. Moreover, we identified Kcnq1ot1 as the upstream of miR-30e-5p/ADAM9 axis and verified that Kcnq1ot1 aggrandized ADAM9 expression in Ang-II-treated cardiomyocytes through absorbing miR-30e-5p. Furthermore, rescue assays confirmed that ADAM9 up-regulation abrogated the repressive effect of Kcnq1ot1 depletion on Ang-II-induced cardiac hypertrophy.