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Specialized medical Conundrum: The 39-Year-Old With Persistent Retained Merchandise associated with Pregnancy and Contingency Uterine Arteriovenous Malformation.
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has been identified as the causal agent of COronaVIrus Disease-19 (COVID-19), an atypical pneumonia-like syndrome that emerged in December 2019. While SARS-CoV-2 titers can be measured by detection of viral nucleic acid, this method is unable to quantitate infectious virions. Measurement of infectious SARS-CoV-2 can be achieved by tissue culture infectious dose-50 (TCID50 ), which detects the presence or absence of cytopathic effect in cells infected with serial dilutions of a virus specimen. However, this method only provides a qualitative infectious virus titer. Plaque assays are a quantitative method of measuring infectious SARS-CoV-2 by quantifying the plaques formed in cell culture upon infection with serial dilutions of a virus specimen. As such, plaque assays remain the gold standard in quantifying concentrations of replication-competent lytic virions. Here, we describe two detailed plaque assay protocols to quantify infectious SARS-CoV-2 using different overlay and staining methods. Both methods have several advantages and disadvantages, which can be considered when choosing the procedure best suited for each laboratory. These assays can be used for several research purposes, including titration of virus stocks produced from infected cell supernatant and, with further optimization, quantification of SARS-CoV-2 in specimens collected from infected animals. © 2019 The Authors. Basic Protocol SARS-CoV-2 plaque assay using a solid double overlay method Alternate Protocol SARS-CoV-2 plaque assay using a liquid overlay and fixation-staining method.The characterization of degradation of biodegradable materials for tissue regeneration is classically carried out in three steps in vitro degradation analysis, in vitro cell culture, and in vivo animal experiments. Each step involves an increasing complexity and should serve a more sophisticated material selection, which serves as an orientation to clinical studies and the final application in patients. Recently, the usefulness of degradation analyses is being discussed. In this context, the aim of this work is to increase the importance of in vitro degradation analysis by using flowing media to move closer to the in vivo situation. In the long term, this should lead to a more sensitive biomaterial characterization as well as to a replacement of time-consuming static or quasi-dynamic incubation experiments. The practicability of the novel chamber is demonstrated in context of a degradation study of silica/collagen/calcium phosphate composites in flowing media with physiological (2.4 mM) and lowered (0.5 mM) calcium ion concentrations. This is done by comparison with static and quasi-dynamic incubation experiments. In order to keep all media regimes comparable to each other, for the dynamic experiment, a flow rate was chosen equivalent to the medium exchange in quasi-dynamic incubation. Under flow-through conditions, there is a clearly decreased tendency to lower the calcium concentration, so that a concentration close to the physiological initial situation can be continuously maintained.Backgroud Associations between sex hormones and diabetic vascular complications was recently studied, but the role luteinizing hormone (LH), playing in diabetic kidney disease (DKD) remains uncertain. We aimed to investigate the relationship of LH and DKD in Chinese men and postmenopausal women with type 2 diabetes mellitus (T2DM). Methods Data were collected from 1775 T2DM men and postmenopausal women in hospital. selleck The odds ratios (OR) and corresponding 95% confidence intervals (CI) in relation to LH quartiles were obtained by multiple logistic regression analysis. Results LH levels were significantly higher in patients with macroalbuminuria than those with microalbuminuria, but not in patients with microalbuminuria than those with normoalbuminuria. Consistently, LH in those with estimated glomerular filtration rate (eGFR) less then 60 mL/min/1.73m2 were significantly higher than those with eGFR≥60 mL/min/1.73m2 . The prevalence of macroalbuminuria was obviously increased for subjects of the fourth quartile of LH vs the first to third quartile (20.4% vs 6.2%, 8.0%, 12.2% in men; 25.3% vs 5.5%, 3.8%, 9.3% in postmenopausal women). Multivariate logistic regression demonstrated that subjects within the highest quartile of LH had higher odds of macroalbuminuria than those within the lowest quartile (OR 4.00, 95%CI 1.87-8.55 for men; OR 9.62, 95%CI 3.42-27.08 for postmenopausal women), independent of age, diabetes duration or other metabolic factors. The area under curve (AUC) for detecting macroalbuminuria based on LH was 0.662 for men, and 0.767 for postmenopausal women. Conclusion High LH levels are positively associated with established DKD among Chinese men and postmenopausal women. Elevated LH may be a promising clinical factor for identifying established DKD. This article is protected by copyright. All rights reserved.Objective The importance of the cold-shock Y-box protein-1 (YB-1) for cell homeostasis is well documented by its association with certain cancer entities. Here, we explored the role of YB-1 in T-cell homeostasis and survival, and the potential contribution of YB-1 in the pathogenesis of systemic lupus erythematosus (SLE). Methods We analyzed the expression of YB-1 and apoptosis by qPCR and FACS of CD4+ T-cells from peripheral blood of healthy donors (n=25) and SLE patients (n=25) ex vivo as well as in vitro after a 6-day-stimulation with αCD3- or αCD3/αCD28-coupled microspheres. YB-1 was overexpressed using lentiviral transduction with GFP-YB-1wt and knock-down using specific shRNA (-3 fold reduction, p less then 0.0001), respectively. Results YB-1 expression was significantly lower in apoptosis-prone and in activated T-cells of patients suffering from SLE compared to those of non-apoptotic and of activated T-cells of healthy volunteers (p=0.001). Knock-down of YB-1 demonstrates that reduced expression of YB-1 in T-cells inevitably led to expression of pro-apoptotic molecules and caspase 3 activation (1.6 fold), and subsequently to apoptosis. Furthermore, we revealed that YB-1 promotes survival pathways involving enhanced protein expression of the kinase Akt (2 fold) and Bcl-2 (3 fold), even when Fas was triggered. YB-1-mediated survival was reversed by Akt and PI3K inactivation. In SLE patients rescue of YB-1 expression strongly promotes survival of T-cells and even prevents cell death in those that were extremely apoptosis-prone. Conclusion Our data show that failure of YB-1 upregulation in T-cells of SLE patients leads to enhanced apoptosis. These findings have implications for diagnostics and possibly new treatment strategies for hematopoietic alterations in SLE.
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