Ulrichpridgen3432

To evaluate the effects of Arf6 downregulation on human prostate cancer cells.

The effects of Arf6 downregulation on cell proliferation, migration, invasion and apoptosis were assessed by MTT, BrdU, scratch, Transwell assays and flow cytometry respectively. AKT, p-AKT, ERK1/2, p-ERK1/2 and Rac1 protein expressions were detected by Western blot.

Downregulating Arf6 by siRNA interference suppressed the mRNA and protein expressions of Arf6. The proliferation capacities of siRNA group at 48h, 72h, and 96h were significantly lower than those of control group (P <0.05). The migration distance of siRNA group at 18h was significantly shorter than that of control group (P <0.01). The number of cells penetrating Transwell chamber membrane significantly decreased in siRNA group compared with that of control group (P <0.01). After 24h, negative control and normal control groups had similar apoptotic rates (P >0.05) which were both significantly lower than that of siRNA group (P <0.01). After Arf6 expression was downregulated, p-ERK1/2 and Rac1 protein expressions were significantly lower than those of control group (P <0.05).

Downregulating Arf6 expression can inhibit the proliferation, migration and invasion of prostate cancer cells in vitro, which may be related to ERK1/2 phosphorylation and Rac1 downregulation.

Downregulating Arf6 expression can inhibit the proliferation, migration and invasion of prostate cancer cells in vitro, which may be related to ERK1/2 phosphorylation and Rac1 downregulation.

There is limited data regarding surgeon volume and partial nephrectomy outcomes. The aim of this study is to report trifecta outcomes of robot-assisted partial nephrectomy (RAPN) performed by the low volume surgeon.

Thirty-nine patients with clinical T1-2 renal tumors who underwent RAPN between 2012 and 2018 were included in this study. Trifecta was defined as negative surgical margins, warm ischemia time ≤20 minutes, and no operative complications. Patient demographics, R.E.N.A.L. nephrometry score, operation time, estimated blood loss, warm ischemia time, length of hospital stay, renal functions, and oncological outcomes were analyzed retrospectively. Complications were graded based on the modified Clavien-Dindo classification system.

The median R.E.N.A.L. nephrometry score was 6 (4-10). RAPN was successfully performed in all but one patient. The median operation time was 180 (90-240) minutes. Warm ischemia was performed only by segmental renal artery control in 35 and, by main renal artery control in three patients. The off-clamp technique was used in two patients. The median warm ischemia time was 16 (0-31) minutes. Seven patients had a warm ischemia time of longer than 20 minutes. Three patients had postoperative complications. The surgical margin was positive in one patient. As a result, the trifecta was achieved in 30 of the 39 patients (77%).

RAPN is a safe and effective minimally invasive alternative in the treatment of renal masses. The present study suggests that reasonable trifecta rates can be achieved even by low volume surgeons.

RAPN is a safe and effective minimally invasive alternative in the treatment of renal masses. The present study suggests that reasonable trifecta rates can be achieved even by low volume surgeons.The development of novel methodologies for catalytic enantioselective functionalization reactions enabled by chiral transient directing groups is accompanying in a paradigm shift in the field of asymmetric synthesis. In particular, these highly atom- and step-economic enantioinduction processes commonly proceed either via enantioselective C-H functionalization, or via enantioselective hydroarylation of the pro-chiral substrates generating point, axial or planar chirality. The use of the transient directing group strategy in C-H functionalizations precludes the stoichiometric installations and removal of directing groups and enables efficient, more compatible and economical chemical routes. This minireview highlights asymmetric transition-metal-catalyzed methodologies involving chiral transient directing groups together with the scope, utility and future perspective of the field.This article describes a protocol for detecting and quantifying RNA phosphorothioate modifications in cellular RNA samples. Starting from solid-phase synthesis of phosphorothioate RNA dinucleotides, followed by purification with reversed-phase HPLC, phosphorothioate RNA dinucleotide standards are prepared for UPLC-MS and LC-MS/MS methods. RNA samples are extracted from cells using TRIzol reagent, then digested with a nuclease mixture and analyzed by mass spectrometry. UPLC-MS is employed first to identify RNA phosphorothioate modifications. An optimized LC-MS/MS method is then employed to quantify the frequency of RNA phosphorothioate modifications in a series of model cells. © 2020 Wiley Periodicals LLC. Basic Protocol 1 Synthesis, purification, and characterization of RNA phosphorothioate dinucleotides Basic Protocol 2 Digestion of RNA samples extracted from cells Basic Protocol 3 Detection and quantification of RNA phosphorothioate modifications by mass spectrometry.Ezetimibe is a top-selling hypolipidemic drug for the treatment of cardiovascular diseases. Biosynthesis of (4S)-3-[(5S)-5-(4-fluorophenyl)-5-hydroxypentanoyl]-4-phenyl-1,3-oxazolidin-2-one ((S)-ET-5) using carbonyl reductase has shown advantages including high catalytic efficiency, excellent stereoselectivity, mild reaction conditions, and environmental friendness, and was considered as the key step for ezetimibe production. The regeneration efficiency of the cofactor, nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) is one of the main restricted factor. Recombinant Escherichia coli strain (smCR125) coexpressing carbonyl reductase (CR125) and glucose dehydrogenase were successfully constructed and applied for the production of (S)-ET-5 for the first time. Without extra addition of the coenzyme NADPH, the yield of 99.8% and the enantiomeric excess (e.e.) of 99.9% were achieved under ET-4 concentration of 200 g/L. Using a substrate fed-batch strategy, under the optimal conditions, the substrate ET-4 concentration was increased to 250 g/L with the yield of 98.9% and the e.e. of 99.9% after 12 hr reaction. The space-time yield of 494.5 g L-1 d-1 and the space-time yield per gram biocatalyst of 24.7 g L-1 d-1 g-1 DCW were achieved, which were higher than ever reported for the biosynthesis of the ezetimibe intermediate.Tobacco cessation among those recently diagnosed with cancer is important to improve their prognosis, yet, many cancer survivors continue to smoke. The epidemiology of tobacco use differs by race and ethnicity, and limited cessation research has been conducted in African American (AA) populations. Here, we assess demographic and clinical variables associated with continued smoking in AAs after a cancer diagnosis. The Detroit Research on Cancer Survivors study is a cohort comprised of AA cancer survivors with breast, prostate, lung, and colorectal cancers. Detroit Research on Cancer Survivors data were utilized from survivors who completed their baseline survey within 18 months of cancer diagnosis (n = 1145); 18% (n = 356) reported smoking at the time of cancer diagnosis, and 57% of these (n = 203) continued to smoke after their diagnosis. Logistic regression models were used to assess factors associated with continued smoking. Living with a smoker (odds ratio [OR] = 2.78, 95% confidence interval [CI] 1.64, 4.70), higher cumulative years of smoking (OR = 1.03, 95% CI 1.01, 1.05, for each year), and a prostate cancer diagnosis (OR = 7.35, 95% CI 3.89, 13.89) were all associated with increased odds of continued smoking. Survivors with higher social well-being scores (measured by the Functional Assessment of Cancer Therapy, a quality of life assessment) were more likely to quit smoking after diagnosis (OR = 0.96, 95% CI 0.93, 1.00). These findings highlight the continued need for personalized cessation strategies to be incorporated into treatment plans for cancer survivors.

As many as one-quarter of all residents in nursing facilities have cardiopulmonary resuscitation (CPR) as a documented choice in the medical record, despite the likelihood of limited medical benefit in this setting.

The aim of this study was to understand the perspectives of healthcare providers and nursing facility residents regarding CPR decisions.

We used qualitative interviews to examine the perspectives of residents with a documented decision for CPR in the medical record. We then compared residents' views with those of healthcare providers who routinely conduct advance care planning (ACP) conversations in the nursing facility setting.

Five themes emerged from the interviews (a) Resident versus Provider Concerns, (b) Offering Information versus Avoidance, (c) Lack of Understanding of CPR, (d) Lack of Awareness, and (e) 'Don't Keep Me on Machines'. Residents held misconceptions about CPR and/or exhibited an overall poor understanding of the relationship between their own health status and the likeACP discussions.

ACP communication models and training should be designed not only to explore nursing facility residents' goals, values, and preferences, but also to elicit any underlying differences in perceptions that may affect communication. Healthcare providers can identifying the primary concerns of residents and assist them with integrating or reframing these issues as a part of ACP discussions.This study examines the longer term relationship between public health insurance expansions and health behaviors. I leverage geographic and temporal variation in the implementation of the Affordable Care Act-facilitated Medicaid expansions and provide the first estimates of the expansions' behavioral impacts during their first 5 years. Using national survey data from the 2010 to 2018 Behavioral Risk Factors Surveillance System and a difference-in-differences regression design, I show that the Medicaid expansions increase utilization of certain forms of preventive care, while reducing heavy drinking. I also find suggestive evidence that the expansions reduce smoking and increase the probability of exercise. These results stand in contrast with earlier studies that used only 2 or 3 years of postexpansion data and found no detectable effect of the Medicaid expansions on health behaviors in the short run. My results, combined with evidence from previous studies, suggest that public insurance expansions may not prompt an immediate change in health behaviors, but newly eligible populations do increase investments in healthy behaviors over time. In the long run, Medicaid expansions may help reduce engagement in risky behaviors like drinking and smoking among low-income people.Inflammatory bowel disease (IBD) is a heterogeneic disease with a variety of treatments targeting different mechanisms. A multistate, mechanistic, mathematical model of IBD was developed in part 1 of this two-part article series. In this paper, application of the model to predict response of key clinical biomarkers following different treatment options for Crohn's disease was explored. Five therapies, representing four different mechanisms of action, were simulated in the model and longitudinal profiles of key clinical markers, C-reactive protein and fecal calprotectin were compared with clinical observations. Model simulations provided an accurate match with both central tendency and variability observed in biomarker profiles. We also applied the model to predict biomarker and clinical response in an experimental, combination therapy of existing therapeutic options and provide possible mechanistic basis for the increased response. Overall, we present a validated, modular, mechanistic model construct, which can be applied to explore key biomarkers and clinical outcomes in IBD.