Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

Intercellular adhesion and membrane polarity in rat hepatocytes: Localization of cell-CAM 105 and other domain-specific membrane proteins in situ and in vitro by electron microscopy

Cell-CAM 105 has been identified as a cell adhesion molecule (CAM) based on the ability of monospecific and monovalent anti-cell-CAM 105 antibodies to inhibit the reaggregation of rat hepatocytes. Although one would expect to find CAMs concentrated in the lateral membrane domain where adhesive interactions predominate, immunofluorescence analysis of rat liver frozen sections revealed that cell-CAM 105 was present exclusively in the bile canalicular (BC) domain of the hepatocyte. To more precisely define the in situ localization of cell-CAM 105, immunoperoxidase and electron microscopy were used to analyze intact and mechanically dissociated fixed liver tissue. Results indicate that although cell-CAM 105 is apparently restricted to the BC domain in situ, it can be detected in the pericanalicular region of the lateral membranes when accessibility to lateral membranes is provided by mechanical dissociation. In contrast, when hepatocytes were labeled following incubation in vitro under conditions used during adhesion assays, cell-CAM 105 had redistributed to all areas of the plasma membrane. Immunofluorescence analysis of primary hepatocyte cultures revealed that cell-CAM 105 and two other BC proteins were localized in discrete domains reminscent of BC while cell-CAM 105 was also present in regions of intercellular contact. These results indicate that the distribution of cell-CAM 105 under the experimental conditions used for cell adhesion assays differs from that in situ and raises the possibility that its adhesive function may be modulated by its cell surface distribution. The implications of these and other findings are discussed with regard to a model for BC formation. Analysis of molecular events involved in BC formation would be accelerated if an in vitro model system were available. Although BC formation in culture has previously been observed, repolarization of cell-CAM 105 and two other domain-specific membrane proteins was incomplete. Since DMSO had been used by Isom et al. to maintain liver-specific gene expression in vitro, the effect of this differentiation system on the polarity of these membrane proteins was examined. Based on findings presented here, DMSO apparently prolongs the expression and facilitates polarization of hepatocyte membrane proteins in vitro

Reducing Variability and Increasing Reproducibility

In this video from InnovATEBIO, Lisa Seidman and Jeanette Mowery talk about educating students on the reduction of variability and the increase of reproducibility in biotechnology lab and manufacturing environments. A basic lab skills for a regulated environment course, examples of student activities, categories of variability, reagents, the process of measuring and detecting variability, issues in weighing chemicals, lab research, error propagation, textbooks, and more are explored. The video recording runs 56:40 minutes in length

Basic Laboratory Methods in a Regulated Environment

Biotechnology transforms knowledge that emerges from life science research into technology, the creation of products of value to people. Beginning biotechnology students therefore need to develop a strong foundation in laboratory science that is integrated with an understanding of product quality. This course provides students with a foundation in basic concepts and techniques necessary to work as effective professionals in a biotechnology laboratory or small scale production facility. The course emphasizes metrology (the study of measurements), solution preparation, performing assays, and basic biological separation methods. These fundamental laboratory techniques are essential for student success in later molecular biology, cell culture, bioprocessing, analytical, and other specialized courses. Throughout the course the principles of product quality systems (e.g., Good Manufacturing/Laboratory Practices and ISO 9000) are integrated as students explore documentation, calibration, accuracy and precision, error reduction, trouble-shooting, verification and validation of assays, and other quality concepts. Integrating a "quality-mindset" into their laboratory work is important both for students who plan to work in a biotechnology company and for students who some day aspire to generate meaningful results in a research environment

Laboratory Manual for Biotechnology and Laboratory Science: The Basics

Laboratory Manual for Biotechnology provides students with the basic laboratory skills and knowledge to pursue a career in biotechnology. The manual, written by four biotechnology instructors with over 20 years of teaching experience, incorporates instruction, exercises, and laboratory activities that the authors have been using and perfecting for years. These exercises and activities serve to engage students and help them understand the fundamentals of working in a biotechnology laboratory. Building students' skills through an organized and systematic presentation of materials, procedures, and tasks, the manual will help students explore overarching themes that relate to all biotechnology workplaces. The fundamentals in this manual are critical to the success of research scientists, scientists who develop ideas into practical products, laboratory analysts who analyze samples in forensic, clinical, quality control, environmental, and other testing laboratories

Effect of Antiplatelet Therapy on Survival and Organ Support–Free Days in Critically Ill Patients With COVID-19

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