Meyersthomson7172

A key activity in small-molecule drug discovery is the characterization of compound-target interactions. Surface plasmon resonance (SPR) is a flexible technique for this purpose, with a wide affinity range (micromoles to picomoles), low protein requirements, and the ability to characterize the kinetics of compound binding. However, a key requirement of SPR is the immobilization of the target protein to the surface of the sensor chip. The most commonly used immobilization techniques (covalent immobilization, streptavidin-biotin) are irreversible in nature, which can afford excellent baseline stability but impose limitations throughput for slowly dissociating compounds or unstable targets. Reversible immobilization (e.g., His-tag-Ni-NTA) is possible but typically precludes accurate quantification of slow dissociation kinetics due to baseline drift.Here we present our investigation of three immobilization strategies (dual-His-tagged target protein, His-tagged streptavidin, and switchavidin) that combine the robustness of irreversible immobilization with the flexibility of reversible immobilization. Each has its own advantages and limitations, and while a universal immobilization procedure remains to be found, these strategies add to the immobilization toolbox that enables previously out-of-scope applications. Such applications are highlighted in two examples that greatly increased throughput for the kinetic characterization of potent kinase inhibitors and kinetic profiling of covalent inhibitors.Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a novel etiological agent of coronavirus disease 2019 (COVID-19). Nigella sativa, commonly known as black seed or black cumin, has been a historical and traditional plant since thousands of years. Based on their therapeutic efficacy, the chief components of terpenoids and flavonoids were selected from N. sativa seeds and seed oil. This study was designed to check the antiviral efficacy of N. sativa main phytoconstituents against five potential targets of SARS-CoV-2 using in silico structure-based virtual screening approach. Out of twenty five phytocomponents, ten components showed best binding affinity against two viral proteins viz. N-terminal RNA binding domain (NRBD; PDB ID 6M3M) of nucleocapsid protein and papain-like protease (PL-PRO; PDB ID 6W9C) of SARS-CoV-2 using AutoDock 4.2.6, AutoDock Vina and iGEMDOCK. PASS analyses of all ten phytocomponents using Lipinski's Rule of five showed promising results. Further, druglikeness and toxicity assessment using OSIRIS Data Warrior v5.2.1 software exhibited the feasibility of phytocomponents as drug candidates with no predicted toxicity. Molecular dynamics simulation study of NRBD of SARS-CoV-2 nucleocapsid protein-alpha-spinasterol complex and PL-PRO-cycloeucalenol complex displayed strong stability at 300 K. Both these complexes exhibited constant root mean square deviation (RMSDs) of protein side chains and Cα atoms throughout the simulation run time. Interestingly, PL-PRO and NRBD are key proteins in viral replication, host cell immune evasion and viral assembly. Thus, NRBD and PL-PRO have the potential to serve as therapeutic targets for N. sativa phytoconstituents in drug discovery process against COVID-19.Prior research has revealed sexual minority youth are more likely to carry weapons both outside of and within school. However, to date, no study has examined the degree to which bullying and harassment is associated with weapon carrying among sexual minority youth. We utilized the Youth Risk Behavior Survey to examine the prevalence and likelihood of carrying weapons by sexual identity, adjusting for adverse experiences. From the Youth Risk Behavior Survey 2009-2015, we examined self-report of adverse experiences (e.g., being bullied, skipping school due to fear for personal safety) and performed Logistic regressions to estimate the odds of carrying a weapon and were adjusted to include demographics and adverse experiences. When surveyed by sexual identity, 14.0% of heterosexual, 21.8% of gay/lesbian, 18.5% of bisexual, and 17.4% of "not sure" students reported carrying a weapon in the past 30 days. Odds of carrying a weapon were significantly increased for youth who skipped school due to feeling unsafe at school, had ever been threatened with a weapon in the past year, and had ever been in a physical fight. Compared to heterosexual female peers, sexual minority women had increased odds of carrying a weapon. α-Conotoxin GI Pediatricians should recognize that experiencing bullying and feeling unsafe are associated with weapon carrying, particularly among sexual minorities. Pediatricians and professionals who work with youth should recognize that reported experiences of bullying may not be the most salient indicator of risk for weapon carrying among all youth, and that other fears of or experiences with bullying are crucial to screen for among sexual minorities in particular.Prior studies have demonstrated that sexual minority college students are two to four times more likely to experience sexual victimization (e.g., sexual assault and/or unwanted or uncomfortable sexual experiences) compared to their heterosexual counterparts. However, research that has focused on the detrimental effects of sexual victimization on health outcomes has paid more attention to heterosexual college samples and community-based adults. Understanding how sexual victimization influences mental health and substance use outcomes among lesbian, gay, bisexual, queer, and asexual (LGBQA) emerging adults in college is warranted given that this developmental period represents a critical risk period for trauma exposure, risk behavior, and psychological distress. Thus, the current study tested how sexual victimization was associated with depressive symptoms, post-traumatic stress disorder (PTSD) symptoms, and alcohol use disorder (AUD) symptoms among 234 diverse college students who self-identified as LGBQA. Additionally, social support was tested as a moderator. Findings indicated that sexual victimization was related to greater depressive symptoms, PTSD symptoms, and AUD symptoms. In addition, perceived social support moderated the relation between sexual victimization and depressive symptoms, however, in a direction contrary to hypotheses. In particular, higher sexual victimization was associated with greater depressive symptoms among LGBQA students with higher levels of social support (b = .29, p = .00), and was not significant among LGBQA students with lower levels of social support (b = .13, p = .26). The current study highlights the need to consider the detrimental effects of sexual victimization on health outcomes among LGBQA college students, as well as the mechanisms through which social support may be influencing these relations.