Preparation of acetazolamide composite microparticles by supercritical antisolvent techniques

The possibility of preparation of ophthalmic drug delivery systems using compressed anti-solvent technology was evaluated. and RL 100 were used as drug carriers, acetazolamide was the model drug processed. Compressed anti-solvent experiments were carried out as a semi-continuous or a batch operation from a liquid solution of polymer(s) + solute dissolved in acetone. Both techniques allowed the recovery of composite particles, but the semi-continuous operation yielded smaller and less aggregated populations than the batch operation. The release behaviour of acetazolamide from the prepared microparticles was studied and most products exhibited a slower release than the single drug. Moreover, the release could be controlled to some extent by varying the ratio of the two Eudragit used in the formulation and by selecting one or the other anti-solvent technique. Simple diffusion models satisfactorily described the release profiles. Composites specifically produced by semi-continuous technique have a drug release rate controlled by a diffusion mechanism, whereas for composites produced by the batch operation, the polymer swelling also contributes to the overall transport mechanism

Studies on the selectivity of the SARS-CoV-2 papain-like protease reveal the importance of the P2' proline of the viral polyprotein

The SARS-CoV-2 papain-like protease (PLpro) is an antiviral drug target that catalyzes the hydrolysis of the viral polyproteins pp1a/1ab, so releasing the non-structural proteins (nsps) 1–3 that are essential for the coronavirus lifecycle. The LXGG↓X motif in pp1a/1ab is crucial for recognition and cleavage by PLpro. We describe molecular dynamics, docking, and quantum mechanics/molecular mechanics (QM/MM) calculations to investigate how oligopeptide substrates derived from the viral polyprotein bind to PLpro. The results reveal how the substrate sequence affects the efficiency of PLpro-catalyzed hydrolysis. In particular, a proline at the P2′ position promotes catalysis, as validated by residue substitutions and mass spectrometry-based analyses. Analysis of PLpro catalyzed hydrolysis of LXGG motif-containing oligopeptides derived from human proteins suggests that factors beyond the LXGG motif and the presence of a proline residue at P2′ contribute to catalytic efficiency, possibly reflecting the promiscuity of PLpro. The results will help in identifying PLpro substrates and guiding inhibitor design

Mass spectrometric assays monitoring the deubiquitinase activity of the SARS-CoV-2 papain-like protease inform on the basis of substrate selectivity and have utility for substrate identification

The SARS-CoV-2 papain-like protease (PLpro) and main protease (Mpro) are nucleophilic cysteine enzymes that catalyze hydrolysis of the viral polyproteins pp1a/1ab. By contrast with Mpro, PLpro is also a deubiquitinase (DUB) that accepts post-translationally modified human proteins as substrates. Here we report studies on the DUB activity of PLpro using synthetic Nε-lysine-branched oligopeptides as substrates that mimic post-translational protein modifications by ubiquitin (Ub) or Ub-like modifiers (UBLs), such as interferon stimulated gene 15 (ISG15). Mass spectrometry (MS)-based assays confirm the DUB activity of isolated recombinant PLpro. They reveal that the sequence of both the peptide fragment derived from the post-translationally modified protein and that derived from the UBL affects PLpro catalysis; the nature of substrate binding in the S sites appears to be more important for catalytic efficiency than binding in the S′ sites. Importantly, the results reflect the reported cellular substrate selectivity of PLpro, i.e. human proteins conjugated to ISG15 are better substrates than those conjugated to Ub or other UBLs. The combined experimental and modelling results imply that PLpro catalysis is affected not only by the identity of the substrate residues binding in the S and S′ sites, but also by the substrate fold and the conformational dynamics of the blocking loop 2 of the PLpro:substrate complex. Nε-Lysine-branched oligopeptides thus have potential to help the identification of PLpro substrates. More generally, the results imply that MS-based assays with Nε-lysine-branched oligopeptides have potential to monitor catalysis by human DUBs and hence to inform on their substrate preferences

Nonmesonic Hyperon Weak Decay Spectra in Λ12^{12}_{\Lambda}C}

We study the nonmesonic weak decay (NMWD) $\Lambda N \to nN$ of the ${}^{12}_{\Lambda} C$ hypernucleus induced by the nucleon $N=n,p$ with transition rate $\Gamma_N$. The nuclear process is described by the interplay of two models; one describing the NMWD of hyperon $\Lambda$ in the nuclear environment, and the other taking into account the Final State Interaction (FSI) of the two outgoing nucleons with the residual nucleus. The first one is done in the framework of the Independent-Particle Shell-Model (IPSM), with the decay dynamics represented by the exchange of $\pi+ \eta+ K+\rho+\omega+K^*$ mesons with usual parametrization. For the second one is used a time dependent multicollisional intranuclear cascade schema (implemented in the CRISP code - Collaboration Rio-S\~ao Paulo). The results obtained for inclusive and exclusive kinetic energy spectra, and the angular correlation are compared with recent data from KEK and FINUDA experiments. The calculated ratio $(\Gamma_n / \Gamma_p)^{\partial{\tiny FSI}}$, between the numbers of emitted back-to-back $nn$ and $np$ pairs, is in good agreement with the experimental data.Comment: Proceedings of the International Nuclear Physics Conference 2010, Vancouver, BC - Canada 4-9 Jul 201

Association of sociocultural stressors with bipolar disorder onset in Puerto Rican youth growing up as members of a minoritized ethnic group: results from the Boricua Youth Longitudinal Study

BACKGROUND: The development of bipolar disorder is currently explained by a complex interaction of genetic and environmental factors. Less is known regarding the influence of sociocultural factors. This study aims to evaluate the incidence and impact of sociocultural factors on bipolar disorder onset in two comparable samples of youth growing up in different social settings. METHODS: We leveraged data from two urban population-based cohorts representative of Puerto Rican children growing up in either San Juan (Puerto Rico) or the South Bronx (NYC) and followed up for 17 years. Bipolar disorder diagnoses were based on retrospective self-reports on the World Health Organization Composite International Diagnostic Interview. We used a causal inference approach to estimate associations of sociocultural factors with bipolar disorder onset after adjusting for potential confounders. FINDINGS: We found that South Bronx children, who grew up as a minoritized group, had twice the risk of bipolar disorder onset as young adults, with an incidence rate of 2.22 new cases per 1000 person-years compared to 1.08 new cases in San Juan (incidence rate difference, 1.13; 95% CI, 0.09-1.20). After adjusting for potential confounders, South Bronx children had the same lifetime hazard of bipolar disorder onset compared to San Juan children. However, our analysis demonstrated that caregivers' exposure to societal cultural stress partially explained the increased risk of bipolar disorder onset in the South Bronx, in addition to the potential contribution of genetics. INTERPRETATION: Our results provide evidence that societal cultural stress can increase the risk of lifetime bipolar disorder onset in youth growing up as a minoritized group. Addressing stress in minoritized groups might reduce the risk of bipolar disorder onset. FUNDING: The Boricua Youth Study has been supported by the National Institutes of HealthMH56401, MH098374, DA033172, and AA020191. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the article

‘We Learn Together’—Translanguaging within a Holistic Approach towards Multilingualism in Education

Within two multilingual education projects in the north of the Netherlands a holistic model for multilingualism in education is being tested. This is done through design-based interventions in which in- and pre-service teachers, teacher trainers and researchers co-develop and evaluate multilingual activities for different school types. Results show that through experimenting in a safe environment teachers gradually embraced their pupils’ multilingualism. This contradicts earlier findings on teachers strongly favouring monolingual instruction and viewing migrant languages as a deficit.<br/

Application of New Accident Analysis Methodologies. Volume II: A Users Manual for BEATS

DTFH61-87-C-00015Researchers in the field of accident analysis have long been aware of the problems associated with drawing statistical inference on safety using accident data. Aside from the problems of accessibility and quality, accident data present a real challenge when it comes to statistical analysis. One of the most serious problems in accident analysis is the regression-to-the-mean bias which occurs due to the non-random site selection process in safety measure evaluation studies. This study presents a new empirical Bayes method (EBEST) which adjusts for regression-to-the-mean bias. Three typical applications in accident analysis are considered for regression-to-the-mean bias, namely: (1) the evaluation of safety treatments; (2) the identification of high hazard locations; and (3) the assimilation of information from multiple safety measure studies (meta-analysis). A computer program was developed to execute these analyses as a part of this study. This manuscript describes the EBEST (Empirical Bayes Estimation of Safety and Transportation) methodology and presents examples of how the method works for each of the three accident analysis applications. This volume, Volume II, is a users manual for the BEATS computer program