Viral dissemination and immune activation modulate antiretroviral drug levels in lymph nodes of SIV-infected rhesus macaques.

Introduction and methods: To understand the relationship between immunovirological factors and antiretroviral (ARV) drug levels in lymph nodes (LN) in HIV therapy, we analyzed drug levels in twenty-one SIV-infected rhesus macaques subcutaneously treated with daily tenofovir (TFV) and emtricitabine (FTC) for three months. Results: The intracellular active drug-metabolite (IADM) levels (TFV-dp and FTC-tp) in lymph node mononuclear cells (LNMC) were significantly lower than in peripheral blood mononuclear cells (PBMC) (P≤0.005). Between Month 1 and Month 3, IADM levels increased in both LNMC (P≤0.001) and PBMC (P≤0.01), with a steeper increase in LNMC (P≤0.01). The viral dissemination in plasma, LN, and rectal tissue at ART initiation correlated negatively with IADM levels at Month 1. Physiologically-based pharmacokinetic model simulations suggest that, following subcutaneous ARV administration, ART-induced reduction of immune activation improves the formation of active drug-metabolites through modulation of kinase activity and/or through improved parent drug accessibility to LN cellular compartments. Conclusion: These observations have broad implications for drugs that need to phosphorylate to exert their pharmacological activity, especially in the settings of the pre-/post-exposure prophylaxis and efficacy of antiviral therapies targeting pathogenic viruses such as HIV or SARS-CoV-2 replicating in highly inflammatory anatomic compartments.

Physical Harm Reduction in Domestic Violence: Does Marijuana Make Assaults Safer?

Studies on the effect of marijuana on domestic violence often suffer from endogeneity issues. To examine the effect of marijuana decriminalization and medical marijuana legalization on serious domestic assaults, we conducted a difference-in-differences analysis on a panel dataset on NIBRS-reported assaults in 24 states over the 12 years between 2005 and 2016. Assaults disaggregated according to situation and extent of injury were employed as dependent variables. We found that while the total number of assaults did not change, decriminalization reduced domestic assaults involving serious injuries by 18%. From a harm reduction perspective, these results suggest that while the extensive margin of violence did not change, the intensive margin measured by the seriousness of assaults were substantially affected by decriminalization. This result may be partially explained by reductions in offender alcohol intoxication and weapon-involved assault.

The importance of forensic evidence for decisions on criminal guilt.

Recent studies have found that the general public perceives forensic evidence to be relatively inaccurate and to involve high levels of human judgement. This study examines how important the general public finds forensic evidence by comparing decisions on guilt and punishment in criminal cases that involve forensic versus eyewitness testimony evidence and examining whether a CSI effect exists. Specifically, this experimental survey study utilized a 2 (crime type: murder or rape) × 4 (evidence type: DNA, fingerprint, victim eyewitness testimony, or bystander eyewitness testimony) - 1 (no victim testimony for murder scenario) design, yielding seven vignettes scenarios to which participants were randomly assigned. Results indicate that forensic evidence was associated with more guilty verdicts and higher confidence in a guilty verdict. Forensic evidence did not change the expected sentence length and did not generally affect the ideal sentence length. However, for rape, respondents believed that the defendant should receive a longer sentence when forensic evidence was presented but forensic evidence did not alter likely sentence that respondents expected the defendant to receive. The results of this study did not support a CSI effect. Overall, this study suggests that forensic evidence - particularly DNA - has a stronger influence during the verdict stage than the sentencing stage.

Mapping the immune environment in clear cell renal carcinoma by single-cell genomics.

Clear cell renal cell carcinoma (ccRCC) is one of the most immunologically distinct tumor types due to high response rate to immunotherapies, despite low tumor mutational burden. To characterize the tumor immune microenvironment of ccRCC, we applied single-cell-RNA sequencing (SCRS) along with T-cell-receptor (TCR) sequencing to map the transcriptomic heterogeneity of 25,688 individual CD45+ lymphoid and myeloid cells in matched tumor and blood from three patients with ccRCC. We also included 11,367 immune cells from four other individuals derived from the kidney and peripheral blood to facilitate the identification and assessment of ccRCC-specific differences. There is an overall increase in CD8+ T-cell and macrophage populations in tumor-infiltrated immune cells compared to normal renal tissue. We further demonstrate the divergent cell transcriptional states for tumor-infiltrating CD8+ T cells and identify a MKI67 + proliferative subpopulation being a potential culprit for the progression of ccRCC. Using the SCRS gene expression, we found preferential prediction of clinical outcomes and pathological diseases by subcluster assignment. With further characterization and functional validation, our findings may reveal certain subpopulations of immune cells amenable to therapeutic intervention.

Public beliefs about the accuracy and importance of forensic evidence in the United States.

Recent advances in forensic science, especially the use of DNA technology, have revealed that faulty forensic analyses may have contributed to miscarriages of justice. In this study we build on recent research on the general public's perceptions of the accuracy of 10 forensic science techniques and of each stage in the investigation process. We find that individuals in the United States hold a pessimistic view of the forensic science investigation process, believing that an error can occur about half of the time at each stage of the process. We find that respondents believe that forensics are far from perfect, with accuracy rates ranging from a low of 55% for voice analysis to a high of 83% for DNA analysis, with most techniques being considered between 65% and 75% accurate. Nevertheless, respondents still believe that forensic evidence is a key part of a criminal case, with nearly 30% of respondents believing that the absence of forensic evidence is sufficient for a prosecutor to drop the case and nearly 40% believing that the presence of forensic evidence - even if other forms of evidence suggest that the defendant is not guilty - is enough to convict the defendant.

Thermodynamic analysis of catalysis by the dihydroorotases from hamster and Bacillus caldolyticus, as compared with the uncatalyzed reaction.

Dihydroorotase (DHOase, EC 3.5.2.3) from the extreme thermophile Bacillus caldolyticus has been subcloned, sequenced, expressed, and purified as a monomer. The catalytic properties of this thermophilic DHOase have been compared with another type I enzyme, the DHOase domain from hamster, to investigate how the thermophilic enzyme is adapted to higher temperatures. B. caldolyticus DHOase has higher Vmax and Ks values than hamster DHOase at the same temperature. The thermodynamic parameters for the binding of L-dihydroorotate were determined at 25 degrees C for hamster DHOase (deltaG = -6.9 kcal/mol, deltaH = -11.5 kcal/mol, TdeltaS = -4.6 kcal/mol) and B. caldolyticus DHOase (deltaG = -5.6 kcal/mol, deltaH = -4.2 kcal/mol, TdeltaS = +1.4 kcal/mol). The smaller enthalpy release and positive entropy for thermophilic DHOase are indicative of a weakly interacting Michaelis complex. Hamster DHOase has an enthalpy of activation of 12.3 kcal/mol, similar to the release of enthalpy upon substrate binding, rendering the kcat/Ks value almost temperature independent. B. caldolyticus DHOase shows a decrease in the enthalpy of activation from 12.2 kcal/mol at temperatures from 30 to 50 degrees C to 5.3 kcal/mol for temperatures of 50-70 degrees C. Vibrational energy at higher temperatures may facilitate the transition ES --> ES(double dagger), making kcat/Ks almost temperature independent. The pseudo-first-order rate constant for water attack on L-dihydroorotate, based on experiments at elevated temperature, is 3.2 x 10(-11) s(-1) at 25 degrees C, with deltaH(double dagger) = 24.7 kcal/mol and TdeltaS(double dagger) = -6.9 kcal/mol. Thus, hamster DHOase enhances the rate of substrate hydrolysis by a factor of 1.6 x 10(14), achieving this rate enhancement almost entirely by lowering the enthalpy of activation (delta deltaH(double dagger) = -19.5 kcal/mol). Both the rate enhancement and transition state affinity of hamster DHOase increase steeply with decreasing temperature, consistent with the development of H-bonds and electrostatic interactions in the transition state that were not present in the enzyme-substrate complex in the ground state.