Driving Under the Influence of Cannabis: Impact of Combining Toxicology Testing with Field Sobriety Tests.

BACKGROUND: Cannabis is increasingly used both medically and recreationally. With widespread use, there is growing concern about how to identify cannabis-impaired drivers. METHODS: A placebo-controlled randomized double-blinded protocol was conducted to study the effects of cannabis on driving performance. One hundred ninety-one participants were randomized to smoke ad libitum a cannabis cigarette containing placebo or delta-9-tetrahydrocannabinol (THC) (5.9% or 13.4%). Blood, oral fluid (OF), and breath samples were collected along with longitudinal driving performance on a simulator (standard deviation of lateral position [SDLP] and car following [coherence]) over a 5-hour period. Law enforcement officers performed field sobriety tests (FSTs) to determine if participants were impaired. RESULTS: There was no relationship between THC concentrations measured in blood, OF, or breath and SDLP or coherence at any of the timepoints studied (P > 0.05). FSTs were significant (P < 0.05) for classifying participants into the THC group vs the placebo group up to 188 minutes after smoking. Seventy-one minutes after smoking, FSTs classified 81% of the participants who received active drug as being impaired. However, 49% of participants who smoked placebo (controls) were also deemed impaired at this same timepoint. Combining a 2 ng/mL THC cutoff in OF with positive findings on FSTs reduced the number of controls classified as impaired to zero, 86 minutes after smoking the placebo. CONCLUSIONS: Requiring a positive toxicology result in addition to the FST observations substantially improved the classification accuracy regarding possible driving under the influence of THC by decreasing the percentage of controls classified as impaired.

E-commerce licensing loopholes: a case study of online shopping for tobacco products following a statewide sales restriction on flavoured tobacco in California.

INTRODUCTION: Retailer licensing programmes can be an effective method of enforcing tobacco control laws, but most programmes do not require e-commerce retailers to obtain licenses. California's implementation of a statewide flavour restriction (Senate Bill 793 (SB-793)) in December 2022 enforced through its tobacco retailer licensing programme presented an opportunity to assess whether the exclusion of e-commerce in the definition of 'tobacco retailer' might have resulted in a shift in consumer behaviour towards e-commerce. METHODS: To examine the association between SB-793 implementation and online shopping for tobacco, we collected weekly Google search rates related to online shopping for cigarettes and vaping products in California from January 2018 to May 2023. We compared observed rates of shopping queries after SB-793 implementation to counterfactual expected rates and prediction intervals (PI) calculated from autoregressive iterative moving average models fit to historical trends. Content analysis was performed on the search results to identify websites marketing flavoured vaping products and menthol cigarettes. RESULTS: The week SB-793 was implemented, shopping queries were 194.4% (95% PI 100.8% to 451.5%) and 161.7% (95% PI 81.7% to 367.5%) higher than expected for cigarettes and vapes, respectively. Cigarette shopping queries remained elevated significantly for 11 weeks and vape shopping queries for 6 weeks. All search results contained links to websites that offered flavoured vaping products or menthol cigarettes to Californian consumers. DISCUSSION: These findings raise concerns about potential loopholes in policy enforcement created by the absence of explicit regulations on e-commerce sales in retailer licensing programmes. Strengthening regulations to include e-commerce and monitoring e-commerce compliance are recommended to enhance the impact of laws enforced through retailer licensing programmes.

Improving the value of public RNA-seq expression data by phenotype prediction.

Publicly available genomic data are a valuable resource for studying normal human variation and disease, but these data are often not well labeled or annotated. The lack of phenotype information for public genomic data severely limits their utility for addressing targeted biological questions. We develop an in silico phenotyping approach for predicting critical missing annotation directly from genomic measurements using well-annotated genomic and phenotypic data produced by consortia like TCGA and GTEx as training data. We apply in silico phenotyping to a set of 70 000 RNA-seq samples we recently processed on a common pipeline as part of the recount2 project. We use gene expression data to build and evaluate predictors for both biological phenotypes (sex, tissue, sample source) and experimental conditions (sequencing strategy). We demonstrate how these predictions can be used to study cross-sample properties of public genomic data, select genomic projects with specific characteristics, and perform downstream analyses using predicted phenotypes. The methods to perform phenotype prediction are available in the phenopredict R package and the predictions for recount2 are available from the recount R package. With data and phenotype information available for 70,000 human samples, expression data is available for use on a scale that was not previously feasible.

Integrated Transcriptomic and Proteomic Analysis of Primary Human Umbilical Vein Endothelial Cells.

Understanding the molecular profile of every human cell type is essential for understanding its role in normal physiology and disease. Technological advancements in DNA sequencing, mass spectrometry, and computational methods allow us to carry out multiomics analyses although such approaches are not routine yet. Human umbilical vein endothelial cells (HUVECs) are a widely used model system to study pathological and physiological processes associated with the cardiovascular system. In this study, next-generation sequencing and high-resolution mass spectrometry to profile the transcriptome and proteome of primary HUVECs is employed. Analysis of 145 million paired-end reads from next-generation sequencing confirmed expression of 12 186 protein-coding genes (FPKM ≥0.1), 439 novel long non-coding RNAs, and revealed 6089 novel isoforms that were not annotated in GENCODE. Proteomics analysis identifies 6477 proteins including confirmation of N-termini for 1091 proteins, isoforms for 149 proteins, and 1034 phosphosites. A database search to specifically identify other post-translational modifications provide evidence for a number of modification sites on 117 proteins which include ubiquitylation, lysine acetylation, and mono-, di- and tri-methylation events. Evidence for 11 "missing proteins," which are proteins for which there was insufficient or no protein level evidence, is provided. Peptides supporting missing protein and novel events are validated by comparison of MS/MS fragmentation patterns with synthetic peptides. Finally, 245 variant peptides derived from 207 expressed proteins in addition to alternate translational start sites for seven proteins and evidence for novel proteoforms for five proteins resulting from alternative splicing are identified. Overall, it is believed that the integrated approach employed in this study is widely applicable to study any primary cell type for deeper molecular characterization.

Characterization of genetic loci that affect susceptibility to inflammatory bowel diseases in African Americans.

BACKGROUND & AIMS: Inflammatory bowel disease (IBD) has familial aggregation in African Americans (AAs), but little is known about the molecular genetic susceptibility. Mapping studies using the Immunochip genotyping array expand the number of susceptibility loci for IBD in Caucasians to 163, but the contribution of the 163 loci and European admixture to IBD risk in AAs is unclear. We performed a genetic mapping study using the Immunochip to determine whether IBD susceptibility loci in Caucasians also affect risk in AAs and identify new associated loci. METHODS: We recruited AAs with IBD and without IBD (controls) from 34 IBD centers in the United States; additional controls were collected from 4 other Immunochip studies. Association and admixture loci were mapped for 1088 patients with Crohn's disease, 361 with ulcerative colitis, 62 with IBD type unknown, and 1797 controls; 130,241 autosomal single-nucleotide polymorphisms (SNPs) were analyzed. RESULTS: The strongest associations were observed between ulcerative colitis and HLA rs9271366 (P = 7.5 × 10(-6)), Crohn's disease and 5p13.1 rs4286721 (P = 3.5 × 10(-6)), and IBD and KAT2A rs730086 (P = 2.3 × 10(-6)). Additional suggestive associations (P < 4.2 × 10(-5)) were observed between Crohn's disease and IBD and African-specific SNPs in STAT5A and STAT3; between IBD and SNPs in IL23R, IL12B, and C2orf43; and between ulcerative colitis and SNPs near HDAC11 and near LINC00994. The latter 3 loci have not been previously associated with IBD, but require replication. Established Caucasian associations were replicated in AAs (P < 3.1 × 10(-4)) at NOD2, IL23R, 5p15.3, and IKZF3. Significant admixture (P < 3.9 × 10(-4)) was observed for 17q12-17q21.31 (IZKF3 through STAT3), 10q11.23-10q21.2, 15q22.2-15q23, and 16p12.2-16p12.1. Network analyses showed significant enrichment (false discovery rate <1 × 10(-5)) in genes that encode members of the JAK-STAT, cytokine, and chemokine signaling pathways, as well those involved in pathogenesis of measles. CONCLUSIONS: In a genetic analysis of 3308 AA IBD cases and controls, we found that many variants associated with IBD in Caucasians also showed association evidence with these diseases in AAs; we also found evidence for variants and loci not previously associated with IBD. The complex genetic factors that determine risk for or protection against IBD in different populations require further study.

RNA-Seq optimization with eQTL gold standards.

BACKGROUND: RNA-Sequencing (RNA-Seq) experiments have been optimized for library preparation, mapping, and gene expression estimation. These methods, however, have revealed weaknesses in the next stages of analysis of differential expression, with results sensitive to systematic sample stratification or, in more extreme cases, to outliers. Further, a method to assess normalization and adjustment measures imposed on the data is lacking. RESULTS: To address these issues, we utilize previously published eQTLs as a novel gold standard at the center of a framework that integrates DNA genotypes and RNA-Seq data to optimize analysis and aid in the understanding of genetic variation and gene expression. After detecting sample contamination and sequencing outliers in RNA-Seq data, a set of previously published brain eQTLs was used to determine if sample outlier removal was appropriate. Improved replication of known eQTLs supported removal of these samples in downstream analyses. eQTL replication was further employed to assess normalization methods, covariate inclusion, and gene annotation. This method was validated in an independent RNA-Seq blood data set from the GTEx project and a tissue-appropriate set of eQTLs. eQTL replication in both data sets highlights the necessity of accounting for unknown covariates in RNA-Seq data analysis. CONCLUSION: As each RNA-Seq experiment is unique with its own experiment-specific limitations, we offer an easily-implementable method that uses the replication of known eQTLs to guide each step in one's data analysis pipeline. In the two data sets presented herein, we highlight not only the necessity of careful outlier detection but also the need to account for unknown covariates in RNA-Seq experiments.

SOARinG to New Heights Through a Structured Medical Student Research Program.

Introduction: Since the US Medical Licensing Examination (USMLE) Step 1 became Pass/Fall in 2022, medical students competing for residency spots must distinguish themselves with alternative criteria. Research experiences and output offer valuable skill development and objective metrics to support competitive residency applications. Objective: We describe the methodological development of a structured program to support, enhance, and track medical student research efforts at the University of South Carolina School of Medicine Greenville, explain the implementation of the program, and summarize initial program outcomes. Methods: The Student Opportunities for Academic Achievement Through Research in Greenville (SOARinG) Program was established to serve as a centralized hub for rising second year medical student research. The program matched medical students with mentored research projects scheduled during the summer following first-year coursework. The program included a required weekly seminar series on research basics and current biomedical literature. SOARinG culminated with a student research symposium for which students submitted abstracts and presented a poster or a talk. Quantitative and qualitative program outcomes of student and mentor satisfaction with the program were measured through surveys. Results and Discussion: The program was successfully implemented in summers 2021 and 2022. Most students (80-95%) in each class engaged in mentored summer research projects. Students reported overall satisfaction with research projects and mentor support. Overall, 69% of students rated their overall research experience in the program as extremely good or very good. Each student submitted an abstract and presented at the program's symposium or alternate research venue. Overall, 97% of research mentors reported that students were adequately prepared for summer research and suggested that students would benefit from additional skills-specific research training. Conclusion: The SOARinG Program provided a formalized process for tracking and showcasing medical student research and allowed for increased student participation in research. Additionally, each participating student produced objective research output, thus enhancing future residency applications.

Blood and Oral Fluid Cannabinoid Profiles of Frequent and Occasional Cannabis Smokers.

Increased prevalence of cannabis consumption and impaired driving are a growing public safety concern. Some states adopted per se driving laws, making it illegal to drive with more than a specified blood concentration of ∆9-tetrahydrocannabinol (THC) in a biological fluid (typically blood). Blood THC concentrations decrease significantly (∼90%) with delays in specimen collection, suggesting the use of alternative matrices, such as oral fluid (OF). We characterized 10 cannabinoids' concentrations, including THC metabolites, in blood and OF from 191 frequent and occasional users by liquid chromatography with tandem mass spectrometry for up to 6 h after ad libitum smoking. Subjects self-titrated when smoking placebo, 5.9 or 13.4% THC cannabis. Higher maximum blood THC concentrations (Cmax) were observed in individuals who received the 5.9% THC versus the 13.4% THC plant material. In blood, the Cmax of multiple analytes, including THC and its metabolites, were increased in frequent compared to occasional users, whereas there were no significant differences in OF Cmax. Blood THC remained detectable (≥5 ng/mL) at the final sample collection for 14% of individuals who smoked either the 5.9 or 13.4% THC cigarette, whereas 54% had detectable THC in OF when applying the same cutoff. Occasional and frequent cannabis users' profiles were compared, THC was detectable for significantly longer duration in blood and OF from frequent users. Detection rates between frequent and occasional users at multiple per se cutoffs showed larger differences in blood versus OF. Understanding cannabinoid profiles of frequent and occasional users and the subsequent impact on detectability with current drug per se driving limits is important to support forensic interpretations and the development of scientifically supported driving under the influence of cannabis laws.

The Effect of 12 Hour Shifts, Time of Day, and Sleepiness on Emotional Empathy and Burnout in Medical Students.

Sleepiness decreases alertness and results in decrements in performance. This is especially problematic in the healthcare field due to restricted sleep from shift-work. Sleepiness increases medical errors, but it also affects emotions and interpersonal interactions. Empathy in physicians is a desirable trait which is associated with increased patient recovery rates and patient satisfaction, and decreased use of pain medication. Shift-work may alter empathy in physicians and affect patient outcomes, but the effects of sleepiness on empathy are unknown. Empathy, which is related to burnout, declines during medical school, while incidence of burnout increases. This study assessed the effect of sleepiness from time of day (TOD) and 12 h shifts on empathy and burnout in medical students. Participants were tested on sleepiness and empathy prior to and immediately following a 12 h Emergency Medical Technician shift. Burnout was assessed following each shift to determine if it was affected by sleepiness, empathy, and shift. TOD affected empathy, with empathy highest in the evening. Sleepiness from working 12 h shifts resulted in decreased empathy and increased burnout, with females showing higher rates on the exhaustion component of burnout. This research demonstrates that TOD affects empathy, and sleepiness decreases empathy and increases burnout in medical students.

Shoulder Muscle Activity Dampens Arm Swing Motion When Altering Upper Limb Mass Characteristics During Locomotion.

Weighting the arms during locomotion results in decreased swing motion and increased shoulder muscle activity. To determine the functional relevance of this activity, participants walked on a treadmill with the arms unweighted, or weighted unilaterally or bilaterally. Similar to past work, the weighted arms decreased in swing amplitude and increased their shoulder muscle activity. A close examination of shoulder muscle activities in specific regions of the arm swing cycle suggested these muscles primarily acted eccentrically for all weighting conditions. These findings suggest that the increased shoulder muscle activities when weighting the arms act to dampen the arms when the inertial characteristics of the arms are altered, as opposed to assisting in driving swing of the heavier arms.

The role of mutations associated with familial neurodegenerative disorders on blood-brain barrier function in an iPSC model.

BACKGROUND: Blood-brain barrier dysfunction is associated with many late-stage neurodegenerative diseases. An emerging question is whether the mutations associated with neurodegenerative diseases can independently lead to blood-brain barrier (BBB) dysfunction. Studies from patient-derived induced pluripotent stem cells suggest that mutations associated with neurodegenerative disease are non-cell autonomous, resulting in gain of toxic function in derived neurons and astrocytes. Here we assess whether selected mutations associated with neurodegenerative diseases can contribute to impairment of the blood-brain barrier. METHODS: We assessed barrier function of confluent monolayers of human brain microvascular endothelial cells (hBMECs) derived from induced pluripotent stem cells (iPSC) from three healthy individuals and eight individuals with neurodegenerative disease. We systematically assessed protein and gene expression of BBB biomarkers, transendothelial resistance (TEER), permeability of Lucifer yellow, permeability of D-glucose, permeability of rhodamine 123, the efflux ratio of rhodamine 123, and P-gp inhibition using Tariquidar for confluent monolayers of human brain microvascular endothelial cell (hBMECs). RESULTS: We provide evidence supporting the hypothesis that mutations associated with neurodegenerative disease can independently cause BBB dysfunction. These functional changes are not catastrophic since barrier breakdown would result in BBB impairment during development. Synergistic interactions between non-cell autonomous cerebrovascular dysfunction and the effects of gain-of-toxic function in neurons (e.g. toxic oligomers) are likely to increase disease burden through a positive feedback mechanism. CONCLUSIONS: These results suggest that the accumulation of defects in brain microvascular endothelial cells may ultimately lead to impairment of the BBB. Small changes in barrier function over time could lead to accumulated defects that result in positive feedback to unrelated central nervous system diseases.

Outbreak of sandfly fever in central Iraq, September 2007.

An outbreak of nonspecific febrile illnesses occurred among U.S. Army troops in September 2007 at a remote, newly established, rural-situated patrol base, south of Baghdad, Iraq. Soldiers displayed an acute flu-like syndrome with symptoms of fever, headache, malaise, and myalgia. A total of 14 cases was identified and treated presumptively as query fever. Subsequent convalescent serum specimens confirmed 13 (92.9%) positive for sandfly Sicilian virus and 3 (21.4%) positive for Coxiella burnetii, with two positive for both. One sandfly Sicilian virus case tested positive for Brucella spp. This outbreak emphasizes the potential for multiple simultaneous disease exposures to endemic diseases in nonindigenous military personnel at remote military locations in Iraq. Recommendations include increased theater disease surveillance, medical training, and vector control.

How to share data for collaboration.

Within the statistics community, a number of guiding principles for sharing data have emerged; however, these principles are not always made clear to collaborators generating the data. To bridge this divide, we have established a set of guidelines for sharing data. In these, we highlight the need to provide raw data to the statistician, the importance of consistent formatting, and the necessity of including all essential experimental information and pre-processing steps carried out to the statistician. With these guidelines we hope to avoid errors and delays in data analysis.

Exaggerated CpH methylation in the autism-affected brain.

BACKGROUND: The etiology of autism, a complex, heritable, neurodevelopmental disorder, remains largely unexplained. Given the unexplained risk and recent evidence supporting a role for epigenetic mechanisms in the development of autism, we explored the role of CpG and CpH (H = A, C, or T) methylation within the autism-affected cortical brain tissue. METHODS: Reduced representation bisulfite sequencing (RRBS) was completed, and analysis was carried out in 63 post-mortem cortical brain samples (Brodmann area 19) from 29 autism-affected and 34 control individuals. Analyses to identify single sites that were differentially methylated and to identify any global methylation alterations at either CpG or CpH sites throughout the genome were carried out. RESULTS: We report that while no individual site or region of methylation was significantly associated with autism after multi-test correction, methylated CpH dinucleotides were markedly enriched in autism-affected brains (~2-fold enrichment at p < 0.05 cutoff, p = 0.002). CONCLUSIONS: These results further implicate epigenetic alterations in pathobiological mechanisms that underlie autism.

A Tobacco-Free Medical Campus Policy is Associated With Decreased Secondhand Smoke Exposure and Increased Satisfaction Among Military Medical Employees: Results of a Mixed-Methods Evaluation.

INTRODUCTION: Tobacco control is an ongoing concern for the U.S. Army. Although tobacco use is currently prohibited within all military hospitals and clinics, known as military treatment facilities (MTFs), no such facility had implemented a tobacco-free medical campus (TFMC) policy before 2012. This evaluation examined the effects of one Army installation's TFMC policy implementation at its medical facilities. MATERIALS AND METHODS: Online questionnaires were distributed to medical campus employees, including Active Duty Soldiers, civilians, and contractors, before policy implementation (N = 1,210) and 12 months following policy implementation (N = 1,147). Chi-square analyses, independent t tests, and logistic regression models were utilized to examine pretest/post-test changes in employees' secondhand smoke (SHS) exposure; tobacco use, motivation to quit, and cessation; and health outcomes. Twenty-three focus groups, interviews, and informal discussions with 65 employees and patients were conducted 13 months after initial policy implementation to capture both the intended and unintended policy effects. RESULTS: After controlling for demographic characteristics, the study found that employees had more than twice the odds of exposure to SHS in the workplace at baseline than at 12-month follow-up (odds ratio: 2.06, 95% confidence interval: 1.73-2.46, p < 0.001). Employees also reported a lower prevalence of diagnosis with chronic bronchitis (p < 0.05) at follow up compared to baseline. Although the mean number of sick days taken for respiratory illness decreased over time, results were not significant after controlling for demographic factors. No significant differences existed in tobacco-use prevalence or quit rates among tobacco users over time. Employees reported significantly higher levels of satisfaction with a TFMC policy than the original policy (p < 0.001) though this finding was moderated by smoker status such that smokers reported lower levels of satisfaction with the policy over time. Qualitative findings revealed that the most common policy effect was that the policy caused smokers to change the location of where they used tobacco to off campus. Findings further revealed several unintended policy effects, including safety concerns and greater visibility of smokers in front of the MTF. CONCLUSION: The first Army MTF TFMC policy was associated with reported reductions in SHS exposure and improvements in some short-term health outcomes. The policy had no observed association with tobacco-use prevalence, motivation to quit, or cessation at 12-month follow-up. Focus group participants discussed several positive and negative policy effects. These policies should be expanded and studied in more depth across military installations, and policy makers should plan mitigation strategies to reduce unintended effects. This is an important step in military tobacco control, but additional efforts will be necessary to curb tobacco use within this population.

Cross-tissue integration of genetic and epigenetic data offers insight into autism spectrum disorder.

Integration of emerging epigenetic information with autism spectrum disorder (ASD) genetic results may elucidate functional insights not possible via either type of information in isolation. Here we use the genotype and DNA methylation (DNAm) data from cord blood and peripheral blood to identify SNPs associated with DNA methylation (meQTL lists). Additionally, we use publicly available fetal brain and lung meQTL lists to assess enrichment of ASD GWAS results for tissue-specific meQTLs. ASD-associated SNPs are enriched for fetal brain (OR = 3.55; P < 0.001) and peripheral blood meQTLs (OR = 1.58; P < 0.001). The CpG targets of ASD meQTLs across cord, blood, and brain tissues are enriched for immune-related pathways, consistent with other expression and DNAm results in ASD, and reveal pathways not implicated by genetic findings. This joint analysis of genotype and DNAm demonstrates the potential of both brain and blood-based DNAm for insights into ASD and psychiatric phenotypes more broadly.

Porn again: some final conclusions.

Contributors were asked to respond to seven questions examining various aspects of gay male pornography. Their responses were collated with the hope that the reader may gain additional insight into this topic.

Transcriptome analysis reveals dysregulation of innate immune response genes and neuronal activity-dependent genes in autism.

Recent studies of genomic variation associated with autism have suggested the existence of extreme heterogeneity. Large-scale transcriptomics should complement these results to identify core molecular pathways underlying autism. Here we report results from a large-scale RNA sequencing effort, utilizing region-matched autism and control brains to identify neuronal and microglial genes robustly dysregulated in autism cortical brain. Remarkably, we note that a gene expression module corresponding to M2-activation states in microglia is negatively correlated with a differentially expressed neuronal module, implicating dysregulated microglial responses in concert with altered neuronal activity-dependent genes in autism brains. These observations provide pathways and candidate genes that highlight the interplay between innate immunity and neuronal activity in the aetiology of autism.