Chemical Composition of Electronic Vaping Products from School Grounds in California.

INTRODUCTION: The use of electronic vaping products (EVPs) containing nicotine, marijuana, and/or other substances remains prominent among youth; with EVPs containing nicotine being the most commonly used tobacco product among youth since 2014. However, a detailed understanding of the chemical composition of these products is limited. METHODS: During February 25th-March 15th, 2019, a total of 576 EVPs, including 233 e-cigarette devices (with 43 disposable vape pens) and 343 e-liquid cartridges/pods/bottled e-liquids, were found or confiscated from a convenience sample of 16 public high schools in California. Liquids inside 251 vape pens and cartridges/pods/bottled e-liquids were analyzed using a gas chromatography/mass spectrometry (GC/MS). For comparison, new JUUL pods, the most commonly used e-cigarette among youth during 2018-2019, with different flavorings and nicotine content were purchased and analyzed. RESULTS: For e-cigarette cartridges/pods/bottled e-liquids, nicotine was detected in 204 of 208 (98.1%) samples. Propylene glycol (PG) and vegetable glycerin (VG) were dominant solvents in nicotine-containing EVPs. Among 43 disposable vape pen devices, cannabinoids such as tetrahydrocannabinol (THC) or cannabidiol (CBD) were identified in 39 of 43 (90.1%) samples, of which 3 contained both nicotine and THC. Differences in chemical compositions were observed between confiscated or collected JUULs and purchased JUULs. Measured nicotine was inconsistent with labels on some confiscated or collected bottled e-liquids. CONCLUSIONS: EVPs from 16 participating schools were found to widely contain substances with known adverse health effects among youth, including nicotine and cannabinoids. There was inconsistency between labeled and measured nicotine on the products from schools. IMPLICATIONS: This study measured the main chemical compositions of EVPs found at 16 California public high schools. Continued efforts are warranted, including at the school-level, to educate, prevent and reduce youth use of EVPs.

Implementation of Electronic Health Records in US Nursing Homes.

While electronic health records have emerged as promising tools to help improve quality of care, nursing homes have lagged behind in implementation. This study assessed electronic health records implementation, associated facility characteristics, and potential impact on quality indicators in nursing homes. Using national Centers for Medicare & Medicaid Services and survey data for nursing homes, a cross-sectional analysis was conducted to identify variations between nursing homes that had and had not implemented electronic health records. A difference-in-differences analysis was used to estimate the longitudinal effect of electronic health records on commonly used quality indicators. Data from 927 nursing homes were examined, 49.1% of which had implemented electronic health records. Nursing homes with electronic health records were more likely to be nonprofit/government owned (P = .04) and had a lower percentage of Medicaid residents (P = .02) and higher certified nursing assistant and registered nurse staffing levels (P = .002 and .02, respectively). Difference-in-differences analysis showed greater quality improvements after implementation for five long-stay and two short-stay quality measures (P = .001 and .01, respectively) compared with those who did not implement electronic health records. Implementation rates in nursing homes are low compared with other settings, and better-resourced facilities are more likely to have implemented electronic health records. Consistent with other settings, electronic health records implementation improves quality in nursing homes, but further research is needed to better understand the mechanism for improvement and how it can best be supported.

Obesity as a Determinant of Staphylococcus aureus Colonization Among Inmates in Maximum-Security Prisons in New York State.

Obesity increases a person's susceptibility to a variety of infections, including Staphylococcus aureus infections, which is an important cause of morbidity in correctional settings. Using a cross-sectional design, we assessed the association between obesity and S. aureus colonization, a risk factor for subsequent infection, in New York State maximum-security prisons (2011-2013). Anterior nares and oropharyngeal cultures were collected. Structured interviews and medical records were used to collect demographic, behavioral, and medical data. Body mass index (BMI; weight (kg)/height (m(2))) was categorized as 18.5-24.9, 25-29.9, 30-34.9, or ≥35. The association between BMI and S. aureus colonization was assessed using log-binomial regression. Thirty-eight percent of 638 female inmates and 26% of 794 male inmates had a BMI of 30 or higher. More than 40% of inmates were colonized. Female inmates with a BMI of 25-29.9 (prevalence ratio (PR) = 1.37, 95% confidence interval (CI): 1.06, 1.76), 30-34.9 (PR = 1.52, 95% CI: 1.17, 1.98), or ≥35 (PR = 1.49, 95% CI: 1.13, 1.96) had a higher likelihood of colonization than did those with a BMI of 18.5-24.9 after we controlled for age, educational level, smoking status, diabetes status, and presence of human immunodeficiency virus. Colonization was higher among male inmates with a BMI of 30-34.9 (PR = 1.27, 95% CI: 1.01, 1.61). Our findings demonstrate an association between BMI and S. aureus colonization among female prisoners. Potential contributory biologic and behavioral factors should be explored.

Gene number determination and genetic polymorphism of the gamma delta T cell co-receptor WC1 genes.

BACKGROUND: WC1 co-receptors belong to the scavenger receptor cysteine-rich (SRCR) superfamily and are encoded by a multi-gene family. Expression of particular WC1 genes defines functional subpopulations of WC1(+) γδ T cells. We have previously identified partial or complete genomic sequences for thirteen different WC1 genes through annotation of the bovine genome Btau_3.1 build. We also identified two WC1 cDNA sequences from other cattle that did not correspond to sequences in the Btau_3.1 build. Their absence in the Btau_3.1 build may have reflected gaps in the genome assembly or polymorphisms among animals. Since the response of γδ T cells to bacterial challenge is determined by WC1 gene expression, it was critical to understand whether individual cattle or breeds differ in the number of WC1 genes or display polymorphisms. RESULTS: Real-time quantitative PCR using DNA from the animal whose genome was sequenced ("Dominette") and sixteen other animals representing ten breeds of cattle, showed that the number of genes coding for WC1 co-receptors is thirteen. The complete coding sequences of those thirteen WC1 genes is presented, including the correction of an error in the WC1-2 gene due to mis-assembly in the Btau_3.1 build. All other cDNA sequences were found to agree with the previous annotation of complete or partial WC1 genes. PCR amplification and sequencing of the most variable N-terminal SRCR domain (domain 1 which has the SRCR "a" pattern) of each of the thirteen WC1 genes showed that the sequences are highly conserved among individuals and breeds. Of 160 sequences of domain 1 from three breeds of cattle, no additional sequences beyond the thirteen described WC1 genes were found. Analysis of the complete WC1 cDNA sequences indicated that the thirteen WC1 genes code for three distinct WC1 molecular forms. CONCLUSION: The bovine WC1 multi-gene family is composed of thirteen genes coding for three structural forms whose sequences are highly conserved among individual cattle and breeds. The sequence diversity necessary for WC1 genes to function as a multi-genic pattern recognition receptor array is encoded in the genome, rather than generated by recombinatorial diversity or hypermutation.

Scavenger receptor WC1 contributes to the γδ T cell response to Leptospira.

WC1 molecules are exclusively expressed on the surface of γδ T cells. They belong to the scavenger receptor cysteine-rich (SRCR) superfamily and are encoded by a multi-gene family. WC1 molecules have been grouped on the basis of antibody reactivity. The expression of WC1 molecules from these serologically defined groups is correlated with differences in γδ T cell responses. The expression of receptors within the WC1.1 group correlates with the capacity of γδ T cells to respond to Leptospira antigen. In this study, we used RNA interference to directly investigate the role of WC1 expression in the response to Leptospira borgpetersenii. We found that when three out of thirteen WC1 gene products were downregulated by RNA interference, γδ T cell proliferation and IFN-γ production in response to Leptospira antigen was significantly reduced. Our data demonstrate that specific receptors in the WC1 family directly participate in Leptospira recognition and/or activation of γδ T cells.

Evolution of the CD163 family and its relationship to the bovine gamma delta T cell co-receptor WC1.

BACKGROUND: The scavenger receptor cysteine rich (SRCR) domain is an ancient and conserved protein domain. CD163 and WC1 molecules are classed together as group B SRCR superfamily members, along with Spalpha, CD5 and CD6, all of which are expressed by immune system cells. There are three known types of CD163 molecules in mammals, CD163A (M130, coded for by CD163), CD163b (M160, coded for by CD163L1) and CD163c-alpha (CD163L1 or SCART), while their nearest relative, WC1, is encoded by a multigene family so far identified in the artiodactyl species of cattle, sheep, and pigs. RESULTS: We annotated the bovine genome and identified genes coding for bovine CD163A and CD163c-alpha but found no evidence for CD163b. Bovine CD163A is widely expressed in immune cells, whereas CD163c-alpha transcripts are enriched in the WC1+ gammadelta T cell population. Phylogenetic analyses of the CD163 family genes and WC1 showed that CD163c-alpha is most closely related to WC1 and that chicken and platypus have WC1 orthologous genes, previously classified as among their CD163 genes. CONCLUSION: Since it has been shown that WC1 plays an important role in the regulation of gammadelta T cell responses in cattle, which, like chickens, have a high percentage of gammadelta T cells in their peripheral blood, CD163c-alpha may play a similar role, especially in species lacking WC1 genes. Our results suggest that gene duplications resulted in the expansion of CD163c-alpha-like and WC1-like molecules. This expanded repertoire was retained by species known as "gammadelta T cell high", but homologous SRCR molecules were maintained by all mammals.

Antigenic basis of diversity in the gammadelta T cell co-receptor WC1 family.

WC1 co-receptors are transmembrane glycoproteins with 11 extracellular scavenger receptor cysteine rich (SRCR) domains. They are related to the CD163 family but are uniquely expressed by gammadelta T cells. We recently showed that at least 13 members comprise the WC1 gene family in cattle, a model animal species for studies of gammadelta T cell biology. Since WC1 co-receptors participate in directing functional responses by gammadelta T cells either through the ligands they bind or the signals they transduce, availability of reagents to identify the expression of individual WC1 molecules of this diverse family would be valuable in further elucidating mechanisms of gammadelta T cell responsiveness. Although monoclonal antibodies (mAbs) have been widely used to identify WC1 co-receptors on gammadelta T cells, the locations of the antigenic epitopes recognized are unknown. Here, we mapped the epitopes to particular SRCR domains and evaluated their distribution among WC1 molecules. To do this, cDNA representing the extracellular domains of seven different WC1 genes was expressed in mammalian cells and analyzed for reactivity with anti-WC1 mAbs using ELISA and Western blotting. The study included mAbs that are broadly reactive with WC1(+) gammadelta T cells and those that divide WC1(+) gammadelta T cells into functionally distinct subpopulations. We found that mAb CC15 is a pan-reactive anti-WC1 mAb recognizing an epitope in the closely related SRCR domains 2 and 7 and that this epitope is present in at least domain 2 or 7 of all seven WC1 molecules evaluated here. Five other anti-WC1 mAbs, typified by mAb IL-A29, were found to be broadly reactive, recognizing epitopes in the related SRCR domains 4 and 9 but each having a unique pattern of reactivity with the seven WC1 molecules. Finally, the subpopulation-specific anti-WC1 mAbs, including those that recognize either the archetypal WC1.1 or WC1.2 molecule, were found to react with epitopes in the most variable WC1 domain, i.e. domain 1, of a restricted number of WC1 co-receptors.

Genomic organization and classification of the bovine WC1 genes and expression by peripheral blood gamma delta T cells.

BACKGROUND: WC1 co-receptors are group B scavenger receptor cysteine-rich molecules that are found exclusively on gammadeltaT cells and are thought to be encoded by a multi-gene family. Previous studies have shown gammadeltaT cells that respond to a particular stimulus have unique WC1 molecules expressed. Prior to the onset of the studies described here only one full-length WC1 nucleotide sequence was publicly available, though three WC1 molecules had been distinguished based on monoclonal antibody reactivity. Furthermore, the number of WC1 genes found in the bovine genome and their sequences had not yet been resolved. RESULTS: By annotating the bovine genome Btau_3.1 assembly, here we show the existence of 13 members in the WC1 gene family and their organization within two loci on chromosome 5 including three distinct exon-intron gene structures one of which coded for a potentially more primitive and smaller WC1 molecule that is similar to the swine WC1 gene. We also provide cDNA evidence as verification for many of the annotated sequences and show transcripts for isoforms derived by alternative splicing. CONCLUSION: It is possible that WC1 diversity contributes to functional differences that have been observed between gammadeltaT cell populations. The studies described here demonstrate that WC1 molecules are encoded by a large, multi-gene family whose transcripts undergo extensive alternative splicing. Similar to other non-rearranging immunoreceptors, it is likely that the WC1 gene repertoire underwent expansion in order to keep pace with rapidly changing ligands.

Differential TCR gene usage between WC1- and WC1+ ruminant gammadelta T cell subpopulations including those responding to bacterial antigen.

Ruminant gammadelta T cells are divided into subpopulations based on the presence or absence of WC1 co-receptors (scavenger-receptor-cysteine-rich family members uniquely expressed on gammadelta T cells). Evidence suggests WC1+ are inflammatory while WC1- are regulatory and that they also differ in their tissue distribution. Recently, this paradigm was refined further as cells that produce interferon-gamma and proliferate to autologous antigens, leptospira antigens, or IL-12 were largely found within the WC1+ subpopulation that bears the WC1.1 antigenic epitope but not that bearing the WC1.2 epitope. Here, the T cell receptor gene expression by these different subpopulations (WC1-, WC1.1+, and WC1.2+) was compared using flow cytometrically-purified cells and reverse transcriptase-polymerase chain reaction (RT-PCR). The WC1- gammadelta T cells had transcripts for all 11 possible combinations of the TRG subgroup V and C genes while those in both WC1+ subpopulations were restricted to TRGV3-TRGC5 and TRGV7-TRGC5. In contrast, all three subpopulations expressed transcripts from all four known bovine TRDV genes. Further analysis of the WC1+ gammadelta T cells that proliferated in leptospira antigen-stimulated cultures indicated that they do not represent a unique subpopulation within the larger WC1+ population based on their TCR gene usage. Moreover, sequencing of 65 transcripts showed that their junctional regions were diverse as TRGJ5-1, TRGJ5-2, TRDJ1, and TRDJ3 were used, and CDR3s ranged from 9 to 24 amino acids. The restricted but shared gammadelta TCR gene usage for WC1.1+, WC1.2+, and WC1(+)-antigen-responsive cells leaves open the possibility that the WC1 co-receptor is an important determining element in the activation process and subsequent response.