STIM1-dependent store-operated calcium entry mediates sex differences in macrophage chemotaxis and monocyte recruitment.

Infiltration of monocyte-derived cells to sites of infection and injury is greater in males than in females, due in part, to increased chemotaxis, the process of directed cell movement toward a chemical signal. The mechanisms governing sexual dimorphism in chemotaxis are not known. We hypothesized a role for the store-operated calcium entry (SOCE) pathway in regulating chemotaxis by modulating leading and trailing edge membrane dynamics. We measured the chemotactic response of bone marrow-derived macrophages migrating toward complement component 5a (C5a). Chemotactic ability was dependent on sex and inflammatory phenotype (M0, M1, and M2), and correlated with SOCE. Notably, females exhibited a significantly lower magnitude of SOCE than males. When we knocked out the SOCE gene, stromal interaction molecule 1 (STIM1), it eliminated SOCE and equalized chemotaxis across both sexes. Analysis of membrane dynamics at the leading and trailing edges showed that STIM1 influences chemotaxis by facilitating retraction of the trailing edge. Using BTP2 to pharmacologically inhibit SOCE mirrored the effects of STIM1 knockout, demonstrating a central role of STIM/Orai-mediated calcium signaling. Importantly, by monitoring the recruitment of adoptively transferred monocytes in an in vivo model of peritonitis, we show that increased infiltration of male monocytes during infection is dependent on STIM1. These data support a model in which STIM1-dependent SOCE is necessary and sufficient for mediating the sex difference in monocyte recruitment and macrophage chemotactic ability by regulating trailing edge dynamics.

Introduction of a new method for two-dimensional NMR quantitative analysis in metabolomics studies.

NMR is one of the most important platforms for metabolomic studies. Though 2D NMR has been applied in metabolomics, most applications have mainly focused on metabolite identification whilst limitations causing a bottle-neck for applying high-throughput 2D NMR data for quantity related statistical analysis lies on the data interpretation methods. In this study, instead of using the traditional methods of calculating the 2D NMR data to search for the important features, a new procedure, which applies the high-resolution 1D NMR metabolites chemical shift range to filter the 2D NMR data, was developed. This new method was demonstrated using both a mixture of standard metabolites and a case study on plant extracts using 2D non-uniform sampling (NUS) total correlation spectroscopy (TOCSY) data. As a result, our method successfully filtered out the important features with a high success rate, and the extracted peaks showed high linearity between the calculated intensities and the concentrations of metabolites from a range of 0.05 mM-2 mM. The method was successfully applied to a metabolomics case study which included 18 Begonia samples that showed excellent peak extractions. In summary, our study has provided a practical new 2D NMR data extraction method for use in future metabolomics studies.

Genetic Variation, Not Cell Type of Origin, Underlies the Majority of Identifiable Regulatory Differences in iPSCs.

The advent of induced pluripotent stem cells (iPSCs) revolutionized human genetics by allowing us to generate pluripotent cells from easily accessible somatic tissues. This technology can have immense implications for regenerative medicine, but iPSCs also represent a paradigm shift in the study of complex human phenotypes, including gene regulation and disease. Yet, an unresolved caveat of the iPSC model system is the extent to which reprogrammed iPSCs retain residual phenotypes from their precursor somatic cells. To directly address this issue, we used an effective study design to compare regulatory phenotypes between iPSCs derived from two types of commonly used somatic precursor cells. We find a remarkably small number of differences in DNA methylation and gene expression levels between iPSCs derived from different somatic precursors. Instead, we demonstrate genetic variation is associated with the majority of identifiable variation in DNA methylation and gene expression levels. We show that the cell type of origin only minimally affects gene expression levels and DNA methylation in iPSCs, and that genetic variation is the main driver of regulatory differences between iPSCs of different donors. Our findings suggest that studies using iPSCs should focus on additional individuals rather than clones from the same individual.

Expression Quantitative Trait Locus Mapping Studies in Mid-secretory Phase Endometrial Cells Identifies HLA-F and TAP2 as Fecundability-Associated Genes.

Fertility traits in humans are heritable, however, little is known about the genes that influence reproductive outcomes or the genetic variants that contribute to differences in these traits between individuals, particularly women. To address this gap in knowledge, we performed an unbiased genome-wide expression quantitative trait locus (eQTL) mapping study to identify common regulatory (expression) single nucleotide polymorphisms (eSNPs) in mid-secretory endometrium. We identified 423 cis-eQTLs for 132 genes that were significant at a false discovery rate (FDR) of 1%. After pruning for strong LD (r2 >0.95), we tested for associations between eSNPs and fecundability (the ability to get pregnant), measured as the length of the interval to pregnancy, in 117 women. Two eSNPs were associated with fecundability at a FDR of 5%; both were in the HLA region and were eQTLs for the TAP2 gene (P = 1.3x10-4) and the HLA-F gene (P = 4.0x10-4), respectively. The effects of these SNPs on fecundability were replicated in an independent sample. The two eSNPs reside within or near regulatory elements in decidualized human endometrial stromal cells. Our study integrating eQTL mapping in a primary tissue with association studies of a related phenotype revealed novel genes and associated alleles with independent effects on fecundability, and identified a central role for two HLA region genes in human implantation success.

Reprogramming LCLs to iPSCs Results in Recovery of Donor-Specific Gene Expression Signature.

Renewable in vitro cell cultures, such as lymphoblastoid cell lines (LCLs), have facilitated studies that contributed to our understanding of genetic influence on human traits. However, the degree to which cell lines faithfully maintain differences in donor-specific phenotypes is still debated. We have previously reported that standard cell line maintenance practice results in a loss of donor-specific gene expression signatures in LCLs. An alternative to the LCL model is the induced pluripotent stem cell (iPSC) system, which carries the potential to model tissue-specific physiology through the use of differentiation protocols. Still, existing LCL banks represent an important source of starting material for iPSC generation, and it is possible that the disruptions in gene regulation associated with long-term LCL maintenance could persist through the reprogramming process. To address this concern, we studied the effect of reprogramming mature LCL cultures from six unrelated donors to iPSCs on the ensuing gene expression patterns within and between individuals. We show that the reprogramming process results in a recovery of donor-specific gene regulatory signatures, increasing the number of genes with a detectable donor effect by an order of magnitude. The proportion of variation in gene expression statistically attributed to donor increases from 6.9% in LCLs to 24.5% in iPSCs (P < 10-15). Since environmental contributions are unlikely to be a source of individual variation in our system of highly passaged cultured cell lines, our observations suggest that the effect of genotype on gene regulation is more pronounced in iPSCs than in LCLs. Our findings indicate that iPSCs can be a powerful model system for studies of phenotypic variation across individuals in general, and the genetic association with variation in gene regulation in particular. We further conclude that LCLs are an appropriate starting material for iPSC generation.

Family planning and resilience: associations found in a Population, Health, and Environment (PHE) project in Western Tanzania.

Using 2016 household survey data from Tanzania, we define and measure resilience within the context of Population, Health, and Environment programming and quantify the link between resilience and family planning. We created a multicomponent model using confirmatory factor analysis in a structural equation modeling context. Factor loadings for eight defined latent factors of resilience were statistically significant (p < 0.001). We created a factor called "FP-MCH" reflecting awareness, attitudes, and access to family planning (FP) and health care services and use of maternal and child health care (MCH) facilities. Analysis, with controls, shows that a 1 standard deviation (SD) increase in FP/MCH was associated with a 0.68 SD increase in resilience (p < 0.01), suggesting that the association between FP/MCH and resilience is robust across a range of factors. Analyses showed that the association between FP/MCH is broadly related to the construct of resilience and not through any single component. This study supports the importance of including FP/MCH as part of integrated projects to enhance resilience.

The maternal HLA-G 1597ΔC null mutation is associated with increased risk of pre-eclampsia and reduced HLA-G expression during pregnancy in African-American women.

The non-classical major histocompatibility complex molecule, human leukocyte antigen (HLA)-G, is thought to contribute to maternal immune tolerance and successful placentation during pregnancy. Genetic polymorphisms in HLA-G are known to influence expression levels as well as the relative expression of individual protein isoforms. As diminished or aberrant HLA-G expression patterns may contribute to the development of certain pregnancy complications, we sought to investigate the association between functional HLA-G polymorphisms and the risk of pre-eclampsia (PE) in African-American women. The association between maternal and fetal genotype at six HLA-G polymorphisms and risk of PE was assessed in 372 pregnancies (314 normotensive; 58 pre-eclamptic). We observed an elevated risk of PE (P = 0.00027) in pregnancies where the mother carried the 1597ΔC allele, a null allele that abolishes expression of full-length HLA-G isoforms. Furthermore, the frequency of the maternal 1597ΔC allele was highest in the subset of pre-eclamptic pregnancies that were delivered preterm, suggesting an association between the null allele and the severity of PE. We then replicated the association between higher maternal 1597ΔC allele frequency and increased severity of PE (P = 0.038) in an independent sample of 533 African-American women. Finally, to investigate the mechanistic basis of this association, we measured circulating soluble HLA-G (sHLA-G) concentrations in maternal serum collected during pregnancy in 51 healthy, normotensive African-American control women and found significantly lower levels in women carrying the 1597ΔC allele (P = 0.012). These results demonstrate that maternal HLA-G genotype is significantly associated with risk of PE in African-American women and is predictive of circulating sHLA-G levels during pregnancy.

Unveiling an indole alkaloid diketopiperazine biosynthetic pathway that features a unique stereoisomerase and multifunctional methyltransferase.

The 2,5-diketopiperazines are a prominent class of bioactive molecules. The nocardioazines are actinomycete natural products that feature a pyrroloindoline diketopiperazine scaffold composed of two D-tryptophan residues functionalized by N- and C-methylation, prenylation, and diannulation. Here we identify and characterize the nocardioazine B biosynthetic pathway from marine Nocardiopsis sp. CMB-M0232 by using heterologous biotransformations, in vitro biochemical assays, and macromolecular modeling. Assembly of the cyclo-L-Trp-L-Trp diketopiperazine precursor is catalyzed by a cyclodipeptide synthase. A separate genomic locus encodes tailoring of this precursor and includes an aspartate/glutamate racemase homolog as an unusual D/L isomerase acting upon diketopiperazine substrates, a phytoene synthase-like prenyltransferase as the catalyst of indole alkaloid diketopiperazine prenylation, and a rare dual function methyltransferase as the catalyst of both N- and C-methylation as the final steps of nocardioazine B biosynthesis. The biosynthetic paradigms revealed herein showcase Nature's molecular ingenuity and lay the foundation for diketopiperazine diversification via biocatalytic approaches.

Fluorescence Quenching of Xanthene Dyes during Amide Bond Formation Using DMTMM.

Fluorophore bioconjugation to proteins, nucleic acids, and other important molecules can provide a powerful approach to sensing, imaging, and quantifying chemical and biological processes. One of the most prevalent methods for fluorophore attachment is through the formation of amide bonds, which are often facilitated by coupling agents to activate carboxylic acid moieties for subsequent nucleophilic attack by amines. 4-(4,6-Dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM) is among the most popular of these coupling agents for bioconjugation due to its ability to facilitate amide bond formation in water. After observing quenching of 5-fluoresceinamine (5-FAM)-conjugated oligonucleotides in the presence of DMTMM, we sought to evaluate the magnitude and scope of this challenge by surveying the effect of DMTMM on a range of fluorescent dyes. A higher quenching effect was consistently observed for xanthene dyes compared to that for cyanine dyes. Further analysis of the impact of DMTMM on FAM shows that quenching occurs independently of whether the dye is free in solution or attached to an oligonucleotide or antibody. Furthermore, we found that FAM-conjugated DNA was unable to recover its fluorescence after the removal of DMTMM, and UV-vis and NMR analyses suggest the formation of new products, such as an adduct formed between FAM and the dimethoxytriazine of DMTMM. As such, DMTMM at high concentrations is not recommended for coupling reactions where targets are fluorescently labeled. This research serves as a word of caution to those utilizing xanthene-containing fluorophores in bioconjugation reactions involving DMTMM.

Livelihood transitions and the changing nature of farmer-herder conflict in Sahelian West Africa.

The accommodation of livestock husbandry with crop agriculture is crucial for the future of the West African Sahel. Present trends are leading to greater restrictions on livestock husbandry and a growing convergence of livelihood practices among groups whose identities are tied to herding and farming. Using the cases of four rural communities in Niger, this study adopts an 'access to resources' framework to analyse the causal connections among: rural peoples' livelihood strategies, everyday social relations of production, perceptions of social groups' identities, and the potential for farmer-herder conflict. While the convergence of livelihoods arguably increases the frequency of conflict triggers, it has also, through the expansion of shared common interests and cross-group, production-related relationships, improved the ability of communities to effectively manage these incipient conflicts.

IL-17 and IL-17C Signaling Protects the Intestinal Epithelium against Diisopropyl Fluorophosphate Exposure in an Acute Model of Gulf War Veterans' Illnesses.

Gulf War Veterans' Illnesses (GWI) encompasses a broad range of unexplained symptomology specific to Veterans of the Persian Gulf War. Gastrointestinal (GI) distress is prominent in veterans with GWI and often presents as irritable bowel syndrome (IBS). Neurotoxins, including organophosphorus pesticides and sarin gas, are believed to have contributed to the development of GWI, at least in a subset of Veterans. However, the effects of such agents have not been extensively studied for their potential impact to GI disorders and immunological stability. Here we utilized an established murine model of GWI to investigate deleterious effects of diisopropyl fluorophosphate (DFP) exposure on the mucosal epithelium in vivo and in vitro. In vivo, acute DFP exposure negatively impacts the mucosal epithelium by reducing tight junction proteins and antimicrobial peptides as well as altering intestinal microbiome composition. Furthermore, DFP treatment reduced the expression of IL-17 in the colonic epithelium. Conversely, both IL-17 and IL-17C treatment could combat the negative effects of DFP and other cholinesterase inhibitors in murine intestinal organoid cells. Our findings demonstrate that acute exposure to DFP can result in rapid deterioration of mechanisms protecting the GI tract from disease. These results are relevant to suspected GWI exposures and could help explain the propensity for GI disorders in GWI Veterans.

Climate Solutions Double as Health Interventions.

The climate crisis threatens to exacerbate numerous climate-sensitive health risks, including heatwave mortality, malnutrition from reduced crop yields, water- and vector-borne infectious diseases, and respiratory illness from smog, ozone, allergenic pollen, and wildfires. Recent reports from the Intergovernmental Panel on Climate Change stress the urgent need for action to mitigate climate change, underscoring the need for more scientific assessment of the benefits of climate action for health and wellbeing. Project Drawdown has analyzed more than 80 solutions to address climate change, building on existing technologies and practices, that could be scaled to collectively limit warming to between 1.5° and 2 °C above preindustrial levels. The solutions span nine major sectors and are aggregated into three groups: reducing the sources of emissions, maintaining and enhancing carbon sinks, and addressing social inequities. Here we present an overview of how climate solutions in these three areas can benefit human health through improved air quality, increased physical activity, healthier diets, reduced risk of infectious disease, and improved sexual and reproductive health, and universal education. We find that the health benefits of a low-carbon society are more substantial and more numerous than previously realized and should be central to policies addressing climate change. Much of the existing literature focuses on health effects in high-income countries, however, and more research is needed on health and equity implications of climate solutions, especially in the Global South. We conclude that adding the myriad health benefits across multiple climate change solutions can likely add impetus to move climate policies faster and further.

Toll-like receptor 2 induces pathogenicity in Th17 cells and reveals a role for IPCEF in regulating Th17 cell migration.

Pathogenic Th17 cells drive inflammation in autoimmune disease, yet the molecular programming underlying Th17 cell pathogenicity remains insufficiently understood. Activation of Toll-like receptor 2 (TLR2) increases Th17 cell inflammatory potential, but little is known regarding the mechanistic outcomes of TLR2 signaling in Th17 cells. Here, we demonstrate that TLR2 is comparable to IL-23 in inducing pathogenicity and increasing the migratory capacity of Th17 cells. We perform RNA sequencing of Th17 cells stimulated though the TLR2 pathway and find differential expression of several genes linked with the Th17 genetic program as well as genes not previously associated with pathogenic Th17 cells, including Ipcef1. Enforced expression of Ipcef1 in Th17 cells abolishes the TLR2-dependent increases in migratory capacity and severely impairs the ability of Th17 cells to induce experimental autoimmune encephalomyelitis. This study establishes the importance of the TLR2 signaling pathway in inducing Th17 cell pathogenicity and driving autoimmune inflammation.

Local Government Approaches to Combating COVID-19 Inequities: A Durham County Department of Public Health Perspective.

When a novel coronavirus disease (COVID-19) made major headlines in 2020, it further exposed an existing public health crisis related to inequities within our communities and health care delivery system. Throughout the COVID-19 pandemic, populations of color had higher infection and mortality rates, and even experienced greater disease severity compared to whites. Populations of color often bear the brunt of COVID-19 and other health inequities, due to the multifaceted relationship between systemic racism and the social determinants of health. As this relationship continues to perpetuate health inequities, the local health department is an agency that has the jurisdiction and responsibility to prevent disease and protect the health of the communities they serve. When equity is integrated into a health department's operational infrastructure as a disease prevention strategy, it can elevate the agency's response to public health emergencies. Collecting, reporting, and tracking demographic data that is necessary to identify inequities becomes a priority to facilitate a more robust public health response. The purpose of this paper is to present strategies of how a local health department operationalized equity in various stages of COVID-19 response and apply these methods to future public health emergencies to better serve vulnerable communities.

Photodegradation of ibuprofen and four other pharmaceutical pollutants on natural pigments sensitized TiO2 nanoparticles.

Pharmaceuticals and personal care products (PPCPs) in water system have drawn increasing concerns in recent years. TiO2 -based photodegradation has shown great potential as a low-cost and sustainable technology in water treatment, however, can only use the UV light range of solar radiation which makes the system less efficient. Dyes have been studied to improve the TiO2 system light-harvesting range, but studies on environmental friendly natural dyes are rare. In this study, a screening method using UV-Vis spectra analysis was carried out on a group of 22 different tropical natural plants for the potential applications on dye-sensitized TiO2 in PPCP treatment. As a result, Begonia "Martin's Mystery" significantly increased TiO2 photodegradation efficiency toward ibuprofen treatments which is first time reported in literature as our best knowledge. Moreover, the promising discovery of Begonia application in ibuprofen treatment has been successfully applied to warfarin and famotidine treatment. Similar results were expanded to many other Begonia species which indicate that Begonia extracts could be excellent sensitizers for TiO2 -based photodegradation of PPCPs. Our discovery suggested that the screening process may potentially open a brand-new way for future TiO2 photodegradation studies before the complex and time-consuming detailed mechanism studies. PRACTITIONER POINTS.: Natural dyes were screened as sensitizers for TiO2 photodegradation of ibuprofen. Ibuprofen photodegradation efficiency was increased twice using Begonia "Martin's Mystery." The Begonia applications were extended to warfarin, trimethoprim, and famotidine. Promising results were also observed using five other Begonia species.

Determining the genetic basis of anthracycline-cardiotoxicity by molecular response QTL mapping in induced cardiomyocytes.

Anthracycline-induced cardiotoxicity (ACT) is a key limiting factor in setting optimal chemotherapy regimes, with almost half of patients expected to develop congestive heart failure given high doses. However, the genetic basis of sensitivity to anthracyclines remains unclear. We created a panel of iPSC-derived cardiomyocytes from 45 individuals and performed RNA-seq after 24 hr exposure to varying doxorubicin dosages. The transcriptomic response is substantial: the majority of genes are differentially expressed and over 6000 genes show evidence of differential splicing, the later driven by reduced splicing fidelity in the presence of doxorubicin. We show that inter-individual variation in transcriptional response is predictive of in vitro cell damage, which in turn is associated with in vivo ACT risk. We detect 447 response-expression quantitative trait loci (QTLs) and 42 response-splicing QTLs, which are enriched in lower ACT GWAS [Formula: see text]-values, supporting the in vivo relevance of our map of genetic regulation of cellular response to anthracyclines.

Timing variability in circle drawing and tapping: probing the relationship between event and emergent timing.

R. Ivry, R. M. Spencer, H. N. Zelaznik, and J. Diedrichsen (2002) have proposed a distinction between timed movements in which a temporal representation is part of the task goal (event timing) and those in which timing properties are emergent. The issue addressed in the present experiment was how timing in conditions conducive to emergent timing becomes established. According to what the authors term the transformation hypothesis, timing initially requires an event-based representation when the temporal goal is defined externally (e.g., by a metronome), but over the first few movement cycles, control processes become established that allow timing to become emergent. Different groups of participants (N = 84) executed either 1 timed interval, 4 timed intervals, or 2 timed intervals separated by a pause. They produced the intervals by either circle drawing, a task associated with emergent timing, or tapping, a task associated with event timing. Analyses of movement variability suggested that similar timing processes were used in the 2 tasks only during the 1st interval. Those results are consistent with the transformation hypothesis and lead to the inference that the transition from event-based control to emergent timing can occur rapidly during continuous movements.

Substance abuse prevention: the role of the school nurse across the continuum of care.

As a health care provider, health educator, and school/family/community liaison, the school nurse is in a unique position to act as a change agent for youth substance abuse prevention. This article discusses the roles of the school nurse as they apply to the prevention of substance abuse among school-age children, across a continuum of care model first introduced by the Institute of Medicine (IOM) in 1994. Through careful assessment, identification of substance abuse risk factors, and promoting the enhancement of protective factors of students, both in and out of the school setting, the school nurse can play a vital role in the prevention of substance abuse. Existing tools, including the IOM Mental Health Intervention Spectrum Model, can be easily adapted to nursing practice and may prove helpful in assisting school nurses in the evaluation and implementation of effective prevention interventions in the school setting.

Whole-genome sequencing of individuals from a founder population identifies candidate genes for asthma.

Asthma is a complex genetic disease caused by a combination of genetic and environmental risk factors. We sought to test classes of genetic variants largely missed by genome-wide association studies (GWAS), including copy number variants (CNVs) and low-frequency variants, by performing whole-genome sequencing (WGS) on 16 individuals from asthma-enriched and asthma-depleted families. The samples were obtained from an extended 13-generation Hutterite pedigree with reduced genetic heterogeneity due to a small founding gene pool and reduced environmental heterogeneity as a result of a communal lifestyle. We sequenced each individual to an average depth of 13-fold, generated a comprehensive catalog of genetic variants, and tested the most severe mutations for association with asthma. We identified and validated 1960 CNVs, 19 nonsense or splice-site single nucleotide variants (SNVs), and 18 insertions or deletions that were out of frame. As follow-up, we performed targeted sequencing of 16 genes in 837 cases and 540 controls of Puerto Rican ancestry and found that controls carry a significantly higher burden of mutations in IL27RA (2.0% of controls; 0.23% of cases; nominal p = 0.004; Bonferroni p = 0.21). We also genotyped 593 CNVs in 1199 Hutterite individuals. We identified a nominally significant association (p = 0.03; Odds ratio (OR) = 3.13) between a 6 kbp deletion in an intron of NEDD4L and increased risk of asthma. We genotyped this deletion in an additional 4787 non-Hutterite individuals (nominal p = 0.056; OR = 1.69). NEDD4L is expressed in bronchial epithelial cells, and conditional knockout of this gene in the lung in mice leads to severe inflammation and mucus accumulation. Our study represents one of the early instances of applying WGS to complex disease with a large environmental component and demonstrates how WGS can identify risk variants, including CNVs and low-frequency variants, largely untested in GWAS.