G Protein-Coupled Receptor-Ligand Pose and Functional Class Prediction.

G protein-coupled receptor (GPCR) transmembrane protein family members play essential roles in physiology. Numerous pharmaceuticals target GPCRs, and many drug discovery programs utilize virtual screening (VS) against GPCR targets. Improvements in the accuracy of predicting new molecules that bind to and either activate or inhibit GPCR function would accelerate such drug discovery programs. This work addresses two significant research questions. First, do ligand interaction fingerprints provide a substantial advantage over automated methods of binding site selection for classical docking? Second, can the functional status of prospective screening candidates be predicted from ligand interaction fingerprints using a random forest classifier? Ligand interaction fingerprints were found to offer modest advantages in sampling accurate poses, but no substantial advantage in the final set of top-ranked poses after scoring, and, thus, were not used in the generation of the ligand-receptor complexes used to train and test the random forest classifier. A binary classifier which treated agonists, antagonists, and inverse agonists as active and all other ligands as inactive proved highly effective in ligand function prediction in an external test set of GPR31 and TAAR2 candidate ligands with a hit rate of 82.6% actual actives within the set of predicted actives.

Trust your gut: Establishing confidence in gastrointestinal models - An overview of the state of the science and contexts of use.

The webinar series and workshop titled Trust Your Gut: Establishing Confidence in Gastrointestinal Models - An Overview of the State of the Science and Contexts of Use was co-organized by NICEATM, NIEHS, FDA, EPA, CPSC, DoD, and the Johns Hopkins Center for Alternatives to Animal Testing (CAAT) and hosted at the National Institutes of Health in Bethesda, MD, USA on October 11-12, 2023. New approach methods (NAMs) for assessing issues of gastrointestinal tract (GIT)-related toxicity offer promise in addressing some of the limitations associated with animal-based assessments. GIT NAMs vary in complexity, from two-dimensional monolayer cell line-based systems to sophisticated 3-dimensional organoid systems derived from human primary cells. Despite advances in GIT NAMs, challenges remain in fully replicating the complex interactions and processes occurring within the human GIT. Presentations and discussions addressed regulatory needs, challenges, and innovations in incorporating NAMs into risk assessment frameworks; explored the state of the science in using NAMs for evaluating systemic toxicity, understanding absorption and pharmacokinetics, evaluating GIT toxicity, and assessing potential allergenicity; and discussed strengths, limitations, and data gaps of GIT NAMs as well as steps needed to establish confidence in these models for use in the regulatory setting.

Non-animal methods to assess whether chemicals may be toxic to the human digestive tract promise to complement or improve on animal-based methods. These approaches, which are based on human or animal cells and/or computer models, are faced with their own technical challenges and need to be shown to predict adverse effects in humans. Regulators are tasked with evaluating submitted data to best protect human health and the environment. A webinar series and workshop brought together scientists from academia, industry, military, and regulatory authorities from different countries to discuss how non-animal methods can be integrated into the risk assessment of drugs, food additives, dietary supplements, pesticides, and industrial chemicals for gastrointestinal toxicity.

Erratum: Improving the feasibility of economical proton-boron-11 fusion via alpha channeling with a hybrid fast and thermal proton scheme [Phys. Rev. E 106, 055215 (2022)].

This corrects the article DOI: 10.1103/PhysRevE.106.055215.

Pharmacokinetics and Safety of Firsocostat, an Acetyl-Coenzyme A Carboxylase Inhibitor, in Participants with Mild, Moderate, and Severe Hepatic Impairment.

Firsocostat is an oral, liver-targeted inhibitor of acetyl-coenzyme A carboxylase in development for the treatment of metabolic dysfunction-associated steatohepatitis. Hepatic organic anion transporting polypeptides play a significant role in the disposition of firsocostat with minimal contributions from uridine diphospho-glucuronosyltransferase and cytochrome P450 3A enzymes. This phase 1 study evaluated the pharmacokinetics and safety of firsocostat in participants with mild, moderate, or severe hepatic impairment. Participants with stable mild, moderate, or severe hepatic impairment (Child-Pugh A, B, or C, respectively [n = 10 per cohort]) and healthy matched controls with normal hepatic function (n = 10 per cohort) received a single oral dose of firsocostat (20 mg for mild and moderate hepatic impairment; 5 mg for severe hepatic impairment) with intensive pharmacokinetic sampling over 96 h. Safety was monitored throughout the study. Firsocostat plasma exposure (AUCinf) was 83%, 8.7-fold, and 30-fold higher in participants with mild, moderate, and severe hepatic impairment, respectively, relative to matched controls. Firsocostat was generally well tolerated, and all reported adverse events were mild in nature. Dose adjustment is not necessary for the administration of firsocostat in patients with mild hepatic impairment. However, based on the observed increases in firsocostat exposure, dose adjustment should be considered for patients with moderate or severe hepatic impairment, and additional safety and efficacy data from future clinical trials will further inform dose adjustment.

Digital self-aligned focusing schlieren.

In this Letter, a digital self-aligned focusing schlieren (D-SAFS) system is introduced. This system uses a digital transparent micro liquid crystal display (µLCD), in combination with a linear polarizer, to act on the linear polarization state of light transmitted in both the forward and reverse directions, essentially acting as both the source and cutoff grids. The use of the µLCD display allows for on-the-fly changes to the cutoff pattern type, spatial frequency, and orientation. This eliminates the need to physically access the source/cutoff grid in order to optimize the instrument's sensitivity, which is necessary with a conventional self-aligned focusing schlieren (SAFS) system.

Evaluation of the Effects of Meal Type and Acid-Reducing Agents on the Pharmacokinetics of Cilofexor, a Selective Nonsteroidal Farnesoid X Receptor Agonist.

Cilofexor is a nonsteroidal farnesoid X receptor agonist being developed in combination with firsocostat/semaglutide for the treatment of nonalcoholic steatohepatitis. This phase 1 study evaluated the effects of food and acid-reducing agents (ARAs) on the pharmacokinetics of cilofexor (100- or 30-mg fixed-dose combination with firsocostat) in healthy participants. Cohorts 1 (n = 20, 100 mg) and 2 (n = 30, 30 mg) followed a 3-period, 2-sequence crossover design and evaluated effects of light-fat and high-fat meals. Cohort 3 (n = 30, 100 mg fasting) followed a 2-period, 2-sequence crossover design and evaluated the effects of a 40-mg single dose of famotidine. Cohort 4 (n = 18, 100 mg) followed a 3-period, 2-sequence crossover design and evaluated the effects of a 40-mg once-daily regimen of omeprazole administered under fasting conditions or following a light-fat meal. Administration with light-fat or high-fat meals resulted in no change and an ∼35% reduction in cilofexor AUC, respectively, relative to the fasting conditions. Under fasting conditions, famotidine increased cilofexor AUC by 3.2-fold and Cmax by 6.1-fold, while omeprazole increased cilofexor AUC by 3.1-fold and Cmax by 4.8-fold. With a low-fat meal, omeprazole increased cilofexor exposure to a lesser extent (Cmax 2.5-fold, AUC 2.1-fold) than fasting conditions. This study suggests that caution should be exercised when cilofexor is administered with ARAs under fed conditions; coadministration of cilofexor (100 or 30 mg) with ARAs under fasting conditions is not recommended with the current clinical trial formulations.

Effect of an online educational module incorporating real-time feedback on accuracy of polyp sizing in trainees: a randomized controlled trial.

BACKGROUND: Although polyp size dictates surveillance intervals, endoscopists often estimate polyp size inaccurately. We hypothesized that an intervention providing didactic instruction and real-time feedback could significantly improve polyp size classification. METHODS: We conducted a multicenter randomized controlled trial to evaluate the impact of different components of an online educational module on polyp sizing. Participants were randomized to control (no video, no feedback), video only, feedback only, or video + feedback. The primary outcome was accuracy of polyp size classification into clinically relevant categories (diminutive [1-5mm], small [6-9mm], large [≥10mm]). Secondary outcomes included accuracy of exact polyp size (inmm), learning curves, and directionality of inaccuracy (over- vs. underestimation). RESULTS: 36 trainees from five training programs provided 1360 polyp size assessments. The feedback only (80.1%, P=0.01) and video + feedback (78.9%, P=0.02) groups had higher accuracy of polyp size classification compared with controls (71.6%). There was no significant difference in accuracy between the video only group (74.4%) and controls (P=0.42). Groups receiving feedback had higher accuracy of exact polyp size (inmm) and higher peak learning curves. Polyps were more likely to be overestimated than underestimated, and 29.3% of size inaccuracies impacted recommended surveillance intervals. CONCLUSIONS: Our online educational module significantly improved polyp size classification. Real-time feedback appeared to be a critical component in improving accuracy. This scalable and no-cost educational module could significantly decrease under- and overutilization of colonoscopy, improving patient outcomes while increasing colonoscopy access.

Peripheral immune function and Alzheimer's disease: a living systematic review and critical appraisal.

BACKGROUND: A growing body of literature examines the relationship between peripheral immune function and Alzheimer's Disease (AD) in human populations. Our living systematic review summarizes the characteristics and findings of these studies, appraises their quality, and formulates recommendations for future research. METHODS: We searched the electronic databases PubMed, PsycINFO, and Web of Science, and reviewed references of previous reviews and meta-analyses to identify human studies examining the relationship between any peripheral immune biomarkers and AD up to September 7th, 2023. We examined patterns of reported statistical associations (positive, negative, and null) between each biomarker and AD across studies. Evidence for each biomarker was categorized into four groups based on the proportion of studies reporting different associations: corroborating a positive association with AD, a negative association, a null association, and presenting contradictory findings. A modified Newcastle-Ottawa scale (NOS) was employed to assess the quality of the included studies. FINDINGS: In total, 286 studies were included in this review. The majority were cross-sectional (n = 245, 85.7%) and hospital-based (n = 248, 86.7%), examining relationships between 187 different peripheral immune biomarkers and AD. Cytokines were the most frequently studied group of peripheral immune biomarkers. Evidence supported a positive association with AD for six biomarkers, including IL-6, IL-1ß, IFN-γ, ACT, IL-18, and IL-12, and a negative association for two biomarkers, including lymphocytes and IL-6R. Only a small proportion of included studies (n = 22, 7.7%) were deemed to be of high quality based on quality assessment. INTERPRETATION: Existing research on peripheral immune function and AD exhibits substantial methodological variations and limitations, with a notable lack of longitudinal, population-based studies investigating a broad range of biomarkers with prospective AD outcomes. The extent and manner in which peripheral immune function can contribute to AD pathophysiology remain open questions. Given the biomarkers that we identified to be associated with AD, we posit that targeting peripheral immune dysregulation may present a promising intervention point to reduce the burden of AD.

Telenomuscristatus Johnson (Hymenoptera, Scelionidae): new diagnostic data, distribution records and host associations.

Specimens of an egg parasitoid wasp, Telenomuscristatus Johnson (Platygastroidea, Scelionidae), were reared from stink bug egg masses collected in the wild, in Maryland, United States. The egg masses were identified morphologically as Halyomorphahalys (Stål), Banasa Stål and Euschistus Dallas (Hemiptera, Pentatomidae). Molecular tools were used to further identify the Euschistus egg masses as E.servus (Say) and E.tristigmus (Say). All of these are new host associations for Te.cristatus. We also provide data to contribute to future identification of Te.cristatus: images of the holotype specimen and COI sequences from two disparate localities.

A revised terminology for male genitalia in Hymenoptera (Insecta), with a special emphasis on Ichneumonoidea.

Applying consistent terminology for morphological traits across different taxa is a highly pertinent task in the study of morphology and evolution. Different terminologies for the same traits can generate bias in phylogeny and prevent correct homology assessments. This situation is exacerbated in the male genitalia of Hymenoptera, and specifically in Ichneumonoidea, in which the terminology is not standardized and has not been fully aligned with the rest of Hymenoptera. In the current contribution, we review the terms used to describe the skeletal features of the male genitalia in Hymenoptera, and provide a list of authors associated with previously used terminology. We propose a unified terminology for the male genitalia that can be utilized across the order and a list of recommended terms. Further, we review and discuss the genital musculature for the superfamily Ichneumonoidea based on previous literature and novel observations and align the terms used for muscles across the literature.

A clock-dependent brake for rhythmic arousal in the dorsomedial hypothalamus.

Circadian clocks generate rhythms of arousal, but the underlying molecular and cellular mechanisms remain unclear. In Drosophila, the clock output molecule WIDE AWAKE (WAKE) labels rhythmic neural networks and cyclically regulates sleep and arousal. Here, we show, in a male mouse model, that mWAKE/ANKFN1 labels a subpopulation of dorsomedial hypothalamus (DMH) neurons involved in rhythmic arousal and acts in the DMH to reduce arousal at night. In vivo Ca2+ imaging reveals elevated DMHmWAKE activity during wakefulness and rapid eye movement (REM) sleep, while patch-clamp recordings show that DMHmWAKE neurons fire more frequently at night. Chemogenetic manipulations demonstrate that DMHmWAKE neurons are necessary and sufficient for arousal. Single-cell profiling coupled with optogenetic activation experiments suggest that GABAergic DMHmWAKE neurons promote arousal. Surprisingly, our data suggest that mWAKE acts as a clock-dependent brake on arousal during the night, when mice are normally active. mWAKE levels peak at night under clock control, and loss of mWAKE leads to hyperarousal and greater DMHmWAKE neuronal excitability specifically at night. These results suggest that the clock does not solely promote arousal during an animal's active period, but instead uses opposing processes to produce appropriate levels of arousal in a time-dependent manner.

Quantitative evaluation of released nanomaterials from carbon nanotube epoxy nanocomposites during environmental exposure and mechanical treatment.

Carbon nanotubes (CNTs) are promising nanomaterials exhibiting high thermal and electrical conductivities, significant stiffness, and high tensile strength. As a result, CNTs have been utilized as additives to enhance properties of various polymeric materials in a broad range of fields. In this study, we investigated the release of CNTs from CNT epoxy nanocomposites exposed to environmental weathering and mechanical stresses. The presence and amount of CNTs released from degraded polymer nanocomposites is important because CNTs can impact physiological systems in humans and environmental organisms. The weathering experiments in this study included nanocomposite exposure to both UV and a water spray, to simulate sunlight and rain exposure, whereas mechanical stresses were induced by shaking and ultrasonication. CNT release from epoxy nanocomposites was quantified by a 14C-labeling method that enabled measurement of the CNT release rates after different weathering and mechanical treatments. In this study, a sample oxidizer was used prior to liquid scintillation counting, because it was shown to reduce interferences from the presence of polymeric materials and achieve a high recovery (95%). Polymer nanocomposite degradation was confirmed by attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), scanning electron microscopy (SEM), and light microscopy. A continuous release of 14C-labeled nanomaterials was observed after each UV and simulated rain exposure period, with 0.23% (mass/mass) of the total embedded mass of CNTs being released from the CNT nanocomposite during the full weathering process, suggesting that the water spray induced sufficient mechanical stress to eliminate the protective effect of the surface agglomerated CNT network. Importantly, additional mechanical stresses imposed on the weathered nanocomposites by shaking and ultrasonication resulted in further release of approximately 0.27% (mass /mass).

Biophysical phenotype mixtures reveal advantages for tumor muscle invasion in vivo.

Bladder, colon, gastric, prostate, and uterine cancers originate in organs surrounded by laminin-coated smooth muscle. In human prostate cancer, tumors that are organ confined, without extracapsular extension through muscle, have an overall cancer survival rate of up to 97% compared with 32% for metastatic disease. Our previous work modeling extracapsular extension reported the blocking of tumor invasion by mutation of a laminin-binding integrin called α6ß1. Expression of the α6AA mutant resulted in a biophysical switch from cell-ECM (extracellular matrix) to cell-cell adhesion with drug sensitivity properties and an inability to invade muscle. Here we used different admixtures of α6AA and α6WT cells to test the cell heterogeneity requirements for muscle invasion. Time-lapse video microscopy revealed that tumor mixtures self-assembled into invasive networks in vitro, whereas α6AA cells assembled only as cohesive clusters. Invasion of α6AA cells into and through live muscle occurred using a 1:1 mixture of α6AA and α6WT cells. Electric cell-substrate impedance sensing measurements revealed that compared with α6AA cells, invasion-competent α6WT cells were 2.5-fold faster at closing a cell-ECM or cell-cell wound, respectively. Cell-ECM rebuilding kinetics show that an increased response occurred in mixtures since the response was eightfold greater compared with populations containing only one cell type. A synthetic cell adhesion cyclic peptide called MTI-101 completely blocked electric cell-substrate impedance sensing cell-ECM wound recovery that persisted in vitro up to 20 h after the wound. Treatment of tumor-bearing animals with 10 mg/kg MTI-101 weekly resulted in a fourfold decrease of muscle invasion by tumor and a decrease of the depth of invasion into muscle comparable to the α6AA cells. Taken together, these data suggest that mixed biophysical phenotypes of tumor cells within a population can provide functional advantages for tumor invasion into and through muscle that can be potentially inhibited by a synthetic cell adhesion molecule.

Capillary flow-driven immunoassay platform for COVID-19 antigen diagnostics.

Over the last few years, the SARS-CoV-2 pandemic has made the need for rapid, affordable diagnostics more compelling than ever. While traditional laboratory diagnostics like PCR and well-plate ELISA are sensitive and specific, they can be costly and take hours to complete. Diagnostic tests that can be used at the point-of-care or at home, like lateral flow assays (LFAs) are a simple, rapid alternative, but many commercially available LFAs have been criticized for their lack of sensitivity compared to laboratory methods like well-plate ELISAs. The Capillary-Driven Immunoassay (CaDI) device described in this work uses microfluidic channels and capillary action to passively automate the steps of a traditional well-plate ELISA for visual read out. This work builds on prior capillary-flow devices by further simplifying operation and use of colorimetric detection. Upon adding sample, an enzyme-conjugated secondary antibody, wash steps, and substrate are sequentially delivered to test and control lines on a nitrocellulose strip generating a colorimetric response. The end user can visually detect SARS-CoV-2 antigen in 15-20 min by naked eye, or results can be quantified using a smartphone and software such as ImageJ. An analytical detection limit of 83 PFU/mL for SARS-CoV-2 was determined for virus in buffer, and 222 PFU/mL for virus spiked into nasal swabs using image analysis, similar to the LODs determined by traditional well-plate ELISA. Additionally, a visual detection limit of 100 PFU/mL was determined in contrived nasal swab samples by polling 20 untrained end-users. While the CaDI device was used for detecting clinically relevant levels of SARS-CoV-2 in this study, the CaDI device can be easily adapted to other immunoassay applications by changing the reagents and antibodies.

Recent advances and current challenges in endoscopic resection with the full-thickness resection device.

Endoscopic full-thickness resection (EFTR) has emerged as a viable technique in the management of mucosal and subepithelial lesions of the gastrointestinal tract (GIT) not amenable to conventional therapeutic approaches. While various devices and techniques have been described for EFTR, a single, combined full-thickness resection and closure device (full-thickness resection device, FTRD system, Ovesco Endoscopy AG, Tuebingen, Germany) has become commercially available in recent years. Initially, the FTRD system was limited to use in the colorectum only. Recently, a modified version of the FTRD has been released for EFTR in the upper GIT as well. This review provides a broad summary of the FTRD, highlighting recent advances and current challenges.

Association of red blood cell distribution width with hospital admission and in-hospital mortality across all-cause adult emergency department visits.

Objectives: To test the association between the initial red blood cell distribution width (RDW) value in the emergency department (ED) and hospital admission and, among those admitted, in-hospital mortality. Materials and Methods: We perform a retrospective analysis of 210 930 adult ED visits with complete blood count results from March 2013 to February 2022. Primary outcomes were hospital admission and in-hospital mortality. Variables for each visit included demographics, comorbidities, vital signs, basic metabolic panel, complete blood count, and final diagnosis. The association of each outcome with the initial RDW value was calculated across 3 age groups (<45, 45-65, and >65) as well as across 374 diagnosis categories. Logistic regression (LR) and XGBoost models using all variables excluding final diagnoses were built to test whether RDW was a highly weighted and informative predictor for each outcome. Finally, simplified models using only age, sex, and vital signs were built to test whether RDW had additive predictive value. Results: Compared to that of discharged visits (mean [SD]: 13.8 [2.03]), RDW was significantly elevated in visits that resulted in admission (15.1 [2.72]) and, among admissions, those resulting in intensive care unit stay (15.3 [2.88]) and/or death (16.8 [3.25]). This relationship held across age groups as well as across various diagnosis categories. An RDW >16 achieved 90% specificity for hospital admission, while an RDW >18.5 achieved 90% specificity for in-hospital mortality. LR achieved a test area under the curve (AUC) of 0.77 (95% confidence interval [CI] 0.77-0.78) for hospital admission and 0.85 (95% CI 0.81-0.88) for in-hospital mortality, while XGBoost achieved a test AUC of 0.90 (95% CI 0.89-0.90) for hospital admission and 0.96 (95% CI 0.94-0.97) for in-hospital mortality. RDW had high scaled weights and information gain for both outcomes and had additive value in simplified models predicting hospital admission. Discussion: Elevated RDW, previously associated with mortality in myocardial infarction, pulmonary embolism, heart failure, sepsis, and COVID-19, is associated with hospital admission and in-hospital mortality across all-cause adult ED visits. Used alone, elevated RDW may be a specific, but not sensitive, test for both outcomes, with multivariate LR and XGBoost models showing significantly improved test characteristics. Conclusions: RDW, a component of the complete blood count panel routinely ordered as the initial workup for the undifferentiated patient, may be a generalizable biomarker for acuity in the ED.

U.S. Breastfeeding Outcomes at the Intersection: Differences in Duration Among Racial and Ethnic Groups With Varying Educational Attainment in a Nationally Representative Sample.

BACKGROUND: As breastfeeding rates in the United States increase, barriers persist for Black, Latine, and low-socioeconomic status household dyads when compared to White and high-socioeconomic status household dyads. Previous breastfeeding disparities research has almost exclusively considered the influence of race, ethnicity, and socioeconomic status separately, although these attributes are not randomly distributed across the population. RESEARCH AIM: To identify breastfeeding duration patterns by race/ethnicity and educational attainment in a nationally representative U.S. National Immunization Survey sample. METHOD: We conducted a cross-sectional, secondary analysis of the U.S. Centers for Disease Control and Prevention's 2020 National Immunization Survey-Child public-use data. To examine breastfeeding and exclusive breastfeeding durations at the intersection of race/ethnicity and educational attainment, we created a 12-item, cross-classified variable using three educational attainment groups and four race/ethnicity groups. We used linear regressions to test these associations. RESULTS: In all, 83% of the sample breastfed. Mean durations of breastfeeding were 7.5 (SE = 1.95) months and exclusive breastfeeding duration was 4.9 (SE = 0.87) months. In adjusted models, multi-race/other high-educational attainment participants had the longest breastfeeding duration by almost 3 weeks (ß: 19.53, 95% CI [5.27, 33.79]), and Black low-educational attainment participants exclusively breastfed for 1 month less than White high-educational attainment participants (ß:-30.23, 95% CI [-40.87, -19.58]). CONCLUSIONS: Examining race/ethnicity and educational attainment together provides an intersectional understanding of breastfeeding outcomes and can inform targeted, culturally appropriate interventions.

Thiamethoxam soil contaminations reduce fertility of soil-dwelling beetles, Aethina tumida.

There in increasing evidence for recent global insect declines. This is of major concern as insects play a critical role in ecosystem functionality and human food security. Even though environmental pollutants are known to reduce insect fertility, their potential effects on insect fitness remain poorly understood - especially for soil-dwelling species. Here, we show that fertility of soil-dwelling beetles, Aethina tumida, is reduced, on average, by half due to field-realistic neonicotinoid soil contaminations. In the laboratory, pupating beetles were exposed via soil to concentrations of the neonicotinoid thiamethoxam that reflect global pollution of agricultural and natural habitats. Emerged adult phenotypes and reproduction were measured, and even the lowest concentration reported from natural habitats reduced subsequent reproduction by 50%. The data are most likely a conservative estimate as the beetles were only exposed during pupation. Since the tested concentrations reflect ubiquitous soil pollution, the data reveal a plausible mechanism for ongoing insect declines. An immediate reduction in environmental pollutants is urgently required if our aim is to mitigate the prevailing loss of species biodiversity.

Releasing and tracking the distribution of adventive Trissolcus japonicus (Hymenoptera: Scelionidae) in Virginia.

Adventive populations of Trissolcus japonicus (Ashmead), an egg parasitoid of the invasive agricultural pest, brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), have been detected in the United States since 2014. Given its importance as an H. halys biocontrol agent, efforts to redistribute T. japonicus began within some US states. Our surveillance for T. japonicus in northwestern Virginia in 2016-2017 yielded annual detections only in 1 county. Thus, to promote its broader establishment, releases of H. halys egg masses parasitized by T. japonicus from Virginia occurred in 2018 (2 releases) and 2020 (1 release) at 9 sites throughout Virginia's tree fruit production regions. Monitoring of T. japonicus and H. halys, using yellow sticky cards deployed in H. halys host trees and pheromone-baited sticky traps, respectively, was conducted from 2018 to 2022. Annual captures of H. halys adults and nymphs appeared to reflect adequate populations to support T. japonicus establishment across most or all sites. Prerelease monitoring yielded a single T. japonicus at 1 site. By 2022, T. japonicus was detected at or near 7 of the remaining 8 release sites, with first detections varying between 1 and 2 yr from the releases in 2018 and 2020. Captures at most sites were very low, but establishment at several locations was indicated by detections in 2-4 seasons. In 2022, T. japonicus surveillance at 11 additional sites in northwestern Virginia yielded detections at all locations, including those at which it had not been detected in 2016-2017, providing evidence for its range expansion.

Identification of food and nutrient components as predictors of Lactobacillus colonization.

A previous double-blind, randomized clinical trial of 42 healthy individuals conducted with Lactobacillus johnsonii N6.2 found that the probiotic's mechanistic tryptophan pathway was significantly modified when the data was stratified based on the individuals' lactic acid bacteria (LAB) stool content. These results suggest that confounding factors such as dietary intake which impact stool LAB content may affect the response to the probiotic treatment. Using dietary intake, serum metabolite, and stool LAB colony forming unit (CFU) data from a previous clinical trial, the relationships between diet, metabolic response, and fecal LAB were assessed. The diets of subject groups with high vs. low CFUs of LAB/g of wet stool differed in their intakes of monounsaturated fatty acids, vegetables, proteins, and dairy. Individuals with high LAB consumed greater amounts of cheese, fermented meats, soy, nuts and seeds, alcoholic beverages, and oils whereas individuals with low LAB consumed higher amounts of tomatoes, starchy vegetables, and poultry. Several dietary variables correlated with LAB counts; positive correlations were determined for nuts and seeds, fish high in N-3 fatty acids, soy, and processed meats, and negative correlations to consumption of vegetables including tomatoes. Using machine learning, predictors of LAB count included cheese, nuts and seeds, fish high in N-3 fatty acids, and erucic acid. Erucic acid alone accurately predicted LAB categorization, and was shown to be utilized as a sole fatty acid source by several Lactobacillus species regardless of their mode of fermentation. Several metabolites were significantly upregulated in each group based on LAB titers, notably polypropylene glycol, caproic acid, pyrazine, and chondroitin sulfate; however, none were correlated with the dietary intake variables. These findings suggest that dietary variables may drive the presence of LAB in the human gastrointestinal tract and potentially impact response to probiotic interventions.