Communication trends over time in new-onset refractory status epilepticus (NORSE): Interim analysis from the NORSE/FIRES Family Registry.

The new-onset refractory status epilepticus (NORSE)/febrile infection-related epilepsy syndrome (FIRES) Family Registry contributes to a systematic effort to collect clinical and epidemiological information on individuals affected by NORSE/FIRES. We explore diagnostic and prognostic information provided to patients and their families, their satisfaction with the communication, and utilisation of palliative care services during acute hospitalization. Communication about the diagnosis of NORSE/FIRES to families has improved since the publication of consensus definitions in 2018, with families being more likely to be told about NORSE/FIRES after 2018. Families rate the quality of prognostic information as being moderate. Palliative care services were involved in a minority of patients. Understanding and characterizing the prevalence and satisfaction of diagnostic and prognostic conversations is important for improving overall care, the quality of physician-patient-family relationships, and the recovery process for those affected by NORSE/FIRES.

Lipid profiling identifies modifiable signatures of cardiometabolic risk in children and adolescents with obesity.

Pediatric obesity is a progressive, chronic disease that can lead to serious cardiometabolic complications. Here we investigated the peripheral lipidome in 958 children and adolescents with overweight or obesity and 373 with normal weight, in a cross-sectional study. We also implemented a family-based, personalized program to assess the effects of obesity management on 186 children and adolescents in a clinical setting. Using mass spectrometry-based lipidomics, we report an increase in ceramides, alongside a decrease in lysophospholipids and omega-3 fatty acids with obesity metabolism. Ceramides, phosphatidylethanolamines and phosphatidylinositols were associated with insulin resistance and cardiometabolic risk, whereas sphingomyelins showed inverse associations. Additionally, a panel of three lipids predicted hepatic steatosis as effectively as liver enzymes. Lipids partially mediated the association between obesity and cardiometabolic traits. The nonpharmacological management reduced levels of ceramides, phospholipids and triglycerides, indicating that lowering the degree of obesity could partially restore a healthy lipid profile in children and adolescents.

Probing the protonation and reduction of heptavalent neptunium with computational guidance.

Influence of pH on the speciation and stability of heptavalent neptunium is poorly understood although it is frequently invoked in the literature to explain experimental observations. The present study employs Density Functional Theory (DFT) methodology to assess the thermodynamic feasibility of protonation reactions for the Np(VII) anion complex and the impact on its reduction to Np(VI). This theoretical framework is then explored experimentally through the titration and systematic protonation of Np(VII) in solution and solid-state samples while monitoring them spectroscopically. Computational results reveal that protonation reactions with the axial OH- ligands of the Np(VII) anionic complex, [NpO4(OH)2]3-, are more thermodynamically favorable than the equatorial oxo ligands. In addition, DFT studies indicated that up to four sequential protonation reactions may be feasible before reduction becomes thermodynamically favorable. Experimental results also uncover that protonation leads to distinct changes in the observable vibrational signals and UV-Vis absorption features. Overall, we observed that the protonation of [NpO4(OH)2]3- in solution and in the solid-state occurs before reduction to the Np(VI)O22+ species.

Why Colloidal Syntheses of Bimetallic Nanoparticles Cannot be Generalized.

Introducing one general synthesis to form bimetallic nanoparticles (NPs) could accelerate the discovery of NPs for promising energy applications. Although colloidal syntheses can provide precise structural and morphological control of bimetallic NPs, the complex chemical nature of multicomponent syntheses challenges the realization of such synthetic simplicity. Common synthetic issues are frequently ascribed to the variation in metal ion precursor reactivities and complex chemical interactions between the different metal surfaces and capping agents employed. However, no systematic studies have shown how these factors compete to ultimately assign the factor limiting the mixing and formation of bimetallic NPs. Here, we provide a parametric investigation of how the intrinsic standard reduction potentials (E0red) of the metal ions and cocapping agents influence the formation of bimetallic AuCu, AuPd, and PdCu NPs. Using a combination of in situ X-ray total scattering along with transmission electron microscopy and nuclear magnetic resonance spectroscopy, we illustrate the multifunctional role of the cocapping agents through interactions with both the metal ion precursors and NP surfaces to stabilize metastable structures. Additionally, we demonstrate how system-specific side reactions and the local metal ion coordination environment can be used to selectively tune the formation kinetics, structure, and morphology of bimetallic NPs. Ultimately, these insights show that the chemical interactions rather than the intrinsic E0red are responsible for the formation of bimetallic NPs. Broadly, these insights should aid the synthetic design of tailored multimetallic NPs.

Med13 is required for efficient P-body recruitment and autophagic degradation of Edc3 following nitrogen starvation.

The Cdk8 kinase module (CKM), a conserved, detachable unit of the Mediator complex, plays a vital role in regulating transcription and communicating stress signals from the nucleus to other organelles. Here, we describe a new transcription-independent role for Med13, a CKM scaffold protein, following nitrogen starvation. In S. cerevisiae, nitrogen starvation triggers Med13 to translocate to the cytoplasm. This stress also induces the assembly of conserved membraneless condensates called processing bodies (P-bodies) that dynamically sequester translationally inactive messenger ribonucleoprotein particles (mRNPs). Cytosolic Med13 co-localizes with P-bodies, where it helps recruit Edc3, a highly conserved decapping activator and P-body assembly factor, into these conserved ribonucleoprotein granules. Moreover, Med13 orchestrates the autophagic degradation of Edc3 through a selective cargo-hitchhiking autophagy pathway that utilizes Ksp1 as its autophagic receptor protein. In contrast, the autophagic degradation of Xrn1, another conserved P-body assembly factor, is Med13 independent. These results place Med13 as a new player in P-body assembly and regulation following nitrogen starvation. They support a model in which Med13 acts as a conduit between P-bodies and phagophores, two condensates that use liquid-liquid phase separation in their assembly.

Evaluating video and hybrid group consultations in general practice: mixed-methods, participatory study protocol (TOGETHER 2).

Background: General practice is facing an unprecedented challenge in managing the consequences of the pandemic. In the midst of a policy drive to balance remote and in-person service provision, substantial workload pressures remain, together with increasing prevalence of long-term conditions, and declining staff numbers and morale. To address these challenges, some practices in the UK have been delivering video and hybrid group consultations (VHGCs) providing clinical care to multiple patients at the same time. Despite positive initial findings and enthusiasm, there are still gaps in our understanding of the influence VHGCs have on patient experience, healthcare utilisation, quality, safety, equity and affordability. Objectives: To generate an in-depth understanding of VHGCs for chronic conditions in general practice, surface assumptions and sociotechnical dynamics, inform practice and extend theorisation. Methods: Mixed-methods, multi-site research study using co-design and participatory methods, from qualitative, quantitative and cost-related perspectives. WP1 includes a national, cross-sectional survey on VHGC provision across the UK. In WP2 we will engage patients and general practice staff in co-design workshops to develop VHGC models with emphasis on digital inclusion and equity. In WP3 we will carry out a mixed-methods process evaluation in up to 10 GP practices across England (5 sites already running VHGCs and 5 comparison sites). Qualitative methods will include interviews, focus groups and ethnographic observation to examine the experiences of patients, carers, clinical and non-clinical NHS staff, commissioners and policy-makers. Quantitative methods will examine the impact of VHGCs on healthcare utilisation in primary and secondary care, patient satisfaction, engagement and activation. We will also assess value for money of group and individual care models from a health economics perspective. Conclusions: We aim to develop transferable learning on sociotechnical change in healthcare delivery, using VHGCs as an exemplar of technology-supported innovation. Findings will also inform the design of a future study.

Aim: To better understand how group consultations for chronic conditions can benefit patients and the health service when delivered via video and/or in-person in general practice. Background: Before the pandemic, group consultations were starting to gain ground in the UK as a new way of delivering clinical care to multiple patients at the same time, with potential benefits resulting from peer support and time efficiencies. When in-person care was restricted due to Covid-19, clinicians started delivering group consultations over video. Despite significant interest, we still know little about how group consultations delivered over video or hybrid models (combining video and in-person sessions) can be best implemented. Research methods: In up to 5 GP practices across England, where group consultations are already being delivered, we will evaluate how these new approaches to clinical care are implemented, and how they may support an inclusive service that engages patients with different needs and preferences. The evaluation will include interviews with patients, carers, NHS staff, policy-makers and commissioners, as well as group discussions and observations, including research led by patients themselves. We will also collect numerical data on the number and type of patients attending, whether they are more satisfied or confident with their self-management, or less likely to need to go to hospital. We will explore costs associated with these new ways of delivering care and will develop comparisons to face-to-face individual appointments. We will also work with comparable clinical sites only delivering one-to-one appointments, to collect numerical data on patient attendance, satisfaction and use of health services. With the involvement of our patient and public involvement (PPI) group, we will bring together our data to develop practical knowledge.

Disrupting the Master Narrative in Academic Biology as LGBTQ+ Ph.D. Students: Learning, Teaching, and Conducting Research.

LGBTQ+ individuals face discrimination and stigma in academic biology. These challenges are likely magnified for graduate students. However, there have been no studies documenting the experiences of LGBTQ+ life sciences graduate students. To address this gap, we conducted an interview study of 22 biology PhD students from 13 universities across the United States who identify as LGBTQ+. We used the master narrative framework to interpret our findings. Master narratives are guidelines that dictate the "expected" and "normal" way one is supposed to navigate life. We considered how graduate students engage with the societal master narrative that treats cisheterosexuality as the norm, as well as the master narrative that expects biology to remain an objective, apolitical space. We found that LGBTQ+ PhD students recognize the anti-LBGTQ+ narratives in academic biology, which can result in instances of discrimination and encourage them to conceal their identities. However, participants pushed back against these master narratives. Graduate students described creating alternative narratives by highlighting how their LGBTQ+ identity has allowed them to become more inclusive instructors and better researchers. Some also purposely reveal their LGBTQ+ identity in academic biology, violating the master narrative that non-science identities should not be discussed in the life sciences.

"I've been really happy since I got that letter!": Longitudinal patient perspectives on lung cancer screening communication.

OBJECTIVE: Experts recommend structured shared decision making when discussing lung cancer screening (LCS) and reporting low-dose computed tomography (LDCT) results. We examined patients' reactions to pre- and post-LDCT results communication processes at three medical centers in the US with established LCS programs. METHODS: Multicenter, qualitative, longitudinal study of patients considering and receiving LCS using data from semi-structured interviews guided by a patient-centered communication model using conventional content analysis. We conducted 61 interviews among 32 patients (sixteen of whom had a nodule on their LDCT) at one month and 12 months after an initial LCS decision making interaction. RESULTS: Participants were mostly satisfied with LCS communication processes pre- and post-LDCT even though guideline concordant shared decision making was rare. Most participants reported no more than mild distress even if the LDCT detected a pulmonary nodule, felt relief after getting the results, and reported the perceived benefits of LCS outweighed their distress. Nearly all participants were satisfied with recommended follow-up plans. They reported that they trusted their clinicians and health care system to provide appropriate care and recommendations. They did not appear to regret their decision since almost all participants planned to get their next LDCT. However, they were at risk of non-adherence to follow-up recommendations since they often relied on the health care system to ensure they received timely follow-up. CONCLUSIONS: Despite receiving guideline discordant decision-making communication, patients seem very satisfied, rarely experience severe distress, and have low decisional regret after LCS decision making and receiving the results of their LDCT.

Salmonella Prevalence and Quantification in Market Hog Lymph Nodes and Tonsils in Several Regions and Seasons of the United States.

Market hog lymph nodes (LNs) can contaminate carcasses with Salmonella, as well as ground and comminuted pork products. The objective of this study was to perform a qualitative and quantitative analysis of LNs from several regions and seasons in the United States to establish a Salmonella prevalence and concentration baseline. Six types of LNs (axillary, mesenteric, subiliac, tracheobronchial, superficial inguinal, prescapular), and tonsils were sampled from market hog carcasses from different regions (east, central, and west) and seasons (winter, spring, and summer/fall). Salmonella was detected and enumerated using BAX®-System-SalQuant® methods and the BAX®-System Real-Time Salmonella Assay. Salmonella prevalence (N = 4,132) was 36% for tonsils, 35% for mesenteric LN, and less than 10% for the other LN types. Of the 601 carcasses tested, 62% were positive for Salmonella, with the highest prevalence occurring during spring in the east (90.9%), and the lowest prevalence occurring during spring in the central region (26.0%). Tonsil prevalence was greatest in the eastern region during spring. Mesenteric LN prevalence was high (>20%) regardless of season or region. Salmonella prevalence in tracheobronchial, subiliac, axillary, and superficial inguinal LNs was generally greatest during the spring or fall and in the eastern region. The median SalQuant® Salmonella concentration was 2.18 log10Salmonella cells/sample. Median SalQuant® concentration for all other sample types fell below the limit of quantification (1 log10Salmonella cells/sample). This longitudinal study can be used by the pork industry for risk assessments and risk-based decision-making.

Induction chemotherapy with cemiplimab in a patient with coexistent vulvar cancer and autoimmune disease: A case report.

There is limited data regarding the use of immunotherapy for patients with vulvar squamous cell carcinoma and coexisting autoimmune disease. Cemiplimab is a PD-1 inhibitor approved for use in patients with locally advanced and metastatic cutaneous squamous cell carcinoma. However, little is known about its efficacy in the setting of vulvar cancer. We present a case of advanced vulvar squamous cell carcinoma treated with induction chemotherapy and immunotherapy with cemiplimab followed by definitive chemoradiation in the setting of multiple autoimmune diseases. She achieved a complete clinical response and experienced no worsening of her autoimmune conditions despite cessation of her immunosuppressants and initiating an immune checkpoint inhibitor. We review existing data on neoadjuvant treatment of vulvar cancer and the use of cemiplimab in genital and inguinal squamous cell carcinomas. Ongoing exploration of cemiplimab's efficacy in vulvar cancer and safety in immunosuppressed patients is critical.

Rethinking IPE duration: a five-year comparative analysis of competency development across two introductory IPE course models.

Best practices have not yet been established in the interprofessional education (IPE) literature to guide the ideal dose and duration of IPE experiences across the curriculum. As such, the content, structure, and delivery format of IPE offerings vary significantly across institutions. The University of Minnesota had the rare opportunity to evaluate learner-perceived collaborative competency outcomes due to the transition of its centrally offered introductory IPE course. Data were collected consistently, longitudinally, and for a high volume of learners and professions across two IPE course models applied within the same academic institution. Retrospective analysis of pre-post interprofessional collaborative competency scores collected for over 5,000 learners from 17 health profession degree programs demonstrated nearly identical self-reported competency attainment for both a 12- and 4-hour introductory IPE offering, assessed using the ICCAS tool over the course of 5-year format transition. If student-reported competency attainment is the same following an introductory IPE experience regardless of the dosage, then academic IPE programs may be better positioned by decreasing their introductory emphasis and instead focus their resources on exploring innovative workplace-based and competency-based IPE strategies in line with contemporary recommendations. Further studies are needed to explore the implications of and next steps in this line of research.

Helicobacter pylori seropositivity associates with hyperglycemia, but not obesity, in Danish children and adolescents.

BACKGROUND: Helicobacter pylori colonizes the human stomach and may affect the inflammatory response, hormone production related to energy regulation, and gastrointestinal microbiota composition. Previous studies have explored a potential association between H. pylori infection and pediatric obesity with varying results. Considering the immunomodulatory effects of early-life infection with H. pylori that can confer beneficial effects, we hypothesized that we would find an inverse relationship between H. pylori seropositivity and obesity among Danish children and adolescents. METHODS: We assessed H. pylori seroprevalence in 713 subjects from an obesity clinic cohort and 990 subjects from a population-based cohort, aged 6 to 19 years, and examined its association with obesity and other cardiometabolic risk factors. RESULTS: No association was found between H. pylori and body mass index standard deviation score (BMI SDS). H. pylori seropositivity was, however, significantly associated with higher fasting plasma glucose levels and the prevalence of hyperglycemia. CONCLUSION: While we did not find an association between H. pylori seropositivity and BMI SDS, we observed a significant association with higher fasting plasma glucose levels and increased prevalence of hyperglycemia, suggesting that H. pylori infection may contribute to impaired glucose regulation in Danish children and adolescents.

End-Tidal CO2 During Enhanced-View Totally Extraperitoneal Hernia Repair: A Comparison of Retrorectus and Intraperitoneal Insufflation.

INTRODUCTION: Although the enhanced-view totally extraperitoneal (eTEP) approach has demonstrated safety, efficacy, and durability for small- to medium-sized hernia repairs, the relationships between retrorectus insufflation, intraoperative respiratory stability, and end-tidal CO2 (ETCO2) levels has not been appraised. METHODS: We conducted a retrospective chart review of patients undergoing elective robotic-assisted ventral hernia repairs at our quaternary academic center from July 2018 through December 2021. Patients were grouped by repair technique, either eTEP or robotic transversus abdominis release (r-TAR). Baseline demographics, intraoperative anesthesia records, and perioperative outcomes were reviewed. Anesthesia data were collected at intubation and 30-min time intervals thereafter. Operative time, length of stay, patient-controlled anesthesia use, and perioperative complications were compared. RESULTS: In total, 205 patients underwent an eTEP repair and 97 patients underwent an r-TAR repair. Intraoperatively, eTEP repairs had significantly higher ETCO2 at the beginning of the case (times 1-4, P < 0.05), and a higher peak ETCO2 (P < 0.05) when compared to r-TAR repairs. This difference in ETCO2 desisted as the case progressed, with a subsequent increase in respiratory rate (times 2-6, P < 0.05) in the eTEP procedures. The eTEP group demonstrated significantly shorter operative times, decreased patient-controlled anesthesia use, and a shorter length of stay. There was no significant difference in postoperative intensive care unit admission or respiratory distress. CONCLUSIONS: This study demonstrates that retrorectus insufflation during eTEP hernia repairs correlated with higher levels of ETCO2 compared to r-TAR repairs yet was not associated with any meaningful difference in perioperative outcomes. Communication of these respiratory differences with anesthesia is needed for proper ventilation adjustments.

Intermittent ozone inhalation during house dust mite-induced sensitization primes for adverse asthma phenotype.

The ability of air pollution to induce acute exacerbation of asthma is well documented. However, the ability of ozone (O3), the most reactive gaseous component of air pollution, to function as a modulator during sensitization is not well established. C57BL/6 J male mice were intranasally sensitized to house dust mite (HDM) (40 µg/kg) for 3 weeks on alternate days in parallel with once-a-week O3 exposure (1 ppm). Mice were euthanized 24 h following the last HDM challenge. Lung lavage, histology, lung function (both forced oscillation and forced expiration-based), immune cell profiling, inflammation (pulmonary and systemic), and immunoglobulin production were assessed. Compared to HDM alone, HDM + O3 leads to a significant increase in peribronchial inflammation (p < 0.01), perivascular inflammation (p < 0.001) and methacholine-provoked large airway hyperreactivity (p < 0.05). Serum total IgG and IgE and HDM-specific IgG1 were 3-5 times greater in HDM + O3 co-exposure compared to PBS and O3-exposed groups. An increase in activated/mature lung total and monocyte-derived dendritic cells (p < 0.05) as well as T-activated, and T memory lymphocyte subset numbers (p < 0.05) were noted in the HDM + O3 group compared to HDM alone group. Concurrent O3 inhalation and HDM sensitization also caused significantly greater (p < 0.05) lung tissue interleukin-17 pathway gene expression and mediator levels in the serum. Redox imbalance was manifested by impaired lung antioxidant defense and increased oxidants. O3 inhalation during allergic sensitization coalesces in generating a significantly worse TH17 asthmatic phenotype.

Effect of Semaglutide on Physical Function, Body Composition, and Biomarkers of Aging in Older Adults With Overweight and Insulin Resistance: Protocol for an Open-Labeled Randomized Controlled Trial.

BACKGROUND: Older adults with type 2 diabetes mellitus (T2DM) or prediabetes are at increased risk of adverse changes in body composition, physical function, and aging-related biomarkers compared to those with normal glucose tolerance. Semaglutide is a glucagon-like peptide 1 receptor agonist that has been approved for T2DM and chronic weight management. Although semaglutide is effective for weight loss and T2DM management, its effects on lean body mass, physical function, and biomarkers of aging are understudied in older adults. OBJECTIVE: This study aims to compare the effects of lifestyle counseling with and that without semaglutide on body composition, physical function, and biomarkers of aging in older adults. METHODS: This is an open-label randomized controlled trial. A total of 20 adults (aged 65 years and older) with elevated BMI (27-40 kg/m2) and prediabetes or well-controlled T2DM (hemoglobin A1c 5.7%-7.5%) are recruited, stratified by sex, and randomized 1:1 to one of 2 groups (semaglutide plus lifestyle counseling vs lifestyle counseling alone) and followed up for 5 months. Those in the semaglutide group are titrated to 1 mg weekly, as tolerated, for 12 weeks. Lifestyle counseling is given by registered dietitians and based on the Diabetes Prevention Program Lifestyle Change Program. Our primary outcomes include changes in lean mass, physical function, and biomarkers of aging. Body composition is measured by dual-energy x-ray absorptiometry and includes total fat mass and lean mass. Physical function is measured by 6-minute walk distance, grip strength, and short physical performance battery. Biomarkers of aging are measured in blood, skeletal muscle, and abdominal adipose tissue to include C-reactive protein, interleukin-6, tumor necrosis factors α, and ß galactosidase staining. RESULTS: The study was funded in December 2021 with a projected data collection period from spring 2023 through summer 2024. CONCLUSIONS: Despite the elevated risk of adverse changes in body composition, physical function, and biomarkers of aging among older adults with glucose intolerance and elevated adiposity, the benefits and risks of commonly prescribed antihyperglycemic or weight loss medications such as semaglutide are understudied. This study aims to fill this knowledge gap to inform clinicians about the potential for additional clinically meaningful, nonglycemic effects of semaglutide. TRIAL REGISTRATION: ClinicalTrials.gov NCT05786521; https://clinicaltrials.gov/study/NCT05786521. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/62667.

Impacts of trace level chromium on formation of superoxide within uranyl triperoxide complexes.

U(VI) peroxides are important within the nuclear fuel cycle, but reactive oxygen species (ROS) can form upon oxidation. Herein, we identified the spectral signatures of a U(VI) diperoxosuperoxide complex (KUPS-1) and observed that the transformation of U(VI) triperoxide (KUT-1) to superoxide forms occurred with trace-level Cr. U(VI) superoxide complexes were identified in EPR solution spectra without the use of spin-traps.

miRNA-mediated control of gephyrin synthesis drives sustained inhibitory synaptic plasticity.

Activity-dependent protein synthesis is crucial for long-lasting forms of synaptic plasticity. However, our understanding of translational mechanisms controlling GABAergic synapses is limited. One distinct form of inhibitory long-term potentiation (iLTP) enhances postsynaptic clusters of GABAARs and the primary inhibitory scaffold, gephyrin, to promote sustained synaptic strengthening. While we previously found that persistent iLTP requires mRNA translation, the mechanisms controlling plasticity-induced gephyrin translation remain unknown. We identify miR153 as a novel regulator of Gphn mRNA translation which controls gephyrin protein levels and synaptic clustering, ultimately impacting inhibitory synaptic structure and function. iLTP induction downregulates miR153, reversing its translational suppression of Gphn mRNA and promoting de novo gephyrin protein synthesis and synaptic clustering during iLTP. Finally, we find that reduced miR153 expression during iLTP is driven by an excitation-transcription coupling pathway involving calcineurin, NFAT and HDACs, which also controls the miRNA-dependent upregulation of GABAARs. Together, we delineate a miRNA-dependent post-transcriptional mechanism that controls the expression of the key synaptic scaffold, gephyrin, and may converge with parallel miRNA pathways to coordinate gene upregulation to maintain inhibitory synaptic plasticity.

Age-Related Effects on Facial Emotion Recognition in Schoolchildren: An ERP Study.

The ability to recognize emotions in others is crucial for social interaction and develops during childhood. We studied the effects of age on emotional facial recognition in schoolchildren using ERP components. Children aged 6, 8, and 10 completed identity, sex, and emotion recognition (happiness, anger, sadness) tasks. The oldest group had the highest accuracy and fastest reaction times. Only the LPP component showed age-related differences, with lower amplitudes in older children. LPP showed higher amplitude during emotion recognition, which may be associated with motivational evaluation. ERP accounted for the temporal dynamics of facial processing, which involve cognitive and emotional processes.

Set Up and Utilization of a Three-Dimensional In Vitro Bioreactor System for Human Intestinal Studies and Microbial Co-Cultures.

The study of human intestinal physiology and host-microbe interactions is crucial for understanding gastrointestinal health and disease. Traditional two-dimensional cell culture models lack the complexity of the native intestinal environment, limiting their utility in studying intestinal biology. Here, we present a detailed protocol for the set up and utilization of a three-dimensional (3D) in vitro bioreactor system for human intestinal studies and bacterial co-culture. This article outlines the design and assembly of the bioreactor system, scaffold fabrication, bacterial culture techniques, analysis methods, and troubleshooting tips. By providing step-by-step instructions, the goal is to enable other laboratories to utilize physiologically relevant tissue models of the human intestine, incorporating key features, such as nutrient flow, multiple human cell types, 3D architecture, and microbial communities. The incorporation of commensal bacteria into the bioreactor system allows for the investigation of complex host-microbe interactions, providing insight into gastrointestinal health and pathology. This article serves as a comprehensive resource for scientists seeking to advance their understanding of intestinal biology toward the development of novel therapeutic strategies for gastrointestinal disorders. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Scaffold design Basic Protocol 2: Intestinal cell culture: Caco2 cells Basic Protocol 3: Intestinal cell culture: organoids Basic Protocol 4: Bioreactor design and set up Basic Protocol 5: Bacteria in 3D bioreactor set up Basic Protocol 6: Bacteria and drug dosing.

Associations of prostate tumor immune landscape with vigorous physical activity and prostate cancer progression.

BACKGROUND: Vigorous physical activity has been associated with lower risk of fatal prostate cancer. However, mechanisms contributing to this relationship are not understood. METHODS: We studied 117 men with prostate cancer in the University of North Carolina Cancer Survivorship Cohort (UNC CSC) who underwent radical prostatectomy, and 101 radiation-treated prostate cancer patients in FASTMAN. Structured questionnaires administered in UNC CSC assessed physical activity. In both studies, digital image analysis of H&E-stained tissues was applied to quantify Tumor Infiltrating Lymphocytes (TILs) in segmented regions. Nanostring gene expression profiling in UNC CSC and microarray in FASTMAN were performed on tumor tissue and a 50-gene signature utilized to predict immune cell types. RESULTS: Vigorous recreational activity, reported by 34 (29.1%) UNC men, was inversely associated with TILs abundance. Tumors of men reporting any vigorous activity versus none showed lower gene expression-predicted abundance of Th, exhausted CD4 T cells and macrophages. T cell subsets, including Treg, Th, Tfh, exhausted CD4 T cells, and macrophages were associated with increased risk of biochemical recurrence, only among men with ERG-positive tumors. CONCLUSIONS: Vigorous activity was associated with lower prostate tumor inflammation and immune microenvironment differences. Macrophages and T cell subsets, including those with immunosuppressive roles and those with lower abundance in men reporting vigorous exercise, were associated with worse outcomes in ERG-positive prostate cancer. IMPACT: Our novel findings contribute to our understanding of the role of the tumor immune microenvironment in prostate cancer progression, and may provide insight into how vigorous exercise could affect prostate tumor biology.