Endoplasmic reticulum morphology regulation by RTN4 modulates neuronal regeneration by curbing luminal transport.

Cell functions rely on intracellular transport systems distributing bioactive molecules with high spatiotemporal accuracy. The endoplasmic reticulum (ER) tubular network constitutes a system for delivering luminal solutes, including Ca2+, across the cell periphery. How the ER structure enables this nanofluidic transport system is unclear. Here, we show that ER membrane-localized reticulon 4 (RTN4/Nogo) is sufficient to impose neurite outgrowth inhibition in human cortical neurons while acting as an ER morphoregulator. Improving ER transport visualization methodologies combined with optogenetic Ca2+ dynamics imaging and in silico modeling, we observed that ER luminal transport is modulated by ER tubule narrowing and dilation, proportional to the amount of RTN4. Excess RTN4 limited ER luminal transport and Ca2+ release, while RTN4 elimination reversed the effects. The described morphoregulatory effect of RTN4 defines the capacity of the ER for peripheral Ca2+ delivery for physiological releases and thus may constitute a mechanism for controlling the (re)generation of neurites.

Dynamic stress- and inflammatory-based regulation of psychiatric risk loci in human neurons.

The prenatal environment can alter neurodevelopmental and clinical trajectories, markedly increasing risk for psychiatric disorders in childhood and adolescence. To understand if and how fetal exposures to stress and inflammation exacerbate manifestation of genetic risk for complex brain disorders, we report a large-scale context-dependent massively parallel reporter assay (MPRA) in human neurons designed to catalogue genotype x environment (GxE) interactions. Across 240 genome-wide association study (GWAS) loci linked to ten brain traits/disorders, the impact of hydrocortisone, interleukin 6, and interferon alpha on transcriptional activity is empirically evaluated in human induced pluripotent stem cell (hiPSC)-derived glutamatergic neurons. Of ~3,500 candidate regulatory risk elements (CREs), 11% of variants are active at baseline, whereas cue-specific CRE regulatory activity range from a high of 23% (hydrocortisone) to a low of 6% (IL-6). Cue-specific regulatory activity is driven, at least in part, by differences in transcription factor binding activity, the gene targets of which show unique enrichments for brain disorders as well as co-morbid metabolic and immune syndromes. The dynamic nature of genetic regulation informs the influence of environmental factors, reveals a mechanism underlying pleiotropy and variable penetrance, and identifies specific risk variants that confer greater disorder susceptibility after exposure to stress or inflammation. Understanding neurodevelopmental GxE interactions will inform mental health trajectories and uncover novel targets for therapeutic intervention.

Gene expression and splicing QTL analysis of blood cells in African American participants from the Jackson Heart Study.

Most gene expression and alternative splicing quantitative trait loci (eQTL/sQTL) studies have been biased toward European ancestry individuals. Here, we performed eQTL and sQTL analyses using TOPMed whole-genome sequencing-derived genotype data and RNA-sequencing data from stored peripheral blood mononuclear cells in 1,012 African American participants from the Jackson Heart Study (JHS). At a false discovery rate of 5%, we identified 17,630 unique eQTL credible sets covering 16,538 unique genes; and 24,525 unique sQTL credible sets covering 9,605 unique genes, with lead QTL at P < 5e-8. About 24% of independent eQTLs and independent sQTLs with a minor allele frequency > 1% in JHS were rare (minor allele frequency < 0.1%), and therefore unlikely to be detected, in European ancestry individuals. Finally, we created an open database, which is freely available online, allowing fast query and bulk download of our QTL results.

Quantification of nisin concentration from fluorescence-based antimicrobial activity assay using Bayesian calibration.

Bacteriocins are ribosomally synthesized peptides with the innate ability to kill or inhibit growth of other bacteria. In recent years, bacteriocins have received increased interest, as their antimicrobial activity enhances food safety and shelf life by combatting pathogens such as Listeria monocytogenes. They also have application potential as an active pharmaceutical compound to combat multidrug-resistant pathogens. As new bacteriocins continue to be discovered, accelerated workflows for screening, identification, and process development have been developed. However, antimicrobial activity measurement is often still limited with regards to quantification and throughput. Here, we present the use of a non-linear calibration model to infer nisin concentrations in cultivation supernatants of Lactococcus lactis ssp. lactis B1629 using readouts of pHluorin2 fluorescence-based antimicrobial activity assays.

Exploring the implementation of national health and social care standards in Ireland: A qualitative descriptive study.

BACKGROUND: National health and social care standards are complex, quality improvement interventions. Standards typically describe a process and/or outcome of safe, quality, person-centred care according to best evidence. Currently, there are 11 national standards that apply to diverse services in Ireland including residential centres, acute hospitals and rehabilitation and community inpatient healthcare services. A better understanding of contextual factors influencing implementation will inform decision-making when selecting implementation strategies to enhance the implementation of standards. AIM: To explore experiences of implementing national health and social care standards and secondly, to identify enablers and barriers to implementation with stakeholders from across multiple levels of the health system. DESIGN: A qualitative descriptive study. METHODS: We conducted six focus groups and eight individual interviews from October to November 2021 with stakeholders at system level (n = 14), organizational level (n = 14) and individual level (n = 10). Focus groups and interviews were audio-recorded, transcribed verbatim and analysed using reflexive thematic analysis. RESULTS: Six themes were generated; (1) Top-down, bottom-up, a team approach: everybody together, we are all involved, we are all responsible, (2) Support tools: accessible tools and bite-size material pertaining to standards will support us to implement standards, (3) Empower with knowledge: increase awareness and understanding of standards, make them relatable in practice so we can make sense of them, (4) A system-wide malaise: we do not have the bandwidth to implement standards, (5) Follow the leader: we need a lead person at every level to inspire implementation, (6) A bi-directional influence: we know inspections drive quality improvements but we still feel trepidation around inspection outcomes. CONCLUSION: Key enablers identified related to teamwork, support tools, leadership and inspections. Key barriers related to workforce issues, a lack of awareness of standards and fear of inspection outcomes. Our findings can be incorporated into strategies to support implementation of standards, ultimately for the benefit of service-users. IMPLICATIONS FOR PRACTICE: The enablers and barriers described in this study reflect the importance of organizational factors in the implementation of standards. Interdisciplinary teams can infer from these findings, which enablers and barriers apply to their own context. These findings can inform decision-making when selecting strategies that can be effective in supporting the implementation of standards. REPORTING METHOD: We have adhered to the Standards for Reporting Qualitative Research (SRQR) guidelines. PATIENT OR PUBLIC CONTRIBUTION: No patient or public contribution.

Quantitative outcomes of a type 2 single arm hybrid effectiveness implementation pilot study for hypertension-HIV integration in Botswana.

BACKGROUND: Successful HIV treatment programs have turned HIV into a chronic condition, but noncommunicable diseases such as hypertension jeopardize this progress. Hypertension control rates among people with HIV (PWH) are low owing to gaps in patient awareness, diagnosis, effective treatment, and management of both conditions at separate clinic visits. Integrated management, such as in our study, InterCARE, can enhance HIV-hypertension integration and blood pressure (BP) control. METHODS: Our pilot study was conducted in two Botswana HIV clinics between October 2021 and November 2022. Based on our formative work, we adopted three main strategies; Health worker training on HTN/cardiovascular disease (CVD) management, adaptation of HIV Electronic Health Record (EHR) for HTN/CVD care, and use of treatment partners to support PWH with hypertension for implementation. We employed the Reach, Effectiveness, Adoption, Implementation, Maintenance (RE-AIM) framework to assess implementation effectiveness and outcomes for BP control at baseline, 6 and 12 months. HIV viral load (VL) suppression was also measured to assess impact of integration on HIV care. RESULTS: We enrolled 290 participants; 35 (12.1%) were lost to follow-up, leaving 255 (87.9%) at 12-months. Median age was 54 years (IQR 46-62), and 77.2% were females. Our interventions significantly improved BP control to < 140/90 mmHg (or < 130/80 mmHg if diagnosis of diabetes or chronic kidney disease), from 137/290 participants, 47.2% at baseline to 206/290 participants, 71.0%, at 12 months (p < 0.001). Among targeted providers, 94.7% received training, with an associated significant increase in counseling on exercise, diet, and medication (all p < 0.001) but EHR use for BP medication prescribing and cardiovascular risk factor evaluation showed no adoption. In the intention-to-treat analysis, HIV VL suppression at 12 months decreased (85.5% vs 93.8%, p = 0.002) due to loss to follow-up but the per protocol analysis showed no difference in VL suppression between baseline and 12 months (97.3% vs 93.3%, p = 0.060). CONCLUSION: The InterCARE pilot study demonstrated that low-cost practical support measures involving the integration of HIV and hypertension/CVD management could lead to improvements in BP control. These results support the need for a large implementation and effectiveness trial. TRIAL REGISTRATION: ClinicalTrials.gov NCT05414526. Registered 18th May 2022.

Crimean-Congo hemorrhagic fever survivors elicit protective non-neutralizing antibodies that target 11 overlapping regions on glycoprotein GP38.

Crimean-Congo hemorrhagic fever virus can cause lethal disease in humans yet there are no approved medical countermeasures. Viral glycoprotein GP38, exclusive to Nairoviridae, is a target of protective antibodies and is a key antigen in preclinical vaccine candidates. Here, we isolate 188 GP38-specific antibodies from human survivors of infection. Competition experiments show that these antibodies bind across 5 distinct antigenic sites, encompassing 11 overlapping regions. Additionally, we show structures of GP38 bound with 9 of these antibodies targeting different antigenic sites. Although these GP38-specific antibodies are non-neutralizing, several display protective efficacy equal to or better than murine antibody 13G8 in two highly stringent rodent models of infection. Together, these data expand our understanding regarding this important viral protein and may inform the development of broadly effective CCHFV antibody therapeutics.

Longitudinal evidence over 2 years of the pandemic shows that poor mental health in people living with obesity may be underestimated.

It is well-documented that people living with obesity are at greater risk of poorer mental health outcomes. The aim of our study was twofold: First, to examine the longitudinal trajectories of depression and anxiety in people living with obesity over two years across eight waves of a UK national COVID-19 survey (March 2020-March 2022) using smoothing-splines mixed-effects models. Second, to investigate participation effects via a missingness analysis to check whether survey attrition over time was related to participant characteristics. Trajectory models showed that those living with overweight and obesity consistently reported significantly higher rates of anxiety and depression compared to those in normal weight categories over two years. Our missingness analysis revealed that depression and anxiety predicted the likelihood of responding to the survey over time, whereby those reporting higher rates of depression and anxiety were less likely to respond to the survey. Our findings add to the literature surrounding the (long-term) link between living with obesity and poor mental health. Notably, our results suggest that people who have poorer mental health were less likely to participate in the survey. Thus, we conclude that it is likely that longitudinal population survey studies potentially underreport mental health problems over time and therefore the realistic impact of obesity on mental health outcomes may be underestimated.

Role of Gubernaculum testis inervation during the process of testicular migration in human fetuses.

PURPOSE: The gubernaculum seems to be the most important anatomical structure in the testicular migration process. The objective of this paper is to review current literature regarding the role of gubernaculum testis nerves in testicular migration. We conducted a comprehensive literature review about the gubernaculum testis innervation. A PubMed database search was performed in April 2024, focusing on gubernaculum testis and cryptorchidism and genitofemoral nerve (GFN) and calcitonin gene-related peptide (CGRP) gene. The gubernaculum has its own nerve supply, the GFN, descending on the anteromedial surface of the psoas muscle from L1-L2 segments. The second phase of testicular descent is regulated by androgens and CGRP, released from the sensory nucleus of the GFN. The GFN doesn't directly play a role in testicular migration but there is a theory that shows a regulatory function of this nerve in hormonal action during this process. The gubernaculum testis has important structural alterations during the testicular migration and the genitofemoral nerve and CGRP gene are of great importance in this process. The genitofemoral nerve provides motor innervation to the cremaster muscle and gubernaculum, which helps regulate the position of the testes within the scrotum.

Homicides of American Indians/Alaska Natives in urban versus rural areas: United States National Violent Death Reporting System, 2003-2020.

BACKGROUND: Missing and Murdered Indigenous People is a historic and contemporary issue that has gained national attention. In 2021, homicide was the eighth leading cause of death among American Indian/Alaska Native (AIAN) persons aged between 1 and 54 years old, and homicide is the sixth leading cause of death among all AIAN males aged 1-54 years old. AIM: These data will build knowledge around AIAN homicides and to identify circumstances that can aid in comprehensive Missing and Murdered Indigenous People prevention efforts. METHODS: AIAN homicide data came from Centers for Disease Control and Prevention's National Violent Death Reporting System, a state/jurisdiction-based surveillance system that collects detailed information about characteristics and circumstances of violent deaths. We examined data from 2003 to 2020 (all available years) from participating states/jurisdictions. We also assessed sociodemographic characteristics of victims and suspects, incident characteristics and differences across dichotomised urban/rural status. The study was conducted in 2022. RESULTS: The National Violent Death Reporting System provided data on 2959 AIAN homicides from 2003 to 2020 (54.2% urban and 45.8% rural). Significant differences based on the two locations included type of weapon used, the location of the injury, race of the primary suspect, the victim's relationship to the suspect and select circumstances precipitating the homicide including crimes precipitating the homicide and homicides stemming from intimate partner violence. OUTCOMES: These findings provide crucial information to strengthen public health efforts for prevention.

Intermediate-sized follicular thyroid cancer surgical trends before and after the 2015 American thyroid association guideline changes.

In 2015, the ATA updated the guidelines to advocate for a lobectomy for tumors <1.0 â€‹cm and total thyroidectomy for tumors >4.0 â€‹cm. Treatment for tumors of intermediate size 1.0-4.0 â€‹cm is dependent on high-risk characteristics. There is limited research comparing the impact of the updated ATA guidelines on clinical practice on intermediate-sized tumors. In this study, the impact of the 2015 ATA guidelines on the surgical treatment of intermediated-sized FTC will be evaluated using the Surveillance, Epidemiology, and End Results (SEER) database. A total of 9983 patients were included; 7769 patients (74.1 â€‹%) were diagnosed pre-ATA guidelines and 2709 patients (25.9 â€‹%) post-ATA guidelines. The mean rate of lobectomy for intermediate-sized tumors was 22.1 â€‹% which increased to 33.4 â€‹% post-ATA updates. The results of the logistic regression showed the rate of lobectomy increased significantly in the post-ATA changes period (p â€‹< â€‹0.001). Future research could benefit from evaluating how these trends impact patient outcome measures.

Immunomodulatory soluble HLA-G and HLA-E are associated with rapidly deteriorating CLAD and HCMV-Viremia after Lung-Transplantation.

Plasma-soluble (s)HLA-G and sHLA-E are immunoregulatory proteins that balance the activation of NKG2A+ immune cells. In lung-transplant recipients (LTRs), dysregulated NKG2A+ natural killer cell responses may result in high-level HCMV replication as well as chronic lung allograft dysfunction (CLAD), and especially the development of rapidly deteriorating CLAD is associated with high mortality. We thus analyzed the kinetics and function of sHLA-G and sHLA-E in follow-up samples of N=76 LTRs to evaluate whether these immunoregulatory proteins are associated with the risk for CLAD and high-level HCMV replication. Here, we demonstrate that rapidly deteriorating CLAD LTRs are hallmarked by continually low (<107 ng/mL) sHLA-G levels. In contrast, high sHLA-E levels were associated with the following development of high-level (>1000 copies/mL) HCMV episodes. Thus, sHLA-G and sHLA-E may serve as novel biomarkers for the development of rapidly deteriorating CLAD and high-level HCMV replication in LTRs.

Cordyceps cateniannulata: An endophyte of coffee, a parasite of coffee leaf rust and a pathogen of coffee pests.

Here, we report on a Cordyceps species entering into a multi-trophic, multi-kingdom association. Cordyceps cateniannulata, isolated from the stem of wild Coffea arabica in Ethiopia, is shown to function as an endophyte, a mycoparasite and an entomopathogen. A detailed polyphasic taxonomic study, including a multilocus phylogenetic analysis, confirmed its identity. An emended description of C. cateniannulata is provided herein. Previously, this species was known as a pathogen of various insect hosts in both the Old and New World. The endophytic status of C. cateniannulata was confirmed by re-isolating it from inoculated coffee plants. Inoculation studies have further shown that C. cateniannulata is a mycoparasite of Hemileia vastatrix, as well as an entomopathogen of major coffee pests; infecting and killing Hypothenemus hampei and Leucoptera coffeella. This is the first record of C. cateniannulata from Africa, as well as an endophyte and a mycoparasite. The implications for its use as a biocontrol agent are discussed.

Metabolic Regulation in the Induction of Trained Immunity.

The innate immune system exhibits features of memory, termed trained immunity, which promote faster and more robust responsiveness to heterologous challenges. Innate immune memory is sustained through epigenetic modifications, affecting gene accessibility, and promoting a tailored gene transcription for an enhanced immune response. Alterations in the epigenetic landscape are intertwined with metabolic rewiring. Here, we review the metabolic pathways that underscore the induction and maintenance of trained immunity, including glycolysis, oxidative phosphorylation, the tricarboxylic acid cycle, and amino acid and lipid metabolism. The intricate interplay of these pathways is pivotal for establishing innate immune memory in distinct cellular compartments. We explore in particular the case of resident lung alveolar macrophages. We propose that leveraging the memory of the innate immune system may present therapeutic potential. Specifically, targeting the metabolic programs of innate immune cells is an emerging strategy for clinical interventions, either to boost immune responses in immunosuppressed conditions or to mitigate maladaptive activation in hyperinflammatory diseases.

Exploring the role of salon professionals in identifying sex trafficking and violence victims in Indiana.

BACKGROUND: Sex trafficking victims often have touchpoints with salons for waxing, styling, and other body modification services required by traffickers. Recently, some states have administered laws requiring salon professionals to receive intimate partner violence (IPV)-related training, with even fewer states mandating training on identifying sex trafficking. This study aimed to understand how salon professionals have witnessed evidence of violence, including IPV and sex trafficking, in the workplace and to explore the differences in their approach to each type of victim. METHODS: In-depth interviews were conducted with salon professionals (N = 10) and law enforcement professionals/policymakers (N = 5). Content and thematic analysis techniques were used. RESULTS: Salon professionals typically identified potential violence through signs such as bruises, odd behavior, and client disclosures, prompting them to engage in cautious conversations. Yet, few were trained to identify and intervene. Often, they responded to suspected violence by talking with the client, sharing concerns with salon leadership, directly intervening on the client's behalf, or contacting the police. Law enforcement and salon professionals had suggestions about improving salon professionals' recognition of and response to violence, including training on victim-focused resources, creating a safe environment, and building relationships with law enforcement. They also suggested strengthening community partnerships to increase resource advocacy and reporting. CONCLUSIONS: One-on-one salon services may provide a unique opportunity to intervene and identify victims of violence, especially when empowered through additional training and collaborative partnerships with community-oriented policing initiates. Implementing training and community-based initiatives could aid salon professionals in gaining greater confidence in knowing what to do when serving a client who is a victim of IPV or sex trafficking.

Determination of the Course of Cyan Fluorescence of Pseudomonas aeruginosa with a Handheld Bacterial Imaging Device.

Chronic wound infections are of clinical concern as they often lead to high rates of mortality and morbidity. A point-of-care handheld bacterial fluorescence imaging has been designed to detect the auto-fluorescent characteristics of most clinically relevant species of bacteria. This device causes most species of bacteria to exhibit red fluorescence due to the production of exoproduct porphyrins. One of the most significant contributors to the pathogenicity of chronic wounds is the pathogen Pseudomonas aeruginosa, and interestingly, this organism exhibits an additional unique cyan fluorescence signature. There is an over 90% positive predictive value that, when a chronic wound exhibits cyan fluorescence with the bacterial fluorescence imaging device, the wound will harbor P. aeruginosa. This project seeks to understand what genetic factor(s) contribute to the cyan phenotype observed.

Behavioral Responses of Imidacloprid-Dosed Farmland Birds to a Simulated Predation Risk.

Sublethal exposure to imidacloprid and other neonicotinoid insecticides may affect the neurological functions of birds. As such, behavior may be compromised. Here, we tested experimentally the effects of 1 and 6 mg/kg bw of imidacloprid on the antipredator behavioral responses of the red-legged partridge (Alectoris rufa) to simulated predator threats. Sixty-six partridges were challenged in groups or individually to intra- and interspecific alarm calls, to a raptor silhouette (aerial predation risk), and to a fox model (terrestrial predation risk). Antipredator behaviors were recorded as active (escape, active vigilance) and passive (passive vigilance, crouching, and freezing) responses. Latency in response to the stimuli, percentage of individuals who responded, response duration, speed of active responses, and vocalizations were measured. In experiments with partridges in the group, crouching against simulated predation risk lasted less time in birds treated with 6 mg a.i./kg bw than in control birds. In the experiments with individual partridges, passive vigilance against the intraspecific alarm lasted longer in birds treated with 6 mg a.i./kg bw than in control birds. The observed hyperreactivity to the predatory threat after a sublethal imidacloprid exposure can have consequences on survival under field conditions, where predation is a main driver of population dynamics.

Proximity driven plastid-nucleus relationships are facilitated by tandem plastid-ER dynamics.

Peri-nuclear clustering (PNC) of chloroplasts has largely been described in senescent and pathogen- or ROS- stressed cells. Stromules, tubular plastid extensions are also observed under similar conditions. Coincident observations of PNC and stromules associate the two phenomena in facilitating retrograde signaling between chloroplasts and the nucleus. However, PNC incidence in non-stressed cells under normal growth and developmental conditions, when stromules are usually not observed, remains unclear. Using transgenic Arabidopsis expressing different organelle-targeted fluorescent proteins we show that PNC is a dynamic subcellular phenomenon that continues in the absence of light and is not dependent on stromule formation. PNC is facilitated by tandem plastid-ER dynamics created through membrane contact sites between the two organelles. While PNC increases upon ER-membrane expansion, some plastids may remain in the peri-nuclear region due to their localization in ER-lined nuclear indentions. Moreover, some PNC plastids may sporadically extend stromules into ER-lined nuclear grooves. Our findings strongly suggest that PNC is not an exclusive response to stress caused by pathogens, high light or exogenous-H2O2 treatment and does not require stromule formation. However, morphological and behavioural alterations in ER and concomitant changes in tandem, plastid-ER dynamics play a major role in facilitating the phenomenon.

Fallopian Tube-Derived High-Grade Serous Cancers Influence Ovarian Production of Norepinephrine and Generate Specific Metabolomic Signatures.

High-grade serous ovarian cancer is the most common and lethal gynecologic malignancy, which is often attributed to the lack of available screenings, allowing the disease to progress unnoticed until it is diagnosed at more aggressive stages. As such, identifying signals in the tumor microenvironment involved in the primary metastasis of tumorigenic fallopian tube epithelial (FTE) cells to the ovary could provide new avenues for prevention, diagnostics, or therapeutic intervention. Since our previous work identified that the interaction of tumorigenic FTE and the ovary causes the release of norepinephrine (NE) from the ovary, we intended to determine the effects of ovarian NE on signaling and invasion of tumorigenic FTE models and high-grade serous ovarian cancer cell lines. We demonstrate that NE does not universally enhance migration, invasion, or adhesion by using multiple cell types but does alter specific oncogenic protein expression in certain models. In vivo, we found that blocking NE signaling via slow-release propranolol pellets significantly increased survival time in mice injected intraperitoneally with murine FTE cells engineered to stably express shRNA for PTEN and an activated KRAS expression construct. Finally, we identified that the metabolome released from the ovary is variable depending upon which cell type it is cocultured with, suggesting that distinct driver mutations in fallopian tube epithelial tumor models and early lesions can alter specific metabolomes within the surrounding ovarian microenvironment. These metabolomes provide the next frontier for evaluating local signals of the tumor microenvironment that facilitate ovarian spread of FTE lesions.