Muscle quality index comparisons between Hispanics and non-Hispanic Caucasians using dual energy X-ray absorptiometry and handgrip strength.

BACKGROUND & AIMS: Muscle quality index (MQI) can be computed in various ways. Also, many studies have evaluated MQI in older adults and non-Hispanic populations. The aim of this study was to compare various muscle quality indexes between Hispanics and non-Hispanic Caucasians when stratifying grip strength and appendicular lean mass measurements. METHODS: 235 participants (aged 25.5 ± 9.5 for males and 26.4 ± 9.9 for females) completed a dual energy X-ray absorptiometry (DXA) scan to assess appendicular lean mass (ALM). Handgrip strength (HGS) was assessed using a handheld dynamometer. MQI was computed using four different models: 1). MQIRA: ALM and HGS of right arm and hand, respectively; 2). MQILA: ALM and HGS of left arm and hand, respectively; 3). MQIARMS: ALM and HGS of both arms and hands, respectively; and 4). MQITOTAL: ALM of upper and lower-limbs and HGS of left and right hand. RESULTS: Hispanic males and females exhibited lower HGS compared to Caucasians with effect sizes ranging from trivial (d = 0.17) to moderate (d = 0.80). Females demonstrated higher MQI values compared to males for MQIARMS (d = 0.70), MQIRA (d = 0.75), and MQILA (d = 0.57). However, MQITOTAL yielded a small practical effect (d = 0.33) in favor of males (3.2 ± 0.5 kg/kg vs. 3.1 ± 0.5 kg/kg). After factoring by sex and ethnicity, Hispanic males and females, compared to non-Hispanic Caucasians males and females, showed trivial-to-small practical differences (d values ranging from 0.03 to 0.39). CONCLUSIONS: These results demonstrate MQI models vary across sex, particularly when utilizing models that account for upper extremity strength and ALM (i.e., MQIARMS, MQIRA, and MQILA). Lastly, to establish consistency in future research, the present study recommends using MQI models that account for ALM of upper- and lower-limbs (i.e., MQITOTAL). However, research measuring muscular strength via one upper-limb (e.g., left hand) might consider measuring ALM of the corresponding arm (e.g., left arm) when computing muscle quality (e.g., MQILA).

Long-term Outcomes of Regressed or "Burnt Out" Primary Testicular Germ Cell Tumors.

OBJECTIVE: To review the presentation and long-term oncologic outcomes of patients with regressed ("burnt out") primary testicular germ cell tumors (GCT). Certain testicular GCT can present with complete regression of the primary tumor. It is not well established if this is associated with more aggressive disease or worse oncologic outcomes. METHODS: We queried our prospectively maintained testicular cancer clinical database at a tertiary cancer center and identified patients without prior chemotherapy who had regressed primary GCT at radical orchiectomy from 1990 to 2023. All specimens were reviewed by a genitourinary pathologist at diagnosis. Long-term clinical outcomes were reported by Kaplan-Meier method. RESULTS: Fifty-six patients met inclusion criteria; at diagnosis, 17 had no evidence of extra-testicular disease and 39 had advanced (clinical stage [CS] II+) GCT. All CSx (no viable disease or germ cell neoplasia in situ at orchiectomy, and no evidence of advanced disease) and CS0 patients were managed with surveillance and had 5-year recurrence-free survival (RFS) of 88% (95% CI: 39%, 98%). All patients with CS II+ disease underwent primary treatment with surgery (n = 5) or first-line chemotherapy (n = 34). Two- and 5-year RFS for patients with CSII+ disease was 94% (95% CI: 78%, 98%) and 90% (95% CI: 72%, 97%), respectively. CONCLUSION: Patients with regressed primary testicular GCT often present with advanced disease, possibly due to lack of early clinical signs from the primary tumor. Our analysis shows excellent long-term oncologic outcomes similar to those reported in the literature for patients with viable primary testicular GCT.

Race- and Ethnicity-Related Differences in Heart Failure With Preserved Ejection Fraction Using Natural Language Processing.

Background: Heart failure with preserved ejection fraction (HFpEF) is the predominant form of HF in older adults. It represents a heterogenous clinical syndrome that is less well understood across different ethnicities. Objectives: This study aimed to compare the clinical presentation and assess the diagnostic performance of existing HFpEF diagnostic tools between ethnic groups. Methods: A validated Natural Language Processing (NLP) algorithm was applied to the electronic health records of a large London hospital to identify patients meeting the European Society of Cardiology criteria for a diagnosis of HFpEF. NLP extracted patient demographics (including self-reported ethnicity and socioeconomic status), comorbidities, investigation results (N-terminal pro-B-type natriuretic peptide, H2FPEF scores, and echocardiogram reports), and mortality. Analyses were stratified by ethnicity and adjusted for socioeconomic status. Results: Our cohort consisted of 1,261 (64%) White, 578 (29%) Black, and 134 (7%) Asian patients meeting the European Society of Cardiology HFpEF diagnostic criteria. Compared to White patients, Black patients were younger at diagnosis and more likely to have metabolic comorbidities (obesity, diabetes, and hypertension) but less likely to have atrial fibrillation (30% vs 13%; P < 0.001). Black patients had lower N-terminal pro-B-type natriuretic peptide levels and a lower frequency of H2FPEF scores ≥6, indicative of likely HFpEF (26% vs 44%; P < 0.0001). Conclusions: Leveraging an NLP-based artificial intelligence approach to quantify health inequities in HFpEF diagnosis, we discovered that established markers systematically underdiagnose HFpEF in Black patients, possibly due to differences in the underlying comorbidity patterns. Clinicians should be aware of these limitations and its implications for treatment and trial recruitment.

Liver Tissue Proteins Improve the Accuracy of Plasma Proteins as Biomarkers in Diagnosing Metabolic Dysfunction-Associated Steatohepatitis.

BACKGROUND: Biomarkers for metabolic dysfunction-associated steatohepatitis (MASH) have been considered based on proteomic and lipidomic data from plasma and liver tissue without clinical benefits. This study evaluated proteomics-based plasma and liver tissue biomarkers collected simultaneously from patients with metabolic dysfunction-associated steatotic liver disease (MASLD). METHODS: Liver tissue and plasma samples were collected during liver biopsy to diagnose MASLD. Untargeted proteomics was performed on 64 patients. RESULTS: Twenty plasma proteins were up- or downregulated in patients with MASH compared with those without MASH. The potential biomarkers utilizing the best combinations of these plasma proteins had an area under the receiver operating curve (AUROC) of 0.671 for detecting those with MASH compared with those without it. However, none of the 20 plasma proteins were represented among the significantly regulated liver tissue proteins in patients with MASH. Ten of them displayed a trend and relevance in liver tissue with MASLD progression. These 10 plasma proteins had an AUROC of 0.793 for MASH identification and higher positive and negative predictive values. CONCLUSION: The plasma and liver protein expressions of patients with MASH were not directly comparable. Plasma protein biomarkers that are also expressed in liver tissue can help improve MASH detection.

The Hurdle of Access to Emerging Therapies and Potential Solutions in the Management of Dyslipidemias.

This review explores the many barriers to accessing lipid-lowering therapies (LLTs) for the prevention and management of atherosclerotic cardiovascular disease (ASCVD). Geographical, knowledge, and regulatory barriers significantly impede access to LLTs, exacerbating disparities in healthcare infrastructure and affordability. We highlight the importance of policy reforms, including pricing regulations and reimbursement policies, for enhancing affordability and streamlining regulatory processes. Innovative funding models, such as value-based pricing and outcome-based payment arrangements, have been recommended to make novel LLTs more accessible. Public health interventions, including community-based programs and telemedicine, can be utilized to reach underserved populations and improve medication adherence. Education and advocacy initiatives led by patient advocacy groups and healthcare providers play a crucial role in raising awareness and empowering patients. Despite the barriers to access, novel LLTs present a big opportunity to reduce the burden of ASCVD, emphasizing the need for collaborative efforts among policymakers, healthcare providers, industry stakeholders, and patient advocacy groups to address these barriers to improve access to LLTs globally.

Paclitaxel, Ifosfamide, and Cisplatin as Initial Salvage Chemotherapy for Germ Cell Tumors: Long-Term Follow-Up and Outcomes for Favorable- and Unfavorable-Risk Disease.

PURPOSE: Paclitaxel, ifosfamide, and cisplatin (TIP) is an established salvage regimen for germ cell tumors (GCT) on the basis of a phase II trial, but efficacy on a large patient cohort including patients with unfavorable risk features and long-term outcomes has not been reported. Herein, we report updated treatment efficacy and long-term follow-up with TIP. PATIENTS AND METHODS: Patients with GCT who received TIP after cisplatin-based chemotherapy were eligible. Favorable response (complete response or partial response with negative tumor markers), overall survival (OS) and progression-free survival (PFS) rates, relapse, and toxicity were determined. Disease was reclassified according to the International Prognostic Factor Study Group (IPFSG) score. RESULTS: Of the 104 patients, 87 had favorable risk factors and 17 had at least one unfavorable factor by Memorial Sloan Kettering Cancer Center (MSKCC) criteria. Ten patients were treated for a second gonadal primary GCT. With a median follow-up of 8.9 years, the 5-year PFS and OS rates were 66% (95% CI, 55 to 74) and 69% (95% CI, 59 to 77), respectively. Among 87 patients with favorable-risk disease, 69 (79%) achieved a favorable response with 5-year PFS and OS rates of 67% (95% CI, 56 to 76) and 72% (95% CI, 61 to 80), respectively. Among 17 patients with MSKCC unfavorable-risk disease, 13 (76%) achieved a favorable response with 5-year PFS and OS rates of 59% (95% CI, 33 to 78) and 56% (95% CI, 28 to 76), respectively. After IPFSG reclassification, 5-year PFS and OS rates for patients with ≤intermediate-risk disease were 75% (95% CI, 50 to 89) and 73% (95% CI, 55 to 85), respectively. CONCLUSION: TIP is an effective second-line regimen for patients with GCT. Similar outcomes were observed in patients with favorable- and unfavorable-risk disease. The randomized TIGER trial (ClinicalTrials.gov identifier: NCT02375204) comparing TIP with high-dose chemotherapy will determine the optimal second-line treatment approach.

Combined Effect of Nigella sativa and Kefir on the Live Performance and Health of Broiler Chickens Affected by Necrotic Enteritis.

Coccidiosis and necrotic enteritis (NE) are prevalent poultry ailments worldwide, leading to decreased live performance and elevated mortality rates without antibiotic usage. This study evaluated Nigella sativa (black cumin) seeds (BCS) and kefir as alternatives to antibiotics for broilers. An in vivo study over a 28-day period, using 384 Cobb 500 male broilers organized into six treatment groups as part of a completely randomized block experimental design was conducted. Each treatment group included eight replicates, with each replicate containing eight birds. The treatments included positive control, negative control, antibiotic control, 5% BCS in feed, 20% kefir in drinking water, and a combination of 5% BCS and 20% kefir. NE was induced in broilers by administering ~5000 oocysts of Eimeria maxima orally on day 14, followed by inoculation with about 108 CFU/mL of Clostridium perfringens (Cp) (strain Cp#4) on days 19, 20, and 21. Live performance metrics including feed intake, body weight gain, and feed conversion were assessed in broilers. Additionally, NE disease outcomes such as lesion scores, mortality rates, and Cp populations in cecum were determined during the study. The BCS, kefir, and the combination had no detrimental effect on broiler live performance. BCS-treated and combination groups had lower NE scores (p > 0.05) in comparison to the positive control and exhibited no significant difference (p > 0.05) from antibiotic control. Additionally, treatment groups and antibiotic control were not significantly different (p > 0.05) in mortality, whereas the BCS and kefir combination significantly reduced (p < 0.05) mortality to 14.1% compared to 31.3% for the positive control. C. perfringens vegetative cells significantly decreased (p < 0.05) in treatments with BCS, kefir, and their combination on days 22 and 28 compared to the positive control. On day 22, Cp sores were significantly lower (p < 0.05) for the kefir and combination treatments compared to the positive control. In conclusion, BCS and kefir successfully reduced C. perfringens infection and mortality without any detrimental impact on broiler live performance with the combined treatment being the most effective. These results suggest that BCS and kefir could serve as potential alternatives to antibiotics in managing NE.

JAK inhibition enhances checkpoint blockade immunotherapy in patients with Hodgkin lymphoma.

Unleashing antitumor T cell activity by checkpoint inhibitor immunotherapy is effective in cancer patients, but clinical responses are limited. Cytokine signaling through the Janus kinase (JAK)-signal transducer and activator of transcription (STAT) pathway correlates with checkpoint immunotherapy resistance. We report a phase I clinical trial of the JAK inhibitor ruxolitinib with anti-PD-1 antibody nivolumab in Hodgkin lymphoma patients relapsed or refractory following checkpoint inhibitor immunotherapy. The combination yielded a best overall response rate of 53% (10/19). Ruxolitinib significantly reduced neutrophil-to-lymphocyte ratios and percentages of myeloid suppressor cells but increased numbers of cytokine-producing T cells. Ruxolitinib rescued the function of exhausted T cells and enhanced the efficacy of immune checkpoint blockade in preclinical solid tumor and lymphoma models. This synergy was characterized by a switch from suppressive to immunostimulatory myeloid cells, which enhanced T cell division.

Correction: Comparison of the broncoalveolar lavage fluid proteomics between foals and adult horses.

[This corrects the article DOI: 10.1371/journal.pone.0290778.].

Pool-Based Surfboard Elicits Activation of Posterior Shoulder Muscles During a Surfing Stroke.

ABSTRACT: Pexa, BS, Johnston, CD, Elder, EE, Ford, KR, Patterson, MQ, and Myers, JB. Pool-based surfboard elicits activation of posterior shoulder muscles during a surfing stroke. J Strength Cond Res 38(7): 1300-1304, 2024-Surfboard paddling may activate posterior shoulder muscles, which are critical to baseball pitchers' injury risk and performance. The purpose of this study was to measure posterior shoulder muscle activation during different phases of the surf stroke (propulsion vs. recovery) on a pool-based surfboard. Twenty healthy active adult subjects completed a familiarization and testing session with the pool-based surfboard. During the testing session, electromyography (EMG) sensors were placed on 6 posterior shoulder muscles: latissimus dorsi, infraspinatus, posterior deltoid, upper trapezius, middle trapezius, and lower trapezius. Subjects completed 4 laps in a pool at 3 separate resistances (low, moderate, and heavy) in a randomized order. The peak EMG signal during each phase (propulsion and recovery) was recorded. A 2-way within subject ANOVA (resistance-by-phase) with post hoc Bonferroni's corrections was used to identify differences in EMG activation. There was a significant main effect of phase for the latissimus dorsi (F = 91.3, p < 0.001), upper trapezius (F = 36.5, p < 0.001), middle trapezius (F = 33.8, p < 0.001), and lower trapezius (F = 21.6, p < 0.001). The latissimus dorsi demonstrated higher activation during the propulsion phase (p < 0.001), and all trapezius muscles demonstrated higher activation during the recovery phase (p < 0.001). There was a significant main effect of resistance for the posterior deltoid (F = 3.4, p = 0.043), with higher muscle activation in the low resistance trials compared with the heavy resistance trials (p = 0.036). Recreationally active individuals demonstrate activation of the posterior shoulder when using a pool-based surfboard. This pool-based surfboard may be beneficial to activate the posterior musculature and may be more accessible than standard surfing to baseball athletes.

The balance between antiviral and antibacterial responses during M. tuberculosis infection is regulated by the ubiquitin ligase CBL.

As a first line of host defense, macrophages must be able to effectively sense and respond to diverse types of pathogens, and while a particular type of immune response may be beneficial in some circumstances, it can be detrimental in others. Upon infecting a macrophage, M. tuberculosis (Mtb) induces proinflammatory cytokines that activate antibacterial responses. Surprisingly, Mtb also triggers antiviral responses that actually hinder the ability of macrophages to control Mtb infection. The ubiquitin ligase CBL suppresses these antiviral responses and shifts macrophages toward a more antibacterial state during Mtb infection, however, the mechanisms by which CBL regulates immune signaling are unknown. We found that CBL controls responses to multiple stimuli and broadly suppresses the expression of antiviral effector genes. We then used mass-spectrometry to investigate potential CBL substrates and identified over 46,000 ubiquitylated peptides in Mtb-infected macrophages, as well as roughly 400 peptides with CBL-dependent ubiquitylation. We then performed genetic interaction analysis of CBL and its putative substrates, and identified the Fas associated factor 2 (FAF2) adapter protein as a key signaling molecule protein downstream of CBL. Together, these analyses identify thousands of new ubiquitin-mediated signaling events during the innate immune response and reveal an important new regulatory hub in this response.

Probe-dependent Proximity Profiling (ProPPr) Uncovers Similarities and Differences in Phospho-Tau-Associated Proteomes Between Tauopathies.

Tauopathies represent a diverse group of neurodegenerative disorders characterized by the abnormal aggregation of the microtubule-associated protein tau. Despite extensive research, the precise mechanisms underlying the complexity of different types of tau pathology remain incompletely understood. Here we describe an approach for proteomic profiling of aggregate-associated proteomes on slides with formalin-fixed, paraffin-embedded (FFPE) tissue that utilizes proximity labelling upon high preservation of aggregate morphology, which permits the profiling of pathological aggregates regardless of their size. To comprehensively investigate the common and unique protein interactors associated with the variety of tau lesions present across different human tauopathies, Alzheimer's disease (AD), corticobasal degeneration (CBD), Pick's disease (PiD), and progressive supranuclear palsy (PSP), were selected to represent the major tauopathy diseases. Implementation of our widely applicable Probe-dependent Proximity Profiling (ProPPr) strategy, using the AT8 antibody, permitted identification and quantification of proteins associated with phospho-tau lesions in well-characterized human post-mortem tissue. The analysis revealed both common and disease-specific proteins associated with phospho-tau aggregates, highlighting potential targets for therapeutic intervention and biomarker development. Candidate validation through high-resolution co-immunofluorescence of distinct aggregates across disease and control cases, confirmed the association of retromer complex protein VPS35 with phospho-tau lesions across the studied tauopathies. Furthermore, we discovered disease-specific associations of proteins including ferritin light chain (FTL) and the neuropeptide precursor VGF within distinct pathological lesions. Notably, examination of FTL-positive microglia in CBD astrocytic plaques indicate a potential role for microglial involvement in the pathogenesis of these tau lesions. Our findings provide valuable insights into the proteomic landscape of tauopathies, shedding light on the molecular mechanisms underlying tau pathology. This first comprehensive characterization of tau-associated proteomes across different tauopathies enhances our understanding of disease heterogeneity and provides a resource for future functional investigation, as well as development of targeted therapies and diagnostic biomarkers.

Comparisons between dual-energy X-ray absorptiometry and bioimpedance devices for appendicular lean mass and muscle quality in Hispanic adults.

The purpose of this study was to compare single- and multi-frequency bioimpedance (BIA) devices against dual-energy X-ray absorptiometry (DXA) for appendicular lean mass (ALM) and muscle quality index (MQI) metrics in Hispanic adults. One hundred thirty-one Hispanic adults (18-55 years) participated in this study. ALM was measured with single-frequency bioimpedance analysis (SFBIA), multi-frequency bioimpedance analysis (MFBIA) and DXA. ALMTOTAL (left arm + right arm + left leg + right leg) and ALMARMS (left arm + right arm) were computed for all three devices. Handgrip strength (HGS) was measured using a dynamometer. The average HGS was used for all MQI models (highest left hand + highest right hand)/2. MQIARMS was defined as the ratio between HGS and ALMARMS. MQITOTAL was established as the ratio between HGS and ALMTOTAL. SFBIA and MFBIA had strong correlations with DXA for all ALM and MQI metrics (Lin's concordance correlation coefficient values ranged from 0·86 (MQIMFBIA-ARMS) to 0·97 (Arms LMSFBIA); all P < 0·001). Equivalence testing varied between methods (e.g. SFBIA v. DXA) when examining the different metrics (i.e. ALMTOTAL, ALMARMS, MQITOTAL and MQIARMS). MQIARMS was the only metric that did not differ from the line of identity and had no proportional bias when comparing all the devices against each other. The current study findings demonstrate good overall agreement between SFBIA, MFBIA and DXA for ALMTOTAL and ALMARMS in a Hispanic population. However, SFBIA and MFBIA have better agreement with DXA when used to compute MQIARMS than MQITOTAL.

ERG activates a stem-like proliferation-differentiation program in prostate epithelial cells with mixed basal-luminal identity.

To gain insight into how ERG translocations cause prostate cancer, we performed single cell transcriptional profiling of an autochthonous mouse model at an early stage of disease initiation. Despite broad expression of ERG in all prostate epithelial cells, proliferation was enriched in a small, stem-like population with mixed-luminal basal identity (called intermediate cells). Through a series of lineage tracing and primary prostate tissue transplantation experiments, we find that tumor initiating activity resides in a subpopulation of basal cells that co-express the luminal genes Tmprss2 and Nkx3.1 (called BasalLum) but not in the larger population of classical Krt8+ luminal cells. Upon ERG activation, BasalLum cells give rise to the highly proliferative intermediate state, which subsequently transitions to the larger population of Krt8+ luminal cells characteristic of ERG-positive human cancers. Furthermore, this proliferative population is characterized by an ERG-specific chromatin state enriched for NFkB, AP-1, STAT and NFAT binding, with implications for TF cooperativity. The fact that the proliferative potential of ERG is enriched in a small stem-like population implicates the chromatin context of these cells as a critical variable for unmasking its oncogenic activity.

Emergence of imaging technology beyond the clinical setting: Utilization of mobile health tools for at-home testing.

Body composition assessment plays a pivotal role in understanding health, disease risk, and treatment efficacy. This narrative review explores two primary aspects: imaging techniques, namely ultrasound (US) and dual-energy x-ray absorptiometry (DXA), and the emergence of artificial intelligence (AI) and mobile health apps in telehealth for body composition. Although US is valuable for assessing subcutaneous fat and muscle thickness, DXA accurately quantifies bone mineral content, fat mass, and lean mass. Despite their effectiveness, accessibility and cost remain barriers to widespread adoption. The integration of AI-powered image analysis may help explain tissue differentiation, whereas mobile health apps offer real-time metabolic monitoring and personalized feedback. New apps such as MeThreeSixty and Made Health and Fitness offer the advantages of clinic-based imaging techniques from the comfort of home. These innovations hold the potential for individualizing strategies and interventions, optimizing clinical outcomes, and empowering informed decision-making for both healthcare professionals and patients/clients. Navigating the intricacies of these emerging tools, critically assessing their validity and reliability, and ensuring inclusivity across diverse populations and conditions will be crucial in harnessing their full potential. By integrating advancements in body composition assessment, healthcare can move beyond the limitations of traditional methods and deliver truly personalized, data-driven care to optimize well-being.

Human Keratinocyte Responses to Woodsmoke Chemicals.

Air pollution consists of complex mixtures of chemicals with serious deleterious health effects from acute and chronic exposure. To help understand the mechanisms by which adverse effects occur, the present work examines the responses of cultured human epidermal keratinocytes to specific chemicals commonly found in woodsmoke. Our earlier findings with liquid smoke flavoring (aqueous extract of charred wood) revealed that such extracts stimulated the expression of genes associated with oxidative stress and proinflammatory response, activated the aryl hydrocarbon receptor, thereby inducing cytochrome P4501A1 activity, and induced cross-linked envelope formation, a lethal event ordinarily occurring during terminal differentiation. The present results showed that furfural produced transcriptional responses resembling those of liquid smoke, cyclohexanedione activated the aryl hydrocarbon receptor, and several chemicals induced envelope formation. Of these, syringol permeabilized the cells to the egress of lactate dehydrogenase at a concentration close to that yielding envelope formation, while furfural induced envelope formation without permeabilization detectable in this way. Furfural (but not syringol) stimulated the incorporation of amines into cell proteins in extracts in the absence of transglutaminase activity. Nevertheless, both chemicals substantially increased the amount of cellular protein incorporated into envelopes and greatly altered the envelope protein profile. Moreover, the proportion of keratin in the envelopes was dramatically increased. These findings are consistent with the chemically induced protein cross-linking in the cells. Elucidating mechanisms by which this phenomenon occurs may help understand how smoke chemicals interact with proteins to elicit cellular responses, interpret bioassays of complex pollutant mixtures, and suggest additional sensitive ways to monitor exposures.

Outer membrane vesicles and the outer membrane protein OmpU govern Vibrio cholerae biofilm matrix assembly.

Biofilms are matrix-encased microbial communities that increase the environmental fitness and infectivity of many human pathogens including Vibrio cholerae. Biofilm matrix assembly is essential for biofilm formation and function. Known components of the V. cholerae biofilm matrix are the polysaccharide Vibrio polysaccharide (VPS), matrix proteins RbmA, RbmC, Bap1, and extracellular DNA, but the majority of the protein composition is uncharacterized. This study comprehensively analyzed the biofilm matrix proteome and revealed the presence of outer membrane proteins (OMPs). Outer membrane vesicles (OMVs) were also present in the V. cholerae biofilm matrix and were associated with OMPs and many biofilm matrix proteins suggesting that they participate in biofilm matrix assembly. Consistent with this, OMVs had the capability to alter biofilm structural properties depending on their composition. OmpU was the most prevalent OMP in the matrix, and its absence altered biofilm architecture by increasing VPS production. Single-cell force spectroscopy revealed that proteins critical for biofilm formation, OmpU, the matrix proteins RbmA, RbmC, Bap1, and VPS contribute to cell-surface adhesion forces at differing efficiency, with VPS showing the highest efficiency whereas Bap1 showing the lowest efficiency. Our findings provide new insights into the molecular mechanisms underlying biofilm matrix assembly in V. cholerae, which may provide new opportunities to develop inhibitors that specifically alter biofilm matrix properties and, thus, affect either the environmental survival or pathogenesis of V. cholerae.IMPORTANCECholera remains a major public health concern. Vibrio cholerae, the causative agent of cholera, forms biofilms, which are critical for its transmission, infectivity, and environmental persistence. While we know that the V. cholerae biofilm matrix contains exopolysaccharide, matrix proteins, and extracellular DNA, we do not have a comprehensive understanding of the majority of biofilm matrix components. Here, we discover outer membrane vesicles (OMVs) within the biofilm matrix of V. cholerae. Proteomic analysis of the matrix and matrix-associated OMVs showed that OMVs carry key matrix proteins and Vibrio polysaccharide (VPS) to help build biofilms. We also characterize the role of the highly abundant outer membrane protein OmpU in biofilm formation and show that it impacts biofilm architecture in a VPS-dependent manner. Understanding V. cholerae biofilm formation is important for developing a better prevention and treatment strategy framework.

Extracellular vesicles from mouse trophoblast cells: Effects on neural progenitor cells and potential participants in the placenta-brain axis†.

The fetal brain of the mouse is thought to be dependent upon the placenta as a source of serotonin (5-hydroxytryptamine; 5-HT) and other factors. How factors reach the developing brain remains uncertain but are postulated here to be part of the cargo carried by placental extracellular vesicles (EV). We have analyzed the protein, catecholamine, and small RNA content of EV from mouse trophoblast stem cells (TSC) and TSC differentiated into parietal trophoblast giant cells (pTGC), potential primary purveyors of 5-HT. Current studies examined how exposure of mouse neural progenitor cells (NPC) to EV from either TSC or pTGC affect their transcriptome profiles. The EV from trophoblast cells contained relatively high amounts of 5-HT, as well as dopamine and norepinephrine, but there were no significant differences between EV derived from pTGC and from TSC. Content of miRNA and small nucleolar (sno)RNA, however, did differ according to EV source, and snoRNA were upregulated in EV from pTGC. The primary inferred targets of the microRNA (miRNA) from both pTGC and TSC were mRNA enriched in the fetal brain. NPC readily internalized EV, leading to changes in their transcriptome profiles. Transcripts regulated were mainly ones enriched in neural tissues. The transcripts in EV-treated NPC that demonstrated a likely complementarity with miRNA in EV were mainly up- rather than downregulated, with functions linked to neuronal processes. Our results are consistent with placenta-derived EV providing direct support for fetal brain development and being an integral part of the placenta-brain axis.

Assessing exogenous carbohydrate intake needed to optimize human endurance performance across sex: insights from modeling runners pursuing a sub-2-h marathon.

Carbohydrate (CHO) availability sustains high metabolic demands during prolonged exercise. The adequacy of current CHO intake recommendations, 30-90 g·h-1 dependent on CHO mixture and tolerability, to support elite marathon performance is unclear. We sought to scrutinize the current upper limit recommendation for exogenous CHO intake to support modeled sub-2-h marathon (S2M) attempts across elite male and female runners. Male and female runners (n = 120 each) were modeled from published literature with reference characteristics necessary to complete a S2M (e.g., body mass and running economy). Completion of a S2M was considered across a range of respiratory exchange rates, with maximal starting skeletal muscle and liver glycogen content predicted for elite male and female runners. Modeled exogenous CHO bioavailability needed for male and female runners were 93 ± 26 and 108 ± 22 g·h-1, respectively (P < 0.0001, d = 0.61). Without exogenous CHO, males were modeled to deplete glycogen in 84 ± 7 min, females in 71 ± 5 min (P < 0.0001, d = 2.21) despite higher estimated CHO oxidation rates in males (5.1 ± 0.5 g·h-1) than females (4.4 ± 0.5 g·h-1; P < 0.0001, d = 1.47). Exogenous CHO intakes ≤ 90 g·h-1 are insufficient for 65% of modeled runners attempting a S2M. Current recommendations to support marathon performance appear inadequate for elite marathon runners but may be more suitable for male runners in pursuit of a S2M (56 of 120) than female runners (28 of 120).NEW & NOTEWORTHY This study scrutinizes the upper limit of exogenous carbohydrate (CHO) recommendations for elite male and female marathoners by modeling sex-specific needs across an extreme metabolic challenge lasting ∼2 h, a sub-2-h marathon. Contemporary nutritional guidelines to optimize marathon performance appear inadequate for most elite marathon runners but appear more appropriate for males over their female counterparts. Future research examining possible benefits of exogenous CHO intakes > 90 g·h-1 should prioritize female athlete study inclusion.