Extracellular DNA traps in a ctenophore demonstrate immune cell behaviors in a non-bilaterian.

The formation of extracellular DNA traps (ETosis) is a first response mechanism by specific immune cells following exposure to microbes. Initially characterized in vertebrate neutrophils, cells capable of ETosis have been discovered recently in diverse non-vertebrate taxa. To assess the conservation of ETosis between evolutionarily distant non-vertebrate phyla, we observed and quantified ETosis using the model ctenophore Mnemiopsis leidyi and the oyster Crassostrea gigas. Here we report that ctenophores - thought to have diverged very early from the metazoan stem lineage - possess immune-like cells capable of phagocytosis and ETosis. We demonstrate that both Mnemiopsis and Crassostrea immune cells undergo ETosis after exposure to diverse microbes and chemical agents that stimulate ion flux. We thus propose that ETosis is an evolutionarily conserved metazoan defense against pathogens.

Educational quality of Robotic Whipple videos on YouTube.

BACKGROUND: Videos on Robotic pancreaticoduodenectomy (RPD) may be watched by surgeons learning RPD. This study sought to appraise the educational quality of RPD videos on YouTube. METHODS: One-hundred videos showing RPD or 'Robotic Whipple' were assessed using validated scales (LAP-VEGaS & Consensus Statement Score (CSS)). The association between the scores and the video characteristics (e.g. order of appearance, provider type etc) was assessed. The minimum number of videos required to cumulatively cover the entire LAP-VEGaS and CSS was also noted. RESULTS: The videos were of variable quality; median LAP-VEGaS = 0.67 (0.17-0.94), median CSS = 0.45 (0.29-0.53). There was no association between the educational quality of the videos and their order of appearance, view counts, provider type, length or country of origin. Videos lacked information such as patient consent (100%), potential pitfalls (97%) or surgeon credentials (84%). The first 29 videos cumulatively met all the criteria of CSS and LAP-VEGaS scores except for reporting consent. CONCLUSION: YouTube videos on RPD are of variable quality, without any recognised predictors of quality, and miss important safety information. An impractical number of videos need to be watched to cumulatively fulfil educational criteria. There is a need for high-quality, peer-reviewed videos that adhere to educational principles.

Dysfunctional Bladder Morphology and Functional Impairments Are Identified in the Alzheimer's Disease APPNL-G-F/NL-G-F Murine Model.

BACKGROUND: While symptoms related to lower urinary tract dysfunction (LUTD) are common in individuals with Alzheimer's disease (AD), pathophysiological links between AD and LUTD remain unclear. OBJECTIVE: This study aimed to investigate whether AD neuropathology would cause autonomic dysfunction along the spinal cord-bladder axis, which could result in alterations in bladder muscle kinetics. METHODS: We utilized APPNL-G-F/NL-G-F knock-in (APP KI) and APPwt/wt (wild-type) mice at two different ages, 4- and 10-month-old, to investigate how AD impacts bladder tissue function by immunohistochemistry, western blotting, and pharmacomyography. RESULTS: We showed that the mucosal layer partially separated from the detrusor in 10-month-old APP KI mouse bladders. Although there was no detectable amyloid deposition in the APP KI bladder, we found amyloid plaques in APP KI lumbar spinal cord. Further immunoblot analysis revealed that tyrosine hydroxylase protein levels were significantly reduced in both 4- and 10-month-old bladder tissues, suggesting reduction of norepinephrine synthesis in APP KI mouse bladders. In contrast, the level of ß2 adrenergic receptor was increased in 4-month-old but not 10-month-old APP KI bladders. In bladder strips, the adrenergic agonist isoproterenol induced increased relaxation in 4- but not 10-month-old APP KI bladders. With 10 Hz electrical field stimulation, 10-month-old APP KI bladder strips were more responsive than wild-type controls, with no differences observed in 4-month-old APP KI bladders. CONCLUSIONS: APP KI mice exhibit LUTD, which is likely arising from amyloid pathology in the spinal cord, and results in maturational declines in presynaptic activity combined with compensatory postsynaptic upregulation.

Ovine Forestomach Matrix in the Surgical Management of Complex Volumetric Soft Tissue Defects: A Retrospective Pilot Case Series.

Background: Volumetric soft tissue loss is an urgent surgical issue and can frequently lead to suboptimal outcomes for patients due to significant soft tissue loss, compromised vital structures, and contamination. Ovine forestomach matrix (OFM) has demonstrated clinical success in the surgical management of soft tissue defects, especially in contaminated fields, and provides an effective option for immediate coverage of exposed vital structures before definitive closure. Methods: This retrospective pilot case series (n = 13 defects) evaluated the clinical effectiveness of OFM (graft and/or particulate formats) in the surgical management of contaminated volumetric soft tissue defects. Patients presented with significant soft tissue loss, often with exposed viscera, tendon, bone, or muscle, and were treated with OFM as part of their inpatient surgical management. All patients had at least 1 significant comorbidity with the potential to complicate their healing trajectory. The primary study endpoint was time to 100% granulation tissue coverage (days), and the secondary endpoint was any device-related postoperative complications. Results: A total of 13 volumetric soft tissue defects were evaluated in 10 patients who underwent surgical reconstruction. Mean defect age was 3.5 ± 5.6 weeks, and mean area was 217.3 ± 77.9 cm2. Most defects had exposed structures (85%), and all defects were Centers for Disease Control and Prevention grade 2 or higher. Mean time to 100% granulation tissue formation was 23.4 ± 9.2 days, with a median product application of 1.0. Staged reconstruction was used in 7 of 13 defects, with the remainder (6 of 13) left to heal via secondary intention using standard wound care protocols. There were no major postoperative infections or adverse events (mean follow-up, 7.4 ± 2.4 weeks.). Conclusions: This retrospective pilot case series builds on a growing body of evidence that OFM can be utilized to facilitate the formation of functional, well-vascularized soft tissue in large contaminated volumetric soft tissue defects.

Temporal origin of nestedness in interaction networks.

Nestedness is a common property of communication, finance, trade, and ecological networks. In networks with high levels of nestedness, the link positions of low-degree nodes (those with few links) form nested subsets of the link positions of high-degree nodes (those with many links), leading to matrix representations with characteristic upper triangular or staircase patterns. Recent theoretical work has connected nestedness to the functionality of complex systems and has suggested that it is a structural by-product of the skewed degree distributions often seen in empirical data. However, mechanisms for generating nestedness remain poorly understood, limiting the connections that can be made between system processes and observed network structures. Here, we show that a simple probabilistic model based on phenology-the timing of copresences among interaction partners-can produce nested structures and correctly predict around two-thirds of interactions in two fish market networks and around one-third of interactions in 22 plant-pollinator networks. Notably, the links most readily explained by frequent actor copresences appear to form a backbone of nested interactions, with the remaining interactions attributable to opportunistic interactions or preferences for particular interaction partners that are not routinely available.

Robot-assisted general surgery in Aotearoa New Zealand.

Robot-assisted surgery refers to a surgeon controlling a robotic device that performs an operation. This viewpoint explores the current state of robot-assisted surgery in Aotearoa New Zealand using the da Vinci Surgical System (Intuitive Surgical, Sunnyvale, California, United States), the only currently available robotic surgical system for general surgery in the country. We describe the contemporary progress in Aotearoa New Zealand compared to Australia and globally, and present emerging high-level evidence from randomised controlled trials regarding the utility of the robot-assisted approach for general surgery procedures. From the available evidence, we suggest that the value of robot-assisted general surgery in the public healthcare system arises from its emerging clinical benefits for complex procedures and its potential to engender equitable access and outcomes, particularly for Maori and Pacific peoples, improve education and training and contribute towards quality assurance and workforce development. Therefore, its implementation aligns with the New Zealand Health Strategy's long-term goals and priority areas to achieve pae ora, healthy futures for all.

Symptoms, viral loads, and rebound among COVID-19 outpatients treated with nirmatrelvir/ritonavir compared to propensity score matched untreated individuals.

BACKGROUND: Nirmatrelvir/ritonavir (N/R) reduces severe outcomes among patients with COVID-19; however, rebound after treatment has been reported. We compared symptom and viral dynamics in community-based individuals with COVID-19 who completed N/R and similar untreated individuals. METHODS: We identified symptomatic participants who tested SARS-CoV-2 positive and were N/R eligible from a COVID-19 household transmission study: index cases from ambulatory settings and their households were enrolled, collecting daily symptoms, medication use, and respiratory specimens for quantitative PCR for 10 days, March 2022-May 2023. Participants who completed N/R (treated) were propensity score matched to untreated participants. We compared symptom rebound, viral load (VL) rebound, average daily symptoms, and average daily VL by treatment status measured after N/R completion or, if untreated, seven days after symptom onset. RESULTS: Treated (n=130) and untreated participants (n=241) had similar baseline characteristics. After treatment completion, treated participants had greater occurrence of symptom rebound (32% vs 20%; p=0.009) and VL rebound (27% vs 7%; p<0.001). Average daily symptoms were lower among treated participants compared to untreated participants without symptom rebound (1.0 vs 1.6; p<0.01), but not statistically lower with symptom rebound (3.0 vs 3.4; p=0.5). Treated participants had lower average daily VLs without VL rebound (0.9 vs 2.6; p<0.01), but not statistically lower with VL rebound (4.8 vs 5.1; p=0.7). CONCLUSIONS: Individuals who completed N/R experienced fewer symptoms and lower VL but were more likely to have rebound compared to untreated individuals. Providers should still prescribe N/R, when indicated, and communicate possible increased rebound risk to patients.

A rare anaerobic cause of vertebral osteomyelitis and psoas abscess: A case study.

Parvimonas micra is an obligate anaerobe that forms part of the normal gastrointestinal flora. The advent of matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF) and 16s ribosomal RNA gene sequencing has led to increased detection of many rare anaerobic isolates, including Parvimonas micra. Typical risk factors for Parvimonas micra bacteremia include dental procedures or spinal instrumentation. Here, we report a case of Parvimonas micra spondylodiscitis and psoas abscess in a patient with no obvious antecedent risk factors and explore the challenges in isolation of the organism from tissue samples.

Prefrontal Cortex Codes Representations of Target Identity and Feature Uncertainty.

Many objects in the real world have features that vary over time, creating uncertainty in how they will look in the future. This uncertainty makes statistical knowledge about the likelihood of features critical to attention demanding processes such as visual search. However, little is known about how the uncertainty of visual features is integrated into predictions about search targets in the brain. In the current study, we test the idea that regions prefrontal cortex code statistical knowledge about search targets before the onset of search. Across 20 human participants (13 female; 7 male), we observe target identity in the multivariate pattern and uncertainty in the overall activation of dorsolateral prefrontal cortex (DLPFC) and inferior frontal junction (IFJ) in advance of the search display. This indicates that the target identity (mean) and uncertainty (variance) of the target distribution are coded independently within the same regions. Furthermore, once the search display appears the univariate IFJ signal scaled with the distance of the actual target from the expected mean, but more so when expected variability was low. These results inform neural theories of attention by showing how the prefrontal cortex represents both the identity and expected variability of features in service of top-down attentional control.SIGNIFICANCE STATEMENT Theories of attention and working memory posit that when we engage in complex cognitive tasks our performance is determined by how precisely we remember task-relevant information. However, in the real world the properties of objects change over time, creating uncertainty about many aspects of the task. There is currently a gap in our understanding of how neural systems represent this uncertainty and combine it with target identity information in anticipation of attention demanding cognitive tasks. In this study, we show that the prefrontal cortex represents identity and uncertainty as unique codes before task onset. These results advance theories of attention by showing that the prefrontal cortex codes both target identity and uncertainty to implement top-down attentional control.

Global determinants of insect mitochondrial genetic diversity.

Understanding global patterns of genetic diversity is essential for describing, monitoring, and preserving life on Earth. To date, efforts to map macrogenetic patterns have been restricted to vertebrates, which comprise only a small fraction of Earth's biodiversity. Here, we construct a global map of predicted insect mitochondrial genetic diversity from cytochrome c oxidase subunit 1 sequences, derived from open data. We calculate the mitochondrial genetic diversity mean and genetic diversity evenness of insect assemblages across the globe, identify their environmental correlates, and make predictions of mitochondrial genetic diversity levels in unsampled areas based on environmental data. Using a large single-locus genetic dataset of over 2 million globally distributed and georeferenced mtDNA sequences, we find that mitochondrial genetic diversity evenness follows a quadratic latitudinal gradient peaking in the subtropics. Both mitochondrial genetic diversity mean and evenness positively correlate with seasonally hot temperatures, as well as climate stability since the last glacial maximum. Our models explain 27.9% and 24.0% of the observed variation in mitochondrial genetic diversity mean and evenness in insects, respectively, making an important step towards understanding global biodiversity patterns in the most diverse animal taxon.

Factors Associated with Mortality after Percutaneous Endoscopic Gastrostomy.

Introduction: Percutaneous endoscopic gastrostomy (PEG) is a common procedure performed world-wide on patients with different comorbidities, with many indications and overall low morbidity. However, studies showed an elevated early mortality in patients undergoing PEG placement. In this systematic review, we review the factors associated with early mortality after PEG. Methods: The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were followed. The methodological index for nonrandomized studies (MINORS) score system was used to perform qualitative assessment of all included studies. Recommendations were summarized for predefined key items. Results: The search found 283 articles. A refined total of 21 studies were included; 20 studies cohort studies and 1 case-control study. For the cohort studies, MINORS score ranged from 7 to 12 out of 16. The single case-control study scored 17 out of 24. The number of study patients ranged from 272 to 181,196. Thirty-day mortality rate varied from 2.4% to 23.5%. Albumin, age, body mass index, C-reactive protein, diabetes mellitus, and dementia were the most frequently associated factors to early mortality in patients undergoing PEG placement. Five studies reported procedure related deaths. Infection was the most commonly reported complication of PEG placement. Conclusions: PEG tube insertion is a fast, safe and effective procedure, but is not free of complications and can have a high early mortality rate as demonstrated in this review. Patient selection should be a key factor and the identification of factors associated with early mortality is important in the elaboration of a protocol to benefit patients.

Nutritional requirements of human induced pluripotent stem cells.

The nutritional requirements for human induced pluripotent stem cell (hiPSC) growth have not been extensively studied. Here, building on our prior work that established the suitable non-basal medium components for hiPSC growth, we develop a simplified basal medium consisting of just 39 components, demonstrating that many ingredients of DMEM/F12 are either not essential or are at suboptimal concentrations. This new basal medium along with the supplement, which we call BMEM, enhances the growth rate of hiPSCs over DMEM/F12-based media, supports derivation of multiple hiPSC lines, and allows differentiation to multiple lineages. hiPSCs cultured in BMEM consistently have enhanced expression of undifferentiated cell markers such as POU5F1 and NANOG, along with increased expression of markers of the primed state and reduced expression of markers of the naive state. This work describes titration of the nutritional requirements of human pluripotent cell culture and identifies that suitable nutrition enhances the pluripotent state.

STAT3 signaling in B cells controls germinal center zone organization and recycling.

Germinal centers (GCs), sites of antibody affinity maturation, are organized into dark (DZ) and light (LZ) zones. Here, we show a B cell-intrinsic role for signal transducer and activator of transcription 3 (STAT3) in GC DZ and LZ organization. Altered zonal organization of STAT3-deficient GCs dampens development of long-lived plasma cells (LL-PCs) but increases memory B cells (MBCs). In an abundant antigenic environment, achieved here by prime-boost immunization, STAT3 is not required for GC initiation, maintenance, or proliferation but is important for sustaining GC zonal organization by regulating GC B cell recycling. Th cell-derived signals drive STAT3 tyrosine 705 and serine 727 phosphorylation in LZ B cells, regulating their recycling into the DZ. RNA sequencing (RNA-seq) and chromatin immunoprecipitation sequencing (ChIP-seq) analyses identified STAT3 regulated genes that are critical for LZ cell recycling and transiting through DZ proliferation and differentiation phases. Thus, STAT3 signaling in B cells controls GC zone organization and recycling, and GC egress of PCs, but negatively regulates MBC output.

Signal peptide mimicry primes Sec61 for client-selective inhibition.

Preventing the biogenesis of disease-relevant proteins is an attractive therapeutic strategy, but attempts to target essential protein biogenesis factors have been hampered by excessive toxicity. Here we describe KZR-8445, a cyclic depsipeptide that targets the Sec61 translocon and selectively disrupts secretory and membrane protein biogenesis in a signal peptide-dependent manner. KZR-8445 potently inhibits the secretion of pro-inflammatory cytokines in primary immune cells and is highly efficacious in a mouse model of rheumatoid arthritis. A cryogenic electron microscopy structure reveals that KZR-8445 occupies the fully opened Se61 lateral gate and blocks access to the lumenal plug domain. KZR-8445 binding stabilizes the lateral gate helices in a manner that traps select signal peptides in the Sec61 channel and prevents their movement into the lipid bilayer. Our results establish a framework for the structure-guided discovery of novel therapeutics that selectively modulate Sec61-mediated protein biogenesis.

Cost-effectiveness of point-of-care versus centralised, laboratory-based nucleic acid testing for diagnosis of HIV in infants: a systematic review of modelling studies.

BACKGROUND: Point-of-care (POC) nucleic acid testing for diagnosis of HIV in infants facilitates earlier initiation of antiretroviral therapy (ART) than with centralised (standard-of-care, SOC) testing, but can be more expensive. We evaluated cost-effectiveness data from mathematical models comparing POC with SOC to provide global policy guidance. METHODS: In this systematic review of modelling studies, we searched PubMed, MEDLINE, Embase, the National Health Service Economic Evaluation Database, Econlit, and conference abstracts, combining terms for "HIV" + "infant"/"early infant diagnosis" + "point-of-care" + "cost-effectiveness" + "mathematical models", without restrictions from database inception to July 15, 2022. We selected reports of mathematical cost-effectiveness models comparing POC with SOC for HIV diagnosis in infants younger than 18 months. Titles and abstracts were independently reviewed, with full-text review for qualifying articles. We extracted data on health and economic outcomes and incremental cost-effectiveness ratios (ICERs) for narrative synthesis. The primary outcomes of interest were ICERs (comparing POC with SOC) for ART initiation and survival of children living with HIV. FINDINGS: Our search identified 75 records through database search. 13 duplicates were excluded, leaving 62 non-duplicate articles. 57 records were excluded and five were reviewed in full text. One article was excluded as it was not a modelling study, and four qualifying studies were included in the review. These four reports were from two mathematical models from two independent modelling groups. Two reports used the Johns Hopkins model to compare POC with SOC for repeat early infant diagnosis testing in the first 6 months in sub-Saharan Africa (first report, simulation of 25 000 children) and Zambia (second report, simulation of 7500 children). In the base scenario, POC versus SOC increased probability of ART initiation within 60 days of testing from 19% to 82% (ICER per additional ART initiation range US$430-1097; 9-month cost horizon) in the first report; and from 28% to 81% in the second ($23-1609, 5-year cost horizon). Two reports compared POC with SOC for testing at 6 weeks in Zimbabwe using the Cost-Effectiveness of Preventing AIDS Complications-Paediatric model (simulation of 30 million children; lifetime horizon). POC increased life expectancy and was considered cost-effective compared with SOC (ICER $711-850 per year of life saved in HIV-exposed children). Results were robust throughout sensitivity and scenario analyses. In most scenarios, platform cost-sharing (co-use with other programmes) resulted in POC being cost-saving compared with SOC. INTERPRETATION: Four reports from two different models suggest that POC is a cost-effective and potentially cost-saving strategy for upscaling of early infant testing compared with SOC. FUNDING: Bill & Melinda Gates Foundation, Unitaid, National Institute of Allergy and Infectious Diseases, National Institute of Child Health and Human Development, WHO, and Massachusetts General Hospital Research Scholars.

Effect of transauricular nerve stimulation on perioperative pain: a single-blind, analyser-masked, randomised controlled trial.

BACKGROUND: Activation of central autonomic pathways, including those regulating the arterial baroreflex, might reduce acute pain. We tested the hypothesis that transcutaneous auricular nerve stimulation (TAN) reduces pain after orthopaedic trauma surgery through autonomic modulation. METHODS: A total of 86 participants aged >18 yr were randomly assigned to 50 min of either sham or active bilateral TAN, undertaken before, and again 24 h after, surgery for orthopaedic trauma. The primary outcome was absolute change in pain 24 h postoperatively, comparing the 100 mm visual analogue scale (VAS) before and after TAN. Secondary outcomes included the minimal clinically important difference in pain (>10 mm increase or reduction in VAS) before/after surgery, using intention-to-treat analysis. Holter monitoring, the analysis of which was masked to allocation, quantified autonomic modulation of heart rate. RESULTS: From June 22, 2021 to July 7, 2022, 79/86 participants (49 yr; 45% female) completed TAN before and after surgery. For the primary outcome, the mean reduction in VAS was 19 mm (95% confidence interval [CI]: 12-26) after active TAN (n=40), vs 10 mm (95% CI: 3-17) after sham TAN (n=39; P=0.023). A minimally clinically important reduction in postoperative pain occurred in 31/40 (78%) participants after active TAN, compared with 15/39 (38%) allocated to sham TAN (odds ratio 5.51 [95% CI: 2.06-14.73]; P=0.001). Only active TAN increased heart rate variability (log low-frequency power increased by 0.19 ms2 [0.01-0.37 ms2]). Prespecified adverse events (auricular skin irritation) occurred in six participants receiving active TAN, compared with two receiving sham TAN. CONCLUSION: Bilateral TAN reduces perioperative pain through autonomic modulation. These proof-of-concept data support a non-pharmacological, generalisable approach to improve perioperative analgesia.