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Abstract Although adoptive T cell therapy provides the T cell pool needed for immediate tumor debulking, the infused T cells generally have a narrow repertoire for antigen recognition and limited ability for long-term protection. Here, we present a hydrogel that locally delivers adoptively transferred T cells to the tumor site while recruiting and activating host antigen-presenting cells with GMCSF or FLT3L and CpG, respectively. T cells alone loaded into these localized cell depots provided significantly better control of subcutaneous B16-F10 tumors than T cells delivered through direct peritumoral injection or intravenous infusion. T cell delivery combined with biomaterial-driven accumulation and activation of host immune cells prolonged the activation of the delivered T cells, minimized host T cell exhaustion, and enabled long-term tumor control. These findings highlight how this integrated approach provide both immediate tumor debulking and long-term protection against solid tumors, including against tumor antigen escape.

Abstract Squamous cell carcinoma antigen recognized by T cells 3 (SART3) is an RNA-binding protein with numerous biological functions including recycling small nuclear RNAs to the spliceosome. Here, we identify recessive variants in SART3 in nine individuals presenting with intellectual disability, global developmental delay and a subset of brain anomalies, together with gonadal dysgenesis in 46,XY individuals. Knockdown of the Drosophila orthologue of SART3 reveals a conserved role in testicular and neuronal development. Human induced pluripotent stem cells carrying patient variants in SART3 show disruption to multiple signalling pathways, upregulation of spliceosome components and demonstrate aberrant gonadal and neuronal differentiation in vitro. Collectively, these findings suggest that bi-allelic SART3 variants underlie a spliceosomopathy which we tentatively propose be termed INDYGON syndrome (Intellectual disability, Neurodevelopmental defects and Developmental delay with 46,XY GONadal dysgenesis). Our findings will enable additional diagnoses and improved outcomes for individuals born with this condition.

Introduction In 2018, the American College of Obstetricians and Gynecologists recommended low-dose aspirin to prevent the onset of pre-eclampsia among women who were at high risk. Factors influencing women’s acceptance of this recommendation span multiple sectors and levels. Understanding how these factors interact will help stakeholders design effective population-level intervention strategies. Our study aims to identify and map relationships among factors influencing the medication decisions of pregnant women at risk of hypertensive disorders.Methods and analysis Preferred Reporting Items for Systematic Reviews and Meta-Analyses for Scoping Reviews (PRISMA-ScR) guidelines will be followed for this review. A research librarian developed a comprehensive search strategy to retrieve published and unpublished English studies after 1 January 1980, involving factors that influence pregnant women’s uptake and adherence to medication for gestational hypertensive disorders. This literature includes perceptions, patterns, acceptance, refusal, tendencies, probability and service utilisation. We will search PubMed, Embase, Web of Science and CINAHL. Reference lists of the selected papers will be searched manually to identify more relevant studies. A two-stage independent screening, consisting of title and abstract screening, followed by full-text screening, will be conducted by two independent reviewers to identify eligible articles. Extracted data will be recorded in a customised variable extraction form and input into a Microsoft Access database. The PRISMA-ScR will be used to guide the presentation of the results, which will be presented in a table and causal map to demonstrate the relationships between extracted variables and medication uptake and adherence. A conceptual simulation model will be formulated to validate the logic of the relationships between variables and identify knowledge gaps. Lastly, experts and stakeholders will be invited to critique and comment on the results.Ethics and dissemination This study does not require ethical approval. The full review results will be presented at a relevant conference and submitted to a peer-reviewed scientific journal for publication.

Background: Traumatic spinal cord injury (TSCI) survivors are confronted by both physical and psychosocial barriers when returning to their communities. Therefore, reintegration is an important aspect of their journey back into social life. Objectives: To assess psychosocial reintegration after TSCI in Rwanda. Method: All community-dwelling adults who were registered in the previous epidemiological study were recruited and injury characteristics questionnaire and the Sydney Psychosocial Reintegration Scale version 2 (SPRS-2) were used to collect data through a telephone interview. Results: The study traced 58 participants, 77.6% (n = 45) were male and 56.9% (n = 33) were categorised with paraplegia. Overall, the results show poor community reintegration. The SPRS-2 and domain mean (SD) scores were: overall SPRS-2 of 20.95 (11.56), occupational activity (OA) of 3.68 (4.31), interpersonal relationship (IR) of 7.11(4.31) and living skills (LS) of 7.43 (5.32). Gender significantly influenced overall SPRS-2 (p = 0.011) and two domains: OA (p = 0.005) and LS (p = 0.012). Level of injury was significantly associated with an OA domain score of SPRS-2 (p = 0.002). Gender explained 29% of the variance in the LS domain of SPRS-2, with males reporting better psychosocial reintegration. Conclusion: Gender strongly predicted psychosocial reintegration following a TSCI, which is an indication of the role of social support. Clinical Implications: Traumatic SCI rehabilitation should be holistic to help prepare the person to return to the community. There should be an assessment of an individual’s readiness to return to the community before discharge from the hospital.

Massive sequencing of SARS-CoV-2 genomes has urged novel methods that employ existing phylogenies to add new samples efficiently instead of de novo inference. 'TIPars' was developed for such challenge integrating parsimony analysis with pre-computed ancestral sequences. It took about 21 seconds to insert 100 SARS-CoV-2 genomes into a 100k-taxa reference tree using 1.4 gigabytes. Benchmarking on four datasets, TIPars achieved the highest accuracy for phylogenies of moderately similar sequences. For highly similar and divergent scenarios, fully parsimony-based and likelihood-based phylogenetic placement methods performed the best respectively while TIPars was the second best. TIPars accomplished efficient and accurate expansion of phylogenies of both similar and divergent sequences, which would have broad biological applications beyond SARS-CoV-2. TIPars is accessible from https://tipars.hku.hk/ and source codes are available at https://github.com/id-bioinfo/TIPars.

An estimated 1.5 million Americans suffer from Type I diabetes mellitus, and its incidence is increasing worldwide. Islet allotransplantation offers a treatment, but the availability of deceased human donor pancreases is limited. The transplantation of islets from gene-edited pigs, if successful, would resolve this problem. Pigs are now available in which the expression of the three known xenoantigens against which humans have natural (preformed) antibodies has been deleted, and in which several human ‘protective’ genes have been introduced. The transplantation of neonatal pig islets has some advantages over that of adult pig islets. Transplantation into the portal vein of the recipient results in loss of many islets from the instant blood-mediated inflammatory reaction (IBMIR) and so the search for an alternative site continues. The adaptive immune response can be largely suppressed by an immunosuppressive regimen based on blockade of the CD40/CD154 T cell co-stimulation pathway, whereas conventional therapy (e.g., based on tacrolimus) is less successful. We suggest that, despite the need for effective immunosuppressive therapy, the transplantation of ‘free’ islets will prove more successful than that of encapsulated islets. There are data to suggest that, in the absence of rejection, the function of pig islets, though less efficient than human islets, will be sufficient to maintain normoglycemia in diabetic recipients. Pig islets transplanted into immunosuppressed nonhuman primates have maintained normoglycemia for periods extending more than two years, illustrating the potential of this novel form of therapy.

L 98-59 d is a Super-Earth planet orbiting an M-type star. We performed retrievals on the transmission spectrum of L 98-59 d obtained using NIRSpec G395H during a single transit, from JWST Cycle 1 GTO 1224. The wavelength range of this spectrum allows us to detect the presence of several atmospheric species. We found that the spectrum is consistent with a high mean molecular weight atmosphere. The atmospheric spectrum indicates the possible presence of the sulfur-bearing species H _2 S and SO _2 , which could hint at active volcanism on this planet if verified by future observations. We also tested for signs of stellar contamination in the spectrum and found signs of unocculted faculae on the star. The tentative signs of an atmosphere on L 98-59 d presented in this work from just one transit bodes well for possible molecular detections in the future, particularly as it is one of the best targets among small exoplanets for atmospheric characterization using JWST.

Detecting atmospheres around planets with a radius below 1.6 R _⊕ , commonly referred to as rocky planets, has proven to be challenging. However, rocky planets orbiting M dwarfs are ideal candidates due to their favorable planet-to-star radius ratio. Here, we present one transit observation of the Super-Earth L98-59 d (1.58 R _⊕ and 2.31 M _⊕ ), at the limit of rocky/gas-rich, using the JWST NIRSpec G395H mode covering the 2.8–5.1 μ m wavelength range. The extracted transit spectrum from a single transit observation deviates from a flat line by 2.6 σ –5.6 σ , depending on the data reduction and retrieval setup. The hints of an atmospheric detection are driven by a large absorption feature between 3.3 and 4.8 μ m. A stellar contamination retrieval analysis rejected the source of this feature as being due to stellar inhomogeneities, making the best fit an atmospheric model including sulfur-bearing species, suggesting that the atmosphere of L98-59 d may not be at equilibrium. This result will need to be confirmed by the analysis of the second NIRSpec G395H visit in addition to the NIRISS SOSS transit observation.

Recent mid-infrared observations with JWST's Mid-Infrared Instrument Low Resolution Spectrometer (MIRI LRS) have resulted in the first direct detections of absorption features from silicate clouds in the transmission spectra of two transiting exoplanets, WASP-17 b and WASP-107 b. In this Letter, we measure the mid-infrared (5–12 μ m) dayside emission spectrum of the benchmark hot Jupiter HD 189733 b with MIRI LRS by combining data from two secondary-eclipse observations. We confirm the previous detection of H _2 O absorption at 6.5 μ m from Spitzer's Infrared Spectrograph (IRS) and additionally detect H _2 S as well as an absorption feature at 8.7 μ m in both secondary-eclipse observations. The excess absorption at 8.7 μ m can be explained by the presence of small (∼0.01 μ m) grains of SiO _2 [s] in the uppermost layers of HD 189733 b’s dayside atmosphere. This is the first direct detection of silicate clouds in HD 189733 b’s atmosphere, and the first detection of a distinct absorption feature from silicate clouds on the dayside of any hot Jupiter. We find that models including SiO _2 [s] are preferred by 6–7 σ over clear models and those with other potential cloud species. The high-altitude location of these silicate particles is best explained by formation in the hottest regions of HD 189733 b’s dayside atmosphere near the substellar point. We additionally find that HD 189733 b’s emission spectrum longward of 9 μ m displays residual features not well captured by our current atmospheric models. When combined with other JWST observations of HD 189733 b’s transmission and emission spectra at shorter wavelengths, these observations will provide us with the most detailed picture to date of the atmospheric composition and cloud properties of this benchmark hot Jupiter.

Lyme disease is the leading tick-borne infection in the United States, caused by the pathogenic spirochete Borreliella burgdorferi, formerly known as Borrelia burgdorferi. Diderms, or bacteria with dual-membrane ultrastructure, such as B. burgdorferi, have multiple methods of transporting and integrating outer membrane proteins (OMPs). Most integral OMPs are transported through the β-barrel assembly machine (BAM) complex. This complex consists of the channel-forming OMP BamA and accessory lipoproteins that interact with the five periplasmic, polypeptide transport-associated (POTRA) domains of BamA. Another system, the translocation and assembly module (TAM) system, has also been implicated in OMP assembly and export. The TAM system consists of two proteins, the BamA paralog TamA which has three POTRA domains and the inner membrane protein TamB. TamB is characterized by a C-terminal DUF490 domain that interacts with the POTRA domains of TamA. Interestingly, while TamB is found in almost all diderms, including B. burgdorferi, TamA is found almost exclusively in Proteobacteria. This strongly suggests a TamA-independent role of TamB in most diderms. We previously demonstrated that BamA interacts with TamB in B. burgdorferi and hypothesized that this is facilitated by the BamA POTRA domains interacting with the TamB DUF490 domain. In this study, we utilized protein-protein co-purification assays to empirically demonstrate that the B. burgdorferi TamB DUF490 domain interacts with BamA POTRA2 and POTRA3. We also observed that the DUF490 domain of TamB interacts with the accessory lipoprotein BamB. To examine if the BamA-TamB interaction is more ubiquitous among diderms, we examined BamA-TamB interactions in Salmonella enterica serovar Typhimurium (St). Interestingly, even though St encodes a TamA protein that interacts with TamB, we observed that the TamB DUF490 of St interacts with BamA in this organism. Our combined findings strongly suggest that the TamB-BamA interaction occurs independent of the TamA component of the TAM protein export system.

We present two-dimensional near-infrared temperature maps of the canonical hot Jupiter WASP-43b using a phase-curve observation with JWST NIRSpec/G395H. From the white-light planetary transit, we improve constraints on the planet’s orbital parameters and measure a planet-to-star radius ratio of ${0.15883}_{-0.00053}^{+0.00056}$ . Using the white-light phase curve, we measure a longitude of maximum brightness of ${6.9}_{-0\buildrel{\circ}\over{.} 5}^{+0\buildrel{\circ}\over{.} 5}$ east of the substellar point and a phase-curve offset of ${10.0}_{-0\buildrel{\circ}\over{.} 8}^{+0\buildrel{\circ}\over{.} 8}$ . We also find a ≈4 σ detection of a latitudinal hotspot offset of $-{13.4}_{-1\buildrel{\circ}\over{.} 7}^{+3\buildrel{\circ}\over{.} 2}$ , the first significant detection of a nonequatorial hotspot in an exoplanet atmosphere. We show that this detection is robust to variations within planetary parameter uncertainties, but only if the transit is used to improve constraints, showing the importance of transit observations to eclipse mapping. Maps retrieved from the NRS1 and NRS2 detectors are similar, with hotspot locations consistent between the two detectors at the 1 σ level. Our JWST data show brighter (hotter) nightsides and a dimmer (colder) dayside at the shorter wavelengths relative to fits to Spitzer 3.6 and 4.5 μ m phase curves. Through comparison between our phase curves and a set of general circulation models, we find evidence for clouds on the nightside and atmospheric drag or high metallicity reducing the eastward hotspot offset.

IntroductionThe Arctic sea ice extent in September (when it is at its lowest) has declined 13% Q10 per decade, and the Arctic Ocean is becoming a more Atlantic-influenced system. Rapid climate-forced changes are taking place in many high-latitude marine ecosystems. The Barents Sea is one such high-latitude shelf ecosystem, between approximately 70° and 80°N in the Norwegian Arctic. The purpose of the current study was to estimate zoobenthic blue carbon across multiple habitats within the Barents Sea (trough, basin, shelf, and shallows), potentially providing values to aid ecosystem-based management of these areas under future climate change scenarios.MethodWe tested this by capture and analysis of 947 high-resolution (each 405.7 × 340.6 mm, 12 MB, 5 megapixels) seabed images at 17 sites with latitudinal cline, linked to a collection of corresponding oceanographic data. Biotas within these images were identified to one of the 14 functional groups and the density was calculated. Mean stored carbon per individual was assigned by ash mass (AM) and ash-free dry mass (AFDM) of individuals caught within Agassiz trawl deployments at the same sites.ResultsTrough sites, except for one site (B16), have a low quantity of zoobenthic blue carbon compared with the shallow, shelf, and basin habitats.DiscussionThe results of a previous study focused entirely on trough habitats and are therefore difficult to scale up as the basis for a meaningful estimate of across-habitat zoobenthic blue carbon in the Barents Sea. Compared with the trough and the basin, the shelf and shallow habitats of the Barents Sea are also subjected to more trawling events through demersal fisheries and showed higher zoobenthic blue carbon stock values.

IntroductionWheat stem sawfly (WSS), Cephus cinctus Norton, is a major pest of common bread wheat (Triticum aestivum L.) and other cultivated cereals in North America. Planting of cultivars with solid stems has been the primary management strategy to prevent yield loss due to WSS infestation, however expression of this phenotype can vary depending on environmental conditions and solid stems hinder biological control of WSS via braconid parasitoids Bracon cephi (Gahan) and Bracon lissogaster Muesebeck. In the hollow stems of oat (Avena sativa L.), WSS larvae experience 100% mortality before they reach late instars, but the mechanisms for this observed resistance have not been characterized. ObjectiveThe objective of this study was to explore additional sources of resistance outside of the historic solid stem phenotype.MethodsHere, we use an untargeted metabolomics approach to examine the response of the metabolome of two cultivars of oat and four cultivars of spring wheat to infestation by WSS. Using liquid chromatography-mass spectrometry (LC-MS), differentially expressed metabolites were identified between oat and wheat which were associated with the phenylpropanoid pathway, phospholipid biosynthesis and signaling, the salicylic acid signaling pathway, indole-3-acetic acid (IAA) degradation, and biosynthesis of 1,4-benzoxazin-3-ones (Bxs). Several phospho- and galacto- lipids were found in higher abundance in oat, and with the exception of early stem solidness cultivar Conan, both species experienced a decrease in abundance once infested. In all wheat cultivars except Conan, an increase in abundance was observed for Bxs HMDBOA-glc and DIBOA-β-D-glucoside after infestation, indicating that this pathway is involved in wheat response to infestation in both solid and hollow stemmed cultivars. Differences between species in compounds involved in IAA biosynthesis, degradation and inactivation suggest that wheat may respond to infestation by inactivating IAA or altering the IAA pool in stem tissue.ConclusionWe propose that the species differences found here likely affect the survival of WSS larvae and may also be associated with differences in stem architecture at the molecular level. Our findings suggest pathways to focus on for future studies in elucidating plant response to WSS infestation.

Amyotrophic lateral sclerosis (ALS) is an incurable neurodegenerative disorder. The most devastating variant is bulbar-onset ALS, which portends a median survival of 24 months from the onset of symptoms. Abundant evidence indicates that neuron metabolism and mitochondrial function are impaired in ALS. Metabolic strategies, particularly fasting and ketogenic diet protocols, alter neuron metabolism and mitochondria function in a manner that may mitigate the symptoms of this disorder. We report the case of a 64-year-old man with a 21-month history of progressive, deteriorating bulbar-onset ALS, with an associated pseudobulbar affect, who implemented a time-restricted ketogenic diet (TRKD) for 18 months. During this time, he improved in ALS-related function (7% improvement from baseline), forced expiratory volume (17% improvement), forced vital capacity (13% improvement), depression (normalized), stress levels (normalized), and quality of life (19% improvement), particularly fatigue (23% improvement). His swallowing impairment and neurocognitive status remained stable. Declines were measured in physical function, maximal inspiratory pressure, and maximal expiratory pressure. Weight loss was attenuated and no significant adverse effects occurred. This case study represents the first documented occurrence of a patient with ALS managed with either a fasting or ketogenic diet protocol, co-administered as a TRKD. We measured improved or stabilized ALS-related function, forced expiratory volume, forced vital capacity, swallowing, neurocognitive status, mood, and quality of life. Measurable declines were restricted to physical function, maximal inspiratory pressure, and maximal expiratory pressure. Now over 45 months since symptom onset, our patient remains functionally independent and dedicated to his TRKD.

BackgroundRoad traffic injuries (RTIs) leading to long-term disability present a significant public health challenge, causing immense personal and societal consequences. Every year, 50 million people are hurt, 1.2 million die, 30% are permanently disabled, and 14% cannot return to work due to road traffic accidents. However, in many developing countries, information on the social integration of patients post-RTI remains limited. This study aimed to identify factors contributing to social integration following road traffic-related orthopedic injuries (RTOI) in Rwanda.MethodologyA multicenter, cross-sectional study included 369 adult Road traffic orthopedic injuries (RTOI) victims from five Rwandan referral hospitals. Participants completed the IMPACT-S Questionnaire between 2 June 2022, and 31 August 2022, two years after the injury. It measured social integration in terms of activities and paricipation. We used logistic regression statistical analysis with a significance level of p 15 days (OR = 4.44, p = 0.02).ConclusionThe majority of RTOI victims in Rwanda achieved successful reintegration into society; nevertheless, their mobility and community engagement were more significantly impacted compared to other aspects assessed by the IMPACT-S scale. The study emphasized the importance of early management, effective rehabilitation, and prompt patient discharge from the hospital in facilitating a successful return to everyday life after road traffic-related orthopedic injuries.

IntroductionWe are developing the California Cognitive Assessment Battery (CCAB) to provide neuropsychological assessments to patients who lack test access due to cost, capacity, mobility, and transportation barriers.MethodsThe CCAB consists of 15 non-verbal and 17 verbal subtests normed for telemedical assessment. The CCAB runs on calibrated tablet computers over cellular or Wi-Fi connections either in a laboratory or in participants’ homes. Spoken instructions and verbal stimuli are delivered through headphones using naturalistic text-to-speech voices. Verbal responses are scored in real time and recorded and transcribed offline using consensus automatic speech recognition which combines the transcripts from seven commercial ASR engines to produce timestamped transcripts more accurate than those of any single ASR engine. The CCAB is designed for supervised self-administration using a web-browser application, the Examiner. The Examiner permits examiners to record observations, view subtest performance in real time, initiate video chats, and correct potential error conditions (e.g., training and performance failures, etc.,) for multiple participants concurrently.ResultsHere we describe (1) CCAB usability with older (ages 50 to 89) participants; (2) CCAB psychometric properties based on normative data from 415 older participants; (3) Comparisons of the results of at-home vs. in-lab CCAB testing; (4) We also present preliminary analyses of the effects of COVID-19 infection on performance. Mean z-scores averaged over CCAB subtests showed impaired performance of COVID+ compared to COVID- participants after factoring out the contributions of Age, Education, and Gender (AEG). However, inter-cohort differences were no longer significant when performance was analyzed with a comprehensive model that factored out the influences of additional pre-existing demographic factors that distinguished COVID+ and COVID- cohorts (e.g., vocabulary, depression, race, etc.,). In contrast, unlike AEG scores, comprehensive scores correlated significantly with the severity of COVID infection. (5) Finally, we found that scoring models influenced the classification of individual participants with Mild Cognitive Impairment (MCI, z-scores < –1.50) where the comprehensive model accounted for more than twice as much variance as the AEG model and reduced racial bias in MCI classification.DiscussionThe CCAB holds the promise of providing scalable laboratory-quality neurodiagnostic assessments to underserved urban, exurban, and rural populations.

Animal songs can change within and between populations as the result of different evolutionary processes. When these processes include cultural transmission, the social learning of information or behaviours from conspecifics, songs can undergo rapid evolutions because cultural novelties can emerge more frequently than genetic mutations. Understanding these song variations over large temporal and spatial scales can provide insights into the patterns, drivers and limits of song evolution that can ultimately inform on the species’ capacity to adapt to rapidly changing acoustic environments. Here, we analysed changes in fin whale (Balaenoptera physalus) songs recorded over two decades across the central and eastern North Atlantic Ocean. We document a rapid replacement of song INIs (inter-note intervals) over just four singing seasons, that co-occurred with hybrid songs (with both INIs), and a clear geographic gradient in the occurrence of different song INIs during the transition period. We also found gradual changes in INIs and note frequencies over more than a decade with fin whales adopting song changes. These results provide evidence of vocal learning in fin whales and reveal patterns of song evolution that raise questions on the limits of song variation in this species.

Abstract Using a dataset of 591 capture events between 2001–2019 in California, USA, we examined the impact of capture methods and immobilization drugs on mountain lion (Puma concolor) welfare. The 3 methods used to capture mountain lions were cage traps, trained hounds, and cable restraints. The drugs used to immobilize mountain lions were either tiletamine/zolazepam (Telazol®), ketamine/medetomidine, or ketamine/xylazine. Mortality occurred in 1.4% of captures, with only one mortality out of 310 captures occurring since 2012. We used a logistic regression framework to compare morbidity and vital parameters of mountain lions among the different capture methods and immobilization drugs used. Vomiting (a risk factor for developing aspiration pneumonia) was the most common severe risk factor associated with cage trapping and was only seen with the use of ketamine/medetomidine or ketamine/xylazine. Morbidity scores were not well predicted by any of the variables we accounted for. Animals immobilized with Telazol® were more likely to experience abnormal heart and respiratory rates, as well as high body temperatures, than those immobilized with the other two combinations. Although there are risks associated with each of the capture methods and drug combinations commonly used in mountain lion captures in California, our analyses demonstrated they are all relatively safe when following appropriate animal welfare practices. Our analyses suggested that unaccounted for factors are equally or more important in explaining injury and physiological abnormality rates, and we urge that agencies train personnel in best practices and conservative decision‐making in order to assure that the welfare of the animal takes precedence over collaring. We suggest training on how to choose between several capture techniques, immobilization drugs and monitoring methods and how to reduce both detrimental effects to mountain lions and danger to humans.

The transmission of DNA through extracellular vesicles (EVs) represents a novel genetic material transfer mechanism that may impact genome evolution and tumorigenesis. We aimed to investigate the potential for vertical DNA transmission within maternal endometrial EVs to the pre-implantation embryo and describe any effect on embryo bioenergetics. We discovered that the human endometrium secretes all three general subtypes of EV - apoptotic bodies (ABs), microvesicles (MVs), and exosomes (EXOs) - into the human endometrial fluid (EF) within the uterine cavity. EVs become uniformly secreted into the EF during the menstrual cycle, with the proportion of different EV populations remaining constant; however, MVs contain significantly higher levels of mitochondrial (mt)DNA than ABs or EXOs. During the window of implantation, MVs contain an eleven-fold higher level of mtDNA when compared to cells-of-origin within the receptive endometrium, which possesses a lower mtDNA content and displays the upregulated expression of mitophagy-related genes. Furthermore, we demonstrate the internalization of EV-derived nuclear-encoded (n)DNA/mtDNA by trophoblast cells of murine embryos, which associates with a reduction in mitochondrial respiration and ATP production. These findings suggest that the maternal endometrium suffers a reduction in mtDNA content during the preconceptional period, that nDNA/mtDNA become packaged into secreted EVs that the embryo uptakes, and that the transfer of DNA to the embryo within EVs occurs alongside the modulation of bioenergetics during implantation.

ABSTRACTBetacoronaviruses encode an internal (I) gene via an alternative reading frame within the nucleocapsid gene, called ORF8b for Middle-East respiratory syndrome coronavirus (MERS-CoV) and ORF9b for severe acute respiratory syndrome coronavirus (SARS-CoV) and SARS-CoV-2. Previous reports suggested that proteins 8b and 9b are involved in evading multiple innate immune signaling pathways. However, their roles in mediating pathogenesis in infected animals have not been determined. In this study, we abrogated the expression of protein 8b in MERS-CoV and protein 9b in SARS-CoV-2. Using mouse models of MERS-CoV and SARS-CoV-2 infection, we found that MERS-CoV lacking protein 8b expression was more virulent, while SARS-CoV-2 lacking protein 9b expression was attenuated compared with the respective wild-type viruses. Upon further analysis, we detected increased levels of type I interferon and enhanced infiltration of immune cells to the lungs of mice infected with MERS-CoV lacking protein 8b expression. These data suggest that the I protein of MERS-CoV plays a role in limiting pathogenesis while that of SARS-CoV-2 enhances disease severity.IMPORTANCEThe function of betacoronavirus internal protein has been relatively understudied. The earliest report on the internal protein of mouse hepatitis virus suggested that the internal protein is a structural protein without significant functions in virus replication and virulence. However, the internal proteins of severe acute respiratory syndrome coronavirus (SARS-CoV), Middle-East respiratory syndrome coronavirus, and SARS-CoV-2 have been shown to evade immune responses. Despite the reported functions of the internal protein in these highly pathogenic human coronaviruses, its role in mediating pathogenesis in experimentally infected animals has not been characterized. Our data indicated that despite the similar genomic location and expression strategy of these internal proteins, their effects on virulence are vastly different and virus specific, highlighting the complexity between host-virus interaction and disease outcome.

Summary: Oral squamous cell carcinoma (OSCC), a prevalent and aggressive neoplasm, poses a significant challenge due to poor prognosis and limited prognostic biomarkers. Leveraging highly multiplexed imaging mass cytometry, we investigated the tumor immune microenvironment (TIME) in OSCC biopsies, characterizing immune cell distribution and signaling activity at the tumor-invasive front. Our spatial subsetting approach standardized cellular populations by tissue zone, improving feature reproducibility and revealing TIME patterns accompanying loss-of-differentiation. Employing a machine-learning pipeline combining reliable feature selection with multivariable modeling, we achieved accurate histological grade classification (AUC = 0.88). Three model features correlated with clinical outcomes in an independent cohort: granulocyte MAPKAPK2 signaling at the tumor front, stromal CD4+ memory T cell size, and the distance of fibroblasts from the tumor border. This study establishes a robust modeling framework for distilling complex imaging data, uncovering sentinel characteristics of the OSCC TIME to facilitate prognostic biomarkers discovery for recurrence risk stratification and immunomodulatory therapy development.

IntroductionAstarte borealis holds great potential as an archive of seasonal paleoclimate, especially due to its long lifespan (several decades to more than a century) and ubiquitous distribution across high northern latitudes. Furthermore, recent work demonstrates that the isotope geochemistry of the aragonite shell is a faithful proxy of environmental conditions. However, the exceedingly slow growth rates of A. borealis in some locations (

Abstract Background Nutrient limitations often lead to metabolic stress during cancer initiation and progression. To combat this stress, the enzyme heme oxygenase 1 (HMOX1, commonly known as HO-1) is thought to play a key role as an antioxidant. However, there is a discrepancy between the level of HO-1 mRNA and its protein, particularly in cells under stress. O-linked β-N-acetylglucosamine (O-GlcNAc) modification of proteins (O-GlcNAcylation) is a recently discovered cellular signaling mechanism that rivals phosphorylation in many proteins, including eukaryote translation initiation factors (eIFs). The mechanism by which eIF2α O-GlcNAcylation regulates translation of HO-1 during extracellular arginine shortage (ArgS) remains unclear. Methods We used mass spectrometry to study the relationship between O-GlcNAcylation and Arg availability in breast cancer BT-549 cells. We validated eIF2α O-GlcNAcylation through site-specific mutagenesis and azido sugar N-azidoacetylglucosamine-tetraacylated labeling. We then evaluated the effect of eIF2α O-GlcNAcylation on cell recovery, migration, accumulation of reactive oxygen species (ROS), and metabolic labeling during protein synthesis under different Arg conditions. Results Our research identified eIF2α, eIF2β, and eIF2γ, as key O-GlcNAcylation targets in the absence of Arg. We found that O-GlcNAcylation of eIF2α plays a crucial role in regulating antioxidant defense by suppressing the translation of the enzyme HO-1 during Arg limitation. Our study showed that O-GlcNAcylation of eIF2α at specific sites suppresses HO-1 translation despite high levels of HMOX1 transcription. We also found that eliminating eIF2α O-GlcNAcylation through site-specific mutagenesis improves cell recovery, migration, and reduces ROS accumulation by restoring HO-1 translation. However, the level of the metabolic stress effector ATF4 is not affected by eIF2α O-GlcNAcylation under these conditions. Conclusions Overall, this study provides new insights into how ArgS fine-tunes the control of translation initiation and antioxidant defense through eIF2α O-GlcNAcylation, which has potential biological and clinical implications.

Abstract High vagal nerve activity, reliability measured by HRV, is considered protective in cancer, reducing oxidative stress, inflammation and opposing sympathetic nerve activity. The present monocentric study examines the relationship between HRV, TNM stage, co-morbidity, systemic inflammation and survival in patients who underwent potentially curative resections for colorectal cancer (CRC). Time-domain HRV measures, Standard Deviation of NN-intervals (SDNN) and Root Mean Square of Successive Differences (RMSSD), were examined as categorical (median) and continuous variables. Systemic inflammation was determined using systemic inflammatory grade (SIG) and co-morbidity using ASA. The primary end point was overall survival (OS) and was analysed using Cox regression. There were 439 patients included in the study and the median follow-up was 78 months. Forty-nine percent (n = 217) and 48% (n = 213) of patients were categorised as having low SDNN (

Abstract Background Dihydropyrimidine dehydrogenase (DPD) is a key enzyme in the metabolism of fluoropyrimidines. Variations in the encoding DPYD gene are associated with severe fluoropyrimidine toxicity and up-front dose reductions are recommended. We conducted a retrospective study to evaluate the impact of implementing DPYD variant testing for patients with gastrointestinal cancers in routine clinical practice in a high volume cancer centre in London, United Kingdom. Methods Patients receiving fluoropyrimidine chemotherapy for gastrointestinal cancer prior to, and following the implementation of DPYD testing were identified retrospectively. After November 2018, patients were tested for DPYD variants c.1905+1G>A (DPYD*2A), c.2846A>T (DPYD rs67376798), c.1679T>G (DPYD*13), c.1236G>A (DPYD rs56038477), c.1601G>A (DPYD*4) prior to commencing fluoropyrimidines alone or in combination with other cytotoxics and/or radiotherapy. Patients with a DPYD heterozygous variant received an initial dose reduction of 25–50%. Toxicity by CTCAE v4.03 criteria was compared between DPYD heterozygous variant and wild type carriers. Results Between 1st December 2018 and 31st July 2019, 370 patients who were fluoropyrimidine naïve underwent a DPYD genotyping test prior to receiving a capecitabine (n = 236, 63.8%) or 5FU (n = 134, 36.2%) containing chemotherapy regimen. 33 patients (8.8%) were heterozygous DPYD variant carriers and 337 (91.2%) were wild type. The most prevalent variants were c.1601G > A (n = 16) and c.1236G > A (n = 9). Mean relative dose intensity for the first dose was 54.2% (range 37.5–75%) for DPYD heterozygous carriers and 93.2% (42.9–100%) for DPYD wild type carriers. Overall grade 3 or worse toxicity was similar in DPYD variant carriers (4/33, 12.1%) as compared to wild-type carriers (89/337, 25.7%; P = 0.0924). Conclusions Our study demonstrates successful routine DPYD mutation testing prior to the initiation of fluoropyrimidine chemotherapy with high uptake. In patients with DPYD heterozygous variants with pre-emptive dose reductions, high incidence of severe toxicity was not observed. Our data supports routine DPYD genotype testing prior to commencement of fluoropyrimidine chemotherapy.

Abstract Poor adherence to asthma preventer medication is associated with life-threatening asthma attacks. The quality and outcomes framework mandated primary care annual asthma review does not include adherence monitoring and the effect of poor adherence on lung function in paediatric primary care patients is unknown. The aim was to investigate the link between inhaled corticosteroid (ICS) adherence and spirometry, fraction of exhaled nitric oxide (FeNO) and asthma control in asthmatic school-age children in this cross-sectional observational study involving three Leicestershire general practices. Children 5–16 years on the practice’s asthma registers, were invited for a routine annual asthma review between August 2018 and August 2019. Prescription and clinical data were extracted from practice databases. Spirometry, bronchodilator reversibility (BDR) and FeNO testing were performed as part of the review. 130 of 205 eligible children (63.4%) attended their review. Mean adherence to ICS was 36.2% (SEM 2.1%) and only 14.6% of children had good adherence (≥75% prescriptions issued). We found no differences in asthma exacerbations in the preceding 12 months between the adherence quartiles. 28.6% of children in the lowest and 5.6% in the highest adherence quartile had BDR ≥ 12% but this was not statistically significant (p = 0.55). A single high FeNO value did not predict adherence to ICS. Adherence to ICS in children with asthma in primary care is poor. The link between adherence to ICS and asthma exacerbations, spirometry and FeNO is complex but knowledge of adherence to ICS is critical in the management of children with asthma.

Abstract Background Understanding the relationship between human evolution and environmental changes is the key to lifting the veil on human origin. The hypothesis that environmental changes triggered the divergence of humans from apes (ca. 9.3–6.5 million years ago, Ma) has been poorly tested because of limited continuous environmental data from fossil localities. Lufengpithecus (12.5-6.0 Ma) found on the southeastern margin of the Tibetan Plateau (SEMTP) across the ape–human split provides a good chance for testing this hypothesis. Results Here, we reconstructed the habitats of L. keiyuanensis (12.5–11.6 Ma) with comprehensive vegetation, climate, and potential food web data by palaeobotanical evidence, together with other multidisciplinary data and partly tested the environment-driven hypothesis by revealing the living conditions of Lufengpithecus. Conclusion A detailed comparison of hominoids on different continents reveals their behaviour and fate divergence across the ape–human split against the background of global climate change, i.e., the stable living conditions of SEMTP not only provided a so-called ‘refuge’ for arboreal Lufengpithecus but also acted as a ‘double-edged sword’, preventing their further evolution while vegetation shifts in East Africa probably stimulated the emergence of human bipedalism, and the intense climatic changes in Europe possibly prevented those hominoids from surviving that time interval. Our findings provide interesting insight into the environmental impacts on the behavioural evolution of hominoids.

This study investigates the average genome size of planktonic prokaryotes across tropical and polar oceans and down to the hadal realm. Using hundreds of metagenomes of marine microorganisms, genome size was found to be highest in the perennially cold polar ocean, suggesting that environmental factors influence genome size selection and the ecological strategies of marine microbes.

Recent advances in the treatment of gastric cancer (GC) with chemotherapy, immunotherapy, anti-angiogenic therapy and targeted therapies have yielded some improvement in survival outcomes; however, metastatic GC remains a lethal malignancy and amongst the leading causes of cancer-related mortality worldwide. Importantly, the ongoing molecular characterisation of GCs continues to uncover potentially actionable molecular targets. Among these, aberrant FGFR2-driven signalling, predominantly arising from FGFR2 amplification, occurs in approximately 3–11% of GCs. However, whilst several inhibitors of FGFR have been clinically tested to-date, there are currently no approved FGFR-directed therapies for GC. In this review, we summarise the significance of FGFR2 as an actionable therapeutic target in GC, examine the recent pre-clinical and clinical data supporting the use of small-molecule inhibitors, antibody-based therapies, as well as novel approaches such as proteolysis-targeting chimeras (PROTACs) for targeting FGFR2 in these tumours, and discuss the ongoing challenges and opportunities associated with their clinical development.

Cyclospora cayetanensis is a foodborne parasite that causes cyclosporiasis, an enteric illness in humans. Genotyping methods are used to genetically discriminate between specimens from cyclosporiasis cases and can complement source attribution investigations if the method is sufficiently sensitive for application to food items. A very sensitive targeted amplicon sequencing (TAS) assay for genotyping C. cayetanensis encompassing 52 loci was recently designed. In this study, we analyzed 66 genetically diverse clinical specimens to assess the change in phylogenetic resolution between the TAS assay and a currently employed eight-marker scheme. Of the 52 markers, ≥50 were successfully haplotyped for all specimens, and these results were used to generate a hierarchical cluster dendrogram. Using a previously described statistical approach to dissect hierarchical trees, the 66 specimens resolved into 24 and 27 distinct genetic clusters for the TAS and an 8-loci scheme, respectively. Although the specimen composition of 15 clusters was identical, there were substantial differences between the two dendrograms, highlighting the importance of both inclusion of additional genome coverage and choice of loci to target for genotyping. To evaluate the ability to genetically link contaminated food samples with clinical specimens, C. cayetanensis was genotyped from DNA extracted from raspberries inoculated with fecal specimens. The contaminated raspberry samples were assigned to clusters with the corresponding clinical specimen, demonstrating the utility of the TAS assay for traceback efforts.

While cytochrome P450 (CYP; P450) enzymes are commonly associated with the metabolism of organic xenobiotics and drugs or the biosynthesis of organic signaling molecules, they are also impacted by a variety of inorganic species. Metallic nanoparticles, clusters, ions, and complexes can alter CYP expression, modify enzyme interactions with reductase partners, and serve as direct inhibitors. This commonly overlooked topic is reviewed here, with an emphasis on understanding the structural and physiochemical basis for these interactions. Intriguingly, while both organometallic and coordination compounds can act as potent CYP inhibitors, there is little evidence for the metabolism of inorganic compounds by CYPs, suggesting a potential alternative approach to evading issues associated with rapid modification and elimination of medically useful compounds.

Pharmaceutical opioids are intravenously or orally administered analgesics. While they are effective in relieving chronic and acute pain, their narrow window of therapeutic use contributes to the high occurrence of abuse. The associated abuse of this family of drugs can be correlated to the increase in dependency, overdose, and death of users. The negative effects of opioids extend beyond the physical and psychological effects experienced by the user to their unregulated synthesis and sale, which contribute to socioeconomic challenges and are a biproduct of this global public health epidemic. From clinical to point-of-care applications, the detection and real-time monitoring of this family of drug is critical in the fight to decrease abuse and improve use in clinical settings. Chromatographic separations and chromatography–mass spectrometry are traditional methods of opioid analyses, but the high cost, long analysis time, and absence of portability highlight the need for the development of fast, in situ, point-of-care analysis, or of community drug monitoring services. This review highlights recent electrochemical and optical (FTIR, Raman, colorimetric, and fluorescent) advances and biosensors for pharmaceutical and illicit opioid analysis. Specifically, an emphasis is placed on the detection of opioids and their metabolites in biological samples and in vitro cellular assays for clinical diagnosis and forensic applications. The challenges and prospects of the role of electrochemical sensors, biosensors, and optical sensors for opioid analysis in promoting clinical diagnosis, forensic study, point-of-care, and community drug monitoring services to reduce harm are also provided.

Motion: For The Outdated Significance of Morphological Assessment in Embryo Selection and the Rise of Advanced Technologies in Reproductive Medicine This symposium lecture presents a compelling debate, shedding light on the diminishing relevance of traditional static morphological assessment as a primary criterion for embryo. The speaker aims to challenge the long-standing reliance on morphological evaluation and advocate for the integration of cutting-edge technologies as a more reliable and objective means of embryo quality assessment. During the debate, the speaker will examine the limitations of morphological assessment, acknowledging its subjective nature, potential bias, and lack of comprehensive insight into embryonic development. The traditional morphological assessment based solely on visual appearance may not adequately predict an embryo’s true potential for successful implantation and healthy pregnancy. A variety of available new technologies will be highlighted during the lecture, showcasing alternatives to morphological embryo assessment. Attendees will gain insight into how these innovative methods can offer more objective and precise measures of embryo quality, enhancing the success rates of assisted reproductive procedures. The symposium will foster an engaging environment, encouraging active participation and thoughtful discussions among attendees from the reproductive medicine community. Participants will have the opportunity to challenge prevailing notions and explore the implications of adopting new technologies in clinical practice. In summary, this debate aims to catalyze a paradigm shift in embryo evaluation, emphasizing the need to move beyond traditional morphological assessment towards embracing advanced technologies, to optimize embryo selection and lead to improved outcomes in IVF. Motion: Against Morphological Assessment of Embryos is Not Outdated In the era of new technologies and artificial intelligence, looking good has never been more important! On-time progression through specific events is requisite for human preimplantation development and provides important visual insights into the health and physiology of the embryo. The formation of 2 distinct pronuclei, the times and patterns of cleavage, and how the cells compact are all key markers of development. Following compaction, the formation of the blastocyst is not only a temporally regulated event, but also an energetically demanding process, one that reflects the metabolic activity (a biomarker of viability) of the embryo. Further, at the blastocyst stage it is possible to visualise and quantitate the formation of both the inner cell mass (ICM) and trophectoderm, with all parameters of blastocyst development linked to transfer outcomes. The development of time-lapse incubation, which can be described as embryo morphology over time, has led to the incorporation of several key morphological features to support embryo selection, further reflecting the significance of embryo morphology. Interestingly, AI algorithms used for embryo selection reflect predominantly those morphological parameters that are known to affect transfer outcomes, such as blastocyst quality. While only specific molecular tests can accurately determine the precise chromosomal composition of an embryo, key morphological features and events can also be used to tag those embryos at greatest risk of being genetically abnormal. In conclusion, key morphological developmental events indirectly inform us of the physiology of the embryo (degree of blastocyst expansion reflects metabolic activity for example), and hence this is why morphological grading systems (such as the Gardner Grade for blastocysts) have been shown to be effective in embryo selection for over the past 25 years, and further have proven to be as useful as AI or biomarker analysis for the majority of patients.

Natural killer (NK) cells are cytotoxic innate immune cells, able to recognize and eliminate virus-infected as well as cancer cells. Metabolic reprogramming is crucial for their activity as they have enhanced energy and nutritional demands for their functions during an infection. Fatty acids (FAs) represent an important source of cellular energy and are essential for proliferation of immune cells. However, the precise role of FAs for NK cells activity in retrovirus infection was unknown. Here we show that activated NK cells increase the expression of the FA uptake receptor CD36 and subsequently the uptake of FAs upon acute virus infection. We found an enhanced flexibility of NK cells to utilize FAs as source of energy compare to naïve NK cells. NK cells that were able to generate energy from FAs showed an augmented target cell killing and increased expression of cytotoxic parameters. However, NK cells that were unable to generate energy from FAs exhibited a severely decreased migratory capacity. Our results demonstrate that NK cells require FAs in order to fight acute virus infection. Susceptibility to severe virus infections as it is shown for people with malnutrition may be augmented by defects in the FA processing machinery, which might be a target to therapeutically boost NK cell functions in the future.

BackgroundEstimations show that at least one in every 3 people in the world needs rehabilitation at some point in the course of their illness or injury. Access to rehabilitation services is an essential part of the continuum of care and is integral to achieving universal health coverage. However, most of the world’s population living in low- and middle-income countries, especially in the sub-Saharan African region, does not have access to adequate rehabilitation services. Wider adoption of digital solutions offers opportunities to support and enhance access to rehabilitation services in sub-Saharan Africa. A region where there is a greater burden and need for these services. There is also little published research about digital rehabilitation in sub-Saharan Africa, as it is an underexamined topic in the region. ObjectiveThis scoping review aims to provide a comprehensive picture of the current evidence of digital interventions in rehabilitation implemented in any health, social, educational, or community setting in the sub-Saharan Africa region. MethodsWe will conduct a scoping review using Arksey and O’Malley’s methodological framework and follow the Joanna Briggs Institute methodology for scoping reviews. We will develop search strategies for a selected number of web-based databases, search for peer-reviewed scientific publications until September 2023, and screen the reference lists of relevant articles. We will include research articles if they describe or report the use of digital interventions in the rehabilitation of patients with any health problem or disability in sub-Saharan Africa. For selected articles, we will extract data using a customized data extraction form and use thematic analysis to compare the findings across studies. ResultsThe preliminary database search in MEDLINE (EBSCO) was completed in May 2023. The research team will conduct a search of relevant articles in the autumn. The results will be synthesized and reported under the key conceptual categories of this review, and we expect the final scoping review to be ready for submission in early 2024. ConclusionsWe expect to find gaps in the research and a lack of detailed information about digital rehabilitation interventions in sub-Saharan Africa, as well as potential areas for further study. We will identify opportunities to inform the development of digital rehabilitation interventions. International Registered Report Identifier (IRRID)PRR1-10.2196/48952

Methane emission by terrestrial invertebrates is restricted to millipedes, termites, cockroaches, and scarab beetles. The arthropod-associated archaea known to date belong to the orders Methanobacteriales, Methanomassiliicoccales, Methanomicrobiales, and Methanosarcinales, and in a few cases also to non-methanogenic Nitrososphaerales and Bathyarchaeales. However, all major host groups are severely undersampled, and the taxonomy of existing lineages is not well developed. Full-length 16S rRNA gene sequences and genomes of arthropod-associated archaea are scarce, reference databases lack resolution, and the names of many taxa are either not validly published or under-classified and require revision. Here, we investigated the diversity of archaea in a wide range of methane-emitting arthropods, combining phylogenomic analysis of isolates and metagenome-assembled genomes (MAGs) with amplicon sequencing of full-length 16S rRNA genes. Our results allowed us to describe numerous new species in hitherto undescribed taxa among the orders Methanobacteriales (Methanacia, Methanarmilla, Methanobaculum, Methanobinarius, Methanocatella, Methanoflexus, Methanorudis, and Methanovirga, all gen. nova), Methanomicrobiales (Methanofilum and Methanorbis, both gen. nova), Methanosarcinales (Methanofrustulum and Methanolapillus, both gen. nova), Methanomassiliicoccales (Methanomethylophilaceae fam. nov., Methanarcanum, Methanogranum, Methanomethylophilus, Methanomicula, Methanoplasma, Methanoprimaticola, all gen. nova), and the new family Bathycorpusculaceae (Bathycorpusculum gen. nov.). Reclassification of amplicon libraries from this and previous studies using this new taxonomic framework revealed that arthropods harbor only CO2 and methyl-reducing hydrogenotrophic methanogens. Numerous genus-level lineages appear to be present exclusively in arthropods, suggesting long evolutionary trajectories with their termite, cockroach, and millipede hosts, and a radiation into various microhabitats and ecological niches provided by their digestive tracts (e.g., hindgut compartments, gut wall, or anaerobic protists). The distribution patterns among the different host groups are often complex, indicating a mixed mode of transmission and a parallel evolution of invertebrate and vertebrate-associated lineages.

Oxalate is a small compound found in certain plant-derived foods and is a major component of calcium oxalate (CaOx) kidney stones. Individuals that consume oxalate enriched meals have an increased risk of forming urinary crystals, which are precursors to CaOx kidney stones. We previously reported that a single dietary oxalate load induces nanocrystalluria and reduces monocyte cellular bioenergetics in healthy adults. The purpose of this study was to extend these investigations to identify specific oxalate-mediated mechanisms in monocytes and macrophages. We performed RNA-Sequencing analysis on monocytes isolated from healthy subjects exposed to a high oxalate (8 mmol) dietary load. RNA-sequencing revealed 1,198 genes were altered and Ingenuity Pathway Analysis demonstrated modifications in several pathways including Interleukin-10 (IL-10) anti-inflammatory cytokine signaling, mitochondrial metabolism and function, oxalic acid downstream signaling, and autophagy. Based on these findings, we hypothesized that oxalate induces mitochondrial and lysosomal dysfunction in monocytes and macrophages via IL-10 and reactive oxygen species (ROS) signaling which can be reversed with exogenous IL-10 or Mitoquinone (MitoQ; a mitochondrial targeted antioxidant). We exposed monocytes and macrophages to oxalate in an in-vitro setting which caused oxidative stress, a decline in IL-10 cytokine levels, mitochondrial and lysosomal dysfunction, and impaired autophagy in both cell types. Administration of exogenous IL-10 and MitoQ attenuated these responses. These findings suggest that oxalate impairs metabolism and immune response via IL-10 signaling and mitochondrial ROS generation in both monocytes and macrophages which can be potentially limited or reversed. Future studies will examine the benefits of these therapies on CaOx crystal formation and growth in vivo.

Purpose: Clinical trials comparing the efficacy of adjuvant chemotherapy (CT) and chemo radiation therapy (CTRT) for stomach adenocarcinoma have reported equivocal results. Hence, the current retrospective cohort study assessed the long-term survival and recurrence outcomes of these therapies, to generate evidence in a real-world scenario. Methods and Materials: Pathologically confirmed patients with stomach adenocarcinoma aged ≥18 years who underwent gastrectomy and D2 lymph nodal dissection at a tertiary cancer hospital from January 2010 to October 2017 were enrolled. Hospital-based follow-up was performed until December 2021. Data were gathered from electronic medical records, supplemented by telephonic interviews for patients who could not come for physical follow-up. CT-alone and CTRT cohorts were compared in terms of survival and recurrence outcomes. Results: The analysis included 158 patients (mean age, 56.42 years; 63.9% male; CT-alone cohort, 69; CTRT cohort, 89). Patients in the CTRT cohort had significantly worse tumor characteristics at baseline (29.2% had the diffuse type of tumor, 94.4% had stage II or III, 68.5% had lympho-vascular space invasion, and 85.4% had lymph node involvement). Recurrence was observed in 13 (19.7%) of the 76 followed-up patients. Although locoregional recurrence was higher in the CT-alone cohort (7 vs 2), distant metastasis was higher in the CTRT cohort (3 vs 1). The overall 5-year survival was 67.0% (SE, 5.0%) and 5-year recurrence-free survival (RFS) was 75.0% (SE, 5.0%). On multivariate Cox regression, no variable was significantly associated with the overall survival, whereas age, positive lymph nodes without extracapsular extension, and lymph node-negative were significantly associated with RFS. The CTRT cohort had significantly (84.0%) higher RFS (hazard ratio, 0.161; 95% CI, 0.056-0.464; P < .001). Conclusions: Patients who received adjuvant CTRT after D2 dissection showed similar overall survival but significantly higher RFS than the CT-alone cohort, despite having worse baseline tumor characteristics.

The volume and composition of a thin layer of liquid covering the airway surface defend the lung from inhaled pathogens and debris. Airway epithelia secrete Cl– into the airway surface liquid through cystic fibrosis transmembrane conductance regulator (CFTR) channels, thereby increasing the volume of airway surface liquid. The discovery that pulmonary ionocytes contain high levels of CFTR led us to predict that ionocytes drive secretion. However, we found the opposite. Elevating ionocyte abundance increased liquid absorption, whereas reducing ionocyte abundance increased secretion. In contrast to other airway epithelial cells, ionocytes contained barttin/Cl– channels in their basolateral membrane. Disrupting barttin/Cl– channel function impaired liquid absorption, and overexpressing barttin/Cl– channels increased absorption. Together, apical CFTR and basolateral barttin/Cl– channels provide an electrically conductive pathway for Cl– flow through ionocytes, and the transepithelial voltage generated by apical Na+ channels drives absorption. These findings indicate that ionocytes mediate liquid absorption, and secretory cells mediate liquid secretion. Segregating these counteracting activities to distinct cell types enables epithelia to precisely control the airway surface. Moreover, the divergent role of CFTR in ionocytes and secretory cells suggests that cystic fibrosis disrupts both liquid secretion and absorption.

The role of aberrant glycosylation in pancreatic ductal adenocarcinoma (PDAC) remains an under-investigated area of research. In this study, we determined that ST6 β-galactoside α2,6 sialyltransferase 1 (ST6GAL1), which adds α2,6-linked sialic acids to N-glycosylated proteins, was upregulated in patients with early-stage PDAC and was further increased in advanced disease. A tumor-promoting function for ST6GAL1 was elucidated using tumor xenograft experiments with human PDAC cells. Additionally, we developed a genetically engineered mouse (GEM) model with transgenic expression of ST6GAL1 in the pancreas and found that mice with dual expression of ST6GAL1 and oncogenic KRASG12D had greatly accelerated PDAC progression compared with mice expressing KRASG12D alone. As ST6GAL1 imparts progenitor-like characteristics, we interrogated ST6GAL1’s role in acinar to ductal metaplasia (ADM), a process that fosters neoplasia by reprogramming acinar cells into ductal, progenitor-like cells. We verified ST6GAL1 promotes ADM using multiple models including the 266-6 cell line, GEM-derived organoids and tissues, and an in vivo model of inflammation-induced ADM. EGFR is a key driver of ADM and is known to be activated by ST6GAL1-mediated sialylation. Importantly, EGFR activation was dramatically increased in acinar cells and organoids from mice with transgenic ST6GAL1 expression. These collective results highlight a glycosylation-dependent mechanism involved in early stages of pancreatic neoplasia.

Background: Limb loss limits functioning and restricts participation in various environments. Persons with lower limb amputations (PLLA) experience challenges ranging from self-care and independence to psychological disorders that negatively impact their functioning. Objectives: To assess the functioning and the level of disability of PLLA with or without prostheses in Rwanda. Method: A descriptive, cross-sectional study was conducted among PLLAs aged 18 years and above in 10 districts of Rwanda. A total of 247 participants were purposively selected to fill the questionnaires. Descriptive and inferential statistics using t-test and binary logistic regression were performed to analyse data using Statistical Package for Social Sciences (SPSS) (version 21.0). Results: Out of 247 PLLA, 99 (40.1%) had prostheses and remaining 148 (59.9%) did not. Majority of PLLA without prostheses reported having more difficulties in mobility (s.d. 3.98), participation (s.d. 5.18) and life activities (s.d. 3.87). The majority of PLLA reported mild and moderate functioning in the domains of cognitive (odds ratio [OR] 8.842, 5.384 with 95% confidence interval [CI]) mobility (OR 16.154, 2.485 with 95% CI) and participation (OR 13.299, 15.282 with 95% CI). Conclusion: Persons without prostheses demonstrated reduced level of functioning and high levels of disability compared to those with prostheses in all domains. However, the mobility, self-activities and the participation domains were the mainly affected. Contribution: The study helps to understand the needs of the PLLA and emphasises that not only having prostheses can improve functioning but also emphasises the psychosocial aspects to reduce disability.

Laboratory experiments measured sediment deposition and turbulent kinetic energy (TKE) in bare and vegetated channels. The model vegetation represented a mangrove pneumatophore canopy. Three solid volume fractions were considered (ϕ= 0.01, 0.02, and 0.04). For the same channel-averaged velocity, the vegetated region had elevated near-bed TKE compared to the bare region. Net deposition in both regions was measured by adding a sediment slurry of 11-micron solid glass spheres to the flume and collecting the deposited sediment from the flume baseboards after a 4-hr experiment. The elevated near-bed TKE in the vegetated region resulted in lower deposition compared to the bare region. A model for deposition probability written in terms of near-bed TKE (TKE model) more accurately predicted the measured deposition than a model based on bed shear stress (τb model). Application of the model to field conditions suggested that, by inhibiting deposition, vegetation-generated TKE facilitates the delivery of sediment farther into the mangrove forest than would be achieved without vegetation-generated TKE.

Background Anthracycline‐induced cardiomyopathy is a leading cause of premature death in childhood cancer survivors, presenting a need to understand the underlying pathogenesis. We sought to examine differential blood‐based mRNA expression profiles in anthracycline‐exposed childhood cancer survivors with and without cardiomyopathy. Methods and Results We designed a matched case‐control study (Children's Oncology Group‐ALTE03N1) with mRNA sequencing on total RNA from peripheral blood in 40 anthracycline‐exposed survivors with cardiomyopathy (cases) and 64 matched survivors without (controls). DESeq2 identified differentially expressed genes. Ingenuity Pathway Analyses (IPA) and Gene Set Enrichment Analyses determined the potential roles of altered genes in biological pathways. Functional validation was performed by gene knockout in human‐induced pluripotent stem cell‐derived cardiomyocytes using CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR‐associated protein 9) technology. Median age at primary cancer diagnosis for cases and controls was 8.2 and 9.7 years, respectively. Thirty‐six differentially expressed genes with fold change ≥±2 were identified; 35 were upregulated. IPA identified “hepatic fibrosis” and “iron homeostasis” pathways to be significantly modulated by differentially expressed genes, including toxicology functions of myocardial infarction, cardiac damage, and cardiac dilation. Leading edge analysis from Gene Set Enrichment Analyses identified lactate dehydrogenase A (LDHA) and cluster of differentiation 36 (CD36) genes to be significantly upregulated in cases. Interleukin 1 receptor type 1, 2 (IL1R1, IL1R2), and matrix metalloproteinase 8, 9 (MMP8, MMP9) appeared in multiple canonical pathways. LDHA‐knockout human‐induced pluripotent stem cell‐derived cardiomyocytes showed increased sensitivity to doxorubicin. Conclusions We identified differential mRNA expression profiles in peripheral blood of anthracycline‐exposed childhood cancer survivors with and without cardiomyopathy. Upregulation of LDHA and CD36 genes suggests metabolic perturbations in a failing heart. Dysregulation of proinflammatory cytokine receptors IL1R1 and IL1R2 and matrix metalloproteinases, MMP8 and MMP9 indicates structural remodeling that accompanies the clinical manifestation of symptomatic cardiotoxicity.

IMPORTANCE:. Extubation failure (EF) after pediatric cardiac surgery is associated with increased morbidity and mortality. OBJECTIVES:. We sought to describe the risk factors associated with early (< 48 hr) and late (48 hr ≤ 168 hr) EF after pediatric cardiac surgery and the clinical implications of these two types of EF. DESIGN, SETTING, AND PARTICIPANTS:. Retrospective cohort study using prospectively collected clinical data for the Pediatric Cardiac Critical Care Consortium (PC4) Registry. Pediatric patients undergoing Society of Thoracic Surgeons benchmark operation or heart transplant between 2013 and 2018 available in the PC4 Registry were included. MAIN OUTCOMES AND MEASURES:. We analyzed demographics and risk factors associated with EFs (primary outcome) including by type of surgery. We identified potentially modifiable risk factors. Clinical outcomes of mortality and length of stay (LOS) were reported. RESULTS:. Overall 18,278 extubations were analyzed. Unplanned extubations were excluded from the analysis. The rate of early EF was 5.2% (948) and late EF was 2.5% (461). Cardiopulmonary bypass time, ventilator duration, airway anomaly, genetic abnormalities, pleural effusion, and diaphragm paralysis contributed to both early and late EF. Extubation during day remote from shift change and nasotracheal route of initial intubation was associated with decreased risk of early EF. Extubation in the operating room was associated with an increased risk of early EF but with decreased risk of late EF. Across all operations except arterial switch, EF portrayed an increased burden of LOS and mortality. CONCLUSION AND RELEVANCE:. Both early and late EF are associated with significant increase in LOS and mortality. Study provides potential benchmarking data by type of surgery. Modifiable risk factors such as route of intubation, time of extubation as well as treatment of potential contributors such as diaphragm paralysis or pleural effusion can serve as focus areas for reducing EFs.

Introduction: A common neurosurgical condition, chronic subdural haematoma (cSDH) typically affects older people with other underlying health conditions. The care of this potentially vulnerable cohort is often, however, fragmented and suboptimal. In other complex conditions, multidisciplinary guidelines have transformed patient experience and outcomes, but no such framework exists for cSDH. This paper outlines a protocol to develop the first comprehensive multidisciplinary guideline from diagnosis to long-term recovery with cSDH. Methods: The project will be guided by a steering group of key stakeholders and professional organisations and will feature patient and public involvement. Multidisciplinary thematic working groups will examine key aspects of care to formulate appropriate, patient-centered research questions, targeted with evidence review using the GRADE framework. The working groups will then formulate draft clinical recommendations to be used in a modified Delphi process to build consensus on guideline contents. Conclusions: We present a protocol for the development of a multidisciplinary guideline to inform the care of patients with a cSDH, developed by cross-disciplinary working groups and arrived at through a consensus-building process, including a modified online Delphi.

Abstract Background Montessori education is the oldest and most widely implemented alternative education in the world, yet its effectiveness has not been clearly established. Objectives The primary objective of this review was to examine the effectiveness of Montessori education in improving academic and nonacademic outcomes compared to traditional education. The secondary objectives were to determine the degree to which grade level, Montessori setting (public Montessori vs. private Montessori), random assignment, treatment duration, and length of follow‐up measurements moderate the magnitude of Montessori effects. Search Methods We searched for relevant studies in 19 academic databases, in a variety of sources known to publish gray literature, in Montessori‐related journals, and in the references of studies retrieved through these searches. Our search included studies published during or before February 2020. The initial search was performed in March 2014 with a follow‐up search in February 2020. Selection Criteria We included articles that compared Montessori education to traditional education, contributed at least one effect size to an academic or nonacademic outcome, provided sufficient data to compute an effect size and its variance, and showed sufficient evidence of baseline equivalency–through random assignment or statistical adjustment–of Montessori and traditional education groups. Data Collection and Analysis To synthesize the data, we used a cluster‐robust variance estimation procedure, which takes into account statistical dependencies in the data. Otherwise, we used standard methodological procedures as specified in the Campbell Collaboration reporting and conduct standards. Main Results Initial searches yielded 2012 articles, of which 173 were considered in detail to determine whether they met inclusion/exclusion criteria. Of these, 141 were excluded and 32 were included. These 32 studies yielded 204 effect sizes (113 academic and 91 nonacademic) across 132,249 data points. In the 32 studies that met minimum standards for inclusion, including evidence of baseline equivalence, there was evidence that Montessori education outperformed traditional education on a wide variety of academic and nonacademic outcomes. For academic outcomes, Hedges' g effect sizes, where positive values favor Montessori, ranged from 0.26 for general academic ability (with high quality evidence) to 0.06 for social studies. The quality of evidence for language (g = 0.17) and mathematics (g = 0.22) was also high. The effect size for a composite of all academic outcomes was 0.24. Science was the only academic outcome that was deemed to have low quality of evidence according to the GRADE approach. Effect sizes for nonacademic outcomes ranged from 0.41 for students' inner experience of school to 0.23 for social skills. Both of these outcomes were deemed as having low quality of evidence. Executive function (g = 0.36) and creativity (g = 0.26) had moderate quality of evidence. The effect size for a composite of all nonacademic outcomes was 0.33. Moderator analyses of the composite academic and nonacademic outcomes showed that Montessori education resulted in larger effect sizes for randomized studies compared to nonrandomized studies, for preschool and elementary settings compared to middle school or high school settings, and for private Montessori compared to public Montessori. Moderator analyses for treatment duration and duration from intervention to follow‐up data collection were inconclusive. There was some evidence for a lack of small sample‐size studies in favor of traditional education, which could be an indicator of publication bias. However, a sensitivity analysis indicated that the findings in favor of Montessori education were nonetheless robust. Authors' Conclusions Montessori education has a meaningful and positive impact on child outcomes, both academic and nonacademic, relative to outcomes seen when using traditional educational methods.