TRPV4 and chloride channels mediate volume sensing in trabecular meshwork cells.

Aqueous humor drainage from the anterior eye determines intraocular pressure (IOP) under homeostatic and pathological conditions. Swelling of the trabecular meshwork (TM) alters its flow resistance but the mechanisms that sense and transduce osmotic gradients remain poorly understood. We investigated TM osmotransduction and its role in calcium and chloride homeostasis using molecular analyses, optical imaging and electrophysiology. Anisosmotic conditions elicited proportional changes in TM cell volume, with swelling, but not shrinking, evoking elevations in intracellular calcium concentration [Ca2+]TM. Hypotonicity-evoked calcium signals were sensitive to HC067047, a selective blocker of TRPV4 channels, whereas the agonist GSK1016790A promoted swelling under isotonic conditions. TRPV4 inhibition partially suppressed hypotonicity-induced volume increases and reduced the magnitude of the swelling-induced membrane current, with a substantial fraction of the swelling-evoked current abrogated by Cl- channel antagonists DIDS and niflumic acid. The transcriptome of volume-sensing chloride channel candidates in primary human was dominated by ANO6 transcripts, with moderate expression of ANO3, ANO7, ANO10 transcripts and low expression of LTTRC genes that encode constituents of the volume-activated anion channel. Imposition of 190 mOsm but not 285 mOsm hypotonic gradients increased conventional outflow in mouse eyes. TRPV4-mediated cation influx thus works with Cl- efflux to sense and respond to osmotic stress, potentially contributing to pathological swelling, calcium overload and intracellular signaling that could exacerbate functional disturbances in inflammatory disease and glaucoma.

Phosphorylation of the nuclear poly(A) binding protein (PABPN1) during mitosis protects mRNA from hyperadenylation and maintains transcriptome dynamics.

Polyadenylation controls mRNA biogenesis, nucleo-cytoplasmic export, translation and decay. These processes are interdependent and coordinately regulated by poly(A)-binding proteins (PABPs), yet how PABPs are themselves regulated is not fully understood. Here, we report the discovery that human nuclear PABPN1 is phosphorylated by mitotic kinases at four specific sites during mitosis, a time when nucleoplasm and cytoplasm mix. To understand the functional consequences of phosphorylation, we generated a panel of stable cell lines inducibly over-expressing PABPN1 with point mutations at these sites. Phospho-inhibitory mutations decreased cell proliferation, highlighting the importance of PABPN1 phosphorylation in cycling cells. Dynamic regulation of poly(A) tail length and RNA stability have emerged as important modes of gene regulation. We therefore employed long-read sequencing to determine how PABPN1 phospho-site mutants affected poly(A) tails lengths and TimeLapse-seq to monitor mRNA synthesis and decay. Widespread poly(A) tail lengthening was observed for phospho-inhibitory PABPN1 mutants. In contrast, expression of phospho-mimetic PABPN1 resulted in shorter poly(A) tails with increased non-A nucleotides, in addition to increased transcription and reduced stability of a distinct cohort of mRNAs. Taken together, PABPN1 phosphorylation remodels poly(A) tails and increases mRNA turnover, supporting the model that enhanced transcriptome dynamics reset gene expression programs across the cell cycle.

Non-Tobacco Nicotine Dependence and Rates of Postoperative Complications in Total Knee Arthroplasty: A Propensity-Matched Comparison.

INTRODUCTION: Tobacco use elevates the incidence of postoperative complications and remains a key modifiable risk factor of perioperative surgical optimization. It remains unclear whether non-tobacco nicotine dependence confers an increased risk of surgical complications. This study evaluates postoperative complications in patients with non-tobacco nicotine dependence for total knee arthroplasty (TKA). METHODS: We queried the TriNetX health database using Current Procedural Terminology and International Classification of Diseases, 10th Revision (ICD-10) codes and identified two cohorts for evaluation. Cohort A was defined as patients who had a TKA; had a dependence on nicotine; did not have nicotine dependence to cigarettes, chewing tobacco, other tobacco products; and were between the ages of 35 and 90 years. Cohort B was defined as patients who had a TKA but did not have a dependence on nicotine or a personal history of nicotine dependence and were between the ages of 35 and 90 years. RESULTS: This study analyzed a total of 10,594 non-tobacco nicotine-dependent patients and 175,079 non-dependent patients who underwent TKA. In the analysis of propensity-matched cohorts, non-tobacco nicotine-dependent patients demonstrated an increased rate of various postoperative complications within 90 days. Dependent patients saw a significantly increased risk of infection after a procedure (P < 0.001), deep vein thrombosis (P < 0.001), pulmonary embolism (P < 0.001), sepsis (P = 0.0065), and prosthetic joint infection (P = 0.0361) and a higher 3-year revision rate (P = 0.0084). DISCUSSION: Non-tobacco nicotine dependence demonstrated an increased associated risk of postoperative surgical complications for patients undergoing TKA. Orthopaedic surgeons should consider evaluating non-tobacco nicotine dependence within their surgical optimization protocol. LEVEL OF EVIDENCE: Level III, Prognostic.

Unifying community-wide whole-brain imaging datasets enables robust automated neuron identification and reveals determinants of neuron positioning in C. elegans.

We develop a data harmonization approach for C. elegans volumetric microscopy data, still or video, consisting of a standardized format, data pre-processing techniques, and a set of human-in-the-loop machine learning based analysis software tools. We unify a diverse collection of 118 whole-brain neural activity imaging datasets from 5 labs, storing these and accompanying tools in an online repository called WormID (wormid.org). We use this repository to train three existing automated cell identification algorithms to, for the first time, enable accuracy in neural identification that generalizes across labs, approaching human performance in some cases. We mine this repository to identify factors that influence the developmental positioning of neurons. To facilitate communal use of this repository, we created open-source software, code, web-based tools, and tutorials to explore and curate datasets for contribution to the scientific community. This repository provides a growing resource for experimentalists, theorists, and toolmakers to (a) study neuroanatomical organization and neural activity across diverse experimental paradigms, (b) develop and benchmark algorithms for automated neuron detection, segmentation, cell identification, tracking, and activity extraction, and (c) inform models of neurobiological development and function.

[Navicular bone fractures in horses: Prognosis after conservative and surgical treatment]. / Strahlbein-Frakturen beim Pferd: Prognose nach konservativer und chirurgischer Therapie.

INTRODUCTION: The aim of this retrospective study is to describe and compare conservative and surgical treatment of navicular fractures in horses. An attempt is made to identify critical points that can favorably influence the prognosis of this orthopedic disease. All horses treated for a navicular fracture at the Equine Clinic, Vetsuisse Faculty, University of Zurich between 2005 and 2017 were included in this study. The severity of lameness at initial examination, radiographic assessment, hoof conformation, treatment (conservative or surgical), complications and outcome were determined from the medical records. Conservative and surgical treatment consisted of stable rest, a controlled exercise program and therapeutic orthopedic shoeing. During surgical treatment, fracture reduction was also carried out with a cortical screw. Computer assisted surgery were used in five horses and computer tomography in three horses. Follow-up examinations included clinical and radiographic examinations. The outcome was divided into three categories: 1 = very good; 2 = good; 3 = poor. Twelve horses met the inclusion criteria; Four horses were treated conservatively and eight were treated surgically. After conservative treatment, two horses (50 %) had a very good outcome and two (50 %) had a good outcome. After surgical treatment, four horses (50 %) had a very good outcome and four (50 %) had a poor outcome. Overall, 67 % of horses had a very good or good outcome, although radiographic signs of bone healing was not present in any of these cases. Various complications were identified, such as the fracture of a screw, the fragmentation of the small navicular bone fragment, the development of osteoarthrosis in the coffin joint and progressive podotrochosis. This study shows that the prognosis of navicular fractures is generally cautious and that degenerative changes to the navicular apparatus worsen the prognosis. In the present study, surgical treatment did not improve the prognosis of navicular fractures despite the intrasurgical use of three-dimensional imaging. However, technical advances could reduce the complication rate in the future.

INTRODUCTION: Le but de cette étude rétrospective était de décrire et de comparer le traitement conservateur et chirurgical des fractures de l'os naviculaires chez le cheval. Une tentative est faite pour identifier les points critiques qui peuvent influencer favorablement le pronostic de cette maladie orthopédique. Tous les chevaux traités pour une fracture de l'os naviculaire à la Clinique équine de la Faculté Vetsuisse de l'Université de Zurich entre 2005 et 2017 ont été inclus dans cette étude. La gravité de la boiterie lors de l'examen initial, l'évaluation radiographique, la conformation du sabot, le traitement (conservateur ou chirurgical), les complications et le résultat ont été déterminés à partir des dossiers médicaux. Le traitement conservateur et chirurgical consistait en un repos au box, un programme d'exercice contrôlé et un ferrage orthopédique thérapeutique. Au cours du traitement chirurgical, une réduction de la fracture a également été effectuée à l'aide d'une vis corticale. La chirurgie assistée par ordinateur a été utilisée sur cinq chevaux et la tomographie assistée par ordinateur sur trois chevaux. Les examens de suivi comprenaient des examens cliniques et radiographiques. Les résultats ont été divisés en trois catégories: 1 = très bon; 2 = bon; 3 = mauvais. Douze chevaux répondaient aux critères d'inclusion ; quatre chevaux ont été traités de manière conservatrice et huit ont été traités chirurgicalement. Après un traitement conservateur, deux chevaux (50 %) ont eu un très bon résultat et deux (50 %) un bon résultat. Après le traitement chirurgical, quatre chevaux (50 %) ont eu un très bon résultat et quatre (50 %) un mauvais résultat. Dans l'ensemble, 67 % des chevaux ont eu un très bon ou un bon résultat, bien que les signes radiographiques de guérison osseuse n'aient été présents dans aucun de ces cas. Diverses complications ont été identifiées, telles que la fracture d'une vis, la fragmentation du petit fragment de l'os naviculaire, le développement d'une arthrose dans l'articulation interphalangienne distale et une podotrochlose progressive. Cette étude montre que le pronostic des fractures de l'os naviculaire est généralement réservé et que les modifications dégénératives de l'appareil naviculaire aggravent le pronostic. Dans la présente étude, le traitement chirurgical n'a pas amélioré le pronostic des fractures du naviculaire malgré l'utilisation intra-chirurgicale de l'imagerie tridimensionnelle. Cependant, les progrès techniques pourraient réduire le taux de complications dans le futur.

Laburnicotides A-F: Acyclic N-Acetyl Oligopeptides from the Nematode-Cyst-Associated Fungus Laburnicola nematophila.

Nematode-associated fungi revealed the potential to produce a broad spectrum of chemical scaffolds. In this study, a mycelial extract of Laburnicola nematophila, a fungal strain derived from the cereal cyst nematode Heterodera filipjevi, was chemically explored and afforded six unprecedentedly reported acylic N-acetyl oligopeptides, laburnicotides A-F (1-6). Structure elucidation of the isolated compounds was established based on comprehensive 1D and 2D NMR spectroscopic analyses together with the acquired HR-ESI-MS spectrometric data. The absolute configuration of amino acid residues in 1-6 was established by performing advanced Marfey's derivatization method. All isolated compounds were assessed for their cytotoxic, antimicrobial, antiviral, and nematicidal activities with no potential activity observed.

Valproic Acid Causes Redox-Regulated Post-Translational Protein Modifications That Are Dependent upon P19 Cellular Differentiation States.

Valproic acid (VPA) is a common anti-epileptic drug and known neurodevelopmental toxicant. Although the exact mechanism of VPA toxicity remains unknown, recent findings show that VPA disrupts redox signaling in undifferentiated cells but has little effect on fully differentiated neurons. Redox imbalances often alter oxidative post-translational protein modifications and could affect embryogenesis if developmentally critical proteins are targeted. We hypothesize that VPA causes redox-sensitive post-translational protein modifications that are dependent upon cellular differentiation states. Undifferentiated P19 cells and P19-derived neurons were treated with VPA alone or pretreated with D3T, an inducer of the nuclear factor erythroid 2-related factor 2 (NRF2) antioxidant pathway, prior to VPA exposure. Undifferentiated cells treated with VPA alone exhibited an oxidized glutathione redox couple and increased overall protein oxidation, whereas differentiated neurons were protected from protein oxidation via increased S-glutathionylation. Pretreatment with D3T prevented the effects of VPA exposure in undifferentiated cells. Taken together, our findings support redox-sensitive post-translational protein alterations in undifferentiated cells as a mechanism of VPA-induced developmental toxicity and propose NRF2 activation as a means to preserve proper neurogenesis.

A joint physics and radiobiology DREAM team vision - Towards better response prediction models to advance radiotherapy.

Radiotherapy developed empirically through experience balancing tumour control and normal tissue toxicities. Early simple mathematical models formalized this practical knowledge and enabled effective cancer treatment to date. Remarkable advances in technology, computing, and experimental biology now create opportunities to incorporate this knowledge into enhanced computational models. The ESTRO DREAM (Dose Response, Experiment, Analysis, Modelling) workshop brought together experts across disciplines to pursue the vision of personalized radiotherapy for optimal outcomes through advanced modelling. The ultimate vision is leveraging quantitative models dynamically during therapy to ultimately achieve truly adaptive and biologically guided radiotherapy at the population as well as individual patient-based levels. This requires the generation of models that inform response-based adaptations, individually optimized delivery and enable biological monitoring to provide decision support to clinicians. The goal is expanding to models that can drive the realization of personalized therapy for optimal outcomes. This position paper provides their propositions that describe how innovations in biology, physics, mathematics, and data science including AI could inform models and improve predictions. It consolidates the DREAM team's consensus on scientific priorities and organizational requirements. Scientifically, it stresses the need for rigorous, multifaceted model development, comprehensive validation and clinical applicability and significance. Organizationally, it reinforces the prerequisites of interdisciplinary research and collaboration between physicians, medical physicists, radiobiologists, and computational scientists throughout model development. Solely by a shared understanding of clinical needs, biological mechanisms, and computational methods, more informed models can be created. Future research environment and support must facilitate this integrative method of operation across multiple disciplines.

The education of experience: Mixed methods evidence demonstrates the benefit of multiparity.

PROBLEM: Existing quantitative data is conflicting concerning whether multiparous birthing parents (individuals with an older child(ren)) experience an easier postpartum transition compared to primiparous birthing parents (first time parents). AIM: This convergent mixed methods study leverages the depth of qualitative inquiry to seek a clearer understanding of the way in which acquired parenting experience contributes to observed quantitative differences in outcomes between parity groups. This work can serve as a first step in planning for supportive interventions that effectively address the postpartum needs of both parity groups. METHODS: Thirty birthing parents (43.3% multiparous; 46.7% racial minorities) completed measures of postpartum functioning, perceived stress, anxiety symptoms, and depression symptoms as well as an interview inquiring about factors impacting postpartum functioning. Scores on postpartum functioning and emotional wellbeing were compared between parity groups, and these findings were merged with the qualitative data on firsthand parenting experience to clarify how acquired experience impacts functioning and emotional wellbeing during the postpartum transition. FINDINGS: Primiparous parents reported significantly: worse postpartum functioning, higher perceived stress, higher levels of depression symptoms, and higher levels of anxiety symptoms. Participants' qualitative report of how acquired parenting experience impacts wellbeing suggests that experience grants parents skills, knowledge, and the opportunity to disconfirm maladaptive cognitions about parenting which allows for increased comfort and confidence in the parental role. CONCLUSIONS: The practical and psychological resources gained from acquiring parenting experience during one's first postpartum period appear to be brought forward into subsequent pregnancies and protect against threats to functioning and emotional wellbeing.

Repair of leukemia-associated single nucleotide variants via interallelic gene conversion.

CRISPR-Cas9 is a useful tool for inserting precise genetic alterations through homology-directed repair (HDR), although current methods rely on provision of an exogenous repair template. Here, we tested the possibility of repairing heterozygous single nucleotide variants (SNVs) using the cell's own wild-type allele rather than an exogenous template. Using high-fidelity Cas9 to perform allele-specific CRISPR across multiple human leukemia cell lines as well as in primary hematopoietic cells from patients with leukemia, we find high levels of reversion to wild-type in the absence of exogenous template. Moreover, we demonstrate that bulk treatment to revert a truncating mutation in ASXL1 using CRISPR-mediated interallelic gene conversion (IGC) is sufficient to prolong survival in a human cell line-derived xenograft model (median survival 33 days vs 27.5 days; p = 0.0040). These results indicate that IGC can be applied to numerous types of leukemia and can meaningfully alter cellular phenotypes at scale. Because our method targets single-base mutations, rather than larger variants targeted by IGC in prior studies, it greatly expands the pool of risk-increasing genetic lesions which could potentially be targeted by IGC. This technique may reduce cost and complexity for experiments modeling phenotypic consequences of SNVs. The principles of SNV-specific IGC demonstrated in this proof-of-concept study could be applied to investigate the phenotypic effects of targeted clonal reduction of leukemogenic SNV driver mutations.

Phaseolus vulgaris STP13.1 is an H+-coupled monosaccharide transporter, present in source leaves and seed coats, with higher substrate affinity at depolarized potentials.

Sugar transport proteins (STPs) are high-affinity H+-coupled hexose symporters. Recently, the contribution of STP13 to bacterial and fungal pathogen resistance across multiple plant species has garnered significant interest. Quantitative PCR analysis of source leaves, developing embryos, and seed coats of Phaseolus vulgaris L. (common bean) revealed that PvSTP13.1 was expressed in source leaves and seed coats throughout seed development. In contrast, PvSTP13.1 transcripts were detected at exceedingly low levels in developing embryos. To characterize the transport mechanism, PvSTP13.1 was expressed in Xenopus laevis oocytes, and inward-directed currents were analyzed using two-electrode voltage clamping. PvSTP13.1 was shown to function as an H+-coupled monosaccharide symporter exhibiting a unique high affinity for hexoses and aldopentoses at depolarized membrane potentials. Specifically, of the 31 assessed substrates, which included aldohexoses, deoxyhexoses, fructose, 3-O-methyl-D-glucose, aldopentoses, polyols, glycosides, disaccharides, trisaccharides, and glucuronic acid, PvSTP13.1 displayed the highest affinity (K 0.5) for glucose (43 µM), mannose (92 µM), galactose (145 µM), fructose (224 µM), xylose (1.0 mM), and fucose (3.7 mM) at pH 5.6 at a depolarized membrane potential of -40 mV. The results presented here suggest PvSTP13.1 contributes to retrieval of hexoses from the apoplasmic space in source leaves and coats of developing seeds.

Enigmatic persistence of aerobic methanotrophs in oxygen-limiting freshwater habitats.

Methanotrophic bacteria mitigate emissions of the potent greenhouse gas methane (CH4) from a variety of anthropogenic and natural sources, including freshwater lakes, which are large sources of CH4 on a global scale. Despite a dependence on dioxygen (O2) for CH4 oxidation, abundant populations of putatively aerobic methanotrophs have been detected within microoxic and anoxic waters and sediments of lakes. Experimental work has demonstrated active aerobic methanotrophs under those conditions, but how they are able to persist and oxidize CH4 under O2 deficiency remains enigmatic. In this review, we discuss possible mechanisms that underpin the persistence and activity of aerobic methanotrophs under O2-limiting conditions in freshwater habitats, particularly lakes, summarize experimental evidence for microbial oxidation of CH4 by aerobic bacteria under low or no O2, and suggest future research directions to further explore the ecology and metabolism of aerobic methanotrophs in O2-limiting environments.

Severe epistaxis after posterior nasal nerve ablation requiring surgical intervention: A multi-center case series.

KEY POINTS: Severe epistaxis occurs in 2% of PNN ablation cases, independent of method or device type. Major epistaxis requiring intervention after PNN ablation can occur on average 20 days post-procedure.

Barriers and facilitators to consuming pulses: a qualitative exploration including effects of trying recipes at home.

Pulses are a healthy, sustainable, low cost food, but consumption levels are low for a variety of reasons, including practical and cooking concerns. This work aimed to explore barriers and facilitators towards pulse consumption and increasing consumption, and the potential value of cooking suggestions and recipes for changing these perceptions. Two qualitative studies were undertaken. In Study 1, 21 participants (10 males, 11 females, of a range of ages, cooking responsibilities, and experiences with pulses) were interviewed both before and after receiving cooking suggestions and recipes. In Study 2, 12 participants (2 males, 10 females, as above) were interviewed once after trying recipes. Interviews were audio-recorded, transcribed, and analysed using thematic analysis. Seven themes described barriers and facilitators towards pulse consumption: 'Enjoyment and Sensory properties'; 'Benefits and Recommendations'; 'Practical Concerns'; 'Cooking Concerns'; 'Compatibility with current diet'; 'Personal Influences'; and 'External Influences'. Some similar themes also referred to increasing consumption: 'Willingness'; 'Awareness, Knowledge of Benefits'; 'Knowledge of Cooking and Practical Concerns'; and 'Compatibility with current diet'. Cooking suggestions and recipe use resulted in themes on 'Awareness'; 'Willingness, Trying New Things'; 'Small Changes'; and facilitators associated with 'Enjoyment, Sensory Properties, Practical Concerns, Benefits' and 'Knowledge, Cooking Ideas and Confidence, Incorporation, Cooking Solutions'. Barriers related to 'Risk and Preconceptions'; 'Awareness, but' inaction and additional considerations were also found. Our findings demonstrate a positive role for pulse consumption for increased experience, familiarity, and confidence with preparing, cooking, and consuming these healthy and sustainable foods.

Desulfoferula mesophilus gen. nov. sp. nov., a mesophilic sulfate-reducing bacterium isolated from a brackish lake sediment.

A novel sulfate-reducing bacterium (strain 12FAKT) was isolated from sediment sampled from a brackish lake in Japan. Respiratory growth was observed with formate and pyruvate as an electron donor. Sulfate, thiosulfate, elemental sulfur and dimethyl sulfoxide were utilized as an electron acceptor. The isolate grew over a temperature range of 18-42 °C (optimum 35-37 °C), a NaCl concentration range of 50-450 mM (optimum 150-300 mM) and a pH range of 6.6-7.5. The 12FAKT genome consists of a circular chromosome with a length of 4.5 Mbp and G + C content of 63.6%. Based on 16S rRNA gene sequence similarity, the closest cultured relative was Desulfarculus baarsii 2st14T (92.2%). Genome-based phylogenetic analysis placed strain 12FAKT within the family Desulfarculaceae but did not affiliate the strain with any existing genus. Taken together, we propose a novel species of a novel genus, Desulfoferula mesophilus gen. nov. sp. nov. with the type strain 12FAKT (= DSM 115219T = JCM 39399T).

Massive ossifying fibroma of mandible: A case report and review of the literature.

Key Clinical Message: Benign fibro-osseous neoplasm. Massive size is rarely reported. May be confused with other pathologies such as fibrous dysplasia or osteosarcoma. Aggressive nature and high recurrence pose management challenges. Treated by surgery. Abstract: Ossifying fibroma is a benign fibro-osseous neoplasm. It can affect both mandible and maxilla. Precise diagnosis can be challenging due to significant overlap of clinicopathological features with other neoplasms. Case reports with massive tumor sizes as presented in our case are uncommon. Huge tumor size can cause alarm for other pathologies such as osteosarcoma. The radiological tests should reassure the attending practitioner and histological examination confirm the diagnosis. The aim of the present report is to discuss a case of a giant ossifying fibroma in a 13-year-old male child. He presented with a progressive mandibular mass for 4 years. Clinical, radiological, and pathological characteristics and surgical treatment approaches are further discussed. This is one of the rare cases of massive ossifying fibroma in the English literature.