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INTRODUCTION: MRI-targeted biopsy improves detection of significant prostate cancer (csPCa) and grade prediction. The aim of this study was to identify factors improving the diagnostic performance of targeted biopsies (TB) in detecting csPCa. METHODS: Retrospective monocenter study of patients who underwent a radical prostatectomy (RP) for prostate cancer (PCa) and diagnosed by transrectal combined biopsies (CB) with elastic MRI/ultrasound fusion. We evaluate the diagnostic performance of standardized (SB), targeted (TB) and CB for csPCa, including sensitivity, specificity, and ROC curve. Univariables and logistic regression analysis were performed to analyze factors improving the diagnostic performance of TB in detecting csPCa on final histopathology. RESULTS: Two hundred four men underwent RP after CB with suspicious lesions (PI-RADS≥3) on MRI were included. csPCa was significantly associated with prostate volume, PSA density, a lesion index in the peripheral zone, with a diameter ≥ 7mm. TB were positives for 174 patients (85.3%). Prostate volume, PSA density, radiological coherence, previous biopsies, and a number of biopsies ≥ 3 were significantly associated with a cancer detection. csPCa on TB, a prostate volume <60ml, an index lesion ≥ 7mm and a peripheral zone location were significant predictive factors for diagnostic of csPCa on final histopathology. Area under the ROC curve values, sensitivities and specificities of CB and TB (adjusted model) were 0.78 [0.72-0.84], 77.3 [70.3-83.4], 78.1 [60-90.7], and 0.85 [0.79-0.90], 83.7 [77.3-88.9] and 75 [56.6-88.5] respectively. CONCLUSION: This study confirms the benefit of CB and suggests that TB for a selected population could be as effective as CB.
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Instability of transgene expression is a major challenge for the biopharmaceutical industry, which can impact yields and regulatory approval. Some tRNA genes (tDNAs) can resist epigenetic silencing, the principal mechanism of expression instability, and protect adjacent genes against the spread of repressive heterochromatin. We have taken two naturally occurring clusters of human tDNAs and tested their ability to reduce epigenetic silencing of transgenes integrated into the genome of Chinese hamster ovary (CHO) cells. We find sustained improvements in productivity both in adherent CHO-K1 cells and in an industrially relevant CHO-DG44 expression system (Apollo X, FUJIFILM Diosynth Biotechnologies). We conclude that specific tDNA clusters offer potential to mitigate the widespread problem of production instability.
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Cricetulus , ARN de Transferencia , Transgenes , Células CHO , Animales , ARN de Transferencia/genética , Humanos , Cricetinae , Epigénesis Genética/genética , Silenciador del Gen , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMEN
BACKGROUND: The chemical space is comprised of a vast number of possible structures, of which an unknown portion comprises the human and environmental exposome. Such samples are frequently analyzed using non-targeted analysis via liquid chromatography (LC) coupled to high-resolution mass spectrometry often employing a reversed phase (RP) column. However, prior to analysis, the contents of these samples are unknown and could be comprised of thousands of known and unknown chemical constituents. Moreover, it is unknown which part of the chemical space is sufficiently retained and eluted using RPLC. RESULTS: We present a generic framework that uses a data driven approach to predict whether molecules fall 'inside', 'maybe' inside, or 'outside' of the RPLC subspace. Firstly, three retention index random forest (RF) regression models were constructed that showed that molecular fingerprints are able to predict RPLC retention behavior. Secondly, these models were used to set up the dataset for building an RPLC RF classification model. The RPLC classification model was able to correctly predict whether a chemical belonged to the RPLC subspace with an accuracy of 92% for the testing set. Finally, applying this model to the 91 737 small molecules (i.e., ≤1 000 Da) in NORMAN SusDat showed that 19.1% fall 'outside' of the RPLC subspace. SIGNIFICANCE AND NOVELTY: The RPLC chemical space model provides a major step towards mapping the chemical space and is able to assess whether chemicals can potentially be measured with an RPLC method (i.e., not every RPLC method) or if a different selectivity should be considered. Moreover, knowing which chemicals are outside of the RPLC subspace can assist in reducing potential candidates for library searching and avoid screening for chemicals that will not be present in RPLC data.
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Patients receiving mechanical ventilation in the intensive care unit (ICU) frequently develop contractile weakness of the diaphragm. Consequently, they may experience difficulty weaning from mechanical ventilation, which increases mortality and poses a high economic burden. Because of a lack of knowledge regarding the molecular changes in the diaphragm, no treatment is currently available to improve diaphragm contractility. We compared diaphragm biopsies from ventilated ICU patients (N = 54) to those of non-ICU patients undergoing thoracic surgery (N = 27). By integrating data from myofiber force measurements, x-ray diffraction experiments, and biochemical assays with clinical data, we found that in myofibers isolated from the diaphragm of ventilated ICU patients, myosin is trapped in an energy-sparing, super-relaxed state, which impairs the binding of myosin to actin during diaphragm contraction. Studies on quadriceps biopsies of ICU patients and on the diaphragm of previously healthy mechanically ventilated rats suggested that the super-relaxed myosins are specific to the diaphragm and not a result of critical illness. Exposing slow- and fast-twitch myofibers isolated from the diaphragm biopsies to small-molecule compounds activating troponin restored contractile force in vitro. These findings support the continued development of drugs that target sarcomere proteins to increase the calcium sensitivity of myofibers for the treatment of ICU-acquired diaphragm weakness.
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Diafragma , Contracción Muscular , Miosinas , Respiración Artificial , Músculos Respiratorios , Humanos , Animales , Miosinas/metabolismo , Diafragma/metabolismo , Diafragma/fisiopatología , Músculos Respiratorios/metabolismo , Ratas , Masculino , Unidades de Cuidados Intensivos , Persona de Mediana Edad , Femenino , Anciano , Hibernación/fisiología , Actinas/metabolismoRESUMEN
Armadillo repeat-containing proteins (ARMCs) are a large family found throughout eukaryotes, which play prominent roles in cell adhesion, signaling and cytoskeletal regulation. The ARMC6 protein is highly conserved in primates, including humans, but to date does not have a clear function beyond initial hints of a link to cancer and telomerase activity. We report here in vitro experiments showing ARMC6 binding to DNA promoter sequences from several cancer-related genes (e.g., EGFR, VEGF and c-MYC), and also to the telomeric RNA repeat (TERRA). ARMC6 binding activity appears to recognize G-quadruplex motifs, which are being increasingly implicated as structure-based protein binding sites in chromosome maintenance and repair. In vivo investigation of ARMC6 function revealed that when this protein is overexpressed in human cell lines, there is different expression of genes connected with oncogenic pathways and those implicated in downstream non-canonical telomerase pathways (e.g., VEGF, hTERT, c-MYC, ESM1, MMP3). ARMC6 is already known to interact with human shelterin protein TRF2 and telomerase. The protein binds G-quadruplex structures and does so preferentially to RNA over DNA. As such, this protein may be an example of how a non-canonical nucleic acid structural motif allows mediation between gene regulation and telomeric chromatin rearrangement pathways.
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G-Cuádruplex , Regiones Promotoras Genéticas , Telómero , Humanos , Telómero/metabolismo , Proteínas del Dominio Armadillo/metabolismo , Proteínas del Dominio Armadillo/genética , Telomerasa/metabolismo , Telomerasa/genética , Neoplasias/genética , Neoplasias/metabolismo , Unión Proteica , Regulación Neoplásica de la Expresión Génica , ARN/metabolismo , ARN/genética , Línea Celular Tumoral , Sitios de Unión , Proteínas de Unión al ADN , Factores de TranscripciónRESUMEN
Most eukaryotic organisms employ a telomerase complex for the maintenance of chromosome ends. The core of this complex is composed of telomerase reverse transcriptase (TERT) and telomerase RNA (TR) subunits. The TERT reverse transcriptase (RT) domain synthesises telomeric DNA using the TR template sequence. The other TERT domains contribute to this process in different ways. In particular, the TERT RNA-binding domain (TRBD) interacts with specific TR motif(s). Using a yeast 3-hybrid system, we show the critical role of Arabidopsis thaliana (At) TRBD and embryophyta-conserved KRxR motif in the unstructured linker preceding the TRBD domain for binding to the recently identified AtTR subunit. We also show the essential role of the predicted P4 stem and pseudoknot AtTR structures and provide evidence for the binding of AtTRBD to pseudoknot and KRxR motif stabilising interaction with the P4 stem structure. Our results thus provide the first insight into the core part of the plant telomerase complex.
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Proteínas de Arabidopsis , Arabidopsis , Telomerasa , Telomerasa/genética , Telomerasa/metabolismo , Telomerasa/química , Arabidopsis/genética , Arabidopsis/enzimología , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/química , ARN/metabolismo , ARN/genética , Técnicas del Sistema de Dos Híbridos , ARN de Planta/genética , ARN de Planta/metabolismo , Conformación de Ácido Nucleico , Unión ProteicaRESUMEN
Background: Cerebral amyloid angiopathy (CAA) is a cerebral small vessel disease in which amyloid-ß accumulates in vessel walls. CAA is a leading cause of symptomatic lobar intracerebral hemorrhage and an important contributor to age-related cognitive decline. Recent work has suggested that vascular dysfunction may precede symptomatic stages of CAA, and that spontaneous slow oscillations in arteriolar diameter (termed vasomotion), important for amyloid-ß clearance, may be impaired in CAA. Methods: To systematically study the progression of vascular dysfunction in CAA, we used the APP23 mouse model of amyloidosis, which is known to develop spontaneous cerebral microbleeds mimicking human CAA. Using in vivo 2-photon microscopy, we longitudinally imaged unanesthetized APP23 transgenic mice and wildtype littermates from 7 to 14 months of age, tracking amyloid-ß accumulation and vasomotion in individual pial arterioles over time. MRI was used in separate groups of 12-, 18-, and 24-month-old APP23 transgenic mice and wildtype littermates to detect microbleeds and to assess cerebral blood flow and cerebrovascular reactivity with pseudo-continuous arterial spin labeling. Results: We observed a significant decline in vasomotion with age in APP23 mice, while vasomotion remained unchanged in wildtype mice with age. This decline corresponded in timing to initial vascular amyloid-ß deposition (â¼8-10 months of age), although was more strongly correlated with age than with vascular amyloid-ß burden in individual arterioles. Declines in vasomotion preceded the development of MRI-visible microbleeds and the loss of smooth muscle actin in arterioles, both of which were observed in APP23 mice by 18 months of age. Additionally, evoked cerebrovascular reactivity was intact in APP23 mice at 12 months of age, but significantly lower in APP23 mice by 24 months of age. Conclusions: Our findings suggest that a decline in spontaneous vasomotion is an early, potentially pre-symptomatic, manifestation of CAA and vascular dysfunction, and a possible future treatment target.
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Wastewater treatment plants (WWTPs) are a point source of nutrients, emit greenhouse gases (GHGs), and produce large volumes of excess sludge. The use of aquatic organisms may be an alternative to the technical post-treatment of WWTP effluent, as they play an important role in nutrient dynamics and carbon balance in natural ecosystems. The aim of this study was therefore to assess the performance of an experimental wastewater-treatment cascade of bioturbating macroinvertebrates and floating plants in terms of sludge degradation, nutrient removal and lowering GHG emission. To this end, a full-factorial experiment was designed, using a recirculating cascade with a WWTP sludge compartment with or without bioturbating Chironomus riparius larvae, and an effluent container with or without the floating plant Azolla filiculoides, resulting in four treatments. To calculate the nitrogen (N), phosphorus (P) and carbon (C) mass balance of this system, the N, P and C concentrations in the effluent, biomass production, and sludge degradation, as well as the N, P and C content of all compartments in the cascade were measured during the 26-day experiment. The presence of Chironomus led to an increased sludge degradation of 44% compared to 25% in the control, a 1.4 times decreased transport of P from the sludge and a 2.4 times increased transport of N out of the sludge, either into Chironomus biomass or into the water column. Furthermore, Chironomus activity decreased methane emissions by 92%. The presence of Azolla resulted in a 15% lower P concentration in the effluent than in the control treatment, and a CO2 uptake of 1.13 kg ha-1 day-1. These additive effects of Chironomus and Azolla resulted in an almost two times higher sludge degradation, and an almost two times lower P concentration in the effluent. This is the first study that shows that a bio-based cascade can strongly reduce GHG and P emissions simultaneously during the combined polishing of wastewater sludge and effluent, benefitting from the additive effects of the presence of both macrophytes and invertebrates. In addition to the microbial based treatment steps already employed on WWTPs, the integration of higher organisms in the treatment process expands the WWTP based ecosystem and allows for the inclusion of macroinvertebrate and macrophyte mediated processes. Applying macroinvertebrate-plant cascades may therefore be a promising tool to tackle the present and future challenges of WWTPs.
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Chironomidae , Gases de Efecto Invernadero , Aguas del Alcantarillado , Aguas Residuales , Chironomidae/metabolismo , Animales , Gases de Efecto Invernadero/metabolismo , Gases de Efecto Invernadero/análisis , Aguas Residuales/química , Fósforo/metabolismo , Fósforo/análisis , Nitrógeno/metabolismo , Nitrógeno/análisis , Eliminación de Residuos Líquidos/métodos , Carbono/metabolismo , Carbono/análisis , Biodegradación Ambiental , Purificación del Agua/métodos , Nutrientes/metabolismo , Nutrientes/análisis , Metano/metabolismo , Metano/análisisRESUMEN
Concentrations of chemicals in river water provide crucial information for assessing environmental exposure and risks from fertilisers, pesticides, heavy metals, illicit drugs, pathogens, pharmaceuticals, plastics and perfluorinated substances, among others. However, using concentrations measured along waterways (e.g., from grab samples) to identify sources of contaminants and understand their fate is complicated by mixing of chemicals downstream from diverse diffuse and point sources (e.g., agricultural runoff, wastewater treatment plants). To address this challenge, a novel inverse modelling approach is presented. Using waterway network topology, it quantifies locations and concentrations of contaminant sources upstream by inverting concentrations measured in water samples. It is computationally efficient and quantifies uncertainty. The approach is demonstrated for 13 contaminants of emerging concern (CECs) in an urban stream, the R. Wandle (London, UK). Mixing (the forward problem) was assumed to be conservative, and the location of sources and their concentrations were treated as unknowns to be identified. Calculated CEC source concentrations, which ranged from below detection limit (a few ng/L) up to 1µg/L, were used to predict concentrations of chemicals downstream. Using this approach, >90% of data were predicted within observational uncertainty. Principal component analysis of calculated source concentrations revealed signatures of two distinct chemical sources. First, pharmaceuticals and insecticides were associated with a subcatchment containing a known point source of treated effluent from a wastewater treatment plant. Second, illicit drugs and salicylic acid were associated with multiple sources, interpreted as input from untreated sewage including Combined Sewer Overflows (CSOs), misconnections, runoff and direct disposal throughout the catchment. Finally, a simple algorithmic approach that incorporates network topology was developed to design sampling campaigns to improve resolution of source apportionment. Inverse modelling of contaminant measurements can provide objective means to apportion sources in waterways from spot samples in catchments on a large scale.
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BACKGROUND: With an increase in robot-assisted surgery across all specialties, adequate training and credentialing strategies need to be identified to ensure patients safety. The meta-analysis assesses the transferability of technical surgical skills between laparoscopic surgery, open surgery, and robot-assisted surgery. DESIGN: A systematic search was conducted in Medline, Cochrane Central Register of Controlled Trials, and Web of Science. Outcomes were categorized into time, process, product, and composite outcome measures and pooled separately using Hedges'g (standardized mean difference [SMD]). Subgroup analyses were performed to assess the effect of study design, virtual reality platforms and task difficulty. RESULTS: Out of 14,120 screened studies, 30 were included in the qualitative synthesis and 26 in the quantitative synthesis. Technical surgical skill transfer was demonstrated from laparoscopic to robot-assisted surgery (composite: SMD 0.40, 95%-confidence interval [CI] [0.19; 0.62], time: SMD 0.62, CI [0.33; 0.91]) and vice versa (composite: SMD 0.66, CI [0.33; 0.99], time [basic skills]: SMD 0.36, CI [0.01; 0.72]). No skill transfer was seen from open to robot-assisted surgery with limited available data. CONCLUSION: Technical surgical skills can be transferred from laparoscopic to robot-assisted surgery and vice versa. Robot-assisted and laparoscopic surgical skills training and credentialing should not be regarded separately, but a reasonable combination could shorten overall training times and increase efficiency. Previous experience in open surgery should not be considered as an imperative prerequisite for training in robot-assisted surgery. Recommendations for studies assessing skill transfer are proposed to increase comparability and significance of future studies. PROSPERO REGISTRATION NUMBER: PROSPERO CRD42018104507.
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Competencia Clínica , Laparoscopía , Procedimientos Quirúrgicos Robotizados , Laparoscopía/educación , Procedimientos Quirúrgicos Robotizados/educación , HumanosRESUMEN
Sargassum spp. strandings in the tropical Atlantic harm local ecosystems due to toxic sulfide levels. We conducted a mesocosm experiment to test the efficacy of iron(III) (hydr)oxides in (a) mitigating sulfide toxicity in mangroves resulting from Sargassum and (b) reducing potentially enhanced greenhouse gas emissions. Our results show that iron addition failed to prevent mangrove mortality caused by highly toxic sulfide concentrations, which reached up to 15,000 µmol l-1 in 14 days; timely removal may potentially prevent mangrove death. Sargassum-impacted mesocosms significantly increased methane, nitrous oxide, and carbon dioxide emissions, producing approximately 1 g CO2-equivalents m-2 h-1 during daylight hours, thereby shifting mangroves from sinks to sources of greenhouse gasses. However, iron addition decreased methane emissions by 62 % and nitrous oxide emissions by 57 %. This research reveals that Sargassum strandings have multiple adverse effects related to chemical and ecological dynamics in mangrove ecosystems, including greenhouse gas emissions.
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Metano , Óxido Nitroso , Sargassum , Sulfuros , Humedales , Hierro , Contaminantes Químicos del Agua/toxicidad , Gases de Efecto Invernadero/análisisRESUMEN
The classification of personality disorder (PD) is undergoing a paradigm shift in which categorically defined specific PDs are being replaced by dimensionally defined maladaptive trait domains. To bridge the classificatory approaches, this study attempts to use items from the categorical PD model in DSM-IV to measure the maladaptive trait domains described in DSM-5 Section III/ICD-11. A general population sample comprising 1228 participants completed the Screening Questionnaire of the Structured Clinical Interview for DSM-IV Axis II (SCID-II-SQ), the Personality Inventory for DSM-5 (PID-5), and the anankastia scale of the Personality Inventory for ICD-11 (PiCD). Using item response theory models and a psychometric linking technique, SCID-II-SQ items were evaluated for their contribution to measuring maladaptive trait domains. The best discriminating items were then selected to derive proxy scales. We found that convergent validity of these proxy scales was in a similar range to that of other self-report measures for PD, except for the proxy scale for PiCD anankastia. However, only the proxy scale for negative affectivity showed acceptable reliability that would allow its application in research settings. Future studies should seek to establish a common metric between specific PDs and maladaptive trait domains using self-report measures with higher specificity or semi-structured interviews.
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Manual Diagnóstico y Estadístico de los Trastornos Mentales , Trastornos de la Personalidad , Psicometría , Humanos , Femenino , Masculino , Trastornos de la Personalidad/diagnóstico , Adulto , Psicometría/normas , Persona de Mediana Edad , Encuestas y Cuestionarios/normas , Reproducibilidad de los Resultados , Inventario de Personalidad/normas , Adulto Joven , Entrevista Psicológica/normas , Adolescente , AncianoRESUMEN
Biopharmaceutical manufacture is transitioning from batch to integrated and continuous biomanufacturing (ICB). The common framework for most ICB, potentially enables a global biomanufacturing ecosystem utilizing modular and multi-function manufacturing equipment. Integrating unit operation hardware and software from multiple suppliers, complex supply chains enabled by multiple customized single-use flow paths, and large volume buffer production/storage make this ICB vision difficult to achieve with commercially available manufacturing equipment. Thus, we developed SymphonX™, a downstream processing skid with advanced buffer management capabilities, a single disposable generic flow path design that provides plug-and-play flexibility across all downstream unit operations and a single interface to reduce operational risk. Designed for multi-product and multi-process cGMP facilities, SymphonX™ can perform stand-alone batch processing or ICB. This study utilized an Apollo™ X CHO-DG44 mAb-expressing cell line in a steady-state perfusion bioreactor, harvesting product continuously with a cell retention device and connected SymphonX™ purification skids. The downstream process used the same chemistry (resins, buffer composition, membrane composition) as our historical batch processing platform, with SymphonX™ in-line conditioning and buffer concentrates. We used surge vessels between unit operations, single-column chromatography (protein A, cation and anion exchange) and two-tank batch virus inactivation. After the first polishing step (cation exchange), we continuously pooled product for 6 days. These 6 day pools were processed in batch-mode from anion exchange to bulk drug substance. This manufacturing scale proof-of-concept ICB produced 0.54 kg/day of drug substance with consistent product quality attributes and demonstrated successful bioburden control for unit-operations undergoing continuous operation.
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Mindreading ability-also referred to as cognitive empathy or mentalizing-is typically conceptualized as a relatively stable dimension of individual differences in the ability to make accurate inferences about the mental states of others. This construct is primarily assessed using self-report questionnaires and task-based performance measures. However, the validity of these measures has been questioned: According to rival interpretations, mindreading tasks may capture general cognitive ability, whereas mindreading self-reports may capture perceived rather than actual mindreading ability. In this preregistered multimethod study involving 700 participants from the U.S. general population, we tested the validity of mindreading measures by examining the nomological network of self-reports and task-based methods using structural equation modeling. Specifically, we contrasted the empirical associations with theoretical predictions that assume mindreading measures are valid versus invalid. More consistent with rival interpretations, mindreading tasks showed a negligible latent correlation with mindreading self-reports (.05) and a large one with general cognitive ability (.85), whereas mindreading self-reports were specifically associated with perceived performance in mindreading tasks (.29). Also more consistent with rival interpretations, neither mindreading self-reports nor task-based measures showed positive unique associations with psychosocial functioning when controlling for general cognitive ability and general positive self-evaluation. Instead, negative unique associations emerged for both methods, although this effect was not robust for tasks. Overall, the results cast doubt on the validity of commonly used mindreading measures and support their rival interpretations. (PsycInfo Database Record (c) 2024 APA, all rights reserved).
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Autoinforme , Teoría de la Mente , Humanos , Femenino , Masculino , Adulto , Persona de Mediana Edad , Adulto Joven , Reproducibilidad de los Resultados , Empatía , Anciano , Adolescente , Percepción Social , PsicometríaRESUMEN
OBJECTIVE: To provide a composite endpoint in pancreatic surgery. SUMMARY BACKGROUND DATA: Single endpoints in prospective and randomized studies have become impractical due to their low frequency and the marginal benefit of new interventions. METHODS: Data from prospective studies were used to develop (n=1273) and validate (n=544) a composite endpoint based on postoperative pancreatic fistula, post-pancreatectomy hemorrhage as well as reoperation and reinterventions. All patients had pancreatectomies of different extents. The association of the developed PAncreatic surgery Composite Endpoint (PACE) with prolonged length of hospital stay (LOS) >75th percentile and mortality was assessed. A single-institution database was used for external validation (n = 2666). Sample size calculations were made for single outcomes and the composite endpoint. RESULTS: In the internal validation cohort, the PACE demonstrated an AUC of 78.0%, a sensitivity of 90.4% and a specificity of 67.6% in predicting a prolonged LOS. In the external cohort, the AUC was 76.9%, the sensitivity 73.8% and the specificity 80.1%. The 90-day mortality rate was significantly different for patients with a positive versus a negative PACE both in the development and internal validation cohort (5.1% vs 0.9%; P< 0.001), as well as in the external validation cohort (8.5% vs 1.2%, P< 0.001). The PACE enabled sample size reductions of up to 80.5% compared to single outcomes. CONCLUSION: The PACE performed well in predicting prolonged hospital stays and can be used as a standardized and clinically relevant endpoint for future prospective trials enabling lower sample sizes and therefore improved feasibility compared to single outcome parameters.
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Terrestrial wetland ecosystems challenge biodiversity-ecosystem function theory, which generally links high species diversity to stable ecosystem functions. An open question in ecosystem ecology is whether assemblages of co-occurring peat mosses contribute to the stability of peatland ecosystem processes. We conducted a two-species (Sphagnum cuspidatum, Sphagnum medium) replacement series mesocosm experiment to evaluate the resistance, resilience, and recovery rates of net ecosystem CO2 exchange (NEE) under mild and deep water table drawdown. Our results show a positive effect of mild water table drawdown on NEE with no apparent role for peat moss mixture. Our study indicates that the carbon uptake capacity by peat moss mixtures is rather resilient to mild water table drawdown, but seriously affected by deeper drought conditions. Co-occurring peat moss species seem to enhance the resilience of the carbon uptake function (i.e. ability of NEE to return to pre-perturbation levels) of peat moss mixtures only slightly. These findings suggest that assemblages of co-occurring Sphagnum mosses do only marginally contribute to the stability of ecosystem functions in peatlands under drought conditions. Above all, our results highlight that predicted severe droughts can gravely affect the sink capacity of peatlands, with only a small extenuating role for peat moss mixtures.
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Ecosistema , Sphagnopsida , Ecología , Biodiversidad , CarbonoRESUMEN
Chinese hamster ovary (CHO) cells are widely used for production of biologics including therapeutic monoclonal antibodies. Cell death in CHO cells is a significant factor in biopharmaceutical production, impacting both product yield and quality. Apoptosis has previously been described as the major form of cell death occurring in CHO cells in bioreactors. However, these studies were undertaken when less was known about non-apoptotic cell death pathways. Here, we report the occurrence of non-apoptotic cell death in an industrial antibody-producing CHO cell line during fed-batch culture. Under standard conditions, crucial markers of apoptosis were not observed despite a decrease in viability towards the end of the culture; only by increasing stress within the system did we observe caspase activation indicative of apoptosis. In contrast, markers of parthanatos and ferroptosis were observed during standard fed-batch culture, indicating that these non-apoptotic cell death pathways contribute to viability loss under these conditions. These findings pave the way for targeting non-conventional cell death pathways to improve viability and biologic production in CHO cells.
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Técnicas de Cultivo Celular por Lotes , Reactores Biológicos , Cricetinae , Animales , Cricetulus , Células CHO , ApoptosisRESUMEN
The recent global outbreak of mpox, caused by monkeypox virus (MPV) emerged in Europe in 2022 and rapidly spread to over 40 countries. The Americas are currently facing the highest impact, reporting over 50,000 cases by early 2023. In this study, we analyzed 880 MPV isolates worldwide to gain insights into the evolutionary patterns and initial introduction events of the virus in Mexico. We found that MPV entered Mexico on multiple occasions, from the United Kingdom, Portugal, and Canada, and subsequently spread locally in different regions of Mexico. Additionally, we show that MPV has an open pangenome, highlighting the role of gene turnover in shaping its genomic diversity, rather than single-nucleotide polymorphisms (SNPs), which do not contribute significantly to genome diversity. Although the genome contains multiple SNPs in coding regions, these remain under purifying selection, suggesting their evolutionary conservation. One notable exception is amino acid position 63 of the protein encoded by the Cop-A4L gene, which is intricately related to viral maturity, which we found to be under strong positive selection. Ancestral state reconstruction indicated that the ancestral state at position 63 corresponds to the amino acid valine, which is present only in isolates of clade I. However, the isolates from the current outbreak contained threonine at position 63. Our findings contribute new information about the evolution of monkeypox virus.
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Mpox , Humanos , Monkeypox virus/genética , México/epidemiología , Filogenia , Aminoácidos/genética , Brotes de EnfermedadesRESUMEN
Structurally complex habitats, such as mangrove forests, allow for rich assemblages of species that benefit from the provided space, volume and substrate. Changes in habitat complexity can affect species abundance, diversity and resilience. In this study, we explored the effects of habitat complexity on food web networks in four developmental stages of mangrove forests with differing structural complexities: climax > degrading > colonizing > bare, by analyzing food web structure, stable isotopes and habitat complexity. We found that food webs became gradually more biodiverse (species richness: +119 %), complex (link density: +39 %), and robust (connectance: -35 %) in climax versus bare stages with increasing complexity of the mangrove forest (i.e., number of trees, leaf cover, and pneumatophore densities). This study shows that habitat complexity drives food web network structure in dynamic mangrove forests. We recommend restoration practitioners to use this food web network approach to quantify habitat restoration successes complementary to traditional biodiversity metrics.