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African swine fever virus (ASFV) is the causative agent of a contagious disease affecting wild and domestic swine. The function of B169L protein, as a potential integral structural membrane protein, remains to be experimentally characterized. Using state-of-the-art bioinformatics tools, we confirm here earlier predictions indicating the presence of an integral membrane helical hairpin, and further suggest anchoring of this protein to the ER membrane, with both terminal ends facing the lumen of the organelle. Our evolutionary analysis confirmed the importance of purifying selection in the preservation of the identified domains during the evolution of B169L in nature. Also, we address the possible function of this hairpin transmembrane domain (HTMD) as a class IIA viroporin. Expression of GFP fusion proteins in the absence of a signal peptide supported B169L insertion into the ER as a Type III membrane protein and the formation of oligomers therein. Overlapping peptides that spanned the B169L HTMD were reconstituted into ER-like membranes and the adopted structures analyzed by infrared spectroscopy. Consistent with the predictions, B169L transmembrane sequences adopted α-helical conformations in lipid bilayers. Moreover, single vesicle permeability assays demonstrated the assembly of lytic pores in ER-like membranes by B169L transmembrane helices, a capacity confirmed by ion-channel activity measurements in planar bilayers. Emphasizing the relevance of these observations, pore-forming activities were not observed in the case of transmembrane helices derived from EP84R, another ASFV protein predicted to anchor to membranes through a α-helical HTMD. Overall, our results support predictions of viroporin-like function for the B169L HTMD.IMPORTANCEAfrican swine fever (ASF), a devastating disease affecting domestic swine, is widely spread in Eurasia, producing significant economic problems in the pork industry. Approaches to prevent/cure the disease are mainly restricted to the limited information concerning the role of most of the genes encoded by the large (160-170 kba) virus genome. In this report, we present the experimental data on the functional characterization of the African swine fever virus (ASFV) gene B169L. Data presented here indicates that the B169L gene encodes for an essential membrane-associated protein with a viroporin function.
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BACKGROUND: To maximize the availability of suitable grafts and ensure effective management, several reports have demonstrated successful outcomes when using kidney grafts with urolithiasis. This multicenter study reports on the management and long-term outcomes of kidney transplantation using renal grafts with lithiasis. METHODS: Retrospective data from three Spanish hospitals were analyzed for kidney transplants involving grafts with nephrolithiasis performed between December 2009 and August 2023. The study included adult patients, excluding those with incomplete records. It evaluated stone characteristics, complications, and outcomes in recipients and in living kidney donors. RESULTS: Out of 38 analyzed kidney transplants, 57.9% were cadaveric and 42.1% were from living kidney donors. Most diagnoses were incidental during donor evaluation, with an average stone size of 7.06 mm. After follow-up (median 26 months), all recipients but one had functioning grafts, and there were no stone recurrences in both recipients and living kidney donors. Conservative management was adopted in 28 cases, while 10 cases required ex-vivo flexible ureterorenoscopy for stone removal. Following conservative management, 5 patients needed additional treatments for stone-related events. CONCLUSIONS: Kidneys with lithiasis can be considered for transplantation in selected cases, resulting in good functional outcomes with no stone recurrence in recipients or living donors.
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Jump actions are common in several sports, and their performance is related to a myriad of biomechanical and physiological factors, with links to athletic performance and imbalances. Currently, a valid, field-based, easy-to-use tool to assess the quality of an explosive jump movement, similar to the required sports movements, is unavailable. Thus, the present study aimed to design and validate a field-based, easy-to-use tool that can be used to assess the quality of movement during an explosive single-leg countermovement jump (SL-CMJ). Ten experts participated in the content validation process for the checklist, including checking item relevance, definition accuracy, and scoring adequacy. Content validity was measured using the Aikens V format. The checklist included the items "Foot orientation," "Knee valgus/varus," "Internal/external hip flexed orientation," "Pelvis tilt," "Thorax tilt," "Thorax rotation," "Foot pronation/supination," "Asymmetrical hip," and "Lumbo-pelvic association". The items achieved a 0.60-0.99 in relevance, 0.70-1.00 in definition accuracy, and 0.80-0.83 in scoring adequacies in the Aikens V proof. The results from the context validation process suggest that the tool may be appropriate to assess athletes' quality of explosive movement. Furthermore, the results derived from such assessments may help to design better and safer training interventions.
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OBJECTIVE: To analyse the adverse events (AEs) associated with apalutamide and the impact of a multidisciplinary team (MDT) protocol on its management at a tertiary care hospital in a real-world setting. METHODS: This was an observational, prospective, cohort study based on real-world evidence at the Hospital Clínic de Barcelona. Includes patients diagnosed with metastatic hormone-sensitive prostate cancer (mHSPC) or high-risk nonmetastatic castration-resistant prostate cancer (nmCRPC) and who started treatment with apalutamide between May 2019 and March 2023 in a real-world clinical setting. RESULTS: Of the 121 patients treated with apalutamide, 52.1% experienced an AE, 19.8% experienced temporarily interruption or a reduction in the dose of apalutamide, and 13.2% discontinued treatment due to AEs. Without MDT protocol (49 patients), 24.5% of patients had to temporarily interrupt or reduce the dose of apalutamide due to AEs, with a median time from the start of treatment of 10.1 months, and 24.5% discontinued apalutamide due to AEs, with a median time from the start of treatment of 3.1 months. Meanwhile, whit MDT protocol (72 patients), 16.7% of patients had to temporarily interrupt or reduce the dose of apalutamide due to AEs, with a median time from the start of treatment of 1.6 months, and 5.6% discontinued apalutamide due to AEs, with a median time from the start of treatment of 4 months. The risk reduction associated with treatment discontinuation was statistically significant (p-value = 0.003). CONCLUSIONS: This study highlights the importance of MDT management of AEs associated with apalutamide to reduce treatment discontinuation.
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Neoplasias de la Próstata Resistentes a la Castración , Neoplasias de la Próstata , Tiohidantoínas , Humanos , Masculino , Tiohidantoínas/efectos adversos , Tiohidantoínas/uso terapéutico , Tiohidantoínas/administración & dosificación , Anciano , Estudios Prospectivos , Neoplasias de la Próstata/tratamiento farmacológico , Persona de Mediana Edad , Neoplasias de la Próstata Resistentes a la Castración/tratamiento farmacológico , Neoplasias de la Próstata Resistentes a la Castración/patología , Grupo de Atención al Paciente , Anciano de 80 o más Años , Estudios de Cohortes , Antineoplásicos/efectos adversos , Antineoplásicos/uso terapéutico , Antineoplásicos/administración & dosificaciónRESUMEN
Currently, there are no reliable prognostic factors to determine which upper tract urothelial carcinoma (UTUC) patients will progress after radical nephroureterectomy (RNU). We aim to evaluate whether liquid-biopsy-based biomarkers (circulating tumor cells (CTCs), cell-free DNA (cfDNA), and circulating tumor DNA (ctDNA)) were able to predict clinical outcomes in localized UTUC patients undergoing RNU. Twenty patients were prospectively enrolled between 2021 and 2023. Two blood samples were collected before RNU and three months later. CTCs and cfDNA were isolated and evaluated using the IsoFlux system and Quant-iT PicoGreen dsDNA kit, respectively. Droplet digital PCR was performed to determine ctDNA status. Cox regression analysis was performed on CTCs, cfDNA, and ctDNA at two different follow-up time points to examine their influence on tumor progression and cancer-specific survival (CSS). During a median follow-up of 18 months, seven (35%) patients progressed and three (15%) died. Multivariate analysis demonstrated that cfDNA levels three months after RNU are a significant predictor of tumor progression (HR = 1.085; p = 0.006) and CSS (HR = 1.168; p = 0.029). No associations were found between CTC enumeration and ctDNA status with any of the clinical outcomes evaluated. The evaluation of cfDNA levels in clinical practice could improve the disease management of UTUC patients.
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Carcinoma de Células Transicionales , Ácidos Nucleicos Libres de Células , Neoplasias de la Vejiga Urinaria , Humanos , Carcinoma de Células Transicionales/diagnóstico , Carcinoma de Células Transicionales/genética , Pronóstico , Biomarcadores , Biopsia LíquidaRESUMEN
TRP channels are important pharmacological targets in physiopathology. TRPV2 plays distinct roles in cardiac and neuromuscular function, immunity, and metabolism, and is associated with pathologies like muscular dystrophy and cancer. However, TRPV2 pharmacology is unspecific and scarce at best. Using in silico similarity-based chemoinformatics we obtained a set of 270 potential hits for TRPV2 categorized into families based on chemical nature and similarity. Docking the compounds on available rat TRPV2 structures allowed the clustering of drug families in specific ligand binding sites. Starting from a probenecid docking pose in the piperlongumine binding site and using a Gaussian accelerated molecular dynamics approach we have assigned a putative probenecid binding site. In parallel, we measured the EC50 of 7 probenecid derivatives on TRPV2 expressed in Pichia pastoris using a novel medium-throughput Ca2+ influx assay in yeast membranes together with an unbiased and unsupervised data analysis method. We found that 4-(piperidine-1-sulfonyl)-benzoic acid had a better EC50 than probenecid, which is one of the most specific TRPV2 agonists to date. Exploring the TRPV2-dependent anti-hypertensive potential in vivo, we found that 4-(piperidine-1-sulfonyl)-benzoic acid shows a sex-biased vasodilator effect producing larger vascular relaxations in female mice. Overall, this study expands the pharmacological toolbox for TRPV2, a widely expressed membrane protein and orphan drug target.
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CONTEXT: Metabolic dysfunction-associated steatotic liver disease (MASLD) is characterized by the intracellular lipid accumulation in hepatocytes. Excess caloric intake and high-fat diets are considered to significantly contribute to MASLD development. OBJECTIVE: To evaluate the hepatic and serum fatty acid (FA) composition in patients with different stages of MASLD, and their relationship with FA dietary intake and MASLD-related risk factors. METHODS: This was a case-control study in patients with obesity undergoing bariatric surgery at a university hospital between January 2020 and December 2021. Participants were distributed in 3 groups: no MASLD (n = 26), steatotic liver disease (n = 33), and metabolic dysfunction-associated steatohepatitis (n = 32). Hepatic and serum FA levels were determined by gas chromatography-mass spectrometry. Nutritional status was evaluated using validated food frequency questionnaires. The hepatic expression of genes involved in FA metabolism was analyzed by reverse transcription quantitative polymerase chain reaction. RESULTS: The hepatic, but not serum, FA profiles were significantly altered in patients with MASLD compared with those without MASLD. No differences were observed in FA intake between the groups. Levels of C16:0, C18:1, and the C18:1/C18:0 ratio were higher, while C18:0 levels and C18:0/C16:0 ratio were lower in patients with MASLD, being significantly different between the 3 groups. Hepatic FA levels and ratios correlated with histopathological diagnosis and other MASLD-related parameters. The expression of genes involved in the FA metabolism was upregulated in patients with MASLD. CONCLUSION: Alterations in hepatic FA levels in MASLD patients were due to enhancement of de novo lipogenesis in the liver.
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Ácidos Grasos , Hígado Graso , Lipidómica , Hígado , Obesidad , Humanos , Masculino , Femenino , Estudios de Casos y Controles , Adulto , Persona de Mediana Edad , Hígado/metabolismo , Hígado/patología , Obesidad/metabolismo , Obesidad/complicaciones , Ácidos Grasos/metabolismo , Hígado Graso/metabolismo , Hígado Graso/patología , Metabolismo de los Lípidos , Cirugía BariátricaRESUMEN
In volleyball, the effect of different factors on serve performance has usually been analyzed with traditional statistical techniques such as logistic regression or discriminant analysis. Purpose: In this study, two of the main models used in unsupervised machine learning (cluster and principal component analysis) were applied to achieve these objectives: (a) to create groups of players considering their serve coefficient, age, height, and team ranking, and (b) to identify which variables related to the serve (type and performance), the players (role, age, and height), and the teams (ranking, match location, and quality of opposition) most explained the total variance of the data during an entire women's volleyball season. Method: A total of 20,936 serves were analyzed during the 132 matches played in the 2017-2018 season in the Liga Iberdrola (women Spanish first division). The variables were related to the serving action (type of serve and performance), the players' traits (player role, age, and height), and the teams' characteristics (final ranking, match location, quality of opposition, and tournament). Results: Cluster analysis showed five groups of players differing in age, serve coefficient, team ranking, and height. Principal component analysis showed how the first five components explained 72.12% of the total variance. From these components, serve coefficient, team ranking, match location, quality of opposition, and player role each contributed more than 10%. Conclusions: These findings can help coaches to improve talent selection and players' development according to competition demands.
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Aprendizaje Automático no Supervisado , Voleibol , Humanos , Femenino , Análisis por Conglomerados , Análisis Discriminante , Estaciones del AñoRESUMEN
BACKGROUND: Fixed-dose combinations of antiretroviral drugs are commonly used to treat HIV infection and therapeutic monitoring is not part of routine clinical practice. However, drug concentrations monitoring might have role in different clinical scenarios as well as for research purposes. This study aimed to develop and validate UHPLC-MS/MS procedures for measuring total and unbound concentrations of bictegravir, dolutegravir, darunavir and doravirine in human plasma. MATERIAL AND METHODS: Equilibrium dialysis preceded sample preparation (based on protein precipitation) for measuring unbound antiretroviral concentrations. Chromatographic separations were achieved on an Acquity®-UPLC® HSS™-T3 column (50 mm × 2.1 mm; 1.8 µm) using a non-linear water/acetonitrile gradient containing 0.1 % formic acid at a 0.5 mL/min flow rate. Antiretrovirals were detected by tandem mass spectrometry in positive electrospray ionisation and multiple reaction monitoring modes. RESULTS: No significant interferences or carry-over were observed. Imprecisions, absolute relative biases, normalised matrix effects and recoveries were ≤15.0 %, ≤11.1 %, (94.7-104.1)% and (96.7-105.5)%, respectively. Non-linear measuring intervals were observed between (25-10,000) µg/L for total/plasma dialysate concentrations and linearity schemes (1.00-100) µg/L for buffer dialysate concentrations. CONCLUSIONS: The UHPLC-MS/MS procedures developed could be used for research purposes and therapeutic drug monitoring of antiretrovirals in routine clinical practice.
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Infecciones por VIH , Espectrometría de Masas en Tándem , Humanos , Espectrometría de Masas en Tándem/métodos , Darunavir , Cromatografía Líquida de Alta Presión/métodos , Infecciones por VIH/tratamiento farmacológico , Diálisis Renal , Soluciones para DiálisisRESUMEN
The aim of this study was to evaluate whether the rally length in high-level Spanish volleyball was longer in women than in men. A total of 1,786 rallies were observed: 792 for women and 994 for men. The recorded variables were match (quarter-final 1, quarter-final 2, semi-final 1, semi-final 2, final), gender (men, women), rally length (seconds), pseudo-rally (no, yes), and terminal event (ball out of sight, ball in/out, fault). Different non-parametric statistical techniques were used to compare the rally length between groups or subsets of data, i.e., the Kruskal-Wallis H test, the Mann-Whitney U test, quantile regression, and survival analysis. The mean and median rally length was significantly and slightly longer in women than in men. The rally length difference between genders was barely 1 s in quantile 0.5 or median, while in quantile 0.95, it was just over 4 s. In women, the probability of ending the rally at 3.9, 5.1, 10.2, and 43.9 s (at 4.4, 6.3, 11.6, and 43.9 s without pseudo-rallies) was 25%, 50%, 75%, and 100%, respectively. In men, the probability of ending the rally at 3.2, 4.3, 7.9, and 29.1 s (at 3.9, 4.8, 8.8, and 29.1 s without pseudo-rallies) was 25%, 50%, 75%, and 100%, respectively. These temporal thresholds can help volleyball coaches to train their players in a coherent manner.
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Circulating tumor DNA (ctDNA) has recently emerged as a real-time prognostic and predictive biomarker for monitoring cancer patients. Here, we aimed to ascertain whether tumor-agnostic ctDNA testing would be a feasible strategy to monitor disease progression and therapeutic response in muscle-invasive bladder cancer (MIBC) patients after radical cystectomy (RC). Forty-two MIBC patients who underwent RC were prospectively included. Blood samples from these patients were collected at different follow-up time points. Two specific mutations (TERT c.1-124C>T and ATM c.1236-2A>T) were analyzed in the patients' plasma samples by droplet digital PCR to determine their ctDNA status. During a median follow-up of 21 months, 24% of patients progressed in a median of six months. ctDNA status was identified as a prognostic biomarker of tumor progression before RC and 4 and 12 months later (HR 6.774, HR 3.673, and HR 30.865, respectively; p < 0.05). Lastly, dynamic changes in ctDNA status between baseline and four months later were significantly associated with patient outcomes (p = 0.045). In conclusion, longitudinal ctDNA analysis using a tumor-agnostic approach is a potential tool for monitoring MIBC patients after RC. The implementation of this testing in a clinical setting could improve disease management and patients' outcomes.
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Ácidos Nucleicos Libres de Células , ADN Tumoral Circulante , Neoplasias de la Vejiga Urinaria , Humanos , ADN Tumoral Circulante/genética , Neoplasias de la Vejiga Urinaria/diagnóstico , Neoplasias de la Vejiga Urinaria/genética , Neoplasias de la Vejiga Urinaria/patología , ADN de Neoplasias , Biomarcadores , Músculos/patología , Biomarcadores de Tumor/genética , MutaciónRESUMEN
BACKGROUND: In ARCHES, treatment intensification of androgen deprivation therapy (ADT) with enzalutamide versus placebo improved clinical outcomes in metastatic hormone-sensitive prostate cancer (mHSPC). Understanding the benefits and tolerability of enzalutamide for men aged ≥75 yr may inform disease management. OBJECTIVE: To determine whether age is associated with clinical outcomes in mHSPC. DESIGN, SETTING, AND PARTICIPANTS: A post hoc analysis of the multinational, double-blind, randomized, placebo-controlled, phase 3 ARCHES trial in 1150 men with mHSPC (median follow-up [mo]: <75 yr, 44.6; ≥75 yr, 44.3) was performed. INTERVENTION: Randomization 1:1 to enzalutamide (160 mg/d) plus ADT or placebo plus ADT; stratification by disease volume and prior docetaxel use. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Overall survival (OS), radiographic progression-free survival (rPFS), safety, and other secondary endpoints were compared between age groups (<75 and ≥75 yr) and treatment arms (Cox proportional hazard models). RESULTS AND LIMITATIONS: Men aged <75 versus ≥75 yr had longer OS (enzalutamide plus ADT: hazard ratio [HR] 0.66; 95% confidence interval [CI] 0.47-0.91; p = 0.02; placebo plus ADT: HR 0.81; 95% CI 0.60-1.09; p = 0.13) and rPFS (enzalutamide plus ADT: HR 0.78; 95% CI 0.58-1.04; p = 0.12; placebo plus ADT: HR 0.98; 95% CI 0.74-1.30; p = 0.007). Enzalutamide improved OS (<75 yr: HR 0.61; 95% CI 0.47-0.79; ≥75 yr: HR 0.76; 95% CI 0.54-1.09) and secondary efficacy endpoints without evidence of statistical heterogeneity, and was generally well tolerated in both age groups, with minimal quality-of-life impact. Older versus younger patients experienced more frequent dose interruptions (20.2% vs 10.9%) and treatment-emergent adverse events (95.2% vs 89.1%). Post hoc examination and small sample size preclude definitive conclusions. CONCLUSIONS: Enzalutamide plus ADT improved efficacy outcomes and was generally well tolerated despite shorter treatment exposure in older patients, indicating enzalutamide's utility in patients with mHSPC aged <75 and ≥75 yr. PATIENT SUMMARY: Enzalutamide is a drug approved to treat men with prostate cancer. In this report, we compared patients aged <75 and ≥75 yr treated with enzalutamide plus androgen deprivation therapy to determine whether age affected how long they lived without the cancer spreading to other parts of their body. We found that, although younger patients had more favorable survival outcomes, enzalutamide was associated with longer survival and reduced disease spread in both age groups.
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Background and aims: The spatial and temporal genetic heterogeneity of bladder cancer (BC) makes challenging to find specific drivers of metastatic disease, thus preventing to determine those BC patients at high risk of tumor progression. Our aim was to identify DNA mutations providing aggressive behavior to bladder tumors and analyze them in patients' cell-free DNA (cfDNA) during their follow-up after radical cystectomy (RC) in order to monitor tumor evolution. Methods: Six BC patients who underwent RC and presented disease progression during their follow-up were included. Next-generation sequencing was used to determine somatic mutations in several primary tumor and metastatic specimens from each patient. Shared DNA mutations between primary bladder tumor and metastatic sites were identified in cfDNA samples through droplet digital PCR. Results: Besides BC genetic heterogeneity, specific mutations in at least one of these genes -TERT, ATM, RB1, and FGFR3- were found in primary tumors and their metastases in all patients. These mutations were also identified in the patients' cfDNA at different follow-up time points. Additionally, the dynamic changes of these mutations in cfDNA allowed us to determine tumor evolution in response to treatment. Conclusion: The analysis of BC mutations associated with poor prognosis in plasma cfDNA could be a valuable tool to monitor tumor evolution, thus improving the clinical management of BC patients.
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El proceso madurativo tiene una gran influencia sobre los factores antropométricos y las capacidades físicas del atleta, y por tanto, sobre el proceso de selección de talentos deportivos. El objetivo de este trabajo fue analizar el estado madurativo y las características antropométricas junto con la comparación de estos datos por sexo en una muestra de 39 jugadores jóvenes de voleibol dentro de un proceso de selección de talentos (19 damas de 14,88±1,05 años y 13 varones de 15,1 años). Se obtuvieron datos de edad cronológica, altura, peso, altura sentado, altura de la madre y padre, la edad pico de crecimiento, el "timing" o periodo de tiempo por encima o por debajo de la edad pico, altura al final del proceso madurativo, el porcentaje actual de altura máxima, los centímetros restantes y el estado madurativo (pre-púber, púber o pos-púber). Los jugadores mostraron una mayor altura en el momento de las mediciones (179,92±6,87 vs 171,05±4,80; p<0,001), así como una mayor altura final calculada (189,46±3,73 vs 178,52±5,17; p<0,001), en comparación a las jugadoras. El pico en la velocidad de crecimiento también fue superior en los jugadores (14,56±0,44 vs 12,60±0,57; p<0,001), aunque su timing era inferior al de las jugadoras (0,531±1,19 vs 2,27±0,64; p<0,001). Esto se debió a un mayor porcentaje de jugadores masculinos en estados puberales, incluyendo un jugador en estadio pre-puberal, mientras que fue abundante la presencia de jugadoras en estado pospuberal. Estos datos reflejan la gran cantidad de jugadores que tienden a estar en periodos avanzados de maduración en procesos de selección de talentos. Por tanto, entrenadores y seleccionadores deben contemplar estas variables para evitar sesgos en el proceso de identificación del talento deportivo.
SUMMARY: The maturation process has a great influence on the anthropometric factors and physical capabilities of the athlete, and therefore, on the selection process of sporting talents. The objective of this work was to analyze the maturational state and anthropometric characteristics together with the comparison of these data by sex in a sample of 39 young volleyball players within a talent selection process (19 ladies of 14.88±1. 05 years old and 13 males aged 15.1 years). Data were obtained on chronological age, height, weight, sitting height, height of the mother and father, peak age of growth, timing or period of time above or below the peak age, height at the end of the process. maturation, the current percentage of maximum height, the remaining centimeters and the maturation status (pre-pubertal, pubertal or post-pubertal). The players showed a greater height at the time of the measurements (179.92±6.87 vs. 171.05±4.80; p<0.001), as well as a greater final calculated height (189.46±3.73 vs. 178.52±5.17; p<0.001), compared to the female players. The peak in growth speed was also higher in male players (14.56±0.44 vs 12.60±0.57; p<0.001), although their timing was lower than that of female players (0.531±1.19 vs 2.27±0.64; p<0.001). This was due to a higher percentage of male players in pubertal states, including one player in a pre-pubertal stage, while the presence of female players in a post-pubertal stage was abundant. These data reflect the large number of players who tend to be in advanced periods of maturation in talent selection processes. Therefore, coaches and selectors must consider these variables to avoid biases in the process of identifying sporting talent.
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Humanos , Masculino , Femenino , Niño , Adolescente , Antropometría , Aptitud Física , Voleibol , Factores Sexuales , Estudios TransversalesRESUMEN
BACKGROUND: Physical testing is crucial for athlete monitoring, talent identification, optimizing training, and tailoring programs to enhance game-performance in elite competitions. HYPOTHESIS: Load-velocity (L-V) relationship variables discriminate between elite and junior volleyball players, correlate with volleyball-specific performance, and are generalizable across lower- and upper-body exercises. STUDY DESIGN: Cross-sectional study. LEVEL OF EVIDENCE: Level 3. METHODS: A total of 9 elite and 11 junior volleyball players were assessed for the L-V relationship (load-axis intercept [L0], velocity-axis intercept [v0], and area under the L-V relationship line [Aline]) during the countermovement jump (CMJ) and bench press throw (BPT) exercises. Block and spike jump height, as well as standing and jumping spike speed were assessed 24 hours later. RESULTS: Elite players presented greater magnitude in the L-V variables (P ≤ 0.03; effect size [ES] ≥ 1.06) and higher volleyball-specific performance (P ≤ 0.03; ES ≥ 1.09) than juniors (except for CMJ v0 and Aline). The L-V relationship variables were significantly associated with the block and spike jump height and jumping spike speed only in elite players (r ≥ 0.703 and P ≤ 0.04 in 11 out of 18 correlations). No significant associations were observed between CMJ and BPT for any L-V relationship variable (r ≤ 581; P ≥ 0.08, except for Aline in junior players). CONCLUSION: The L-V relationship is a practical procedure to assess volleyball players' maximal mechanical capacities, which are associated with volleyball-specific performance in elite players. However, these data should not be used interchangeably between playing standards or exercises. CLINICAL RELEVANCE: This information might help strength and conditioning coaches to prescribe more effective training programs that focus on developing the specific physical capacities necessary for players to potentially advance to elite status.
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Bacterial competition is a significant driver of toxin polymorphism, which allows continual compensatory evolution between toxins and the resistance developed to overcome their activity. Bacterial Rearrangement hot spot (Rhs) proteins represent a widespread example of toxin polymorphism. Here, we present the 2.45 Å cryo-electron microscopy structure of Tse5, an Rhs protein central to Pseudomonas aeruginosa type VI secretion system-mediated bacterial competition. This structural insight, coupled with an extensive array of biophysical and genetic investigations, unravels the multifaceted functional mechanisms of Tse5. The data suggest that interfacial Tse5-membrane binding delivers its encapsulated pore-forming toxin fragment to the target bacterial membrane, where it assembles pores that cause cell depolarisation and, ultimately, bacterial death.
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Toxinas Bacterianas , Dermatitis , Humanos , Microscopía por Crioelectrón , Toxinas Bacterianas/genética , Membranas , Proteínas Bacterianas/genética , Secuencia de Bases , Membrana CelularRESUMEN
OBJECTIVES: To evaluate the efficacy and safety of the two-pill regimen bictegravir/emtricitabine/tenofovir alafenamide (BIC/FTC/TAF) plus darunavir/cobicistat as a switching strategy in heavily treatment-experienced people living with HIV (PLWH). METHODS: Multicentre, prospective, single-arm pilot clinical trial. Participants were virologically suppressed adults receiving a stable antiretroviral regimen of at least three pills from at least three drug families due to previous virological failures and/or toxicities with no documented resistance to integrase strand transfer inhibitors or darunavir (≥15 points, Stanford). Clinical and laboratory assessments were performed at 0, 4, 12, 24, 36 and 48 weeks. HIV-1 proviral DNA was amplified and sequenced by Illumina at baseline. Plasma bictegravir concentrations were determined in 22 patients using UHPLC-MS/MS. The primary study endpoint was viral load (VL)<â50 copies/mL at Week 48 (ITT). RESULTS: We enrolled 63 participants (92% men) with median baseline CD4 count of 515 cells/mm3 (IQR: 334.5-734.5), 24 years on ART (IQR: 15.9-27.8). The median number of pills was 4 (range: 3-10). At baseline, proviral DNA was amplified in 39 participants: 33/39 had resistance mutations. Three participants discontinued owing to toxicity. At 48 weeks, 95% had VLâ<â50 copies/mL by ITT and 100% by PP analysis. A modest increase was observed in the bictegravir plasma concentration, and a significant decrease in estimated glomerular filtration rate was observed only at Week 4, probably related to interaction with renal transporters. CONCLUSIONS: Our data suggest that BIC/FTC/TAFâ+âdarunavir/cobicistat is an effective, well-tolerated regimen that may improve convenience and, potentially, long-term success in stable heavily pre-treated PLWH.
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Fármacos Anti-VIH , Infecciones por VIH , Adulto , Femenino , Humanos , Masculino , Adenina/uso terapéutico , Alanina/uso terapéutico , Fármacos Anti-VIH/efectos adversos , Antirretrovirales/uso terapéutico , Cobicistat/uso terapéutico , Darunavir/uso terapéutico , ADN/uso terapéutico , Emtricitabina/uso terapéutico , Infecciones por VIH/tratamiento farmacológico , Estudios Prospectivos , Espectrometría de Masas en TándemRESUMEN
The envelope (E) protein is a small polypeptide that can form ion channels in coronaviruses. In SARS coronavirus 2 (SARS-CoV-2), the agent that caused the recent COVID-19 pandemic, and its predecessor SARS-CoV-1, E protein is found in the endoplasmic reticulum-Golgi intermediate compartment (ERGIC), where virion budding takes place. Several reports claim that E protein promotes the formation of "cation-selective channels". However, whether this term represents specificity to certain ions (e.g., potassium or calcium) or the partial or total exclusion of anions is debatable. Herein, we discuss this claim based on the available data for SARS-CoV-1 and -2 E and on new experiments performed using the untagged full-length E protein from SARS-CoV-2 in planar lipid membranes of different types, including those that closely mimic the ERGIC membrane composition. We provide evidence that the selectivity of the E-induced channels is very mild and depends strongly on lipid environment. Thus, despite past and recent claims, we found no indication that the E protein forms cation-selective channels that prevent anion transport, and even less that E protein forms bona fide specific calcium channels. In fact, the E channel maintains its multi-ionic non-specific neutral character even in concentrated solutions of Ca2+ ions. Also, in contrast to previous studies, we found no evidence that SARS-CoV-2 E channel activation requires a particular voltage, high calcium concentrations or low pH, in agreement with available data from SARS-CoV-1 E. In addition, sedimentation velocity experiments suggest that the E channel population is mostly pentameric, but very dynamic and probably heterogeneous, consistent with the broad distribution of conductance values typically found in electrophysiological experiments. The latter has been explained by the presence of proteolipidic channel structures.
Asunto(s)
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/metabolismo , Proteínas del Envoltorio Viral/química , Calcio , Pandemias , Iones , LípidosRESUMEN
Dynorphin A (DynA) is an endogenous neuropeptide that besides acting as a ligand of the κ-opioid receptor, presents some non-opioid pathophysiological properties associated to its ability to induce cell permeability similarly to cell-penetrating peptides (CPPs). Here, we use electrophysiology experiments to show that amphiphilic DynA generates aqueous pores in neutral membranes similar to those reported previously in charged membranes, but we also find other events thermodynamically incompatible with voltage-driven ion channel activity (i.e. non-zero currents with no applied voltage in symmetric salt conditions, reversal potentials that exceed the theoretical limit for a given salt concentration gradient). By comparison with current traces generated by other amphiphilic molecule known to spontaneously cross membranes, we hypothesize that DynA could directly translocate across neutral bilayers, a feature never observed in charged membranes following the same electrophysiological protocol. Our findings suggest that DynA interaction with the cellular membrane is modulated by the lipid charge distribution, enabling either passive ionic transport via membrane remodeling and pore formation or by peptide direct internalization independent of cellular transduction pathways.
Asunto(s)
Dinorfinas , Membrana Dobles de Lípidos , Membrana Dobles de Lípidos/química , Dinorfinas/farmacología , Dinorfinas/análisis , Dinorfinas/química , Membrana Celular/metabolismo , Péptidos/química , Canales Iónicos/metabolismoRESUMEN
BACKGROUND: The aim of this study was to determine the differences in the incidence, epidemiology, clinical characteristics and risk factors of infections in living donor kidney transplant recipients using robotic-assisted kidney transplantation (RAKT) and open approach. METHODS: We conducted a retrospective observational study from January 2016 to December 2019. For the risk factor analysis, a matched case-control study (1:1 ratio) was performed (robotic vs open). Control subjects were matched for living donor and time of transplantation. The data included de novo immunosuppressive regimen, delayed graft function, urological complications, acute allograft rejection and incidence, clinical features, microbiological findings and outcomes of infections. RESULTS: Ninety-four RAKT and 84 controls were included. There were no differences between groups in terms of age, gender, BMI, median days of hospitalization, immunosuppressive regimen, need for surgical urologic procedures post-transplantation, presence of urinary leak or acute allograft rejection. Thirty-five percent of all recipients analyzed presented an infection, mostly asymptomatic bacteriuria (49%), symptomatic urinary tract infection (31%) and surgical site infection (10%). Pseudomonas aeruginosa was the most frequent isolated microorganism in 67%, followed by E. coli (20%), Enterococcus faecalis (17%) and Klebsiella pneumoniae (10%). Eight percent of the microorganisms were multidrug resistant. The open kidney transplantation group presented more infections compared to RAKT (43 vs 27%, p = 0.04). After multivariate analysis, need for surgical urologic procedure post-transplantation (OR 6.2, 95% CI 1.1-35), BMI ≥ 30 (OR 3.6, 95% CI 1.5-9) and acute allograft rejection (OR 3.2, 95% CI 1.2-8.5) were associated with infection, whereas RAKT (OR 0.5, 95% CI 0.2-0.9) and the use of JJ catheter (OR 0.36, 95% CI 0.17-0.72) were protective factors. CONCLUSIONS: Infection is a frequent event in patients receiving a living donor kidney transplant. Acute allograft rejection, need for surgical urologic procedure post-transplantation and BMI were associated with infection, whereas robotic surgery was a protective factor in living donor kidney transplantation.