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Cerebral oedema is associated with morbidity and mortality after traumatic brain injury (TBI)1. Noradrenaline levels are increased after TBI2-4, and the amplitude of the increase in noradrenaline predicts both the extent of injury5 and the likelihood of mortality6. Glymphatic impairment is both a feature of and a contributor to brain injury7,8, but its relationship with the injury-associated surge in noradrenaline is unclear. Here we report that acute post-traumatic oedema results from a suppression of glymphatic and lymphatic fluid flow that occurs in response to excessive systemic release of noradrenaline. This post-TBI adrenergic storm was associated with reduced contractility of cervical lymphatic vessels, consistent with diminished return of glymphatic and lymphatic fluid to the systemic circulation. Accordingly, pan-adrenergic receptor inhibition normalized central venous pressure and partly restored glymphatic and cervical lymphatic flow in a mouse model of TBI, and these actions led to substantially reduced brain oedema and improved functional outcomes. Furthermore, post-traumatic inhibition of adrenergic signalling boosted lymphatic export of cellular debris from the traumatic lesion, substantially reducing secondary inflammation and accumulation of phosphorylated tau. These observations suggest that targeting the noradrenergic control of central glymphatic flow may offer a therapeutic approach for treating acute TBI.
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Edema Encefálico , Lesiones Traumáticas del Encéfalo , Sistema Glinfático , Norepinefrina , Animales , Ratones , Antagonistas Adrenérgicos/farmacología , Antagonistas Adrenérgicos/uso terapéutico , Edema Encefálico/complicaciones , Edema Encefálico/tratamiento farmacológico , Edema Encefálico/metabolismo , Edema Encefálico/prevención & control , Lesiones Traumáticas del Encéfalo/complicaciones , Lesiones Traumáticas del Encéfalo/tratamiento farmacológico , Lesiones Traumáticas del Encéfalo/metabolismo , Modelos Animales de Enfermedad , Sistema Glinfático/efectos de los fármacos , Sistema Glinfático/metabolismo , Inflamación/complicaciones , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Inflamación/prevención & control , Vasos Linfáticos/metabolismo , Norepinefrina/metabolismo , Fosforilación , Receptores Adrenérgicos/metabolismoRESUMEN
Cells of vertebrate and invertebrate organisms express proteins specialized in membrane channel-based cell-cell communication that are absent in unicellular organisms. We recently described the prediction of some members of the large-pore channel family in kinetoplastids, consisting of proteins called unnexins, which share several structural features with innexin and pannexin proteins. Here, we demonstrated that the unnexin1 protein (Unx1) is delivered to the cell membrane, displaying a topology consisting of four transmembrane domains with C and N termini on the cytoplasmic side and form large-pore channels that are permeable to small molecules. Low extracellular Ca2+/Mg2+ levels or extracellular alkalinization, but not mechanical stretching, increases channel activity. The Unx1 channel mediates the influx of Ca2+ and does not form intercellular dye coupling between HeLa Unx1 transfected cells. Unx1 channel function was further evidenced by its ability to mediate ionic currents when expressed in Xenopus oocytes. Downregulation of Unx1 mRNA with morpholine contains Trypanosoma cruzi invasion. Phylogenetic analysis revealed the presence of Unx1 homologs in other protozoan parasites, suggesting a conserved function for these channel parasites in other protists. Our data demonstrate that Unx1 forms large-pore membrane channels, which may serve as a diffusional pathway for ions and small molecules that are likely to be metabolic substrates or waste products, and signaling autocrine and paracrine molecules that could be involved in cell invasion. As morpholinos-induced downregulation of Unx1 reduces the infectivity of trypomastigotes, the Unx1 channels might be an attractive target for developing trypanocide drugs.
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Subunidades de Proteína , Filogenia , Membrana Celular , Citoplasma , MorfolinosRESUMEN
Ca2+ signalling plays a crucial role in determining lymphatic muscle cell excitability and contractility through its interaction with the Ca2+-activated Cl- channel anoctamin 1 (ANO1). In contrast, the large-conductance (BK) Ca2+-activated K+ channel (KCa) and other KCa channels have prominent vasodilatory actions by hyperpolarizing vascular smooth muscle cells. Here, we assessed the expression and contribution of the KCa family to mouse and rat lymphatic collecting vessel contractile function. The BK channel was the only KCa channel consistently expressed in fluorescence-activated cell sorting-purified mouse lymphatic muscle cell lymphatic muscle cells. We used a pharmacological inhibitor of BK channels, iberiotoxin, and small-conductance Ca2+-activated K+ channels, apamin, to inhibit KCa channels acutely in ex vivo isobaric myography experiments and intracellular membrane potential recordings. In basal conditions, BK channel inhibition had little to no effect on either mouse inguinal-axillary lymphatic vessel (MIALV) or rat mesenteric lymphatic vessel contractions or action potentials (APs). We also tested BK channel inhibition under loss of ANO1 either by genetic ablation (Myh11CreERT2-Ano1 fl/fl, Ano1ismKO) or by pharmacological inhibition with Ani9. In both Ano1ismKO MIALVs and Ani9-pretreated MIALVs, inhibition of BK channels increased contraction amplitude, increased peak AP and broadened the peak of the AP spike. In rat mesenteric lymphatic vessels, BK channel inhibition also abolished the characteristic post-spike notch, which was exaggerated with ANO1 inhibition, and significantly increased the peak potential and broadened the AP spike. We conclude that BK channels are present and functional on mouse and rat lymphatic muscle cells but are otherwise masked by the dominance of ANO1. KEY POINTS: Mouse and rat lymphatic muscle cells express functional BK channels. BK channels make little contribution to either rat or mouse lymphatic collecting vessel contractile function in basal conditions across a physiological pressure range. ANO1 limits the peak membrane potential achieved in the action potential and sets a plateau potential limiting the voltage-dependent activation of BK. BK channels are activated when ANO1 is absent or blocked and slightly impair contractile strength by reducing the peak membrane potential achieved in the action potential spike and accelerating the post-spike repolarization.
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Potenciales de Acción , Anoctamina-1 , Canales de Potasio de Gran Conductancia Activados por el Calcio , Vasos Linfáticos , Animales , Anoctamina-1/metabolismo , Anoctamina-1/genética , Canales de Potasio de Gran Conductancia Activados por el Calcio/metabolismo , Canales de Potasio de Gran Conductancia Activados por el Calcio/fisiología , Ratones , Ratas , Potenciales de Acción/fisiología , Masculino , Vasos Linfáticos/fisiología , Vasos Linfáticos/metabolismo , Ratones Endogámicos C57BL , Ratones Noqueados , Contracción Muscular/fisiología , Ratas Sprague-Dawley , Femenino , Miocitos del Músculo Liso/fisiología , Miocitos del Músculo Liso/metabolismo , Miocitos del Músculo Liso/efectos de los fármacosRESUMEN
Recent works on electric bubbles (including the experimental demonstration of electric skyrmions) constitute a breakthrough akin to the discovery of magnetic skyrmions some 15 years ago. So far research has focused on obtaining and visualizing these objects, which often appear to be immobile (pinned) in experiments. Thus, critical aspects of magnetic skyrmions-e.g., their quasiparticle nature, Brownian motion-remain unexplored (unproven) for electric bubbles. Here we use predictive atomistic simulations to investigate the basic dynamical properties of these objects in pinning-free model systems. We show that it is possible to find regimes where the electric bubbles can present long lifetimes (â¼ns) despite being relatively small (diameter <2 nm). Additionally, we find that they can display stochastic dynamics with large and highly tunable diffusion constants. We thus establish the quasiparticle nature of electric bubbles and put them forward for the physical effects and applications (e.g., in token-based probabilistic computing) considered for magnetic skyrmions.
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Nanostructured ferroelectrics display exotic multidomain configurations resulting from the electrostatic and elastic boundary conditions they are subject to. While the ferroelectric domains appear frozen in experimental images, atomistic second-principles studies suggest that they may become spontaneously mobile upon heating, with the polar order melting in a liquidlike fashion. Here, we run molecular dynamics simulations of model systems (PbTiO_{3}/SrTiO_{3} superlattices) to study the unique features of this transformation. Most notably, we find that the multidomain state loses its translational and orientational orders at different temperatures, resembling the behavior of liquid crystals and yielding an intermediate hexaticlike phase. Our simulations reveal the mechanism responsible for the melting and allow us to characterize the stochastic dynamics in the hexaticlike phase: we find evidence that it is thermally activated, with domain reorientation rates that grow from tens of gigahertzs to terahertzs in a narrow temperature window.
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INTRODUCTION: To date, there have been no studies conducted on the development of interosseous muscles (IO) in the human hand. This study aimed to investigate the development of these muscles in order to clarify their terminal insertions and their relationship with the metacarpophalangeal joints. METHODS: Serial sections of 25 human specimens (9 embryos and 16 fetuses) between the 7th and 14th week of development, sourced from the Collection of the Department of Anatomy and Embryology at UCM Faculty of Medicine, were analysed bilaterally using a conventional optical microscope. RESULTS: Our findings revealed that, during the 7th week of development, the metacarpophalangeal interzone mesenchyme extended into the extensor apparatus of the fingers. Furthermore, we observed that the joint capsule and the tendon of the IO derive from the articular interzone mesenchyme. By the end of the 7th week, corresponding to Carnegie stage 21, the myotendinous junction appeared, initiating cavitation of the metacarpophalangeal joint. During the fetal period, the terminal insertions of the IO were identified: both the dorsal interosseous (DI) and palmar interosseous (PI) muscles insert into the metacarpophalangeal joint capsule and establish a connection with the volar plate located at the base of the proximal phalanx and the extensor apparatus. Some muscle fibres also attach to the joint capsule at the level of the proximal synovial cul-de-sac. The functional implications of these findings are discussed within this work. CONCLUSION: This study provides the first detailed description of the development of the interosseous muscles in the human hand.
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Circumstances that precipitate interactions among species that have never interacted during their evolutionary histories create ideal conditions for the generation of zoonoses. Zoonotic diseases have caused some of the most devastating epidemics in human history. Contact among species that come from different ecosystems or regions creates the risk of zoonoses. In certain situations, humans are generating and promoting conditions that contribute to the creation of infectious diseases and zoonoses. These conditions lead to interactions between wildlife species that have hitherto not interacted under normal circumstances. I call for recognition of the zoonotic potential that novel and unwanted interactions have; identification of these new interactions that are occurring among wild animals, domestic animals, and humans; and efforts to stop these kinds of interactions because they can give rise to zoonotic outbreaks. Live animal markets, the exotic pet trade, illegal wildlife trade, human use and consumption of wild animals, invasive non-native species, releasing of exotic pets, and human encroachment in natural areas are among the activities that cause the most interactions among wild species, domestic species, and humans. These activities should not occur and must be controlled efficiently to prevent future epidemic zoonoses. Society must develop a keen ability to identify these unnatural interactions and prevent them. Controlling these interactions and efficiently addressing their causal factors will benefit human health and, in some cases, lead to positive environmental, ethical, and socioeconomic outcomes. Until these actions are taken, humanity will face future zoonoses and zoonotic pandemic.
Evitar interacciones novedosas e indeseadas entre especies para disminuir el riesgo de zoonosis Resumen Las circunstancias que promueven interacciones entre especies que nunca han interactuado durante sus historias evolutivas crean condiciones ideales para la generación de zoonosis. Las enfermedades zoonóticas han causado algunas de las epidemias más devastadoras en la historia de la humanidad. El contacto entre especies que provienen de diferentes ecosistemas o regiones crea el riesgo de zoonosis. En determinadas situaciones, los seres humanos estamos generando y promoviendo condiciones que contribuyen a la creación de enfermedades infecciosas y zoonosis. Estas condiciones conducen a interacciones entre especies silvestres que hasta ahora no habían interactuado en circunstancias normales. Hago un llamado para que se reconozca el potencial zoonótico que tienen las interacciones nuevas y no deseadas; que se identifiquen estas nuevas interacciones que provocamos entre animales silvestres, animales domésticos y humanos; y esforzarnos para detener este tipo de interacciones porque pueden dar lugar a brotes zoonóticos. Los mercados de animales vivos, el comercio de mascotas exóticas, el comercio ilegal de vida silvestre, el uso y consumo humano de animales silvestres, las especies invasoras no nativas, la liberación de mascotas exóticas y la invasión humana en áreas naturales, se encuentran entre las actividades que causan la mayor cantidad de interacciones entre especies silvestres, especies domésticas y humanos. Estas actividades no deberían ocurrir y deben controlarse eficientemente para prevenir futuras zoonosis epidémicas. La sociedad debe desarrollar una gran capacidad para identificar estas interacciones antinaturales y prevenirlas. Controlar estas interacciones y abordar eficientemente sus factores causales beneficiará la salud humana y, en algunos casos, conducirá a resultados ambientales, éticos y socioeconómicos positivos. Mientras estas medidas no se tomen, la humanidad enfrentará futuras zoonosis y pandemias zoonóticas.
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Animales Salvajes , Zoonosis , Zoonosis/prevención & control , Zoonosis/epidemiología , Animales , Humanos , Conservación de los Recursos Naturales , Animales Domésticos , Especies IntroducidasRESUMEN
BACKGROUND: The objective of this study was to analyze the effects of sedation administration on clinical parameters, comfort status, intubation requirements, and the pediatric intensive care unit (PICU) length of stay (LOS) in children with acute respiratory failure (ARF) receiving noninvasive ventilation (NIV). METHODS: Thirteen PICUs in Spain participated in a prospective, multicenter, observational trial from January to December 2021. Children with ARF under the age of five who were receiving NIV were included. Clinical information and comfort levels were documented at the time of NIV initiation, as well as at 3, 6, 12, 24, and 48 h. The COMFORT-behavior (COMFORT-B) scale was used to assess the patients' level of comfort. NIV failure was considered to be a requirement for endotracheal intubation. RESULTS: A total of 457 patients were included, with a median age of 3.3 months (IQR 1.3-16.1). Two hundred and thirteen children (46.6%) received sedation (sedation group); these patients had a higher heart rate, higher COMFORT-B score, and lower SpO2/FiO2 ratio than did those who did not receive sedation (non-sedation group). A significantly greater improvement in the COMFORT-B score at 3, 6, 12, and 24 h, heart rate at 6 and 12 h, and SpO2/FiO2 ratio at 6 h was observed in the sedation group. Overall, the NIV success rate was 95.6%-intubation was required in 6.1% of the sedation group and in 2.9% of the other group (p = 0.092). Multivariate analysis revealed that the PRISM III score at NIV initiation (OR 1.408; 95% CI 1.230-1.611) and respiratory rate at 3 h (OR 1.043; 95% CI 1.009-1.079) were found to be independent predictors of NIV failure. The PICU LOS was correlated with weight, PRISM III score, respiratory rate at 12 h, SpO2 at 3 h, FiO2 at 12 h, NIV failure and NIV duration. Sedation use was not found to be independently related to NIV failure or to the PICU LOS. CONCLUSIONS: Sedation use may be useful in children with ARF treated with NIV, as it seems to improve clinical parameters and comfort status but may not increase the NIV failure rate or PICU LOS, even though sedated children were more severe at technique initiation in the present sample.
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Unidades de Cuidado Intensivo Pediátrico , Ventilación no Invasiva , Insuficiencia Respiratoria , Humanos , Ventilación no Invasiva/métodos , Ventilación no Invasiva/estadística & datos numéricos , Estudios Prospectivos , Femenino , Masculino , Lactante , Unidades de Cuidado Intensivo Pediátrico/estadística & datos numéricos , Unidades de Cuidado Intensivo Pediátrico/organización & administración , Insuficiencia Respiratoria/terapia , España , Preescolar , Hipnóticos y Sedantes/uso terapéutico , Hipnóticos y Sedantes/administración & dosificación , Sedación Consciente/métodos , Sedación Consciente/estadística & datos numéricosRESUMEN
PURPOSE: Associations between health-related parameters and lung function remain unclear in childhood. The study aims to evaluate the relationship between physical fitness and anthropometric parameters with the lung function of healthy scholar-aged children. METHOD: A total of 418 children aged 7 years old participated in this study. The associations of physical fitness (handgrip strength, standing broad jump, and 800-m run) and anthropometric (waist circumference and body mass index) parameters with lung function (forced vital capacity and forced expiratory volume in 1 s) were analyzed using a mixed-linear regression model. RESULTS: Girls had significantly lower forced vital capacity values (P = .006) and physical fitness (P < .030) compared to boys. On mixed-linear regression analyses, waist circumference (P = .003) was independently associated with forced vital capacity, explaining 34.6% of its variance, while handgrip strength (P = .042) and waist circumference (P = .010) were independently associated with forced expiratory volume in 1 second, accounting together for 26.5% of its variance in 7-year-old healthy children. CONCLUSIONS: Handgrip strength and waist circumference were associated with lung function in healthy children highlighting the influence of upper body muscular strength and trunk dimension on lung function. Our results corroborate the need to promote physical fitness during childhood to protect against lung complications in later on in life.
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AIM AND OBJECTIVES: To analyse the levels of anxiety, depression, post-traumatic stress, and burnout among nursing professionals working in the Imbabura region of Ecuador during the COVID-19 pandemic and identify the contributing socio-occupational factors. BACKGROUND: The high demand for care of COVID-19 patients led to increased work pressure on nurses, owing to increased demands for care and shortages of medical supplies and protective equipment. DESIGN: A cross-sectional study was conducted from September to December 2022 using a self-administered questionnaire addressed to nursing professionals who cared for COVID-19 patients. METHODS: The questionnaire included socio-demographic characteristics, the Spanish adaptation of Hospital Anxiety and Depression Scale (HADS-Spanish), Impact of Event Scale-Revised (IES-R) for the evaluation of post-traumatic stress disorder (PTSD), and the Spanish adaptation of the Maslach Burnout Inventory-Human Services Survey (MBI-HSS-Spanish) for burnout assessment. Univariate and multivariate analyses were performed. RESULTS: Of the 782 participants, 88.6% had a high level of burnout (MBI-HSS-Spanish scale score > 27). Female nurses, nurses with eight-hour work shifts, and older professionals exhibited high levels of anxiety and depression. Prolonged working hours in COVID-19 patient care services were found to be a risk factor for burnout and post-traumatic stress. CONCLUSIONS: Participating nurses presented with a high level of chronic work stress and exhibited signs of anxiety and depression during the period under consideration. Providing nurses with psychological support measures and performing liaison consultations will alleviate the psychological burden on nurses. RELEVANCE TO CLINICAL PRACTICE: The study has shown that accounting for the environments where the emotional impact is greatest and how to reduce it would not only reduce anxiety, depression, and burnout in nurses but also improve the quality of care, not only in pandemic. PATIENT OR PUBLIC CONTRIBUTION: Nurses contributed to the conduct of the study by participating in the data collection via questionaries.
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PURPOSE: Depending on its axis, pronation varies from the radius rotation around the steady ulna to the reciprocal adduction of the radius and abduction of the ulna. While there is no question that pronator teres is a central pronation agonist, anconeus's role is not settled. The current investigation comparing palpation and ultrasonography in these two muscles during pronation along the axis capitulum-second digit evolved from a serendipitous finding in a clinical anatomy seminar. METHODS: Single-hand palpation and two-transducer ultrasonography over anconeus and pronator teres were used on ten normal subjects to investigate their contraction during pronation around the capitulum-second digit axis. These studies were done independently and blind to the results of the other. The statistical analysis between palpation and ultrasonography was performed with Cohen's kappa coefficient and the χ2 test. RESULTS: On palpation, on resisted full pronation, anconeus contracted in 8/10 subjects and pronator teres in 10/10 subjects. Without resistance, the corresponding ratios were 5/10 and 9/10. On two-transducer ultrasonography, the comparable ratios were 7/10 and 10/10, and 3/10 and 10/10. A fair concordance (Cohen's kappa = 0.21) between palpation and ultrasonography in detecting the simultaneous status of anconeus and pronator teres during resisted full pronation. Anatomic dissection illustrated the elements involved. CONCLUSIONS: Plain palpation confirmed by ultrasonography showed the simultaneous contraction of anconeus and pronator teres during resisted pronation in most of the studied subjects. The study suggests that palpation can be helpful in directly studying muscle activity during movement.
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Voluntarios Sanos , Músculo Esquelético , Palpación , Pronación , Ultrasonografía , Humanos , Pronación/fisiología , Ultrasonografía/métodos , Masculino , Adulto , Femenino , Músculo Esquelético/diagnóstico por imagen , Músculo Esquelético/fisiología , Músculo Esquelético/anatomía & histología , Palpación/métodos , Adulto Joven , Contracción Muscular/fisiologíaRESUMEN
Maturity status and relative age are two of the determining factors in talent development. The aim of the study was to analyze the influence of biological maturity status and relative age on physical performance in young male and female handball players. The sample included 48 males (14.11 ± 1.17 years) and 41 females (14.25 ± 1.64 years) players from one Spanish professional handball academy. Anthropometric data (height, sitting height, body mass and self-reported biological parent heights) and physical performance data (CMJ, DJ, 20 m speed, T-test and throwing velocity) were collected. Biological maturity status was determined as the percentage of predicted adult height, while relative age was estimated in birth quartiles based on biennial age grouping (Q1-Q8). The results showed a positive correlation between maturity status and CMJ in male players (p < 0.01). Differences in CMJ performance according to maturity status were identified (p < 0.05), with higher jump heights being recorded especially in early maturing boys (p < 0.01) and first lines and wings (p < 0.05). The variance in CMJ test scores could be explained by the maturity status by 42.90% in U-15 (p < 0.05) and 72.60% in U-16 male players (p < 0.001). By contrast, no differences were found in girls (p > 0.05). Moreover, no relationships were found between relative age and indices of physical performance (p > 0.05). Overall, maturity status had greater impacts on the tests of physical performance than relative age. Stakeholders should monitor the maturity status of young handball players to avoid physical performance biases that do not allow them to develop their sporting potential.
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The direct auration of arenes is a key step in numerous gold-catalyzed reactions. Although reported more than 100â years ago, understanding of its underlying mechanism has been hampered by the difficulties in the isolation of relevant intermediates given the propensity of gold(III) species to undergo reductive elimination. Here, we report the synthesis and isolation of a new family of intriguing zwitterionic [C(sp3)^C(sp2)]-auracyclopentanes, as well as of their alkyl-gold(III) precursors and demonstrate their value as mechanistic probes to study the C(sp2)-Au bond-forming event. Experimental investigations employing Kinetic Isotope Effects (KIE), Hammett plot, and Eyring analysis provided important insights into the formation of the auracycle. The data suggest a SEAr mechanism wherein the slowest step might be the π-coordination between the arene and the gold(III) center, en route to the Wheland intermediate. We also show that these auracyclopentanes can work as catalysts in several gold-promoted transformations.
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Synucleinopathies are a group of neurodegenerative diseases without effective treatment characterized by the abnormal aggregation of alpha-synuclein (aSyn) protein. Changes in levels or in the amino acid sequence of aSyn (by duplication/triplication of the aSyn gene or point mutations in the encoding region) cause familial cases of synucleinopathies. However, the specific molecular mechanisms of aSyn-dependent toxicity remain unclear. Increased aSyn protein levels or pathological mutations may favor abnormal protein-protein interactions (PPIs) that could either promote neuronal death or belong to a coping response program against neurotoxicity. Therefore, the identification and modulation of aSyn-dependent PPIs can provide new therapeutic targets for these diseases. To identify aSyn-dependent PPIs we performed a proximity biotinylation assay based on the promiscuous biotinylase BioID2. When expressed as a fusion protein, BioID2 biotinylates by proximity stable and transient interacting partners, allowing their identification by streptavidin affinity purification and mass spectrometry. The aSyn interactome was analyzed using BioID2-tagged wild-type (WT) and pathological mutant E46K aSyn versions in HEK293 cells. We found the 14-3-3 epsilon isoform as a common protein interactor for WT and E46K aSyn. 14-3-3 epsilon correlates with aSyn protein levels in brain regions of a transgenic mouse model overexpressing WT human aSyn. Using a neuronal model in which aSyn cell-autonomous toxicity is quantitatively scored by longitudinal survival analysis, we found that stabilization of 14-3-3 protein-proteins interactions with Fusicoccin-A (FC-A) decreases aSyn-dependent toxicity. Furthermore, FC-A treatment protects dopaminergic neuronal somas in the substantia nigra of a Parkinson's disease mouse model. Based on these results, we propose that the stabilization of 14-3-3 epsilon interaction with aSyn might reduce aSyn toxicity, and highlight FC-A as a potential therapeutic compound for synucleinopathies.
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Sinucleinopatías , alfa-Sinucleína , Ratones , Humanos , Animales , alfa-Sinucleína/metabolismo , Proteínas 14-3-3/genética , Proteínas 14-3-3/metabolismo , Células HEK293 , Ratones Transgénicos , Neuronas Dopaminérgicas/metabolismoRESUMEN
OBJECTIVE: To develop an experimental method for routine isolation and short-term culture of primary lymphatic endothelial cells from specific collecting vessels. METHODS: Lymphatic endothelial cell tubes (LECTs) were isolated from micro-dissected collecting vessels. LECTs were allowed to attach and grow for ~3 weeks before being passaged. Non-purified cultures were partially characterized by immunofluorescence and RT-PCR at passages 1-2. RESULTS: The method was validated in cultures of primary lymphatic endothelial cells (LECs) from male and female mice. After 1 or 2 passages, >60% of the LECs maintained expression of Prox1. Expression of 22 different genes was assessed using RT-PCR. Prox1, Vegfr3, eNos, Cdh5, Pecam1, Cx43, Cx37, and Cx47, among others, were expressed in these short-term cultured LECs, while Myh11, Cnn1, Desmin, and Cd11b were not detected. Prox1 expression, as determined by western blotting, was similar in cultured LECs from age-matched male and female mice. Confocal imaging of intracellular calcium in cultures of primary LECs from Cdh5-GCaMP8 mice demonstrated that a functional phenotype was maintained, similar to lymphatic endothelial cells in freshly isolated vessels. CONCLUSIONS: This method provides an innovative tool for routine isolation and study of primary LECs from specific collecting lymphatic vessels from any mouse, and in fact, from other species.
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Células Endoteliales , Vasos Linfáticos , Femenino , Masculino , Animales , Ratones , Células Endoteliales/metabolismo , Vasos Linfáticos/metabolismo , FenotipoRESUMEN
Colorectal cancer (CRC) is one of the most common cancers worldwide. Its etiopathogenesis is complex, mainly influenced by genetic instability caused by the accumulation of mutations. The XRCC1 gene, which is involved in DNA repair, has been associated with CRC through the R194W (C194T) and R399Q (G399A) polymorphisms, but the results are inconsistent. Here, we analyzed the association of these polymorphisms with sporadic CRC in a northeastern Mexican population, including 155 male CRC patients and 155 male controls. Genotyping was performed using the RFLP method. An association with CRC was found for the 399A allele (G vs A; OR = 1.48 (1.03-2.13), P=0.034) and for the 399AA genotype in a codominant model (AA vs GG; OR = 3.11 (1.06-9.10), P=0.031). In contrast, there were no significant differences between CRC patients and controls for the C194T polymorphism (C vs T; OR = 0.82 (0.52-1.31), P=0.41). These results are consistent with many similar studies, but further research is needed to verify whether the XRCC1 R194W and R399Q polymorphisms play a role in CRC etiology. The functional significance of these polymorphisms is unclear, but some studies suggest that they influence DNA repair capacity and, thus, cancer risk.
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Neoplasias Colorrectales , Proteínas de Unión al ADN , Proteína 1 de Reparación por Escisión del Grupo de Complementación Cruzada de las Lesiones por Rayos X , Humanos , Masculino , Estudios de Casos y Controles , Neoplasias Colorrectales/epidemiología , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Proteínas de Unión al ADN/genética , Predisposición Genética a la Enfermedad , Genotipo , Polimorfismo Genético , Polimorfismo de Nucleótido Simple/genética , Proteína 1 de Reparación por Escisión del Grupo de Complementación Cruzada de las Lesiones por Rayos X/genéticaRESUMEN
INTRODUCTION: Acute plastic deformation refers to a traumatic bending or bowing without a detectable cortical defect. CASE PRESENTATION AND DISCUSSION: We describe a rare case from an individual that was exhumed from the Hispano-Mudejar necropolis in Uceda (Guadalajara, Spain) dated between the 13th and 14th centuries AD. The case corresponds to an adult woman, with a bowing involvement of the left ulna and radius. After making the differential diagnosis with various pathologies likely to present with this alteration, we reached the diagnosis of acute plastic deformation of the forearm through external and radiological examination and comparison with the healthy contralateral forearm. CONCLUSIONS: Acute plastic deformation is a rare traumatic injury, not described until the last century and only rarely described in palaeopathological contexts. We contribute a new case, the first being sufficiently documented, contributing to the knowledge and diagnosis of this type of trauma in the ancient bone, while deepening the knowledge of the living conditions of the medieval Mudejar population of Uceda.
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Traumatismos del Antebrazo , Antebrazo , Adulto , Femenino , Humanos , Antebrazo/patología , Traumatismos del Antebrazo/patología , Historia Medieval , Radio (Anatomía)/lesiones , Radio (Anatomía)/patología , Cúbito/lesiones , Cúbito/patologíaRESUMEN
The calculation of relative free energies (ΔΔG) for charge-changing mutations at protein-protein interfaces through alchemical methods remains challenging due to variations in the system's net charge during charging steps, the possibility of mutated and contacting ionizable residues occurring in various protonation states, and undersampling issues. In this study, we present a set of strategies, collectively termed TIRST/TIRST-H+, to address some of these challenges. Our approaches combine thermodynamic integration (TI) with the prediction of pKa shifts to calculate ΔΔG values. Moreover, special sets of restraints are employed to keep the alchemically transformed molecules separated. The accuracy of the devised approaches was assessed on a large and diverse data set comprising 164 point mutations of charged residues (Asp, Glu, Lys, and Arg) to Ala at the protein-protein interfaces of complexes with known three-dimensional structures. Mean absolute and root-mean-square errors ranging from 1.38 to 1.66 and 1.89 to 2.44 kcal/mol, respectively, and Pearson correlation coefficients of â¼0.6 were obtained when testing the approaches on the selected data set using the GPU-TI module of Amber18 suite and the ff14SB force field. Furthermore, the inclusion of variable protonation states for the mutated acid residues improved the accuracy of the predicted ΔΔG values. Therefore, our results validate the use of TIRST/TIRST-H+ in prospective studies aimed at evaluating the impact of charge-changing mutations to Ala on the stability of protein-protein complexes.
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Proteínas , Estudios Prospectivos , Proteínas/genética , Proteínas/química , Entropía , Termodinámica , MutaciónRESUMEN
In this work, we studied the optical properties of Dy-doped Gd2O3 nanoparticles (NPs) before and after their APTES functionalisation. We obtained luminescent Dy@Gd2O3 NPs (0.5, 1, and 5% mol) using a modified polyol method. Our work describes their detailed structural analysis using FT-IR, XRD, HRTEM, TGA and XAS techniques. The results show that these systems present a crystalline structure with a body-centred cubic cell and particle sizes of 10 nm. The dopant position was inferred as substitutional, through XAS analysis at the M4,5-edges of Gd and Dy and K-edge of O, and in C2 sites, based on photoluminescence studies. There was sensitization of the luminescence by the matrix as shown by the emission increase of the hypersensitive transition (6F9/2 â 6H13/2, 572 nm) and also a broadband appears around 510 nm attributed to defects in Gd2O3. An enhanced emissive lifetime of 398 µs was found for the sample doped at 1%. We functionalised the Dy@Gd2O3 (at 1%) NPs with 3-aminopropiltrietoxisilane (APTES) for further application as a biomarker sensor. We found that these NPs conserved their luminescence after adding the surface agent (avoiding quenching effects) making them potential materials for biosensing.
RESUMEN
The aim of this work was to develop a comparative study between Bacillus paralicheniformis TB197 and B. subtilis ATCC 21332 strains in terms of growth, cyclic lipopeptide production, nematicidal activity, and active lipopeptide characteristics. Crude lipopeptide extracts (CLEs) from their fermentation broths were obtained, and their nematicidal activity (NA) was estimated as the mean lethal dose (LD50), employing Caenorhabditis elegans. Using a bioguided approach, CLE components were fractionated by semipreparative thin layer chromatography, and active lipopeptides were characterized by mass spectrometry. Both strains produced similar concentrations of CLEs (p ≥ 0.05) (0.99 ± 0.11 and 1.14 ± 0.15 mg/mL by TB197 and ATCC 21332, respectively). The estimated LD50 values of CLEs from the TB197 and ATCC 21332 strains were 3.88 and 8.15 mg/mL, respectively, showing that the NA of the TB197 strain CLE was 2.1-fold higher (p ≤ 0.05). Mass spectrometry revealed that strain TB197 synthesizes several families of lipopeptides, namely, fengycin A (C14-C17), fengycin B (C16-C17), surfactin (C15-C17), and lichenysin (C12, C13, C14, and C16), from which fengycins and lichenysins possess the highest NA (100 and 60% mortality in C. elegans larvae, respectively), while the ATCC 21332 strain produces mainly surfactin (C13-C17) (NA 63% mortality). The main differences found in this study were that the TB197 strain has a higher tolerance to inhibition by the product, and the lipopeptides they synthesize have a higher nematicidal activity due to the diversity of families compared to ATCC 21332. Likewise, it was shown that more polar lipopeptides (fengycins) are more effective at causing mortality in C. elegans larvae. KEY POINTS: ⢠The nematicidal activity of lipopeptides from TB197 is higher than from ATCC 21332 ⢠TB197 produces surfactin, lichenysin, and fengycin, while ATCC 21332 mainly produces surfactin ⢠The most polar lipopeptides (fengycins) cause more mortality in C. elegans L2.