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Oxidation of phosphite (HPO32-) to phosphate (HPO42-) releases electrons at a very low redox potential (E0'= -690 mV) which renders phosphite an excellent electron donor for microbial energy metabolism. To date, two pure cultures of strictly anaerobic bacteria have been isolated that run their energy metabolism on the basis of phosphite oxidation, the Gram-negative Desulfotignum phosphitoxidans (DSM 13687) and the Gram-positive Phosphitispora fastidiosa (DSM 112739). Here, we describe the key enzyme for dissimilatory phosphite oxidation in these bacteria. The enzyme catalyzed phosphite oxidation in the presence of adenosine monophosphate (AMP) to form adenosine diphosphate (ADP), with concomitant reduction of oxidized nicotinamide adenine dinucleotide (NAD+) to reduced nicotinamide adenine dinucleotide (NADH). The enzyme of P. fastidiosa was heterologously expressed in Escherichia coli. It has a molecular mass of 35.2 kDa and a high affinity for phosphite and NAD+. Its activity was enhanced more than 100-fold by addition of ADP-consuming adenylate kinase (myokinase) to a maximal activity between 30 and 80 mU x mg protein-1. A similar NAD-dependent enzyme oxidizing phosphite to phosphate with concomitant phosphorylation of AMP to ADP is found in D. phosphitoxidans, but this enzyme could not be heterologously expressed. Based on sequence analysis, these phosphite-oxidizing enzymes are related to nucleotide-diphosphate-sugar epimerases and indeed represent AMP-dependent phosphite dehydrogenases (ApdA). A reaction mechanism is proposed for this unusual type of substrate-level phosphorylation reaction.
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NAD , Fosfitos , NAD/metabolismo , Fosfitos/metabolismo , Oxirredução , Monofosfato de Adenosina/metabolismo , Difosfato de Adenosina/metabolismo , FosfatosRESUMO
OBJECTIVES: The aim of this study was to develop machine learning-based models for predicting acute cerebral infarction (ACI) in patients. METHODS: We extracted the data of ACI patients and non-ACI patients (as control) from two hospitals. The Lasso algorithm was employed to select the most crucial features associated with ACI. Five machine learning algorithms-based models were trained, which was performed with 10-fold cross-validation. Then, the area under the receiver operating characteristic curve (AUC), accuracy, and F1-score were calculated in the training models. Accordingly, the training models with excellent performance was selected as the final predictive model. The relative importance of variables was analyzed and ranked. RESULTS: A total of 150 patients were diagnosed with ACI (50.00%), with a higher proportion of males (70.67% vs. 44.00%) compared to the non-ACI patients. The logistic regression model exhibited a good performance in predicting ACI in the training set, as evidenced by its highest AUC, accuracy, sensitivity, and F1-score. Furthermore, feature importance analysis showed that blood glucose, gender, smoking history, serum homocysteine, folic acid, and C-reactive protein were the top six crucial variables of the logistic regression. CONCLUSIONS: In our work, the ACI risk prediction model developed by the logistic regression exhibited excellent performance. This could contribute to the identification of risk variables for ACI patients and enables clinicians timely and effective interventions.
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Infarto Cerebral , Aprendizado de Máquina , Humanos , Masculino , Feminino , Infarto Cerebral/epidemiologia , Infarto Cerebral/diagnóstico , Estudos Retrospectivos , Idoso , Pessoa de Meia-Idade , Fatores de Risco , Modelos LogísticosRESUMO
Phosphite is a stable phosphorus compound that, together with phosphate, made up a substantial part of the total phosphorus content of the prebiotic Earth's crust. Oxidation of phosphite to phosphate releases electrons at an unusually low redox potential (-690 mV at pH 7.0). Numerous aerobic and anaerobic bacteria use phosphite as a phosphorus source and oxidise it to phosphate for synthesis of nucleotides and other phosphorus-containing cell constituents. Only two pure cultures of strictly anaerobic bacteria have been isolated so far that use phosphite as an electron donor in their energy metabolism, the Gram-positive Phosphitispora fastidiosa and the Gram-negative Desulfotignum phosphitoxidans. The key enzyme of this metabolism is an NAD+ -dependent phosphite dehydrogenase enzyme that phosphorylates AMP to ADP. These phosphorylating phosphite dehydrogenases were found to be related to nucleoside diphosphate sugar epimerases. The produced NADH is channelled into autotrophic CO2 fixation via the Wood-Ljungdahl (CO-DH) pathway, thus allowing for nearly complete assimilation of the substrate electrons into bacterial biomass. This extremely efficient type of electron flow connects energy and carbon metabolism directly through NADH and might have been important in the early evolution of life when phosphite was easily available on Earth.
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Fosfitos , Fosfitos/química , Fosfitos/metabolismo , Elétrons , NAD/metabolismo , Anaerobiose , Oxirredução , Fósforo/metabolismo , FosfatosRESUMO
A new strictly anaerobic bacterium, strain DYL19T, was enriched and isolated with phosphite as the sole electron donor and CO2 as a single carbon source and electron acceptor from anaerobic sewage sludge sampled at a sewage treatment plant in Constance, Germany. It is a Gram-positive, spore-forming, slightly curved, rod-shaped bacterium which oxidizes phosphite to phosphate while reducing CO2 to biomass and small amounts of acetate. Optimal growth is observed at 30 °C, pH 7.2, with a doubling time of 3 days. Beyond phosphite, no further inorganic or organic electron donor can be used, and no other electron acceptor than CO2 is reduced. Sulphate inhibits growth with phosphite and CO2. The G+C content is 45.95 mol%, and dimethylmenaquinone-7 is the only quinone detectable in the cells. On the basis of 16S rRNA gene sequence analysis and other chemotaxonomic properties, strain DYL19T is described as the type strain of a new genus and species, Phosphitispora fastidiosa gen. nov., sp. nov.
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Peptococcaceae/classificação , Fosfitos , Filogenia , Esgotos , Anaerobiose , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Alemanha , Oxirredução , Peptococcaceae/isolamento & purificação , Fosfitos/metabolismo , Quinonas/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Esgotos/microbiologiaRESUMO
A convenient synthesis of 1,2-dihydroquinazolines via rearrangement of indazolium salts was described. A mechanistic study using isotope labeling experiment revealed that the rearrangement passes through cleavage of N-N bond/ring opening after basic deprotonation of 2-benzyl in indazolium salts to yield intermediate E, which proceeds in an intramolecular N-nucleophilic addition to form the observed product. Computational analyses imply that the pathway of the rearrangement is determined by the energy barriers of the ring-closing process and the stability of the product.
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Despite runners frequently suffering from dermatologic issues during long distance running, there is no compelling evidence quantitatively investigating their underlying injury mechanism. This study aimed to determine the foot morphology and temperature changes during long distance running and reveal the effect of these alterations on the injury risk of bruised toenail by measuring the subjective-perceived hallux comfort and gap length between the hallux and toebox of the shoe. Ten recreational runners participated in the experimental tests before (baseline), immediately after 5 and 10 km of treadmill running (12 km/h), in which the foot morphology was measured by a 3D foot scanner, the foot temperature was detected by an infrared camera, the perceived comfort was recorded by a visual analogue scale, and the gap length in the sagittal plane was captured by a high-speed camera. Ball width became narrower (106.39 ± 6.55 mm) and arch height (12.20 ± 2.34 mm) was reduced greatly after the 10 km run (p < 0.05). Foot temperature increased significantly after 5 and 10 km of running, and the temperature of dorsal hallux (35.12 ± 1.46 °C), dorsal metatarsal (35.92 ± 1.59 °C), and medial plantar metatarsal (37.26 ± 1.34 °C) regions continued to increase greatly from 5 to 10 km of running (p < 0.05). Regarding hallux comfort, the perceived scores significantly reduced after 5 and 10 km of running (2.10 ± 0.99, p < 0.05). In addition, during one running gait cycle, there was a significant increase in gap length at initial contact (39.56 ± 6.45 mm, p < 0.05) for a 10 km run, followed by a notable decrease upon reaching midstance (29.28 ± 6.81 mm, p < 0.05). It is concluded that the reduced ball width and arch height while increased foot temperature during long-distance running would exacerbate foot-shoe interaction, potentially responsible for bruised toenail injuries.
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Contusões , Corrida , Humanos , Unhas , Temperatura , Pé , Extremidade InferiorRESUMO
Background: During competition and training, exercises involving the lungs may occur throughout the sport, and fatigue is a major injury risk factor in sport, before and after fatigue studies of changes in the lungs are relatively sparse. This study is to investigate into how fatigue affects the lower limb's biomechanics during a forward lunge. Methods: 15 healthy young men participate in this study before and after to exposed to a fatigue protocol then we tested the forward lunge to obtain kinematic, kinetic changing during the task, and to estimate the corresponding muscles' strength changes in the hip, knee, and ankle joints. The measurement data before and after the fatigue protocol were compared with paired samples t-test. Results: In the sagittal and horizontal planes of the hip and knee joints, in both, the peak angles and joint range of motion (ROM) increased, whereas the moments in the sagittal plane of the knee joint smaller. The ankle joint's maximum angle smaller after fatigue. Peak vertical ground reaction force (vGRF) and peak contact both significantly smaller after completing the fatigue protocol and the quadriceps mean and maximum muscular strength significantly increased. Conclusion: After completing a fatigue protocol during lunge the hip, knee, and ankle joints become less stable in both sagittal and horizontal planes, hip and knee range of motion becomes greater. The quadriceps muscles are more susceptible to fatigue and reduced muscle force. Trainers should focus more on the thigh muscle groups.
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The Y-Balance Test (YBT) is a reliable tool for assessing the dynamic balance of athletes' lower limbs. This study aimed to compare the effects of the YBT on lower limb biomechanics before and after fatigue. Sixteen adult male recreational athletes were recruited for the study, and motion capture in combination with a force plate was used to collect kinematic, dynamics, and center of pressure (COP) data of the dominant leg during YBT testing before and after fatigue. Based on the research findings, there were significant statistical differences in the distances reached during the YBT in three directions before and after fatigue. After fatigue, there is a significant decrease in the ROM of the hip and knee joints in all three directions. Also, there is a significant increase in hip joint torque in the anterior- and posterior-lateral directions, while a significant decrease in hip and ankle joint torque is observed in the posterior-medial direction. Moreover, there is an increasing trend in positive and negative joint work for the hip, knee, and ankle joints in all three directions after fatigue. The range of COP displacement also increases following fatigue. The decline in YBT scores demonstrates the detrimental impact of fatigue on the dynamic balance of the lower limbs of adult male amateur athletes. We hope that these results can provide information for athletes and coaches to better understand the effects of fatigue on the dynamic balance of lower limbs, so as to carry out targeted lower limb balance training and prevent sports injuries.
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Background: Detecting fatigue at the early stages of a run could aid training programs in making adjustments, thereby reducing the heightened risk of injuries from overuse. The study aimed to investigate the effects of running fatigue on plantar force distribution in the dominant and nondominant feet of amateur runners. Methods: Thirty amateur runners were recruited for this study. Bilateral time-series plantar forces were employed to facilitate automatic fatigue gait recognition using convolutional neural network (CNN) and CNN-based long short-term memory network (ConvLSTM) models. Plantar force data collection was conducted both before and after a running-induced fatigue protocol using a FootScan force plate. The Keras library in Python 3.8.8 was used to train and tune deep learning models. Results: The results demonstrated that more mid-forefoot and heel force occurs during bilateral plantar and less midfoot fore force occurs in the dominant limb after fatigue (p < 0.001). The time of peak forces was significantly shortened at the midfoot and sum region of the nondominant foot, while it was delayed at the hallux region of the dominant foot (p < 0.001). In addition, the ConvLSTM model showed higher performance (Accuracy = 0.867, Sensitivity = 0.874, and Specificity = 0.859) in detecting fatigue gait than CNN (Accuracy = 0.800, Sensitivity = 0.874, and Specificity = 0.718). Conclusions: The findings of this study could offer empirical data for evaluating risk factors linked to overuse injuries in a single limb, as well as facilitate early detection of fatigued gait.
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This study compares foot-ankle temporal kinematics characteristics during planned and unplanned gait termination (PGT and UGT) in subjects with different arch stiffnesses (ASs) based on the statistical nonparametric mapping (SnPM) method. By measuring three-dimensional arch morphological parameters under different loading conditions, 28 healthy male subjects were classified and participated in gait termination (GT) tests to collect metatarsophalangeal (MTP) and ankle-joint kinematics data. The two-way repeated-measures ANOVA using SnPM was employed to assess the impacts of AS on foot-ankle kinematics during PGT and UGT. Our results show that joint angles (MTP and ankle joints) were altered owing to AS and GT factors. The flexible arches hahadve periods of significantly greater MTP and ankle joint angles than those of stiff arches during the stance phase of GT, whereas subjects exhibited significantly smaller ankle and MTP joint angles during UGT. These results add additional insights into the morphological arch biomechanical function, and the comprehensive compensatory adjustment of lower-limb joints during gait stopping caused by unplanned stimulation.
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IMPACT STATEMENT: This study investigated the effect and mechanism of Smad4 in ovarian carcinoma (OC) cell viability and demonstrated that Smad4 acted as a tumor suppressor in OC, which may contribute to the understanding of molecular mechanisms underlying OC occurrence and progression. Smad4 expression was decreased in the OC specimens, but Smad4 recovery in the OC cell lines impaired the survival and viability of OC cells by increasing autophagy and apoptosis. Further investigation showed that Smad4 interacted with the P85 subunit of PI3K and caused deactivation of the PI3K/mTOR pathway. Therefore, Smad4 could be considered as a target in cancer therapy due to its regulatory effect in OC carcinogenesis.
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Carcinoma Epitelial do Ovário/metabolismo , Carcinoma Epitelial do Ovário/patologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteína Smad4/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Apoptose/fisiologia , Autofagia/fisiologia , Linhagem Celular Tumoral , Feminino , HumanosRESUMO
T helper 17 (Th17) cells are regarded as key factors in the pathogenesis of multiple sclerosis (MS). Although the involvement of certain microRNAs (miRNAs) in the development of MS has been reported, their roles in Th17 cell differentiation and MS pathogenesis remain elusive. In this study, we identified that let-7f-5p expression is significantly downregulated in CD4+ T cells from MS patients and during the process of Th17 differentiation. The overexpression of let-7f-5p suppressed Th17 differentiation, whereas the knockdown of let-7f-5p expression enhanced this progress. We then explored the molecular mechanism through which let-7f-5p suppressed Th17 differentiation and identified signal transducer and activator of transcription 3 (STAT3), a pivotal transcription factor of Th17 cells, as a direct target of let-7f-5p. In contrast to the downregulated expression of let-7f-5p, STAT3 and p-STAT3 protein levels were dramatically upregulated and inversely correlated with let-7f-5p in peripheral blood CD4+ T cells from MS patients. In conclusion, let-7f-5p functions as a potential inhibitor of Th17 differentiation in the pathogenesis of MS by targeting STAT3 and may serve as a new therapeutic target.