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[Figure: see text].
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Cardiopatías Congénitas/genética , Acetiltransferasa A N-Terminal/genética , Acetiltransferasa E N-Terminal/genética , Procesamiento Proteico-Postraduccional , Acetilación , Células Cultivadas , Niño , Haploinsuficiencia , Cardiopatías Congénitas/metabolismo , Cardiopatías Congénitas/patología , Humanos , Células Madre Pluripotentes Inducidas/metabolismo , Mutación Missense , Proteoma/metabolismoRESUMEN
Sepsis is a complex infectious syndrome in which neutrophil participation is crucial for patient survival. Neutrophils quickly sense and eliminate the pathogen by using different effector mechanisms controlled by metabolic processes. The mammalian target of rapamycin (mTOR) pathway is an important route for metabolic regulation, and its role in neutrophil metabolism has not been fully understood yet, especially the importance of mTOR complex 2 (mTORC2) in the neutrophil effector functions. In this study, we observed that the loss of Rictor (mTORC2 scaffold protein) in primary mouse-derived neutrophils affects their chemotaxis by fMLF and their microbial killing capacity, but not the phagocytic capacity. We found that the microbicidal capacity was impaired in Rictor-deleted neutrophils because of an improper fusion of granules, reducing the hypochlorous acid production. The loss of Rictor also led to metabolic alterations in isolated neutrophils, increasing aerobic glycolysis. Finally, myeloid-Rictor-deleted mice (LysMRic Δ/Δ) also showed an impairment of the microbicidal capacity, increasing the bacterial burden in the Escherichia coli sepsis model. Overall, our results highlight the importance of proper mTORC2 activation for neutrophil effector functions and metabolism during sepsis.
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Diana Mecanicista del Complejo 2 de la Rapamicina/metabolismo , Neutrófilos/metabolismo , Sepsis/metabolismo , Sepsis/microbiología , Animales , Quimiotaxis/fisiología , Escherichia coli/metabolismo , Femenino , Glucólisis/fisiología , Humanos , Ácido Hipocloroso/metabolismo , Ratones , Ratones Endogámicos C57BL , Fagocitosis/fisiología , Transducción de Señal/fisiologíaRESUMEN
Hypertrophic cardiomyopathy is one of the most common inherited cardiomyopathies and a leading cause of sudden cardiac death in young adults. Despite profound insights into the genetics, there is imperfect correlation between mutation and clinical prognosis, suggesting complex molecular cascades driving pathogenesis. To investigate this, we performed an integrated quantitative multi-omics (proteomic, phosphoproteomic, and metabolomic) analysis to illuminate the early and direct consequences of mutations in myosin heavy chain in engineered human induced pluripotent stem-cell-derived cardiomyocytes relative to late-stage disease using patient myectomies. We captured hundreds of differential features, which map to distinct molecular mechanisms modulating mitochondrial homeostasis at the earliest stages of pathobiology, as well as stage-specific metabolic and excitation-coupling maladaptation. Collectively, this study fills in gaps from previous studies by expanding knowledge of the initial responses to mutations that protect cells against the early stress prior to contractile dysfunction and overt disease.
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Cardiomiopatía Hipertrófica , Células Madre Pluripotentes Inducidas , Adulto Joven , Humanos , Dinámicas Mitocondriales , Multiómica , Proteómica , Cardiomiopatía Hipertrófica/genética , Miocitos Cardíacos/metabolismo , Mutación , Células Madre Pluripotentes Inducidas/metabolismoRESUMEN
ABSTRACT: Filho, MM, Venturini, GRdO, Moreira, OC, Leitão, L, Mira, PA, Castro, JB, Aidar, FJ, Novaes, JdS, Vianna, JM, and Caputo Ferreira, ME. Effects of different types of resistance training and detraining on functional capacity, muscle strength, and power in older women: A randomized controlled study. J Strength Cond Res 36(4): 984-990, 2022-Resistance training (RT) increases muscle strength, power, and functional capacity (FC) of older women. However, these benefits can be lost partially or totally with detraining. This study aimed to compare the effect of 20 weeks of different types of RT and 4 weeks of detraining on muscle strength, power, and FC in older women. Ninety-five older women were randomly divided into 4 experimental groups (strength endurance, power, absolute strength, and relative strength training) and 1 control group (CG). We assessed muscle strength (10RM test) and muscle power of the lower (countermovement jump) and upper limbs (medicine ball pitch). Functional capacity was assessed by the Senior Fitness Test, which comprises the following tests: 30-second arm curl, 30-second chair stand, back scratch, chair sit and reach, 8-foot up and go, and 6-minute walk. The experiment lasted 24 weeks (familiarization: 2 weeks; neural adaptation: 6 weeks; specific training: 12 weeks; and detraining: 4 weeks). Muscle strength, lower and upper limb power (all p < 0.05), 30-second arm curl, 30-second chair stand, 8-foot up and go, 6-minute walk (all p < 0.001), and lower limb flexibility (p = 0.002) improved in all experimental groups after training and CG showed no differences in any of these variables. After detraining, muscle strength, lower and upper limb power (p < 0.05 for all), and FC decreased in comparison to the end of RT (30-second arm curl, 30-second chair stand, 8-foot up and go, 6-minute walk, and lower limb flexibility, p < 0.05 for all). Although the FC of the subjects has been reduced after 4 weeks of detraining, it was maintained at higher levels in comparison to baseline. These results suggested that older women can be submitted to different types of RT to achieve improvements in general fitness.
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Entrenamiento de Fuerza , Anciano , Ejercicio Físico/fisiología , Femenino , Humanos , Fuerza Muscular/fisiología , Aptitud Física/fisiología , Entrenamiento de Fuerza/métodos , CaminataRESUMEN
BACKGROUND: Hypertrophic cardiomyopathy (HCM) is caused by pathogenic variants in sarcomere protein genes that evoke hypercontractility, poor relaxation, and increased energy consumption by the heart and increased patient risks for arrhythmias and heart failure. Recent studies show that pathogenic missense variants in myosin, the molecular motor of the sarcomere, are clustered in residues that participate in dynamic conformational states of sarcomere proteins. We hypothesized that these conformations are essential to adapt contractile output for energy conservation and that pathophysiology of HCM results from destabilization of these conformations. METHODS: We assayed myosin ATP binding to define the proportion of myosins in the super relaxed state (SRX) conformation or the disordered relaxed state (DRX) conformation in healthy rodent and human hearts, at baseline and in response to reduced hemodynamic demands of hibernation or pathogenic HCM variants. To determine the relationships between myosin conformations, sarcomere function, and cell biology, we assessed contractility, relaxation, and cardiomyocyte morphology and metabolism, with and without an allosteric modulator of myosin ATPase activity. We then tested whether the positions of myosin variants of unknown clinical significance that were identified in patients with HCM, predicted functional consequences and associations with heart failure and arrhythmias. RESULTS: Myosins undergo physiological shifts between the SRX conformation that maximizes energy conservation and the DRX conformation that enables cross-bridge formation with greater ATP consumption. Systemic hemodynamic requirements, pharmacological modulators of myosin, and pathogenic myosin missense mutations influenced the proportions of these conformations. Hibernation increased the proportion of myosins in the SRX conformation, whereas pathogenic variants destabilized these and increased the proportion of myosins in the DRX conformation, which enhanced cardiomyocyte contractility, but impaired relaxation and evoked hypertrophic remodeling with increased energetic stress. Using structural locations to stratify variants of unknown clinical significance, we showed that the variants that destabilized myosin conformations were associated with higher rates of heart failure and arrhythmias in patients with HCM. CONCLUSIONS: Myosin conformations establish work-energy equipoise that is essential for life-long cellular homeostasis and heart function. Destabilization of myosin energy-conserving states promotes contractile abnormalities, morphological and metabolic remodeling, and adverse clinical outcomes in patients with HCM. Therapeutic restabilization corrects cellular contractile and metabolic phenotypes and may limit these adverse clinical outcomes in patients with HCM.
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Miosinas Cardíacas/genética , Cardiomiopatía Hipertrófica/metabolismo , Mutación Missense/genética , Miocitos Cardíacos/fisiología , Cadenas Pesadas de Miosina/genética , Sarcómeros/metabolismo , Adenosina Trifosfatasas , Animales , Cardiomiopatía Hipertrófica/genética , Células Cultivadas , Metabolismo Energético , Humanos , Células Madre Pluripotentes Inducidas/citología , Ratones , Simulación de Dinámica Molecular , Relajación Muscular , Contracción Miocárdica , Miocitos Cardíacos/citología , Conformación Proteica , Sarcómeros/genéticaRESUMEN
Atherosclerotic plaque development is closely associated with the hemodynamic forces applied to endothelial cells (ECs). Among these, shear stress (SS) plays a key role in disease development since changes in flow intensity and direction could stimulate an atheroprone or atheroprotective phenotype. ECs under low or oscillatory SS (LSS) show upregulation of inflammatory, adhesion, and cellular permeability molecules. On the contrary, cells under high or laminar SS (HSS) increase their expression of protective and anti-inflammatory factors. The mechanism behind SS regulation of an atheroprotective phenotype is not completely elucidated. Here we used proteomics and metabolomics to better understand the changes in endothelial cells (human umbilical vein endothelial cells) under in vitro LSS and HSS that promote an atheroprone or atheroprotective profile and how these modifications can be connected to atherosclerosis development. Our data showed that lipid metabolism, in special cholesterol metabolism, was downregulated in cells under LSS. The low-density lipoprotein receptor (LDLR) showed significant alterations both at the quantitative expression level as well as regarding posttranslational modifications. Under LSS, LDLR was seen at lower concentrations and with a different glycosylation profile. Finally, modulating LDLR with atorvastatin led to the recapitulation of a HSS metabolic phenotype in EC under LSS. Altogether, our data suggest that there is significant modulation of lipid metabolism in endothelial cells under different SS intensities and that this could contribute to the atheroprone phenotype of LSS. Statin treatment was able to partially recover the protective profile of these cells.
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Aterosclerosis/metabolismo , Hemodinámica , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Metabolismo de los Lípidos , Lipidómica/métodos , Mecanotransducción Celular , Proteómica/métodos , Aterosclerosis/tratamiento farmacológico , Aterosclerosis/patología , Aterosclerosis/fisiopatología , Atorvastatina/farmacología , Células Cultivadas , Colesterol/metabolismo , Glicosilación , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Humanos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacología , Metabolismo de los Lípidos/efectos de los fármacos , Mecanotransducción Celular/efectos de los fármacos , Fenotipo , Placa Aterosclerótica , Procesamiento Proteico-Postraduccional , Receptores de LDL/metabolismo , Flujo Sanguíneo Regional , Estrés MecánicoRESUMEN
INTRODUCTION: Metabolomics allows exploration of novel biomarkers and provides insights on metabolic pathways associated with disease. To date, metabolomics studies on CKD have been largely limited to Caucasian populations and have mostly examined surrogate end points. OBJECTIVE: In this study, we evaluated the role of metabolites in predicting a primary outcome defined as dialysis need, doubling of serum creatinine or death in Brazilian macroalbuminuric DKD patients. METHODS: Non-targeted metabolomics was performed on plasma from 56 DKD patients. Technical triplicates were done. Metabolites were identified using Agilent Fiehn GC/MS Metabolomics and NIST libraries (Agilent MassHunter Work-station Quantitative Analysis, version B.06.00). After data cleaning, 186 metabolites were left for analyses. RESULTS: During a median follow-up time of 2.5 years, the PO occurred in 17 patients (30.3%). In non-parametric testing, 13 metabolites were associated with the PO. In univariate Cox regression, only 1,5-anhydroglucitol (HR 0.10; 95% CI 0.01-0.63, p = .01), norvaline and L-aspartic acid were associated with the PO. After adjustment for baseline renal function, 1,5-anhydroglucitol (HR 0.10; 95% CI 0.02-0.63, p = .01), norvaline (HR 0.01; 95% CI 0.001-0.4, p = .01) and aspartic acid (HR 0.12; 95% CI 0.02-0.64, p = .01) remained significantly and inversely associated with the PO. CONCLUSION: Our results show that lower levels of 1,5-anhydroglucitol, norvaline and L-aspartic acid are associated with progression of macroalbuminuric DKD. While norvaline and L-aspartic acid point to interesting metabolic pathways, 1,5-anhydroglucitol is of particular interest since it has been previously shown to be associated with incident CKD. This inverse biomarker of hyperglycemia should be further explored as a new tool in DKD.
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Albuminuria/metabolismo , Desoxiglucosa/química , Nefropatías Diabéticas/metabolismo , Metabolómica , Albuminuria/sangre , Biomarcadores/sangre , Biomarcadores/metabolismo , Brasil , Creatinina/sangre , Creatinina/metabolismo , Nefropatías Diabéticas/sangre , Método Doble Ciego , Cromatografía de Gases y Espectrometría de Masas , HumanosRESUMEN
Vascular smooth muscle cells (VSMCs) are thought to assume a quiescent and homogeneous mechanical behavior after arterial tree development phase. However, VSMCs are known to be molecularly heterogeneous in other aspects and their mechanics may play a role in pathological situations. Our aim was to evaluate VSMCs from different arterial beds in terms of mechanics and proteomics, as well as investigate factors that may influence this phenotype. VSMCs obtained from seven arteries were studied using optical magnetic twisting cytometry (both in static state and after stretching) and shotgun proteomics. VSMC mechanical data were correlated with anatomical parameters and ultrastructural images of their vessels of origin. Femoral, renal, abdominal aorta, carotid, mammary, and thoracic aorta exhibited descending order of stiffness (G, P < 0.001). VSMC mechanical data correlated with the vessel percentage of elastin and amount of surrounding extracellular matrix (ECM), which decreased with the distance from the heart. After 48 h of stretching simulating regional blood flow of elastic arteries, VSMCs exhibited a reduction in basal rigidity. VSMCs from the thoracic aorta expressed a significantly higher amount of proteins related to cytoskeleton structure and organization vs. VSMCs from the femoral artery. VSMCs are heterogeneous in terms of mechanical properties and expression/organization of cytoskeleton proteins along the arterial tree. The mechanical phenotype correlates with the composition of ECM and can be modulated by cyclic stretching imposed on VSMCs by blood flow circumferential stress.
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Arterias/fisiología , Músculo Liso Vascular/fisiología , Miocitos del Músculo Liso/fisiología , Animales , Arterias/metabolismo , Ciclo Celular/fisiología , Colágeno/metabolismo , Elastina/metabolismo , Femenino , Músculo Liso Vascular/metabolismo , Miocitos del Músculo Liso/metabolismo , Proteómica , Sus scrofaRESUMEN
Systemic lupus erythematosus (SLE) is a highly heterogenous autoimmune disease that affects multiple organs, including the heart. The mechanisms by which myocardial injury develops in SLE, however, remain poorly understood. Here we engineered human cardiac tissues and cultured them with IgG fractions containing autoantibodies from SLE patients with and without myocardial involvement. We observed unique binding patterns of IgG from two patient subgroups: (i) patients with severe myocardial inflammation exhibited enhanced binding to apoptotic cells within cardiac tissues subjected to stress, and (ii) patients with systolic dysfunction exhibited enhanced binding to the surfaces of viable cardiomyocytes. Functional assays and RNA sequencing (RNA-seq) revealed that IgGs from patients with systolic dysfunction exerted direct effects on engineered tissues in the absence of immune cells, altering tissue cellular composition, respiration and calcium handling. Autoantibody target characterization by phage immunoprecipitation sequencing (PhIP-seq) confirmed distinctive IgG profiles between patient subgroups. By coupling IgG profiling with cell surface protein analyses, we identified four pathogenic autoantibody candidates that may directly alter the function of cells within the myocardium. Taken together, these observations provide insights into the cellular processes of myocardial injury in SLE that have the potential to improve patient risk stratification and inform the development of novel therapeutic strategies.
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Systemic lupus erythematosus (SLE) is a heterogenous autoimmune disease that affects multiple organs, including the heart. The mechanisms of myocardial injury in SLE remain poorly understood. In this study, we engineered human cardiac tissues and cultured them with IgG from patients with SLE, with and without myocardial involvement. IgG from patients with elevated myocardial inflammation exhibited increased binding to apoptotic cells within cardiac tissues subjected to stress, whereas IgG from patients with systolic dysfunction exhibited enhanced binding to the surface of live cardiomyocytes. Functional assays and RNA sequencing revealed that, in the absence of immune cells, IgG from patients with systolic dysfunction altered cellular composition, respiration and calcium handling. Phage immunoprecipitation sequencing (PhIP-seq) confirmed distinctive IgG profiles between patient subgroups. Coupling IgG profiling with cell surfaceome analysis identified four potential pathogenic autoantibodies that may directly affect the myocardium. Overall, these insights may improve patient risk stratification and inform the development of new therapeutic strategies.
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Chagas disease is caused by the parasite Trypanosoma cruzi (T. cruzi) and, among all the chronic manifestations of the disease, Chronic Chagas Cardiomyopathy (CCC) is the most severe outcome. Despite high burden and public health importance in Latin America, there is a gap in understanding the molecular mechanisms that results in CCC development. Previous studies showed that T. cruzi uses the host machinery for infection and replication, including the repurposing of the responses to intracellular infection such as mitochondrial activity, vacuolar membrane, and lysosomal activation in benefit of parasite infection and replication. One common signaling upstream to many responses to parasite infection is mTOR pathway, previous associated to several downstream cellular mechanisms including autophagy, mitophagy and lysosomal activation. Here, using human iPSC derived cardiomyocytes (hiPSCCM), we show the mTOR pathway is activated in hiPSCCM after T. cruzi infection, and the inhibition of mTOR with rapamycin reduced number of T. cruzi 48 h post infection (hpi). Rapamycin treatment also reduced lysosome migration from nuclei region to cell periphery resulting in less T. cruzi inside the parasitophorous vacuole (PV) in the first hour of infection. In addition, the number of parasites leaving the PV to the cytoplasm to replicate in later times of infection was also lower after rapamycin treatment. Altogether, our data suggest that host's mTOR activation concomitant with parasite infection modulates lysosome migration and that T. cruzi uses this mechanism to achieve infection and replication. Modulating this mechanism with rapamycin impaired the success of T. cruzi life cycle independent of mitophagy.
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Enfermedad de Chagas , Trypanosoma cruzi , Humanos , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/parasitología , Enfermedad de Chagas/parasitología , Trypanosoma cruzi/fisiología , Serina-Treonina Quinasas TOR , Lisosomas/metabolismo , Lisosomas/parasitología , Sirolimus/metabolismoRESUMEN
Ecological conditions in the Amazon rainforests are historically favorable for the transmission of numerous tropical diseases, especially vector-borne diseases. The high diversity of pathogens likely contributes to the strong selective pressures for human survival and reproduction in this region. However, the genetic basis of human adaptation to this complex ecosystem remains unclear. This study investigates the possible footprints of genetic adaptation to the Amazon rainforest environment by analyzing the genomic data of 19 native populations. The results based on genomic and functional analysis showed an intense signal of natural selection in a set of genes related to Trypanosoma cruzi infection, which is the pathogen responsible for Chagas disease, a neglected tropical parasitic disease native to the Americas that is currently spreading worldwide.
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Enfermedad de Chagas , Trypanosoma cruzi , Humanos , Trypanosoma cruzi/genética , Ecosistema , Enfermedad de Chagas/genética , Enfermedad de Chagas/parasitología , Pueblos IndígenasRESUMEN
Introduction: Chagas cardiomyopathy, a disease caused by Trypanosoma cruzi (T. cruzi) infection, is a major contributor to heart failure in Latin America. There are significant gaps in our understanding of the mechanism for infection of human cardiomyocytes, the pathways activated during the acute phase of the disease, and the molecular changes that lead to the progression of cardiomyopathy. Methods: To investigate the effects of T. cruzi on human cardiomyocytes during infection, we infected induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) with the parasite and analyzed cellular, molecular, and metabolic responses at 3 hours, 24 hours, and 48 hours post infection (hpi) using transcriptomics (RNAseq), proteomics (LC-MS), and metabolomics (GC-MS and Seahorse) analyses. Results: Analyses of multiomic data revealed that cardiomyocyte infection caused a rapid increase in genes and proteins related to activation innate and adaptive immune systems and pathways, including alpha and gamma interferons, HIF-1α signaling, and glycolysis. These responses resemble prototypic responses observed in pathogen-activated immune cells. Infection also caused an activation of glycolysis that was dependent on HIF-1α signaling. Using gene editing and pharmacological inhibitors, we found that T. cruzi uptake was mediated in part by the glucose-facilitated transporter GLUT4 and that the attenuation of glycolysis, HIF-1α activation, or GLUT4 expression decreased T. cruzi infection. In contrast, pre-activation of pro-inflammatory immune responses with LPS resulted in increased infection rates. Conclusion: These findings suggest that T. cruzi exploits a HIF-1α-dependent, cardiomyocyte-intrinsic stress-response activation of glycolysis to promote intracellular infection and replication. These chronic immuno-metabolic responses by cardiomyocytes promote dysfunction, cell death, and the emergence of cardiomyopathy.
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Cardiomiopatía Chagásica , Enfermedad de Chagas , Trypanosoma cruzi , Humanos , Trypanosoma cruzi/metabolismo , Miocitos Cardíacos/metabolismo , Enfermedad de Chagas/parasitología , Inmunidad InnataRESUMEN
BACKGROUND: The mechanisms implicated in the genesis of delayed radiofrequency (RF) effects remain unclear, but may be related to extension of the lesion beyond the region of coagulative necrosis. The role of apoptosis in this process has not been previously reported. We assessed whether RF promotes apoptosis in the region surrounding acute ablation lesions in a rat model. METHODS: Wistar rats (n=30; weight 300 g) were anesthesized, the chest was opened, and the heart was exposed. A modified unipolar RF ablation (custom catheter 4.5-mm-tip diameter, 12 Watts, 10 seconds) was undertaken on the left ventricular anterolateral epicardial surface and the chest was closed. After 2 hours, animals were killed for histological (hematoxylin and eosin, TdT-mediated dUTP Nick End-Labeling [TUNEL] assay) and immunohistochemical (anti-BAD and anti-caspase 3 antibodies) analysis (n=18). Additional animals (n=12) were sacrificed at 2 (n=3), 24 (n=3), 48 (n=3), and 72 hours (n=3) after ablation exclusively for anti-BAD Western Blotting analysis. RESULTS: Lesions were characterized by well-defined regions of coagulative necrosis. In 18/18 (100%) animals, TUNEL assay revealed positive luminescent reaction cells in the region surrounding the lesion, extending up to 2 mm from the border zone. However, microscopic evaluation of the nuclei and immunohistochemical and anti-BAD Western Blotting analysis were negative in all (100%) rats. Thus, positive TUNEL reaction in the periphery of the ablation lesion likely reflects nonspecific DNA damage. CONCLUSION: RF ablation does not promote apoptosis in the periphery of the myocardial lesion. This finding may have implications for the elucidation of late lesion extension following RF ablation.
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Apoptosis , Procedimientos Quirúrgicos Cardíacos , Ablación por Catéter/métodos , Miocardio/patología , Animales , Caspasa 3/análisis , Ablación por Catéter/instrumentación , Inmunohistoquímica , Masculino , Ratas , Ratas Wistar , Proteína Letal Asociada a bcl/análisisRESUMEN
BACKGROUND: Physical activity (PA) and physical fitness are key factors for quality of life (QoL) for older women. The aging process promotes the decrease in some capacities such as strength, which affect the activities of daily life. This loss of strength leads to a reduction in balance and an increased risk of falls as well as a sedentary lifestyle. Resistance Training (RT) is an effective method to improve balance and strength but different RT protocols can promote different responses. Power training has a higher impact on the performance of activities of daily life. Therefore, our study aimed to analyze if different RT protocols promote individual responses in balance, QoL and PA levels of older women and which are more effective for the older women. METHODS: Ninety-four older women were divided into four RT groups (relative strength endurance training, SET; Traditional strength training, TRT; absolute strength training, AST; power training, PWT) and one control group (CG). Each RT group performed a specific protocol for 16 weeks. At baseline and after 8 and 16 weeks, we assessed balance through the Berg balance scale; PA levels with a modified Baecke questionnaire and QoL with World Health Organization Quality of Life-BREF (WHOQOL-BREF) and World Health Organization Quality of Life-OLD module (WHOQOL-OLD). RESULTS: Balance improved after 16 weeks (baseline vs. 16 weeks; p < 0.05) without differences between all RT groups. PWT (2.82%) and TRT (3.48%) improved balance in the first 8 weeks (baseline vs. 8 weeks; p < 0.05). PA levels increased in PWT, TRT and AST after 16 weeks (baseline vs. 16 weeks; p < 0.05). CONCLUSION: All RT protocols improved PA levels and QoL after 16 weeks of training. For the improvement of balance, QoL and PA, older women can be subjected to PWT, AST and SET, and not be restricted to TRT.
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Entrenamiento de Fuerza , Anciano , Ejercicio Físico/fisiología , Femenino , Humanos , Fuerza Muscular/fisiología , Aptitud Física , Calidad de Vida , Entrenamiento de Fuerza/métodos , Encuestas y CuestionariosRESUMEN
Chagas disease is a tropical zoonosis caused by Trypanosoma cruzi. After infection, the host present an acute phase, usually asymptomatic, in which an extensive parasite proliferation and intense innate immune activity occurs, followed by a chronic phase, characterized by low parasitemia and development of specific immunity. Most individuals in the chronic phase remain without symptoms or organ damage, a state called indeterminate IND form. However, 20 to 40% of individuals develop cardiac or gastrointestinal complications at any time in life. Cardiomyocytes have an important role in the development of Chronic Chagas Cardiomyopathy (CCC) due to transcriptional and metabolic alterations that are crucial for the parasite survival and replication. However, it still not clear why some infected individuals progress to a cardiomyopathy phase, while others remain asymptomatic. In this work, we used hiPSCs-derived cardiomyocytes (hiPSC-CM) to investigate patterns of infection, proliferation and transcriptional response in IND and CCC patients. Our data show that T. cruzi infection and proliferation efficiency do not differ significantly in PBMCs and hiPSC-CM from both groups. However, RNA-seq analysis in hiPSC-CM infected for 24 hours showed a significantly different transcriptional response to the parasite in cells from IND or CCC patients. Cardiomyocytes from IND showed significant differences in the expression of genes related to antigen processing and presentation, as well as, immune co-stimulatory molecules. Furthermore, the downregulation of collagen production genes and extracellular matrix components was significantly different in these cells. Cardiomyocytes from CCC, in turn, showed increased expression of mTORC1 pathway and unfolded protein response genes, both associated to increased intracellular ROS production. These data point to a differential pattern of response, determined by baseline genetic differences between groups, which may have an impact on the development of a chronic outcome with or without the presentation of cardiac symptoms.
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Cardiomiopatía Chagásica , Enfermedad de Chagas , Células Madre Pluripotentes Inducidas , Trypanosoma cruzi , Enfermedad Crónica , Humanos , Células Madre Pluripotentes Inducidas/metabolismo , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/parasitología , Transcriptoma , Trypanosoma cruzi/metabolismoRESUMEN
Alzheimer's disease (AD) is the most common form of dementia in the elderly, affecting cognitive, intellectual, and motor functions. Different hypotheses explain AD's mechanism, such as the amyloidogenic hypothesis. Moreover, this disease is multifactorial, and several studies have shown that gut dysbiosis and oxidative stress influence its pathogenesis. Knowing that kefir is a probiotic used in therapies to restore dysbiosis and that the bioactive peptides present in it have antioxidant properties, we explored its biotechnological potential as a source of molecules capable of modulating the amyloidogenic pathway and reducing oxidative stress, contributing to the treatment of AD. For that, we used Drosophila melanogaster model for AD (AD-like flies). Identification of bioactive peptides in the kefir sample was made by proteomic and peptidomic analyses, followed by in vitro evaluation of antioxidant and acetylcholinesterase inhibition potential. Flies were treated and their motor performance, brain morphology, and oxidative stress evaluated. Finally, we performed molecular docking between the peptides found and the main pathology-related proteins in the flies. The results showed that the fraction with the higher peptide concentration was positive for the parameters evaluated. In conclusion, these results revealed these kefir peptide-rich fractions have therapeutic potential for AD.
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Enfermedad de Alzheimer , Kéfir , Acetilcolinesterasa/metabolismo , Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/metabolismo , Animales , Antioxidantes/análisis , Brasil , Drosophila melanogaster/metabolismo , Disbiosis , Simulación del Acoplamiento Molecular , Péptidos/química , ProteómicaRESUMEN
BACKGROUND: Chronic Chagas Cardiomyopathy (CCC) usually develops between 10 and 20 years after the first parasitic infection and is one of the leading causes of end-stage heart failure in Latin America. Despite the great inter-individual variability in CCC susceptibility (only 30% of infected individuals ever present CCC), there are no known predictors for disease development in those chronically infected. METHODOLOGY/PRINCIPAL FINDINGS: We describe a new susceptibility locus for CCC through a GWAS analysis in the SaMi-Trop cohort, a population-based study conducted in a Chagas endemic region from Brazil. This locus was also associated with CCC in the REDS II Study. The newly identified locus (rs34238187, OR 0.73, p-value 2.03 x 10-9) spans a haplotype of approximately 30Kb on chromosome 18 (chr18: 5028302-5057621) and is also associated with 80 different traits, most of them blood protein traits significantly enriched for immune-related biological pathways. Hi-C data show that the newly associated locus is able to interact with chromatin sites as far as 10Mb on chromosome 18 in a number of different cell types and tissues. Finally, we were able to confirm, at the tissue transcriptional level, the immune-associated blood protein signature using a multi-tissue differential gene expression and enrichment analysis. CONCLUSIONS/SIGNIFICANCE: We suggest that the newly identified locus impacts CCC risk among T cruzi infected individuals through the modulation of a downstream transcriptional and protein signature associated with host-parasite immune response. Functional characterization of the novel risk locus is warranted.
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Cardiomiopatía Chagásica , Enfermedad de Chagas , Trypanosoma cruzi , Cromatina , Cromosomas Humanos Par 18/genética , Cromosomas Humanos Par 18/metabolismo , Estudio de Asociación del Genoma Completo , Humanos , Trypanosoma cruzi/fisiologíaRESUMEN
OBJECTIVES: Clinical and epidemiological studies have suggested a correlation between vitamin D deficiency (VDD) and high blood pressure (BP). This study aimed to test the hypothesis that high BP induced by VDD is associated with altered expression and covalent modification of apical sodium transporters along the nephron. The contributions of the intrarenal renin-angiotensin system (RAS) and oxidative stress were also investigated. METHODS: Male Wistar rats were fed a vitamin D-free (nâ=â26) or standard diet (nâ=â25) for 30 days. BP was recorded using noninvasive and invasive procedures. The expression levels of total and phosphorylated apical sodium transporters in rat renal cortex and medulla were evaluated by immunoblotting. Intrarenal RAS components were assessed by immunoblotting and ELISA. Renal oxidative stress was analyzed by measuring the concentrations of thiobarbituric acid reactive substances and reduced glutathione. RESULTS: Higher BP levels in VDD rats than controls were accompanied by overexpression and hyperphosphorylation of renal cortical and medullary Na+-K+-2Cl- cotransporter type 2, enhanced levels of phosphorylated Na+/H+ exchanger type 3, and reduced expression levels of total and phosphorylated Na+/Cl- cotransporter. Changes in intrarenal RAS induced by VDD vs. controls included the marked elevation of medullary renin expression, higher expression of cortical angiotensinogen, higher urinary angiotensinogen excretion, and higher cortical and medullary angiotensin II content. VDD rats displayed higher thiobarbituric acid reactive substances/glutathione ratios in the renal cortex and medulla than controls. CONCLUSION: These results suggest that the molecular mechanisms underlying the effects of VDD on BP may include the upregulation of Na+-K+-2Cl- cotransporter type 2 and activation of intrarenal RAS and oxidative stress.
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Hipertensión , Deficiencia de Vitamina D , Animales , Presión Sanguínea , Riñón/metabolismo , Masculino , Ratas , Ratas Wistar , Sodio/metabolismo , Simportadores de Cloruro de Sodio-Potasio , Miembro 3 de la Familia de Transportadores de Soluto 12RESUMEN
In bees from genus Melipona, differential feeding is not enough to fully explain female polyphenism. In these bees, there is a hypothesis that in addition to the environmental component (food), a genetic component is also involved in caste differentiation. This mechanism has not yet been fully elucidated and may involve epigenetic and metabolic regulation. Here, we verified that the genes encoding histone deacetylases HDAC1 and HDAC4 and histone acetyltransferase KAT2A were expressed at all stages of Melipona scutellaris, with fluctuations between developmental stages and castes. In larvae, the HDAC genes showed the same profile of Juvenile Hormone titers-previous reported-whereas the HAT gene exhibited the opposite profile. We also investigated the larvae and larval food metabolomes, but we did not identify the putative queen-fate inducing compounds, geraniol and 10-hydroxy-2E-decenoic acid (10HDA). Finally, we demonstrated that the histone deacetylase inhibitor 10HDA-the major lipid component of royal jelly and hence a putative regulator of honeybee caste differentiation-was unable to promote differentiation in queens in Melipona scutellaris. Our results suggest that epigenetic and hormonal regulations may act synergistically to drive caste differentiation in Melipona and that 10HDA is not a caste-differentiation factor in Melipona scutellaris.