Clarissa P. C. Gomes

Exercise induction of gut microbiota modifications in obese, non-obese and hypertensive rats

BackgroundObesity is a multifactor disease associated with cardiovascular disorders such as hypertension. Recently, gut microbiota was linked to obesity pathogenesisand shown to influence the host metabolism. Moreover, several factors such as host-genotype and life-style have been shown to modulate gut microbiota composition. Exercise is a well-known agent used for the treatment of numerous pathologies, such as obesity and hypertension; it has recently been demonstrated to shape gut microbiota consortia. Since exercise-altered microbiota could possibly improve the treatment of diseases related to dysfunctional microbiota, this study aimed to examine the effect of controlled exercise training on gut microbial composition in Obese rats (n = 3), non-obese Wistar rats (n = 3) and Spontaneously Hypertensive rats (n = 3). Pyrosequencing of 16S rRNA genes from fecal samples collected before and after exercise training was used for this purpose.ResultsExercise altered the composition and diversity of gut bacteria at genus level in all rat lineages. Allobaculum (Hypertensive rats), Pseudomonas and Lactobacillus (Obese rats) were shown to be enriched after exercise, while Streptococcus (Wistar rats), Aggregatibacter and Sutturella (Hypertensive rats) were more enhanced before exercise. A significant correlation was seen in the Clostridiaceae and Bacteroidaceae families and Oscillospira and Ruminococcus genera with blood lactate accumulation. Moreover, Wistar and Hypertensive rats were shown to share a similar microbiota composition, as opposed to Obese rats. Finally, Streptococcus alactolyticus, Bifidobacterium animalis, Ruminococcus gnavus, Aggregatibacter pneumotropica and Bifidobacterium pseudolongum were enriched in Obese rats.ConclusionsThese data indicate that non-obese and hypertensive rats harbor a different gut microbiota from obese rats and that exercise training alters gut microbiota from an obese and hypertensive genotype background.

A Review of Computational Tools in microRNA Discovery

Since microRNAs (miRNAs) were discovered, their impact on regulating various biological activities has been a surprising and exciting field. Knowing the entire repertoire of these small molecules is the first step to gain a better understanding of their function. High throughput discovery tools such as next-generation sequencing significantly increased the number of known miRNAs in different organisms in recent years. However, the process of being able to accurately identify miRNAs is still a complex and difficult task, requiring the integration of experimental approaches with computational methods. A number of prediction algorithms based on characteristics of miRNA molecules have been developed to identify new miRNA species. Different approaches have certain strengths and weaknesses and in this review, we aim to summarize several commonly used tools in metazoan miRNA discovery.

The complexity, function and applications of RNA in circulation

Blood carries a wide array of biomolecules, including nutrients, hormones, and molecules that are secreted by cells for specific biological functions. The recent finding of stable RNA of both endogenous and exogenous origin in circulation raises a number of questions and opens a broad, new field: exploring the origins, functions, and applications of these extracellular RNA molecules. These findings raise many important questions, including: what are the mechanisms of export and cellular uptake, what is the nature and source of their stability, what molecules do they interact with in the blood, and what are the possible biological functions of the circulating RNA? This review summarizes some key recent developments in circulating RNA research and discusses some of the open questions in the field.

Proteomics applied to exercise physiology: a cutting-edge technology.

Exercise research has always drawn the attention of the scientific community because it can be widely applied to sport training, health improvement, and disease prevention. For many years numerous tools have been used to investigate the several physiological adaptations induced by exercise stimuli. Nowadays a closer look at the molecular mechanisms underlying metabolic pathways and muscular and cardiovascular adaptation to exercise are among the new trends in exercise physiology research. Considering this, to further understand these adaptations as well as pathology attenuation by exercise, several studies have been conducted using molecular investigations, and this trend looks set to continue. Through enormous biotechnological advances, proteomic tools have facilitated protein analysis within complex biological samples such as plasma and tissue, commonly used in exercise research. Until now, classic proteomic tools such as one‐ and two‐dimensional polyacrylamide gel electrophoresis have been used as standard approaches to investigate proteome modulation by exercise. Furthermore, other recently developed in gel tools such as differential gel electrophoresis (DIGE) and gel‐free techniques such as the protein labeling methods (ICAT, SILAC, and iTRAQ) have empowered proteomic quantitative analysis, which may successfully benefit exercise proteomic research. However, despite the three decades of 2‐DE development, neither classic nor novel proteomic tools have been convincingly explored by exercise researchers. To this end, this review gives an overview of the directions in which exercise‐proteome research is moving and examines the main tools that can be used as a novel strategy in exercise physiology investigation. J. Cell. Physiol. 227: 885–898, 2012.

Circulating miR-1, miR-133a, and miR-206 levels are increased after a half-marathon run.

Abstract Context: Circulating miRNAs are potential biomarkers that can be important molecules driving cell-to-cell communication. Objective: To investigate circulating muscle-specific miRNAs in recreational athletes. Materials and methods: Three miRNAs from whole plasma before and after a half-marathon were analyzed by qPCR. Results: MiR-1, −133a, and −206 significantly increased after the race. Discussion: Increased levels of miRNAs after exercise point to potential biomarkers and to the possibility of being functional players following endurance training. Conclusion: These miRNAs are potential biomarkers of muscle damage or adaptation to exercise.

NanoUPLC/MSE proteomic analysis reveals modulation on left ventricle proteome from hypertensive rats after exercise training

UNLABELLED NanoUPLC/MS(E) was used to verify the effects of 8weeks of low (SHR-LIT=4) and high (SHR-HIT=4) intensity training over the left ventricle proteome of hypertensive rats (SHR-C=4). Training enhanced the aerobic capacity and reduced the systolic blood pressure in all exercised rats. NanoUPLC/MS(E) identified 250 proteins, with 233 in common to all groups and 16 exclusive to SHR-C, 2 to SHR-LIT, and 2 to the SHR-HIT. Cardiac hypertrophy related proteins appeared only in SHR-C. The SHR-LIT enhanced the abundance of 30 proteins and diminished 6, while SHR-HIT enhanced the abundance of 39 proteins and reduced other 7. The levels of metabolic (β and γ-enolase, adenine phosphoribosultransferase, and cytochrome b-c1), myofibril (myosin light chain 4, tropomyosin α and β-chain), and transporter proteins (hemoglobin, serum albumin, and hemopexin) were increased by both intensities. Transcription regulator and histone variants were enhanced by SHR-LIT and SHR-HIT respectively. SHR-LIT reduced the concentration of myosin binding protein C, while desmin and membrane voltage dependent anion selective channel protein-3 were reduced only by SHR-HIT. In addition, polyubiquitin B and C, and transcription regulators decreased in both intensities. Exercise also increased the concentration of anti-oxidant proteins, peroxiredozin-6 and glutathione peroxidase-1. BIOLOGICAL SIGNIFICANCE Pathologic left ventricle hypertrophy if one of the major outcomes of hypertension being a strong predictor of heart failure. Among the various risk factors for cardiovascular disorders, arterial hypertension is responsible for the highest rates of mortality worldwide. In this way, this present study contribute to the understanding of the molecular mechanisms involved in the attenuation of hypertension and the regression of pathological cardiac hypertrophy induced by exercise training.

The Effects of Acute and Chronic Exercise on Skeletal Muscle Proteome

Skeletal muscle plasticity and its adaptation to exercise is a topic that is widely discussed and investigated due to its primary role in the field of exercise performance and health promotion. Repetitive muscle contraction through exercise stimuli leads to improved cardiovascular output and the regulation of endothelial dysfunction and metabolic disorders such as insulin resistance and obesity. Considerable improvements in proteomic tools and data analysis have broth some new perspectives in the study of the molecular mechanisms underlying skeletal muscle adaptation in response to physical activity. In this sense, this review updates the main relevant studies concerning muscle proteome adaptation to acute and chronic exercise, from aerobic to resistance training, as well as the proteomic profile of natural inbred high running capacity animal models. Also, some promising prospects in the muscle secretome field are presented, in order to better understand the role of physical activity in the release of extracellular microvesicles and myokines activity. Thus, the present review aims to update the fast‐growing exercise‐proteomic scenario, leading to some new perspectives about the molecular events under skeletal muscle plasticity in response to physical activity. J. Cell. Physiol. 232: 257–269, 2017.

Determination of the Maximal Lactate Steady State in Obese Zucker Rats

This study aims to identify the maximum lactate steady state (MLSS) in obese rats in order to provide a more effective tool in the exercise training prescription for this important animal model. To make such determination, obese (Zucker, n=5) (390.0±18.8 g) and lean (Wistar, n=5) (227.3±26.2 g) rats were studied. After adaptation of animals to treadmill, the MLSS was determined by using 3 different velocities (10 m.min⁻¹, 12.5 m.min⁻¹ and 15 m.min⁻¹ for Zucker and 15 m.min⁻¹, 20 m.min⁻¹ and 25 m.min⁻¹ for Wistar). The MLSS was defined as the highest blood lactate concentration that increased up to 1 mmol.L⁻¹ during constant exercise. In obese rats, the MLSS was found in a velocity considerably lower than in lean controls (12.5 m.min⁻¹ and 20 m.min⁻¹), respectively (p<0.05). Therefore, the identification of MLSS in obese Zucker rats is an important tool for exercise prescription and evaluation in obese rat models.

Assessment of maximal lactate steady state during treadmill exercise in SHR

BackgroundSpontaneously hypertensive rats (SHR) are one of the main animal models used for studying the effects of exercise on hypertension. Therefore, the determination of adequate intensity has been essential for secure and optimized exercise prescriptions concerning hypertensive subjects. This study aimed to identify the MLSS in SHR by using a treadmill test to improve the protocols and further prescriptions of exercise intensity.FindingsIn order to carry out this determination, SHR (n = 10) animals (~17.5 weeks; 227.4 ± 29.3 g; 172.4 ± 8.1 mmHg systolic blood pressure) were divided into two groups (G1 n = 5; G2 n = 5). Rats underwent a test with three different velocities to determine the MLSS. The MLSS was considered as the highest effort intensity where the blood lactate did not vary more than 1 mmol.L-1 from the 10th to the 25th minute. The MLSS was reached at a velocity of 20 m.min-1 with 3.8 ± 0.5 mmol.L-1 of lactate for G1. Additionally, the results were validated in G2. However, when the test was applied at 25 m.min-1, there was no stabilization of BLC in G1 and G2.ConclusionsIn this study it was possible to identify the MLSS in SHR rats, which is an excellent evaluation tool to control exercise intensity. These data are of considerable importance in studies using physical exercise as a means of research in hypertension and may lead to the intensity of exercise being prescribed more appropriately.

Comparative Proteomical Analysis of Zygotic Embryo and Endosperm from Coffea arabica Seeds

During coffee seed development, proteins are predominantly deposited in cotyledons and in the endosperm. Reserve proteins of the 11S family are the most abundant globulins in coffee seeds, acting as a nitrogen source during roasting and guaranteeing flavor and aroma. The aim of the present study was to compare the protein profiles of endosperm and zygotic embryos of coffee seeds. Proteins were extracted from whole seed as well as from embryo and endosperm, separately. Total proteins were analyzed by two-dimensional electrophoresis (2-DE) followed by identification by mass spectrometry (MS). The most abundant spots observed in the gels of coffee seeds were excised, digested with trypsin, and identified by MS as subunits of the 11S globulin. Spots with identical pI and molecular masses were also observed in the protein profiles of coffee endosperm and embryo, indicating that 11S protein is also highly expressed in those tissues. Peptide sequence coverage of about 20% of the entire 11S globulin was obtained. Three other proteins were identified in the embryo and endosperm 2-DE profiles as a Cupin superfamily protein, an allergenic protein (Pru ar 1), exclusive to the endosperm 2D map, and a hypothetical protein, observed only in the zygotic embryo profile.