Influence of body composition and cardiorespiratory fitness on plasma HSP72, norepinephrine, insulin, and glucose responses to an acute aerobic exercise bout performed in the fed state.

Being overweight is already considered a metabolic risk factor, which can be overcome by increasing cardiorespiratory fitness (CRF). Acute exercise is known to induce changes in plasma hormones and heat shock proteins release. However, there is a lack of studies investigating the impact of body composition and CRF on these variables following acute aerobic exercise. To assess the influence of body composition and cardiorespiratory fitness on plasma heat shock protein 72 kDa (HSP72), norepinephrine (NE), insulin, and glucose responses to an acute aerobic exercise bout in the fed state. Twenty-four healthy male adults were recruited and allocated into three groups: overweight sedentary (n = 8), normal weight sedentary (n = 8), and normal weight active (n = 8). The volunteers performed an acute moderate exercise session on a treadmill at 70% of VO2 peak. Blood samples were drawn at baseline, immediately post-exercise, and at 1-h post-exercise. The exercise session did not induce changes in HSP72 nor NE but changes in glucose and insulin were affected by body mass index. Also, subjects with elevated CRF maintain reduced NE through exercise. At baseline, the overweight sedentary group showed elevated NE, insulin, and glucose; these last two impacting the HOMA-IR index. Thirty minutes of aerobic exercise at 70% VO2 peak, in the fed state, did not change the levels of plasma NE and HSP72. Elevated body composition seems to impact metabolic profile and increase sympathetic activity. Conversely, subjects with increased cardiorespiratory fitness seem to have attenuated sympathetic activity.

Effects of physical exercise on the functionality of human nucleotidases: A systematic review.

Nucleotidases contribute to the regulation of inflammation, coagulation, and cardiovascular activity. Exercise promotes biological adaptations, but its effects on nucleotidase activities and expression are unclear. The objective of this study was to review systematically the effects of exercise on nucleotidase functionality in healthy and unhealthy subjects. The MEDLINE, EMBASE, Cochrane Library, and Web of Science databases were searched to identify, randomized clinical trials, non-randomized clinical trials, uncontrolled clinical trials, quasi-experimental, pre-, and post-interventional studies that evaluated the effects of exercise on nucleotidases in humans, and was not limited by language and date. Two independent reviewers performed the study selection, data extraction, and assessment of risk of bias. Of the 203 articles identified, 12 were included in this review. Eight studies reported that acute exercise, in healthy and unhealthy subjects, elevated the activities or expression of nucleotidases. Four studies evaluated the effects of chronic training on nucleotidase activities in the platelets and lymphocytes of patients with metabolic syndrome, chronic kidney disease, and hypertension and found a decrease in nucleotidase activities in these conditions. Acute and chronic exercise was able to modify the blood plasma and serum levels of nucleotides and nucleosides. Our results suggest that short- and long-term exercise modulate nucleotidase functionality. As such, purinergic signaling may represent a novel molecular adaptation in inflammatory, thrombotic, and vascular responses to exercise.

Cardiometabolic effects of early v. delayed time-restricted eating plus energetic restriction in adults with overweight and obesity: an exploratory randomised clinical trial.

This exploratory study investigated the effects of early v. delayed time-restricted eating (TRE) plus caloric restriction (CR) on body weight, body composition and cardiometabolic parameters in adults with overweight and obesity. Adults (20-40 years) were randomised to one of three groups for 8 weeks: early time-restricted eating (eTRE; 08.00-16.00) plus CR, delayed time-restricted eating (dTRE; 12.00-20.00) plus CR or only CR (CR; 08.00-20.00). All groups were prescribed a 25 % energy deficit relative to daily energy requirements. Thirteen participants completed the study in the eTRE and CR groups and eleven in the dTRE group (n 37). After the interventions, there was no significant difference between the three groups for any of the outcomes. Compared with baseline, significant decreases were observed in the body weight (eTRE group: -4·2 kg; 95 % CI, -5·6, -2·7; dTRE group: -4·8 kg; 95 % CI, -5·9, -3·7; CR: -4·0 kg; 95 % CI, -5·9, -2·1), fat mass (eTRE group: -2·9 kg; 95 % CI, -3·9, -1·9; dTRE group: -3·6 kg; 95 % CI, -4·6, -2·5; CR: -3·1 kg; 95 % CI, -4·3, -1·8) and fasting glucose levels (eTRE group: -4 mg/dl; 95 % CI, -8, -1; dTRE group: -2 mg/dl; 95 % CI, -8, 3; CR: -3 mg/dl; 95 % CI, -8, 2). In a free-living setting, TRE with a energetic deficit, regardless of the time of day, promotes similar benefits in weight loss, body composition and cardiometabolic parameters. However, given the exploratory nature of our study, further investigation is needed to confirm these findings.

Chronic aerobic exercise prevents high-fructose diet-induced impairment in blood pressure in healthy young adults: a double-blind, randomised clinical trial.

The purpose of the study was to verify the effect of 4 weeks of a high-fructose diet (HFD) associated with aerobic training on the risk factors for cardiometabolic diseases. Twenty-one young adults were randomised into three groups: HFD (HFD: 1 g/kg body weight of fructose/day), high-glucose diet (HGD: 1 g/kg body weight of glucose/day) and high-fructose diet and exercise (HFDE: 1 g/kg body weight of fructose/day + 3 weekly 60-minute sessions of aerobic exercise). Before and after the 4 weeks of the intervention, blood samples were taken and flow-mediated dilatation, insulin resistance index, pancreatic beta cell functional capacity index, insulin sensitivity index and 24-h blood pressure were evaluated. HFD showed an increase in uric acid concentrations (P = 0·040), and HGD and HFDE groups showed no changes in this outcome between pre- and post-intervention; however, the HFDE group showed increased uric acid concentrations from the middle to the end of the intervention (P = 0·013). In addition, the HFD group showed increases in nocturnal systolic blood pressure (SBP) (P = 0·022) and nocturnal diastolic blood pressure (DBP) (P = 0·009). The HGD group exhibited decreases in nocturnal SBP (P = 0·028) and nocturnal DBP (P = 0·031), and the HFDE group showed a decrease in 24-h SBP (P = 0·018). The consumption of 1 g/kg of fructose per day may increase uric acid concentrations and blood pressure in adults. Additionally, aerobic exercises along with fructose consumption attenuate changes in uric acid concentrations and prevent impairment in nocturnal blood pressure.

Post-weaning social isolation impairs purinergic signaling in rat brain.

Early life stressors, such as social isolation (SI), can disrupt brain development contributing to behavioral and neurochemical alterations in adulthood. Purinergic receptors and ectonucleotidases are key regulators of brain development in embryonic and postnatal periods, and they are involved in several psychiatric disorders, including schizophrenia. The extracellular ATP drives purinergic signaling by activating P2X and P2Y receptors and it is hydrolyzed by ectonucleotidases in adenosine, which activates P1 receptors. The purpose of this study was to investigate if SI, a rodent model used to replicate abnormal behavior relevant to schizophrenia, impacts purinergic signaling. Male Wistar rats were reared from weaning in group-housed or SI conditions for 8 weeks. SI rats exhibited impairment in prepulse inhibition and social interaction. SI presented increased ADP levels in cerebrospinal fluid and ADP hydrolysis in the hippocampus and striatum synaptosomes. Purinergic receptor expressions were upregulated in the prefrontal cortex and downregulated in the hippocampus and striatum. A2A receptors were differentially expressed in SI prefrontal cortex and the striatum, suggesting distinct roles in these brain structures. SI also presented decreased ADP, adenosine, and guanosine levels in the cerebrospinal fluid in response to D-amphetamine. Like patients with schizophrenia, uric acid levels were prominently increased in SI rats after D-amphetamine challenge. We suggest that the SI-induced deficits in prepulse inhibition might be related to the SI-induced changes in purinergic signaling. We provide new evidence that purinergic signaling is markedly affected in a rat model relevant to schizophrenia, pointing out the importance of purinergic system in psychiatry conditions.

New insights into cytotoxic mechanisms of bozepinib against glioblastoma.

Glioblastoma (GBM) is the most frequent and aggressive brain tumor in adults and the current treatments only have a modest effect on patient survival. Recent studies show that bozepinib (BZP), a purine derivative, has potential applications in cancer treatment. The aim of this study was to evaluate the effect of BZP against GBM cells, specially concerning the purinergic system. Thus, GBM cells (C6 and U138 cell lines) were treated with BZP and cell viability, cell cycle, and annexin/PI assays, and active caspase-3 measurements were carried out. Besides, the effect of BZP over the purinergic system was also evaluated in silico and in vitro. Finally, we evaluate the action of BZP against important markers related to cancer progression, such as Akt, NF-κB, and CD133. We demonstrate here that BZP reduces GBM cell viability (IC50 = 5.7 ± 0.3 µM and 12.7 ± 1.5 µM, in C6 and U138 cells, respectively), inducing cell death through caspase-dependent apoptosis, autophagosome formation, activation of NF-κB, without any change in cell cycle progression or on the Akt pathway. Also, BZP modulates the purinergic system, inducing an increase in CD39 enzyme expression and activity, while inhibiting CD73 activity and adenosine formation, without altering CD73 enzyme expression. Curiously, one cycle of treatment resulted in enrichment of GBM cells expressing NF-κB and CD133+, suggesting resistant cells selection. However, after another treatment round, the resistant cells were eliminated. Altogether, BZP presented in vitro anti-glioma activity, encouraging further in vivo studies in order to better understand its mechanism of action.

Acute moderate-intensity aerobic exercise promotes purinergic and inflammatory responses in sedentary, overweight and physically active subjects.

NEW FINDING: What is the central question of this study? How does moderate-intensity aerobic exercise affect the behaviour of purinergic enzymes in sedentary, overweight and physically active subjects? What is the relationship between purinergic and inflammatory responses triggered by exercise? What is the main finding and its importance? Moderate-intensity aerobic exercise modifies the activity of purinergic enzymes and the levels of nucleotides and nucleosides. These results are similar in subjects with different biological characteristics. 5'-Nucleotidase activity and adenosine levels are associated with inflammatory responses. This study suggests that a purinergic pathway is related to the inflammatory responses triggered by exercise. ABSTRACT: Purinergic signalling is a mechanism of extracellular communication that modulates events related to exercise, such as inflammation and coagulation. Herein, we evaluated the effects of acute moderate-intensity exercise on the activities of purinergic enzymes and plasma levels of adenine nucleotides in individuals with distinct metabolic characteristics. We analysed the relationship between purinergic parameters, inflammatory responses and cardiometabolic markers. Twenty-four healthy males were assigned to three groups: normal weight sedentary (n = 8), overweight sedentary (n = 8) and normal weight physically active (n = 8). The volunteers performed an acute session of moderate-intensity aerobic exercise on a treadmill at 70% of V̇O2peak ; blood samples were drawn at baseline, immediately post-exercise and at 1 h post-exercise. Immediately post-exercise, all subjects showed increases in ATP, ADP, AMP and p-nitrophenyl thymidine 5'-monophosphate hydrolysis, while AMP hydrolysis remained increased at 1 h after exercise. High-performance liquid chromatography analysis demonstrated lower levels of ATP and ADP at post- and 1 h post-exercise in all groups. Conversely, adenosine and inosine levels increased at post-exercise, but only adenosine remained augmented at 1 h after exercise in all groups. With regard to inflammatory responses, the exercise protocol increased tumour necrosis factor α (TNF-α) and interleukin 8 (IL-8) concentrations in all subjects, but only TNF-α remained elevated at 1 h after exercise. Significant correlations were found between the activity of 5'-nucleotidase, adenosine levels, V̇O2peak , triglyceride, TNF-α and IL-8 levels. Our findings suggest a purinergic signalling pathway that participates, at least partially, in the inflammatory responses triggered by acute moderate-intensity exercise. The response of soluble nucleotidases to acute moderate exercise appears to be similar between subjects of different biological profiles.

Characterization and antiproliferative activity of glioma-derived extracellular vesicles.

Aim: To characterize a method to isolate glioma-derived extracellular vesicles (GEVs) and understand their role in immune system modulation and glioma progression. Materials & methods: GEVs were isolated by differential centrifugation from C6 cell supernatant and characterized by size and expression of CD9, HSP70, CD39 and CD73. The glioma model was performed by injecting C6 glioma cells into the right striatum of Wistar rats in the following groups: controls (C6 cells alone), coinjection (C6 cells + GEVs) and GEVs by intranasal administration followed by immune cells, tumor size and cells proliferation analyses. Results: GEVs presented uniform size (175 nm), expressed CD9, HSP70, CD39, CD73 and produced adenosine. In vivo, we observed a reduction in tumor size, in cell proliferation (Ki-67) and in a regulatory cell marker (FoxP3). Conclusion: GEVs, administered before or at tumor challenge, have antiproliferative properties and reduce regulatory cells in the glioma microenvironment.

P2X7 Purinergic Receptor Is Involved in the Pathophysiology of Mania: a Preclinical Study.

The pathophysiology of bipolar disorder remains incompletely elucidated. The purinergic receptor, P2X7 (P2X7R), plays a central role in neuroinflammation, the establishment, and maintenance of microglial activation and neuronal damage/death, all characteristics of bipolar disorder pathology. The present study aims to explore the participation of the P2X7R in a preclinical pharmacological model of mania. We analyzed the modulatory effects of the P2X7R antagonist, brilliant blue, on behavior, monoamines, gene expression, serum purine levels, and cell typing in a pharmacological model of mania induced by D-amphetamine (AMPH) in mice. Our results corroborate an association between the P2X7 receptor and the preclinical animal model of mania, as demonstrated by the decreased responsiveness to AMPH in animals with pharmacologically blocked P2X7R. This study further suggests a possible dopaminergic mechanism for the action of P2X7 receptor antagonism. Additionally, we observed increased peripheral levels of adenosine, a neuroprotective molecule, and increased central expression of Entpd3 and Entpd1 leading to the hydrolysis of ATP, a danger signal, possibly as an attempt to compensate for the damage induced by AMPH. Lastly, P2X7R antagonism in the AMPH model was found to potentially modulate astrogliosis. Our results support the hypothesis that P2X7R plays a vital role in the pathophysiology of mania, possibly by modulating the dopaminergic pathway and astrogliosis, as reflected in the behavioral changes observed. Taken together, this study suggests that a purinergic system imbalance is associated with the AMPH-induced preclinical animal model of mania. P2X7R may represent a promising molecular therapeutic target for bipolar disorder.

Hydrolysis of ATP, ADP, and AMP is increased in blood plasma of prostate cancer patients.

Prostate cancer is among the major malignancies that affect men around the world. Adenine nucleotides are important signaling molecules that mediate innumerous biological functions in pathophysiological conditions, including cancer. These molecules are degraded by several ectoenzymes named ectonucleotidases that produce adenosine in the extracellular medium. Some of these ecto-enzymes can be found in soluble in the blood stream. Thus, the present study aimed to evaluate the hydrolysis of adenine nucleotides (ATP, ADP, and AMP) in the plasma blood of patients with prostate cancer. Peripheral blood samples were collected, and questionnaires were filled based on the clinical data of the medical records. The nucleotide hydrolysis was performed by Malachite Green method using ATP, ADP, and AMP as substrates. Plasma from prostate cancer patients presented an elevated hydrolysis of all nucleotides evaluated when compared to healthy individuals. NTPDase inhibitor (ARL67156) and the alkaline phosphatase inhibitor (levamisole) did not alter ATP hydrolysis. However, AMP hydrolysis was reduced by the CD73 inhibitor, APCP, and by levamisole, suggesting the action of a soluble form of CD73 and alkaline phosphatase. On microvesicles, it was observed that there was a low expression and activity of CD39 and almost absent of CD73. The correlation of ATP, ADP, and AMP hydrolysis with clinic pathological data demonstrated that patients who received radiotherapy showed a higher AMP hydrolysis than those who did not, and patients with lower clinical stage (CS-IIA) presented an elevated ATP hydrolysis when compared to those with more advanced clinical stages (CS-IIB and CS-III). Patients of all clinical stages presented an elevated AMPase activity. Therefore, we can suggest that the nucleotide hydrolysis might be attributed to soluble ecto-enzymes present in the plasma, which, in a coordinate manner, produce adenosine in the blood stream, favoring prostate cancer progression.

Acute and residual effects of aerobic exercise on fructose-induced postprandial lipemia on lean male subjects.

PURPOSE: The addition of fructose to one or more meals daily may lead to increased postprandial lipemia (PPL). Aerobic exercise has been successful in preventing those increases; however, the duration of exercise effects is still unknown. The aim of this study was to evaluate the acute and residual effects of aerobic exercise and fructose ingestion on PPL. METHODS: Twelve young and sedentary men completed a crossover blinded randomized trial. On day 0, they performed 45 min of aerobic exercise at 60% of VO2peak, or 45 min of resting. On day 1, they received a high-fat meal together with one of the following conditions: (a) a fructose-rich beverage (FRUCT), or (b) exercise performed 13 h before the fructose-rich beverage ingestion (FRUCTEX), or (c) a dextrose-based beverage (DEX). On day 2, all subjects received a high-fat meal plus dextrose. Five blood samples were taken on days 1 and 2, to measure triglycerides (TG), HDL cholesterol, VLDL, total cholesterol (TC), glucose and insulin. RESULTS: On day 1, the delta of the TG peak was higher for FRUCT compared to DEX condition (+ 73.7%; p = 0.019). Total area under the curve (AUC) of TG was lower on the condition FRUCTEX compared to FRUCT (+ 30%; p = 0.001). There was no effect of the beverages or the exercise on VLDL, TC, HDL and non-HDL cholesterol (p > 0.05). There were no differences found in any of the parameters assessed on day 2 (p > 0.05). CONCLUSIONS: Fructose consumption (0.5 g/kg) severely increased postprandial TG on day 1, but not on day 2. Previous exercise performance could lead to ~ 30% reduction on the AUC of postprandial TG in 13 h, but not after 37 h followed by fructose consumption. The regularity of physical exercise practice seems to be essential to promote a constant hypolipemic effect.

Methionine and/or Methionine Sulfoxide Alter Ectoenzymes Activities in Lymphocytes and Inflammatory Parameters in Serum from Young Rats: Acute and Chronic Effects.

In this study we investigated the effect of acute and chronic treatment with Met and/or methionine sulfoxide (MetO) on ectonucleotidases and cholinesterases activities from lymphocytes and purine derivatives compounds, C-protein reactive, interleukin-10, interleukin-6, and tumor necrosis factor-α levels in serum of young rats. Adenosine triphosphate hydrolysis was decreased in lymphocytes 1 h after treatment by MetO and Met + MetO. However, adenosine triphosphate and adenosine diphosphate hydrolysis in lymphocytes was increased in the groups MetO and Met + MetO and adenosine deaminase activity was increased in MetO 3 h after the treatment. Acetylcholinesterase activity was increased in lymphocytes after 3 h and 21 days of treatment by MetO and Met + MetO, while serum butyrycholinesterase activity was decreased after 1 h and 21 days of treatment in the same groups. In chronic treatment, interleukin-6 and tumor necrosis factor-α level were increased, while that interleukin-10 level was decreased by Met, MetO, and Met + MetO when compared to control group. C-protein reactive level was increased by MetO and Met + MetO. Adenosine triphosphate and adenosine monophosphate levels were reduced in all amino acids treated groups, while adenosine diphosphate and hypoxanthine were enhanced by MetO and Met + MetO. Adenosine and xanthine were reduced in the MetO group, whereas inosine levels were decreased in the MetO and Met + MetO groups. These findings help to understand the inflammatory alterations observed in hypermethioninemia.

Altered extracellular ATP, ADP, and AMP hydrolysis in blood serum of sedentary individuals after an acute, aerobic, moderate exercise session.

Nucleotidases participate in the regulation of physiological and pathological events, such as inflammation and coagulation. Exercise promotes distinct adaptations, and can influence purinergic signaling. In the present study, we investigated soluble nucleotidase activities in the blood serum of sedentary young male adults at pre- and post-acute moderate aerobic exercise. In addition, we evaluated how this kind of exercise could influence adenine nucleotide concentrations in the blood serum. Sedentary individuals were submitted to moderate aerobic exercise on a treadmill; blood samples were collected pre- and post-exercise, and serum was separated for analysis. Results showed increases in ATP, ADP, and AMP hydrolysis post-exercise, compared to pre-exercise values. The ecto-nucleotide pyrophosphatase/phosphodiesterase was also evaluated, showing an increased activity post-exercise, compared to pre-exercise. Purine levels were analyzed by HPLC in the blood serum, pre- and post-exercise. Decreased levels of ATP and ADP were found post-exercise, in contrast with pre-exercise values. Conversely, post-exercise levels of adenosine and inosine increased compared to pre-exercise levels. Our results indicate an influence of acute exercise on ATP metabolism, modifying enzymatic behavior to promote a protective biological environment.

Hepatic and seric levels of purines in rats experimentally infected by Fasciola hepatica.

The aim of this study was to evaluate hepatic and seric levels of purines, as well as their breakdown products in rats infected by Fasciola hepatica on days 15 and 87 post-infection (PI). Rats were divided into two groups: uninfected (n = 10) and infected (n = 20). On day 15 (n = 5 for uninfected group and n = 10 for infected group) and 87 PI (n = 5 for uninfected group and n = 10 for infected group), animals were euthanized for sampling to evaluate levels of purines by high-performance liquid chromatography. In serum, ATP increased (P < 0.05) and ADP decreased (P < 0.05) on days 15 and 87 PI, while AMP increased (P < 0.05) only on day 15 PI. Hypoxanthine levels increased (P < 0.05) on days 15 and 87 PI, while adenosine and xanthine levels decreased and increased (P < 0.05), respectively, on day 87 PI. No difference was observed regarding seric inosine and uric acid (P > 0.05). Hepatic ATP, adenosine, and uric acid levels decreased (P < 0.05) on days 15 and 87 PI. AMP levels decreased (P < 0.05) on day 87 PI, while xanthine levels increased (P < 0.05) on day 15 PI in the liver. Also in the liver, hypoxanthine levels increased (P < 0.05) on day 15 PI and decreased (P < 0.05) on day 87 PI. On the other hand, there was no difference on hepatic ADP and inosine levels (P > 0.05). Therefore, it is possible to conclude that F. hepatica infection can change purine levels, which may be associated with an inflammatory process, and these alterations may influence fasciolosis pathogenesis.

Physical training normalizes nucleotide hydrolysis and biochemical parameters in blood serum from streptozotocin-diabetic rats.

Ectonucleotidases and the nucleotide metabolism have been implicated as important regulators in diabetes disease. We evaluated the ectonucleotidase activities and biochemical parameters in blood serum of streptozotocin (STZ)-induced diabetic rats submitted a physical training protocol. We observed a raise in ATP, ADP, AMP and p-Nph-5'-TMP hydrolysis rate and in the levels of cholesterol and triglycerides in rat blood serum, after 30 days of diabetes induction. However, in serum of rats submitted a physical training protocol by forced swimming, both the nucleotide hydrolysis rate and the lipids levels returned to the control values. Considering that diabetes leads to multiple pathophysiological alterations, the modulations observed in ectonucleotidase activities may be part of the events involved in these alterations. Then the physical training is a very important way to control the vascular alterations developed in diabetes.