Trends and challenges in analytical chemistry for multi-analysis of illicit drugs employing wastewater-based epidemiology.

Wastewater-based epidemiology (WBE) for quantification of illicit drug biomarkers (IDBs) in wastewater samples is an effective tool that can provide information about drug consumption. The most commonly quantified IDBs belong to different chemical classes, including cocaine, amphetamine-type stimulants, opioids, and cannabinoids, so the different chemical properties of these molecules pose a challenge in the development of analytical methods for multi-analyte analysis. Recent workflows include the steps of sampling and storage, sample preparation using solid-phase extraction (SPE) or without extraction, and quantification of analytes employing gas or liquid chromatography coupled with mass spectrometry. The greatest difficulty is due to the fact that wastewater samples are complex chemical mixtures containing analytes with different chemical properties, often present at low concentrations. Therefore, in the development of analytical methods, there is the need to simplify and optimize the analytical workflows, reducing associated uncertainties, analysis times, and costs. The present work provides a critical bibliographic survey of studies published from the year 2020 until now, highlighting the challenges and trends of published analytical workflows for the multi-analysis of IDBs in wastewater samples, considering sampling and sample preparation, method validation, and analytical techniques.

Respiratory Viral Shedding in Healthcare Workers Reinfected with SARS-CoV-2, Brazil, 2020.

We documented 4 cases of severe acute respiratory syndrome coronavirus 2 reinfection by non-variant of concern strains among healthcare workers in Campinas, Brazil. We isolated infectious particles from nasopharyngeal secretions during both infection episodes. Improved and continued protection measures are necessary to mitigate the risk for reinfection among healthcare workers.

Gas6 drives Zika virus-induced neurological complications in humans and congenital syndrome in immunocompetent mice.

Zika virus (ZIKV) has the ability to cross placental and brain barriers, causing congenital malformations in neonates and neurological disorders in adults. However, the pathogenic mechanisms of ZIKV-induced neurological complications in adults and congenital malformations are still not fully understood. Gas6 is a soluble TAM receptor ligand able to promote flavivirus internalization and downregulation of immune responses. Here we demonstrate that there is a correlation between ZIKV neurological complications with higher Gas6 levels and the downregulation of genes associated with anti-viral response, as type I IFN due to Socs1 upregulation. Also, Gas6 gamma-carboxylation is essential for ZIKV invasion and replication in monocytes, the main source of this protein, which was inhibited by warfarin. Conversely, Gas6 facilitates ZIKV replication in adult immunocompetent mice and enabled susceptibility to transplacental infection. Our data indicate that ZIKV promotes the upregulation of its ligand Gas6, which contributes to viral infectivity and drives the development of severe adverse outcomes during ZIKV infection.

Motivations and attitudes of Brazilian dairy farmers regarding the use of automated behaviour recording and analysis systems.

In this Research Communication we investigate the motivations of Brazilian dairy farmers to adopt automated behaviour recording and analysis systems (ABRS) and their attitudes towards the alerts that are issued. Thirty-eight farmers participated in the study distributed into two groups, ABRS users (USERS, n = 16) and non-users (NON-USERS, n = 22). In the USERS group 16 farmers accepted being interviewed, answering a semi-structured interview conducted by telephone, and the answers were transcribed and codified. In the NON-USERS group, 22 farmers answered an online questionnaire. Descriptive analysis was applied to coded answers. Most farmers were young individuals under 40 years of age, with undergraduate or graduate degrees and having recently started their productive activities, after a family succession process. Herd size varied with an overall average of approximately 100 cows. Oestrus detection and cow's health monitoring were the main reasons given to invest in this technology, and cost was the most important factor that prevented farmers from purchasing ABRS. All farmers in USERS affirmed that they observed the target cows after receiving a health or an oestrus alert. Farmers believed that they were able to intervene in the evolution of the animals' health status, as the alerts gave a window of three to four days before the onset of clinical signs of diseases, anticipating the start of the treatment.The alerts issued by the monitoring systems helped farmers to reduce the number of cows to be observed and to identify pre-clinically sick and oestrous animals more easily. Difficulties in illness detection and lack of definite protocols impaired the decision making process and early treatment, albeit farmers believed ABRS improved the farm's routine and reproductive rates.

High, but not low, exercise volume shifts the balance of renin-angiotensin system toward ACE2/Mas receptor axis in skeletal muscle in obese rats.

Metabolic syndrome is a cluster of metabolic risk factors that is linked to central obesity, elevated blood pressure, insulin resistance (IR), and dyslipidemia, where the renin-angiotensin system (RAS) may provide a link among them. This study aimed to evaluate volume exercise effects comparing low vs. high volume of chronic aerobic exercise on RAS axes in skeletal muscle in a diet-induced obesity (DIO) rat model. For this, male Wistar-Kyoto rats were fed a standard chow (SC) diet or a high-fat (HF) diet for 32 wk. Animals receiving the HF diet were randomly divided into low exercise volume (LEV, 150 min/wk) and high exercise volume (HEV, 300 min/wk) at the 20th week. After 12 wk of aerobic treadmill training, the body mass and composition, blood pressure, glucose and lipid metabolism, RAS axes, insulin signaling, and inflammatory pathway were performed. HEV slowed the body mass gain, reduced intra-abdominal fat pad and leptin levels, improved total and peripheral body composition and inflammatory cytokine, reduced angiotensin II type 1 receptor expression, and increased Mas receptor protein expression compared with the HF animals. Sedentary groups (SC and HF) presented lower time to exhaustion and maximal velocity compared with the LEV and HEV groups. Both exercise training groups showed reduced resting systolic blood pressure and heart rate, improved glucose tolerance, IR, insulin signaling, and lipid profile. We conclude that the HEV, but not LEV, shifted the balance of RAS toward the ACE2/Mas receptor axis in skeletal muscle, presenting protective effects against the DIO model.

Exercise training dose differentially alters muscle and heart capillary density and metabolic functions in an obese rat with metabolic syndrome.

NEW FINDINGS: What is the central question of this study? Regular exercise is recommended as a non-pharmacological approach for the prevention and treatment of metabolic syndrome. However, the impact of different combinations of intensity, duration and frequency of exercise on metabolic syndrome and microvascular density has not been reported. What is the main finding and its importance? We provide evidence on the impact of aerobic exercise dose on metabolic and microvascular alterations in an experimental model of metabolic syndrome induced by high-fat diet. We found that the exercise frequency and duration were the main factors affecting anthropometric and metabolic parameters and microvascular density in the skeletal muscle. Exercise intensity was related only to microvascular density in the heart. We evaluated the effect of the frequency, duration and intensity of exercise training on metabolic parameters and structural capillary density in obese rats with metabolic syndrome. Wistar-Kyoto rats were fed either a standard commercial diet (CON) or a high-fat diet (HFD). Animals that received the HFD were randomly separated into either a sedentary (SED) group or eight different exercise groups that varied according to the frequency, duration and intensity of training. After 12 weeks of aerobic exercise training, the body composition, aerobic capacity, haemodynamic variables, metabolic parameters and capillary density in the heart and skeletal muscle were evaluated. All the exercise training groups showed reduced resting systolic blood pressure and heart rate and normalized fasting glucose. The minimal amount of exercise (90 min per week) produced little effect on metabolic syndrome parameters. A moderate amount of exercise (150 min per week) was required to reduce body weight and improve capillary density. However, only the high amount of exercise (300 min per week) significantly reduced the amount of body fat depots. The three-way ANOVA showed a main effect of exercise frequency and duration for the improvement of metabolic syndrome and capillary density in skeletal muscle. Exercise intensity was a main factor in reversing microvascular rarefaction in the heart.

Central Sympathetic Modulation Reverses Microvascular Alterations in a Rat Model of High-Fat Diet-Induced Metabolic Syndrome.

OBJECTIVES: The objective of this study was to investigate the role of the SNS on hemodynamic, metabolic, and microvascular alterations in a rat model of HFD-induced MS with salt supplementation. METHODS: In total, 40 adult male Wistar rats were fed normal chow (n = 10) or a HFD (n = 30) for 20 weeks. Thereafter, the HFD group received the centrally acting sympatho-modulatory drugs clonidine (0.1 mg/kg) or rilmenidine (1 mg/kg) or vehicle (n = 10/group) orally by gavage. FCD was evaluated using intravital video microscopy, and the SCD was evaluated using histochemical analysis. RESULTS: The pharmacological modulation of the SNS induced concomitant reductions in SBP, HR and plasma catecholamine levels. These effects were accompanied by a reversal of functional and structural capillary rarefaction in the skeletal muscle in both treated groups and an increase in SCD in the left ventricle only in the rilmenidine group. Improvement of the lipid profile and of glucose intolerance was also obtained only with rilmenidine treatment. CONCLUSIONS: Modulation of sympathetic overactivity results in the reversal of microvascular rarefaction in the skeletal muscle and left ventricle and improves metabolic parameters in an experimental model of MS in rats.

Memory deficits and oxidative stress in cerebral ischemia-reperfusion: neuroprotective role of physical exercise and green tea supplementation.

Ischemic stroke is a major cause of morbidity and mortality all over the world. Among impairments observed in survivors there is a significant cognitive learning and memory deficit. Neuroprotective strategies are being investigated to minimize such deficits after an ischemia event. Here we investigated the neuroprotective potential of physical exercise and green tea in an animal model of ischemia-reperfusion. Eighty male rats were divided in 8 groups and submitted to either transient brain ischemia-reperfusion or a sham surgery after 8 weeks of physical exercise and/or green tea supplementation. Ischemia-reperfusion was performed by bilateral occlusion of the common carotid arteries during 30 min. Later, their memory was evaluated in an aversive and in a non-aversive task, and hippocampus and prefrontal cortex were removed for biochemical analyses of possible oxidative stress effects. Ischemia-reperfusion impaired learning and memory. Reactive oxygen species were increased in the hippocampus and prefrontal cortex. Eight weeks of physical exercise and/or green tea supplementation before the ischemia-reperfusion event showed a neuroprotective effect; both treatments in separate or together reduced the cognitive deficits and were able to maintain the functional levels of antioxidant enzymes and glutathione.

Coping with the experience of frustration throughout life: Sex- and age-specific effects of early life stress on the susceptibility to reward devaluation.

Early life stress may lead to lifelong impairments in psychophysiological functions, including emotional and reward systems. Unpredicted decrease in reward magnitude generates a negative emotional state (frustration) that may be involved with susceptibility to psychiatric disorders. We evaluated, in adolescents and adult rats of both sexes, whether maternal separation (MS) alters the ability to cope with an unexpected reduction of reward later in life. Litters of Wistar rats were divided into controls (non handled - NH) or subjected to MS. Animals were trained to find sugary cereal pellets; later the amount was reduced. Increased latency to reach the reward-associated area indicates higher inability to regulate frustration. The dorsal hippocampus (dHC) and basolateral amygdala (BLA) were evaluated for protein levels of NMDA receptor subunits (GluN2A/GluN2B), synaptophysin, PSD95, SNAP-25 and CRF1. We found that adult MS males had greater vulnerability to reward reduction, together with decreased GluN2A and increased GluN2B immunocontent in the dHC. MS females and adolescents did not differ from controls. We concluded that MS enhances the response to frustration in adult males. The change in the ratio of GluN2A and GluN2B subunits in dHC could be related to a stronger, more difficult to update memory of the aversive experience.

Reemergence of Oropouche virus between 2023 and 2024 in Brazil.

Background: Oropouche virus (OROV; species Orthobunyavirus oropoucheense) is an arthropod-borne virus that has caused outbreaks of Oropouche fever in Central and South America since the 1950s. This study investigates virological factors contributing to the reemergence of Oropouche fever in Brazil between 2023 and 2024. Methods: In this study, we combined OROV genomic, molecular, and serological data from Brazil from 1 January 2015 to 29 June 2024, along with in vitro and in vivo characterization. Molecular screening data included 93 patients with febrile illness between January 2023 and February 2024 from the Amazonas State. Genomic data comprised two genomic OROV sequences from patients. Serological data were obtained from neutralizing antibody tests comparing the prototype OROV strain BeAn 19991 and the 2024 epidemic strain. Epidemiological data included aggregated cases reported to the Brazilian Ministry of Health from 1 January 2014 to 29 June 2024. Findings: In 2024, autochthonous OROV infections were detected in previously non-endemic areas across all five Brazilian regions. Cases were reported in 19 of 27 federal units, with 83.2% (6,895 of 8,284) of infections in Northern Brazil and a nearly 200-fold increase in incidence compared to reported cases over the last decade. We detected OROV RNA in 10.8% (10 of 93) of patients with febrile illness between December 2023 and May 2024 in Amazonas. We demonstrate that the 2023-2024 epidemic was caused by a novel OROV reassortant that replicated approximately 100-fold higher titers in mammalian cells compared to the prototype strain. The 2023-2024 OROV reassortant displayed plaques earlier than the prototype, produced 1.7 times more plaques, and plaque sizes were 2.5 larger compared to the prototype. Furthermore, serum collected in 2016 from previously OROV-infected individuals showed at least a 32-fold reduction in neutralizing capacity against the reassortment strain compared to the prototype. Interpretation: These findings provide a comprehensive assessment of Oropouche fever in Brazil and contribute to a better understanding of the 2023-2024 OROV reemergence. The recent increased incidence may be related to a higher replication efficiency of a new reassortant virus that also evades previous immunity.

Commercial Extruded Plant-Based Diet Lowers Circulating Levels of Trimethylamine N-Oxide (TMAO) Precursors in Healthy Dogs: A Pilot Study.

Elevations in circulating trimethylamine N-oxide (TMAO) and its precursors are observed in humans and dogs with heart failure and are associated with adverse outcomes in people. Dietary intervention that reduces or excludes animal ingredients results in rapid reduction of plasma TMAO and TMAO precursors in people, but the impact of diet in dogs has not been studied. The objective of the current study was to determine the effect of diet on plasma TMAO and 2 of its precursors (choline and betaine) in dogs fed a commercial extruded plant-based diet (PBD) or a commercial extruded traditional diet (TD) containing animal and plant ingredients. Sixteen healthy adult mixed breed dogs from a university colony were enrolled in a randomized, 2-treatment, 2-period crossover weight-maintenance study. Mean (SD) age and body weight of the dogs were 2.9 years (± 1.7) and 14.5 kg (± 4.0), respectively. Eight dogs were female (3 intact, 5 spayed) and 8 dogs were male (4 intact, 4 castrated). Plasma choline, betaine and TMAO were quantified by LC-SID-MRM/MS at baseline, and after 4 weeks on each diet. Choline and betaine were also quantified in the diets. Plasma choline levels were significantly lower (P = 0.002) in dogs consuming a PBD (Mean ± SD, 6.8 µM ± 1.2 µM) compared to a TD (Mean ± SD, 7.8 µM ± 1.6 µM). Plasma betaine levels were also significantly lower (P = 0.03) in dogs consuming a PBD (Mean ± SD, 109.1 µM ± 25.3 µM) compared to a TD (Mean ± SD, 132.4 µM ± 32.5 µM). No difference (P = 0.71) in plasma TMAO was detected in dogs consuming a PBD (Median, IQR, 2.4 µM, 2.1 µM) compared to a TD (Median, IQR, 2.3 µM, 1.1 µM). Betaine content was lower in the PBD than in the TD while choline content was similar in the diets. Our findings indicate consumption of a commercial extruded PBD for 4 weeks reduces circulating levels of the TMAO precursors choline and betaine, but not TMAO, in healthy adult dogs.

Mild to moderate overweight in dogs: is there an impact on routine hematological and biochemical profiles, echocardiographic parameters and cardiac autonomic modulation?

Obesity is considered the most common nutritional disease of dogs. Even though overt obesity is more likely to impair health, even moderately overweight dogs are at greater risk for requiring medication for chronic health problems earlier in life. Although the number of overweight dogs far exceeds the number of obese ones, most of the studies published so far focused on derangements in a mixed overweight/obese population (Body condition score - BCS ≥7/9) rather than in separated groups. This study aimed to evaluate the impact of mild to moderate obesity on routine hematological and biochemical profile and cardiovascular parameters in dogs. Nine healthy lean (BCS =4-5/9) and 24 overweight dogs (BCS = 6-7/9) were enrolled. Complete blood count, serum biochemistry analyses, echocardiographic parameters, and cardiac autonomic function by heart rate variability (HRV) were determined. In our study population, although total protein, globulin and phosphorus concentrations were increased in overweight compared to lean dogs, all complete blood count and biochemical parameters were within reference ranges for both groups. Parameters usually increased in obese dogs, like triglycerides and cholesterol concentrations, were within reference ranges in our overweight population. There were no significant changes in echocardiographic parameters, but HRV had a significant decrease in high frequency (HF) power (P = 0.02), suggesting a depression in parasympathetic activity. Our findings show that mild to moderate overweight dogs do not show the hematological and echocardiographic alterations already reported for mixed overweight/obese populations but might have impaired cardiac autonomic modulation. Although not enough to make conclusions, our data raise the question of whether research studies should place overweight and obese dogs in the same category.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Short-term amino acid, clinicopathologic, and echocardiographic findings in healthy dogs fed a commercial plant-based diet.

Consumer demand for commercially prepared plant-based (PB) dog food is increasing, but studies evaluating the short- or long-term effects of PB diets on canine health are lacking. The objective of this study was to assess the short-term amino acid (AA), clinicopathologic, and echocardiographic findings in 34 client-owned dogs fed a commercial extruded plant-based diet (PBD) in which pea protein was the primary protein source and 4 control dogs fed a commercial extruded traditional diet (TD). Plasma AA and whole blood taurine concentrations were measured in dogs at baseline and after 4 weeks on the PBD or the TD. Hematologic, serum biochemical, and echocardiographic testing were performed at baseline and after 12 weeks on the PBD or the TD. Four dogs in the PBD group did not complete the study. All essential AAs, except methionine, were higher in dogs after 4 weeks on the PBD compared to baseline. Taurine (plasma and whole blood) was also higher after 4 weeks on the PBD compared to baseline. A meaningful difference was detected in whole blood taurine between the PBD group and the control group at 4 weeks (P = .026) with the PBD group being higher. Median hematologic and biochemical results for the PBD group were within normal limits at baseline and at 12 weeks. In the PBD group, left ventricular internal diastolic dimension (LVIDd, P = < .001) and normalized LVIDd (P = .031) were higher 12 weeks post-PBD compared to baseline. There were no meaningful differences in left ventricular internal systolic dimension (LVIDs), normalized LVIDs, or fractional shortening 12 weeks post-PBD. There was no statistical evidence of difference between the 2 groups of dogs for any of the echocardiographic parameters at baseline or at 12 weeks. Essential AA or taurine deficiency was not observed in this cohort of dogs fed a commercial extruded PBD. Additionally, clinically relevant hematologic, serum biochemical and echocardiographic alterations were not detected. Further research is required to determine if long-term static feeding of PB diets can meet and maintain AA and other nutrient targets in dogs.

Reliability of panoramic radiography in predicting proximity of third molars to the mandibular canal: A comparison using cone-beam computed tomography.

PURPOSE: The purpose of this study was to analyze the reliability of 7 panoramic radiographic signs for predicting proximity of the root apices of mandibular third molars to the mandibular canal using cone-beam computed tomography and to correlate these findings with the Pell and Gregory and the Winter classification systems. MATERIALS AND METHODS: An observational, cross-sectional, descriptive study was conducted on 74 patients with bilateral impacted mandibular third molars. Four panoramic radiographic signs were observed in the tooth root (darkening, deflection, and narrowing of the root apices, and bifid apices), and another 3 in the mandibular canal (diversion, narrowing, and interruption of the mandibular canal). Cone-beam computed tomography images were analyzed to identify disruption and diversion of the mandibular canal and root deflection. RESULTS: Binary logistic regression showed that only 4 of the 7 panoramic radiographic signs were able to predict proximity of the root apices of the mandibular third molars to the mandibular canal: darkening of the root, deflection of the root, narrowing of the root, and interruption of the mandibular canal (P<0.05). CONCLUSION: Darkening, deflection, and narrowing of the root, in tandem with the interruption of the mandibular canal on panoramic radiographs, indicate that cone-beam computed tomography should be performed when planning the extraction of impacted mandibular third molars. Proximity between mandibular third molars and the mandibular canal is correlated with the Winter classification.

Workers' occupational health surveillance systems in low- and middle-income countries: a scoping review protocol.

OBJECTIVE: The aim of this scoping review is to describe the characteristics of occupational surveillance systems available in low- and middle-income countries. INTRODUCTION: Well-established workers' health surveillance systems provide important data to guide public health decision-making. A previous review on occupational health surveillance systems in developed countries found that insufficient collection of information and resources affects the work of policymakers and researchers. Identifying the characteristics of existing systems in low- and middle-income countries can assist in decision-making in these countries. INCLUSION CRITERIA: This review will consider studies on occupational health surveillance systems used in both public and private health services in low- and middle-income countries, which describe, implement, analyze, or evaluate surveillance systems on workers' health and working conditions. METHODS: Primary studies, government documents, and other records focusing on information systems of occupational and work-related diseases, occupational accidents, or injuries, published in English, Portuguese, Spanish, or Italian will be retrieved from seven databases, including LILACS, Scopus, PubMed, CINAHL, and Embase, among others. Various sources of gray literature, such as low- and middle-income countries' government websites, will also be searched. After title and abstract screening, relevant articles will be analyzed in full. Data extracted will be presented in tables and narrative form to provide comprehensive information.

Takotsubo syndrome induced by brachytherapy in a patient with endocervical adenocarcinoma.

BACKGROUND: Takotsubo syndrome (TTS), also known as stress cardiomyopathy, apical ballooning syndrome and broken heart syndrome, is characterized by acute-onset chest pain, electrocardiographic (ECG) abnormalities and reversible left ventricular (LV) disfunction in the absence of a culprit obstructive lesion in the coronary arteries; therefore, myocardial infarction is the most important differential diagnosis. Usually induced by emotional/physical stress, its treatment consists in hemodynamic support until complete and spontaneous recovery occurs, which is generally achieved within a few days to weeks. Cervical malignancies are an important public health issue in low/middle-income countries and, in the setting of locally advanced disease, concurrent chemoradiation followed by brachytherapy is considered the standard treatment, harboring curative potential. CASE REPORT: We report a case of a 38-year-old woman who underwent concurrent chemoradiotherapy and developed cardiopulmonary arrest in ventricular fibrillation during a brachytherapy session. Complementary tests disclosed altered ECG and cardiac biomarkers, no evidence of coronary artery obstruction, as well as LV disfunction consistent with TTS on echocardiogram and cardiac MRI. After few days of supportive therapy, complete recovery of heart function was observed. CONCLUSIONS: Especially for cancer patients, who usually experience intense emotional/physical stress intrinsically associated with their diagnosis and aggressive treatments, considering TTS as a differential diagnosis is warranted. Intracavitary brachytherapy procedure may represent a trigger for TTS.

Response to anti-PD1 immunotherapy in patients with metastatic cutaneous sarcoma: case reports and literature review.

Dermal sarcomas represent a group or rare malignancies of mesenchymal origin. Although surgical excision with wide margins can be curative, in the advanced/metastatic setting, treatment options are limited and the benefit from anthracycline-based chemotherapy or targeted agents is usually short-lived. Tumor mutational burden and PD-L1 expression scores can be used as predictive biomarker for response to immunotherapy in some metastatic cancers. The role of immune-checkpoint blockade for sarcoma patients remains investigational. Here we present three cases of dermal sarcomas with high TMB and PD-L1 expression and responses to anti-PD1 agents in two of them.

Aliskiren improves renal morphophysiology and inflammation in Wistar rats with 2K1C renovascular hypertension.

Hypertension is characterized by persistent elevated blood pressure levels, one of the leading causes of death in the world. Renovascular hypertension represents the most common cause of secondary hypertension, and its progress is associated with overactivation of the renin angiotensin aldosterone system (RAAS), causing systemic and local changes. Aliskiren is a renin-inhibiting drug that optimizes RAAS suppression. In this sense, the objective of the present study was to analyze the morphophysiology of the left kidney in Wistar rats with renovascular hypertension after treatment with Aliskiren. Parameters such as systolic blood pressure, urinary creatinine and protein excretion, renal cortex structure and ultrastructure, fibrosis and tissue inflammation were analyzed. Our results showed that the hypertensive animals treated with Aliskiren presented a reestablishment of blood pressure, expression of renin, and renal function, as well as a remodeling of morphological alterations through the reduction of fibrosis. The treatment regulated the laminin expression and decreased pro-inflammatory cytokines, restoring the integrity of the glomerular filtration barrier. Therefore, our findings suggest that Aliskiren has a renoprotective effect acting on the improvement of the morphology, physiology and pathology of the renal cortex of animals with renovascular hypertension.

Modulation of cardiac renin-angiotensin system, redox status and inflammatory profile by different volumes of aerobic exercise training in obese rats.

Overactivation of the classical arm of the renin-angiotensin (Ang) system (RAS) occurs during inflammation, oxidative stress and obesity-induced cardiomyopathy. The activation of the protective arm of RAS may act to counterbalance the deleterious effects of the classical RAS. Although aerobic exercise training (AET) shifts the balance of the RAS towards the protective arm, little is known about the molecular adaptations to different volumes of AET. The aim of this study was to evaluate the impact of AET volume on the modulation of RAS, as well as on cardiac biomarkers of oxidative stress and inflammation, in a diet-induced obesity model. Male Wistar rats were fed either control (CON) or high fat (HF) diet for 32 weeks. At week 20, HF group was subdivided into sedentary, low (LEV, 150 min/week) or high (HEV, 300 min/week) exercise volume. After 12 weeks of exercise, body mass gain, systolic blood pressure and heart rate were evaluated, as well as RAS, oxidative stress and inflammation in the heart. Body mass gain, systolic blood pressure and heart rate were higher in HF group when compared with SC group. Both trained groups restored systolic blood pressure and heart rate, but only HEV reduced body mass gain. Regarding the cardiac RAS, the HF group exhibited favoring of the classical arm and both trained groups shifted the balance towards the counterregulatory protective arm. The HF group had higher B1R expression and lower B2R expression than the control group, and B2R expression was reverted in both trained groups. The HF group also presented oxidative stress. The LEV and HEV groups improved the cardiac redox status by reducing Nox 2 and nitrotyrosine expression, but only the LEV group was able to increase the antioxidant defense by increasing Nrf2 signaling. While the HF group presented higher TNF-α, IL-6 and NFκB expression, and lower IL-10 expression, than the SC group, both training protocols improved the inflammatory profile. Although both trained groups improved the deleterious changes related to obesity cardiomyopathy, it is clear that the molecular mechanisms differ between them. Our results suggest that different exercise volumes might reach different molecular targets, and this could be a relevant factor when using exercise to manage obesity.