Mesenchymal stem cell therapy associated with endurance exercise training: Effects on the structural and functional remodeling of infarcted rat hearts.

We tested the effects of early mesenchymal stem cell (MSC) therapy associated with endurance exercise on the structural and functional cardiac remodeling of rats with myocardial infarctation (MI). Male Wistar rats (40 days old) were divided into 6 groups: control and exercise sham; control and exercise MI; and control and exercise MI MSC. MI was surgically induced and bone marrow-derived MSCs were immediately injected via caudal vein (concentration: 1 × 10(6 )cells). Twenty-four hours later ET groups exercised on a treadmill (5 days/week; 60 min/day; 60% of maximal running velocity) for 12 weeks. Structural and functional changes were determined by echocardiography. Contractility and intracellular global calcium ([Ca(2 +)]i) transient were measured in myocytes from the left ventricular (LV) non-infarcted area. Calcium regulatory proteins were measured by Western blot. MI increased (p < 0.05) heart, ventricular and LV weights and its ratios to body weight; LV internal dimension in diastole (LVID-D) and in systole (LVID-S) and LV free wall in diastole (LVFW-D), but reduced the thickness of interventricular septum in systole (IVS-S), ejection fraction (EF) and fractional shortening (FS). MI augmented (p < 0.05) the times to peak and to half relaxation of cell shortening as well as the amplitude of the [Ca(2 +)]i transient and the times to peak and to half decay. Early MSCs therapy restored LVFW-D, IVS-S and the amplitude and time to half decay of the [Ca(2 +)]i transient. Early endurance exercise intervention increased (p < 0.05) LVFW-S, IVS-S, EF and FS, and reduced the times to peak and to half relaxation of cell shortening, and the amplitude of the [Ca(2 +)]i transient. Exercise training also increased the expression of left ventricular SERCA2a and PLBser16. Nevertheless, the combination of these therapies did not cause additive effects. In conclusion, combining early MSCs therapy and endurance exercise does not potentiate the benefits of such treatments to structural and functional cardiac remodeling in infarcted rats.

Power spectrum analysis of cardiovascular variability during passive heating in conscious rats.

The cardiovascular system plays a direct role in the maintenance of body temperature. Whether passive heating alters cardiovascular autonomic modulation in conscious rats is still unknown. This study investigated the effects of passive heating on systolic blood pressure variability (SBPV) and heart rate variability (HRV) in conscious rats and the involvement of the renin-angiotensin system in the passive heating effects on SBPV and HRV. Fourteen male Wistar rats were randomly assigned to the control group or the losartan treatment group. A catheter was implanted in the left carotid artery to record pulsatile arterial pressure (PAP), and a telemetry sensor was implanted in the abdominal cavity to measure body temperature (Tbody). After recovering from surgery, the animals were subjected to a passive heating protocol (35°C; 30min) in resting conditions, during which Tbody, tail skin temperature and PAP were measured. The mean arterial pressure, systolic and diastolic blood pressure, heart rate, double product (i.e., the product of systolic blood pressure by heart rate), SBPV and HRV were calculated from the PAP. SBPV and HRV were analyzed in terms of both time and frequency domains. Increases in the thermoregulatory and cardiovascular parameters were observed during passive heating in both groups, and those increases were reflected in the higher time and frequency domains of the SBPV. However, passive heating was not effective in altering HRV. Passive heating altered SBPV but not HRV in conscious rats when they were treated with losartan.

Meniscal allograft transplantation in rabbit.

This study evaluated the technique for meniscal allograft transplantation using allografts preserved in glycerin 98% in rabbits. Euthanasia was performed at 70 days to compare the transplanted (TM1 to TM16) versus the contralateral meniscus (OM1 to OM16). Sixteen menisci, 8 transplanted and 8 contralateral, were submitted to gross examination, histomorphometric analysis for identification and quantification of cellular type, and for quantification and distribution of collagen fibers. A revascularization study was conducted in all of the other samples. Lengths of the OM varied from 0.9 to 1.0 cm and two TM were smaller. All TM were completely attached to the synovial membrane, except for one case that presented partial fixation. Both, TM and OM had similar amounts of chondrocytes, fibroblasts and fibrocytes, and at the horns, chondrocytes were predominant. The collagen fibers in TM were well organized throughout the body, and disorganized at the horns. These fibers in OM were organized. The amounts of collagen type I and III, and the vascularization of the perimeniscal tissue and of the edge were similar in OM and TM. These results demonstrated graft integration and thus this transplantation technique and preservation method may be recommended.

Temperature Control of Hypertensive Rats during Moderate Exercise in Warm Environment.

The control of body temperature in Spontaneously Hypertensive Rat (SHR) subjected to exercise in warm environment was investigated. Male SHR and Wistar rats were submitted to moderate exercise in temperate (25°C) and warm (32°C) environments while body and tail skin temperatures, as well as oxygen consumption, were registered. Total time of exercise, workload performed, mechanical efficiency and heat storage were determined. SHR had increased heat production and body temperature at the end of exercise, reduced mechanical efficiency and increased heat storage (p < 0.05). Furthermore, these rats also showed a more intense and faster increase in body temperature during moderate exercise in the warm environment (p < 0.05). The lower mechanical efficiency seen in SHR was closely correlated with their higher body temperature at the point of fatigue in warm environment (p < 0.05). Our results indicate that SHR exhibit significant differences in body temperature control during moderate exercise in warm environment characterized by increased heat production and heat storage during moderate exercise in warm environment. The combination of these responses result in aggravated hyperthermia linked with lower mechanical efficiency. Key PointsThe practice of physical exercise in warm environment has gained importance in recent decades mainly because of the progressive increases in environmental temperature;To the best of our knowledge, these is the first study to analyze body temperature control of SHR during moderate exercise in warm environment;SHR showed increased heat production and heat storage that resulted in higher body temperature at the end of exercise;SHR showed reduced mechanical efficiency;These results demonstrate that when exercising in a warm environment the hypertensive rat exhibit differences in temperature control.

The benefits of endurance training in cardiomyocyte function in hypertensive rats are reversed within four weeks of detraining.

The aim of the present study was to verify the effects of low-intensity endurance training and detraining on the mechanical and molecular properties of cardiomyocytes from spontaneously hypertensive rats (SHRs). Male SHRs and normotensive control Wistar rats at 16-weeks of age were randomly divided into eight groups of eight animals: NC8 and HC8 (normotensive and hypertensive control for 8weeks); NT8 and HT8 (normotensive and hypertensive trained at 50-60% of maximal exercise capacity for 8weeks); NC12 and HC12 (normotensive and hypertensive control for 12weeks); NDT and HDT (normotensive and hypertensive trained for 8weeks and detrained for 4weeks). The total exercise time until fatigue (TTF) was determined by a maximal exercise capacity test. Resting heart rate (RHR) and systolic arterial pressure (SAP) were measured. After the treatments, animals were killed by cervical dislocation and left ventricular myocytes were isolated by enzymatic dispersion. Isolated cells were used to determine intracellular global Ca(2+) ([Ca(2+)]i) transient and cardiomyocyte contractility (1Hz; ~25°C). [Ca(2+)]i regulatory proteins were measured by Western blot, and the markers of pathologic cardiac hypertrophy by quantitative real-time polymerase chain reaction (q-RT-PCR). Exercise training augmented the TTF (NC8, 11.4±1.5min vs. NT8, 22.5±1.4min; HC8, 11.7±1.4min vs. HT8, 24.5±1.3min; P<0.05), reduced RHR (NT8initial, 340±8bpm vs. NT8final, 322±10bpm; HT8initial, 369±8bpm vs. HT8final, 344±10bpm; P<0.05), and SBP in SHR animals (HC8, 178±3mmHg vs. HT8, 161±4mmHg; P<0.05). HC8 rats showed a slower [Ca(2+)]i transient (Tpeak, 83.7±1.8ms vs. 71.7±2.4ms; T50%decay, 284.0±4.3ms vs. 264.0±4.1ms; P<0.05) and cell contractility (Vshortening, 86.1±6.7µm/s vs. 118.6±6.7µm/s; Vrelengthening, 57.5±7.4µm/s vs. 101.3±7.4µm/s; P<0.05), and higher expression of ANF (300%; P<0.05), skeletal α-actin (250%; P<0.05) and a decreased α/ß-MHC ratio (70%; P<0.05) compared to NC8. Exercise training increased [Ca(2+)]i transient (NC8, 2.39±0.06F/F0 vs. NT8, 2.72±0.06F/F0; HC8, 2.28±0.05F/F0 vs. HT8, 2.82±0.05F/F0; P<0.05), and cell contractility (NC8, 7.4±0.3% vs. NT8, 8.4±0.3%; HC8, 6.8±0.3% vs. HT8, 7.8±0.3%; P<0.05). Furthermore, exercise normalized the expression of ANF, skeletal α-actin, and the α/ß-MHC ratio in HT8 rats, augmented the expression of SERCA2a (NC8, 0.93±0.15 vs. NT8, 1.49±0.14; HC8, 0.83±0.13 vs. HT8, 1.32±0.14; P<0.05) and PLBser16 (NC8, 0.89±0.18 vs. NT8, 1.23±0.17; HC8, 0.77±0.17 vs. HT8, 1.32±0.16; P<0.05), and reduced PLBt/SERCA2a (NC8, 1.21±0.19 vs. NT8, 0.50±0.21; HC8, 1.38±0.17 vs. HT8, 0.66±0.21; P<0.05). However, all these adaptations returned to control values within 4weeks of detraining in both SHR and normotensive control animals. In conclusion, low-intensity endurance training induces positive benefits to left ventricular myocyte mechanical and molecular properties, which are reversed within 4weeks of detraining.

Swimming Training Does Not Affect the Recovery of Femoral Midshaft Structural and Mechanical Properties in Growing Diabetic Rats Treated with Insulin.

BACKGROUND: The effects of swimming training associated with insulin treatment on the cortical bone health in young rats with severe type 1 diabetes remain unclear, although there is evidence of such effects on the cancellous bone. This study examined the effects of swimming training combined with insulin therapy on the femoral midshaft structural and mechanical properties in growing rats with type 1 diabetes. METHODS: Male Wistar rats were divided into six groups (n = 10): control sedentary, control exercise, diabetic sedentary, diabetic exercise, diabetic sedentary plus insulin and diabetic exercise plus insulin. Diabetic rats received an injection (60 mg/kg body weight) of streptozotocin (STZ). Exercised animals underwent a swimming program for eight weeks. RESULTS: Diabetes induced by STZ decreased the bone mineral content (BMC) and density (BMD), and cortical thickness and maximum load and tenacity in the femoral midshaft. Insulin treatment partially counteracted the damages induced by diabetes on BMC, BMD and cortical thickness and tenacity. Swimming training did not affect the femoral structural and mechanical properties in diabetic rats. The combination of treatments did not potentiate the insulin effects. In conclusion, swimming training does not affect the benefits of insulin treatment on the femoral midshaft structural and mechanical properties in growing rats with severe type 1 diabetes.

A Recombinant Protein SARS-CoV-2 Candidate Vaccine Elicits High-titer Neutralizing Antibodies in Macaques.

Background: Vaccines that generate robust and long-lived protective immunity against SARS-CoV-2 infection are urgently required. Methods: We assessed the potential of vaccine candidates based on the SARS-CoV-2 spike in cynomolgus macaques (M. fascicularis) by examining their ability to generate spike binding antibodies with neutralizing activity. Antigens were derived from two distinct regions of the spike S1 subunit, either the N-terminal domain or an extended C-terminal domain containing the receptor-binding domain and were fused to the human IgG1 Fc domain. Three groups of 2 animals each were immunized with either antigen, alone or in combination. The development of antibody responses was evaluated through 20 weeks post-immunization. Results: A robust IgG response to the spike protein was detected as early as 2 weeks after immunization with either protein and maintained for over 20 weeks. Sera from animals immunized with antigens derived from the RBD were able to prevent binding of soluble spike proteins to the ACE2 receptor, shown by in vitro binding assays, while sera from animals immunized with the N-terminal domain alone lacked this activity. Crucially, sera from animals immunized with the extended receptor binding domain but not the N-terminal domain had potent neutralizing activity against SARS-CoV-2 pseudotyped virus, with titers in excess of 10,000, greatly exceeding that typically found in convalescent humans. Neutralizing activity persisted for more than 20 weeks. Conclusions: These data support the utility of spike subunit-based antigens as a vaccine for use in humans.

"A recombinant protein SARS-CoV-2 candidate vaccine elicits high-titer neutralizing antibodies in macaques."

Vaccines that generate robust and long-lived protective immunity against SARS-CoV-2 infection are urgently required. We assessed the potential of vaccine candidates based on the SARS-CoV-2 spike in cynomolgus macaques (M. fascicularis) by examining their ability to generate spike binding antibodies with neutralizing activity. Antigens were derived from two distinct regions of the spike S1 subunit, either the N-terminal domain (NTD) or an extended C-terminal domain containing the receptor-binding domain (RBD) and were fused to the human IgG1 Fc domain. Three groups of 2 animals each were immunized with either antigen, alone or in combination. The development of antibody responses was evaluated through 20 weeks post-immunization. A robust IgG response to the spike protein was detected as early as 2 weeks after immunization with either protein and maintained for over 20 weeks. Sera from animals immunized with antigens derived from the RBD were able to prevent binding of soluble spike proteins to the ACE2 receptor, shown by in vitro binding assays, while sera from animals immunized with the NTD alone lacked this activity. Crucially, sera from animals immunized with the RBD but not the NTD had potent neutralizing activity against SARS-CoV-2 pseudotyped virus, with titers in excess of 10,000, greatly exceeding that typically found in convalescent humans. Neutralizing activity persisted for more than 20 weeks. These data support the utility of spike subunit-based antigens as a vaccine for use in humans.

Swimming training potentiates the recovery of femoral neck strength in young diabetic rats under insulin therapy.

OBJECTIVE: To test whether swimming training benefits femoral neck strength in young diabetic rats under insulin therapy. METHODS: A total of 60 male Wistar rats (age: 40 days) were divided equally into the following six groups: control sedentary, control exercise, diabetic sedentary, diabetic exercise, diabetic sedentary plus insulin and diabetic exercise plus insulin. Diabetes was induced with a unique intraperitoneal injection (60 mg/kg body weight) of streptozotocin. Seven days after the injection and after 12 hours of fasting, the animals with blood glucose levels ≥300 mg/dL were considered diabetic. Seven days after the induction of diabetes, the animals in the exercise groups were subjected to progressive swimming training (final week: 90 min/day; 5 days/week; 5% load) for eight weeks. The animals in the insulin groups received a daily dose of insulin (2-4 U/day) for the same period. RESULTS: Severe streptozotocin-induced diabetes reduced the structural properties of the femoral neck (trabecular bone volume, trabecular thickness and collagen fiber content). The femoral neck mechanical properties (maximum load and tenacity) were also impaired in the diabetic rats. Insulin therapy partially reversed the damage induced by diabetes on the structural properties of the bone and mitigated the reductions in the mechanical properties of the bone. The combination of therapies further increased the femoral neck trabecular bone volume (∼30%), trabecular thickness (∼24%), collagen type I (∼19%) and type III (∼13%) fiber contents, maximum load (∼25%) and tenacity (∼14%). CONCLUSIONS: Eight weeks of swimming training potentiates the recovery of femoral neck strength in young rats with severe streptozotocin-induced diabetes under insulin therapy.

Effects of exercise training and stem cell therapy on the left ventricle of infarcted rats.

INTRODUCTION AND OBJECTIVES: Stem cell therapy and aerobic exercise are non-pharmacological therapies following myocardial infarction. The aim of this study was to test whether aerobic exercise training enhances the benefits of mesenchymal stem cell (MSC) therapy on remodeling of the extracellular matrix and fetal gene expression in the left ventricle of infarcted rats. METHODS: Myocardial infarction was surgically induced in six-week old male Wistar rats. Animals were divided into four groups: sedentary control (SC) and sedentary and stem cell treated (SCMSC); exercised (EX) and exercised and stem cell treated (EXMSC). Bone marrow-derived MSCs were immediately transplanted via the tail vein (concentration: 1×106 cells). Exercise training (five days/week, 60 min/day; 60% of maximal running speed) started 24 hours after myocardial infarction and lasted for 12 weeks. RESULTS: Exercise capacity was higher in exercised than in sedentary groups. Animals in the SCMSC, EX and EXMSC groups exhibited better cardiac function than those in SC. Collagen content was lower in the SCMSC, EX and EXMSC groups than in SC and skeletal α-actin expression was lower in EX and EXMSC than in SC. The α/ß-MHC ratio was higher in EX and EXMSC than in SC. The combination of therapies further reduced collagen content in the remote region of the infarct (∼24%) and skeletal α-actin expression (∼30%). CONCLUSION: Aerobic exercise training appears to enhance the beneficial effects of stem cell therapy on remodeling of the extracellular matrix and fetal gene expression in the left ventricle of rats with moderate infarction.

Moderate Continuous Aerobic Exercise Training Improves Cardiomyocyte Contractility in Β1 Adrenergic Receptor Knockout Mice.

BACKGROUND: The lack of cardiac ß1-adrenergic receptors (ß1-AR) negatively affects the regulation of both cardiac inotropy and lusitropy, leading, in the long term, to heart failure (HF). Moderate-intensity aerobic exercise (MCAE) is recommended as an adjunctive therapy for patients with HF. OBJECTIVE: We tested the effects of MCAE on the contractile properties of left ventricular (LV) myocytes from ß1 adrenergic receptor knockout (ß1ARKO) mice. METHODS: Four- to five-month-old male wild type (WT) and ß1ARKO mice were divided into groups: WT control (WTc) and trained (WTt); and ß1ARKO control (ß1ARKOc) and trained (ß1ARKOt). Animals from trained groups were submitted to a MCAE regimen (60 min/day; 60% of maximal speed, 5 days/week) on a treadmill, for 8 weeks. P ≤ 0.05 was considered significant in all comparisons. RESULTS: The ß1ARKO and exercised mice exhibited a higher (p < 0.05) running capacity than WT and sedentary ones, respectively. The ß1ARKO mice showed higher body (BW), heart (HW) and left ventricle (LVW) weights, as well as the HW/BW and LVW/BW than WT mice. However, the MCAE did not affect these parameters. Left ventricular myocytes from ß1ARKO mice showed increased (p < 0.05) amplitude and velocities of contraction and relaxation than those from WT. In addition, MCAE increased (p < 0.05) amplitude and velocities of contraction and relaxation in ß1ARKO mice. CONCLUSION: MCAE improves myocyte contractility in the left ventricle of ß1ARKO mice. This is evidence to support the therapeutic value of this type of exercise training in the treatment of heart diseases involving ß1-AR desensitization or reduction.

Efficacy of sarolaner (Simparic™) against induced infestations of Amblyomma cajennense on dogs.

BACKGROUND: Amblyomma cajennense is the main vector of Rickettsia rickettsii which causes Brazilian spotted fever. This adult tick preferably infests horses and capybaras, but has low host specificity during its immature stages, thus posing a threat to humans and dogs. In this study, the efficacy of sarolaner (Simparic™/Simparica®, Zoetis) when administered once orally to dogs at 2 mg/kg was evaluated against induced infestations of A. cajennense nymphs for up to 35 days after treatment. METHODS: Based on pretreatment tick counts, 20 dogs were randomly allocated to treatment with sarolaner (Simparic™) dosed at 2 mg/kg of body weight or a placebo on Day 0 of the study. Artificial infestations were performed using laboratory raised A. cajennense nymphs on study days -2, 5, 12, 19, 26 and 33. Efficacy was determined at 48 h post-treatment or post-infestation at each time point relative to the counts for dogs that received placebo. RESULTS: There were no adverse reactions to treatment. A single dose of sarolaner (Simparic™) provided 100% efficacy on study days 2, 7 and 14; and ≥ 99.6% on days 21, 28 and 35. Geometric mean live tick counts for sarolaner were significantly lower than those for placebo on all days (P < 0.0001). CONCLUSIONS: Under the conditions of the present study, sarolaner (Simparic™) administered once orally at 2 mg/kg provided 100% efficacy against existing infestations and ≥ 99.6% efficacy within 48 h against weekly challenges of A. cajennense for at least 35 days after treatment.

Swimming training attenuates the morphological reorganization of the myocardium and local inflammation in the left ventricle of growing rats with untreated experimental diabetes.

Diabetic cardiomyopathy is associated with cardiac remodeling, myocardial dysfunction, low-grade inflammation, and reduced cardiac adiponectin in patients with type 1 diabetes mellitus (T1DM). Alternatively, physical exercise is an important strategy for the management of diabetes. This study aimed to investigate the influence of low-intensity swimming training in cardiac cytokines, structural remodeling, and cardiomyocyte contractile dysfunction in growing rats with untreated experimental DM. Thirty-day-old male Wistar rats were divided into four groups (n=14, per group): sedentary control (SC), exercised control (EC), sedentary diabetic (SD), and exercised diabetic (ED). Diabetes was induced by streptozotocin (60 mg kg(-1), i.p.). Animals from exercised groups swam (5 days/week, 90 min/day, loading up to 5% body weight around the animal's chest) for 8 weeks. The left ventricle (LV) was removed for molecular, morphological, and cardiomyocyte mechanical analysis. Diabetic animals presented cardiac remodeling with myocardial histoarchitectural disorganization, fibrosis, and necrosis. The capillary density was lower in diabetic animals. LV cardiomyocytes from diabetic animals exhibited more prolonged time to the peak of contraction and time to half relaxation than those from control animals. The cardiac levels of interleukin 10, nitric oxide, and total and high molecular weight (HMW) adiponectin were significantly decreased in diabetic animals. Exercise training reduced the level of TNF-α, increased capillary density, and attenuated the histopathological parameters assessed in diabetic rats. In conclusion, the cardiac structural remodeling coexists with reduced levels of total and HMW adiponectin, inflammation, and cardiomyocyte contractility dysfunction in experimental DM. More important, low-intensity swimming training attenuates part of these pathological changes, indicating the beneficial role for exercise in untreated T1DM.

Attenuation of Ca2+ homeostasis, oxidative stress, and mitochondrial dysfunctions in diabetic rat heart: insulin therapy or aerobic exercise?

We tested the effects of swimming training and insulin therapy, either alone or in combination, on the intracellular calcium ([Ca(2+)]i) homeostasis, oxidative stress, and mitochondrial functions in diabetic rat hearts. Male Wistar rats were separated into control, diabetic, or diabetic plus insulin groups. Type 1 diabetes mellitus was induced by streptozotocin (STZ). Insulin-treated groups received 1 to 4 UI of insulin daily for 8 wk. Each group was divided into sedentary or exercised rats. Trained groups were submitted to swimming (90 min/day, 5 days/wk, 8 wk). [Ca(2+)]i transient in left ventricular myocytes (LVM), oxidative stress in LV tissue, and mitochondrial functions in the heart were assessed. Diabetes reduced the amplitude and prolonged the times to peak and to half decay of the [Ca(2+)]i transient in LVM, increased NADPH oxidase-4 (Nox-4) expression, decreased superoxide dismutase (SOD), and increased carbonyl protein contents in LV tissue. In isolated mitochondria, diabetes increased Ca(2+) uptake, susceptibility to permeability transition pore (MPTP) opening, uncoupling protein-2 (UCP-2) expression, and oxygen consumption but reduced H2O2 release. Swimming training corrected the time course of the [Ca(2+)]i transient, UCP-2 expression, and mitochondrial Ca(2+) uptake. Insulin replacement further normalized [Ca(2+)]i transient amplitude, Nox-4 expression, and carbonyl content. Alongside these benefits, the combination of both therapies restored the LV tissue SOD and mitochondrial O2 consumption, H2O2 release, and MPTP opening. In conclusion, the combination of swimming training with insulin replacement was more effective in attenuating intracellular Ca(2+) disruptions, oxidative stress, and mitochondrial dysfunctions in STZ-induced diabetic rat hearts.

Acute and chronic observations of complete atrioventricular block in rats.

The mechanisms of production, and gross, microscopic and electrocardiograhic findings of surgically-induced complete heart block (CHB) in the adult rat are presented. This is an effective in vivo model for establishing alternative methods to electronic pacemakers and for providing detailed information aimed at replacement, reduction and refinement of the technique. Sternal thoracotomy was employed to identify the epicardial fat pad by the aortic root, used as a landmark for cauterization of the atrioventricular (AV) node. Stable CHB was produced in 60 rats with a 70% survival rate. The best survival rate was observed in 8-week-old animals weighing 221 ± 27.6 g. Heart rate before cauterization was 387 ± 55 bpm, reduced after cauterization to 126 ± 40 bpm in the survival and to 65 ± 19 bpm in the non-survival groups. At 30 days findings were: elevated left ventricular end-diastolic pressure (21 ± 5.4 mmHg, P < 0.05); maximal rate of rise of left ventricular pressure (LVP) during isovolumetric contraction (2192 ± 235 mmHg/s, P < 0.05); maximal rate of decrease of LVP (-1658 ± 191 mmHg/s, P < 0.05); isovolumetric relaxation constant (5.7 ± 0.8 ms, P < 0.05) with wet-to-dry lung-weight ratio (78.1 ± 0.4, P < 0.05); heart weight/body weight (0.6 ± 0.1, P < 0.05); heart volume (1.8 ± 0.3 mL, P < 0.05); longitudinal diameter (20.2 ± 1.91 mm, P < 0.05); and transversal diameter (17.0 ± 1.4 mm, P < 0.05) with supported dilated cardiomyopathy which culminated in chronic heart failure. CHB hearts had increased preload and replacement of myofibrils by collagen. CHB was achieved reproducibly by cauterization of the rat AV node and/or His bundle. This led to electrophysiological, hemodynamic, and structural remodeling, and could be useful in long-term cardiac remodeling assessments and potential therapy development.

Swimming training potentiates the recovery of femoral neck strength in young diabetic rats under insulin therapy

OBJECTIVE: To test whether swimming training benefits femoral neck strength in young diabetic rats under insulin therapy. METHODS: A total of 60 male Wistar rats (age: 40 days) were divided equally into the following six groups: control sedentary, control exercise, diabetic sedentary, diabetic exercise, diabetic sedentary plus insulin and diabetic exercise plus insulin. Diabetes was induced with a unique intraperitoneal injection (60 mg/kg body weight) of streptozotocin. Seven days after the injection and after 12 hours of fasting, the animals with blood glucose levels ≥300 mg/dL were considered diabetic. Seven days after the induction of diabetes, the animals in the exercise groups were subjected to progressive swimming training (final week: 90 min/day; 5 days/week; 5% load) for eight weeks. The animals in the insulin groups received a daily dose of insulin (2-4 U/day) for the same period. RESULTS: Severe streptozotocin-induced diabetes reduced the structural properties of the femoral neck (trabecular bone volume, trabecular thickness and collagen fiber content). The femoral neck mechanical properties (maximum load and tenacity) were also impaired in the diabetic rats. Insulin therapy partially reversed the damage induced by diabetes on the structural properties of the bone and mitigated the reductions in the mechanical properties of the bone. The combination of therapies further increased the femoral neck trabecular bone volume (∼30%), trabecular thickness (∼24%), collagen type I (∼19%) and type III (∼13%) fiber contents, maximum load (∼25%) and tenacity (∼14%). CONCLUSIONS: Eight weeks of swimming training potentiates the recovery of femoral neck strength in young rats with severe streptozotocin-induced diabetes under insulin therapy.

Differentiation of adipose tissue-derived mesenchymal stem cells into cardiomyocytes.

BACKGROUND: Cardiomyocytes have small potential for renovation and proliferation in vivo. Consequently, the heart muscle has limited capacity of self-renewal. Mesenchymal stem cells (MSC) therapy, as well as MSC differentiated into cardiomyocytes, has been used in the attempt to minimize the effects of ischemic-hypoxic lesions and those affecting the electrical conduction system of the heart. OBJECTIVE: The present study compared three distinct protocols for induced differentiation of MSC into cardiomyocytes aimed at finding a viable method for producing a large number of functional cells expressing cardiomyogenic phenotype. METHODS: Mesenchymal stem cells were obtained from the adipose tissue of young transgenic Lewis rats expressing green fluorescent protein (GFP), and submitted to three distinct differentiation-inducing media: 1) Planat-Bérnard, 2) 5-azacytidine, and 3) Planat-Bérnard + 5-azacytidine; further, these cells were identified based on the expression of cardiac cell markers. RESULTS: All three protocols detected the expression of sarcomeric-alpha-actinin protein in the exoskeleton of cells, expression of connexin-43 in the nuclear and cytoplasmic membrane, and formation of gap junctions, which are necessary for electrical impulse propagation in the myocardium. However, no spontaneous cell contraction was observed with any of the tested protocols. CONCLUSION: Induction with 5-azacytidine provided an effective cadiomyogenic cellular differentiation similar to that obtained with Planat-Bénard media. Therefore, 5-azacytidine was the method of choice for being the simplest, fastest and lowest-cost protocol for cell differentiation.

Ventricular remodeling in growing rats with experimental diabetes: The impact of swimming training.

Diabetic cardiomyopathy is associated with cardiac muscle remodeling, resulting in myocardial dysfunction, whereas exercise training (ET) is a useful nonpharmacological strategy for the therapy of cardiac diseases. This study tested the effects of low-intensity swimming-training on the structural remodeling of the left ventricle (LV) in growing rats with unmanaged experimental diabetes. Thirty-day-old male Wistar rats were divided into four groups (n=5/group): sedentary-control (SC), exercised-control (EC), sedentary-diabetic (SD), and exercised-diabetic (ED). Swimming-training rats exercised 5 days/week, 90min/day, with a load of 5% BW during 8 weeks. Sections of LV were stained with Periodic acid-Schiff, Sirius Red, and Gomori's reticulin. Seven days and 8 weeks after streptozotocin (STZ) induction (60mgkg(-1) BW), blood glucose (BG) in the diabetic groups (SD=581.40±40.48; ED=558.00±48.89) was greater (p<0.05) than in their controls (SC=88.80±21.70; EC=85.60±11.55). Swimming-training reduced BG by 23mg/dL in the diabetics (p>0.05). The LV of diabetic rats had increased interstitial collagen and reticular fibers on the extracellular matrix and presented glycogen accumulation. More importantly, all these adverse tissue changes induced by STZ were attenuated by ET. Together, these findings support the idea of a beneficial role of exercise in the LV remodeling in rats with unmanaged type-1 diabetes mellitus.

Moderate Continuous Aerobic Exercise Training Improves Cardiomyocyte Contractility in Β1 Adrenergic Receptor Knockout Mice / Treinamento Aeróbico Contínuo de Intensidade Moderada Melhora a Contratilidade do Cardiomiócito em Camundongos com Nocaute para o Receptor B1-Adrenérgico

Abstract Background: The lack of cardiac β1-adrenergic receptors (β1-AR) negatively affects the regulation of both cardiac inotropy and lusitropy, leading, in the long term, to heart failure (HF). Moderate-intensity aerobic exercise (MCAE) is recommended as an adjunctive therapy for patients with HF. Objective: We tested the effects of MCAE on the contractile properties of left ventricular (LV) myocytes from β1 adrenergic receptor knockout (β1ARKO) mice. Methods: Four- to five-month-old male wild type (WT) and β1ARKO mice were divided into groups: WT control (WTc) and trained (WTt); and β1ARKO control (β1ARKOc) and trained (β1ARKOt). Animals from trained groups were submitted to a MCAE regimen (60 min/day; 60% of maximal speed, 5 days/week) on a treadmill, for 8 weeks. P ≤ 0.05 was considered significant in all comparisons. Results: The β1ARKO and exercised mice exhibited a higher (p < 0.05) running capacity than WT and sedentary ones, respectively. The β1ARKO mice showed higher body (BW), heart (HW) and left ventricle (LVW) weights, as well as the HW/BW and LVW/BW than WT mice. However, the MCAE did not affect these parameters. Left ventricular myocytes from β1ARKO mice showed increased (p < 0.05) amplitude and velocities of contraction and relaxation than those from WT. In addition, MCAE increased (p < 0.05) amplitude and velocities of contraction and relaxation in β1ARKO mice. Conclusion: MCAE improves myocyte contractility in the left ventricle of β1ARKO mice. This is evidence to support the therapeutic value of this type of exercise training in the treatment of heart diseases involving β1-AR desensitization or reduction.

Resumo Fundamento: A falta de receptores β1-adrenérgicos (β1-AR) cardíacos afeta negativamente a regulação de inotropismo e lusitropismo cardíacos, levando, no longo prazo, a insuficiência cardíaca (IC). Recomenda-se exercício aeróbico contínuo de intensidade moderada (EACM) como adjuvante no tratamento de pacientes com IC. Objetivo: Testar os efeitos do EACM nas propriedades contráteis de miócitos do ventrículo esquerdo (VE) de camundongos com nocaute para o receptor β1-adrenérgico (β1ARKO). Método: Camundongos machos com 4 a 5 meses de idade, wild-type (WT) e β1ARKO foram divididos em grupos: WT controle (WTc) e treinado (WTt); e β1ARKO controle (β1ARKOc) e treinado (β1ARKOt). Os grupos treinados foram submetidos a regime de EACM (60 min/dia; 60% da velocidade máxima, 5 dias/semana) em esteira rolante, por 8 semanas. Adotou-se P ≤ 0,05 como nível de significância em todas as comparações. Resultados: Os animais β1ARKO (β1ARKOc + β1ARKOt) correram uma distância maior do que os animais WT (WTc + WTt) (p < 0,05). Os camundongos β1ARKO apresentaram maiores pesos corporal (PC), do coração (PCo) e do ventrículo esquerdo (PVE), assim como PCo/PC e PVE/PC do que os camundongos WT. Entretanto, o EACM não afetou tais parâmetros. Os miócitos do VE de camundongos β1ARKO apresentaram maiores (p < 0,05) amplitude e velocidades de contração e relaxamento do que os dos camundongos WT. Além disso, o EACM aumentou (p < 0,05) a amplitude e as velocidades de contração e relaxamento nos camundongos β1ARKO. Conclusão: O EACM melhora a contratilidade do miócito do VE de camundongos β1ARKO. Tal achado confirma o valor terapêutico desse tipo de treinamento físico para o tratamento de doenças cardíacas envolvendo dessensibilização ou redução de β1-AR.

CONSUMO DE ÁLCOOL E A INFLUÊNCIA DO EXERCÍCIO FÍSICO NA ATIVIDADE ENZIMÁTICA DE RATOS WISTAR / ALCOHOL CONSUMPTION AND THE INFLUENCE OF PHYSICAL EXERCISE ON ENZYME ACTIVITY OF WISTAR RATS / CONSUMO DE ALCOHOL Y LA INFLUENCIA DEL EJERCICIO FÍSICO EN LA ACTIVIDAD DE LA ENZIMA DE RATAS WISTAR

RESUMO Introdução: Biomarcadores vem sendo utilizados para monitorar o uso do álcool e, atualmente, os mais sensíveis e específicos são enzimas hepáticas, por exemplo, gama glutamiltransferase (GGT), alanina aminotransferase (ALT), aspartato aminotransferase (AST) e fosfatase alcalina (ALP). Objetivo: Verificar, a partir da experimentação animal, as alterações provocadas pelo uso de álcool e pela prática de atividade física nas enzimas hepáticas e pancreáticas. Métodos: Vinte e quatro ratos da linhagem Wistar foram distribuídos aleatoriamente em grupos experimentais, alojados em gaiolas com ambiente controlado, divididos de acordo com os tratamentos recebidos. No tratamento inicial, foi administrado álcool aos grupos álcool sedentário (AS) e álcool exercitado (AE) e, ao final da quarta semana, iniciou-se o programa de treinamento físico em esteira com os grupos AE e controle exercitado (CE). A coleta de sangue foi realizada por punção cardíaca ao final de cada experimento. Na análise estatística, utilizou-se teste de análise de variância (ANOVA) seguido de teste de Tukey e teste de Kruskal-Wallis. Resultados: O grupo AS apresentou valores significativamente mais elevados de ALT e ALP quando comparado aos grupos CE e AE, respectivamente. Não foram observadas diferenças significativas entre os quatro grupos estudados para os parâmetros AST, GGT e amilase. Conclusão: A associação entre consumo de álcool e sedentarismo aumentou a liberação das enzimas ALT e ALP em ratos Wistar; a prática de exercício físico aeróbico após abstinência alcoólica evitou o aumento da liberação de ALP no plasma desses animais.

ABSTRACT Introduction: Biomarkers have been used to monitor the use of alcohol and currently the most sensitive and specific are the liver enzymes, for example, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP). Objective: To determine through animal experiments the changes caused by alcohol and the physical activity in liver and pancreatic enzymes. Methods: Twenty-four Wistar rats were randomly assigned into experimental groups, housed in cages with controlled environment, divided according to the treatment received. In the initial treatment, alcohol was administered to sedentary alcohol (SA) and exercised alcohol (EA) groups and at the end of the fourth week the program of physical training on a treadmill began with the AE and control exercised (CE) groups. Blood collection was performed by cardiac puncture at the end of each experiment. For the statistical analysis we used analysis of variance (ANOVA) followed by Tukey test and Kruskal-Wallis test. Results: The AS group had significantly higher values of ALT and ALP when compared to CE and EA groups, respectively. No significant differences were observed between the four groups for the parameters AST, GGT and amylase. Conclusion: The association between alcohol consumption and physical inactivity increased the release of the enzymes ALT and ALP in Wistar rats; the practice of aerobic exercise after alcohol withdrawal prevented the increased release of ALP in the plasma of these animals.

RESUMEN Introducción: Los biomarcadores se han utilizado para controlar el consumo de alcohol y en la actualidad las enzimas hepáticas son las más sensibles y específicas, por ejemplo, gamma glutamil transferasa (GGT), alanina aminotransferasa (ALT), aspartato aminotransferasa (AST) y fosfatasa alcalina (ALP). Objetivo: Determinar, a partir de experimentos con animales, los cambios provocados por el alcohol y la actividad física en las enzimas hepáticas y pancreáticas. Métodos: Veinticuatro ratones Wistar fueron asignados al azar a los grupos experimentales, fueron alojados en jaulas con ambiente controlado, divididos de acuerdo a los tratamientos recibidos. En el tratamiento inicial, el alcohol se administró a los grupos alcohol sedentario (AS) y alcohol ejercicio (AE) y el final de la cuarta semana, se inició el programa de entrenamiento físico en una caminadora con los grupos AE y el control ejercitado (CE). La recogida de sangre se realizó mediante punción cardiaca al final de cada experimento. En el análisis estadístico se utilizó el análisis de varianza (ANOVA), seguido por la prueba de Tukey y la prueba de Kruskal-Wallis. Resultados: El grupo AS tuvo valores significativamente más elevados de ALT y ALP en comparación con los grupos CE y AE, respectivamente. No se observaron diferencias significativas entre los cuatro grupos para los parámetros de AST, GGT y amilasa. Conclusión: La asociación entre el consumo de alcohol y la falta de actividad física aumenta la liberación de las enzimas ALT y ALP en ratones Wistar; la práctica de ejercicio aeróbico después de la retirada del alcohol impidió el aumento de la liberación de ALP en el plasma de estos animales.