Elevated concentrations of testosterone and its synthetic analogs may induce changes in cardiovascular function. However, the effects of the combination of anabolic/androgenic steroid (AAS) treatment and exercise training on systolic and diastolic cardiac function are poorly understood. In the present study, we aimed to investigate the effects of low-dose steroid treatment (stanozolol) on cardiac contractile parameters when this steroid treatment was combined with exercise training in rats and the effects of chronic steroid treatment on the Frank-Starling (length-tension curves) relationship. Male Wistar rats were randomly assigned to one of four groups: U (untrained), US (untrained and treated with stanozolol 5 mg/kg/week), T (trained, 16 m/min/1 h) and TS (trained and treated with stanozolol 5 mg/kg/week). Continuous exercise training was conducted 5 days/week for 8 consecutive weeks. The speed of the treadmill was gradually increased to a final setting of 16 m/min/1 h. Experiments were divided into two independent series: 1) central hemodynamic analysis for mean arterial blood pressure (MAP) and cardiac output (CO) measurements and 2) isolated papillary muscle preparation in Krebs solution. Stanozolol treatment significantly increased the MAP and the heart size in untrained and trained rats (U 113±2; T 106±2; US 138±8 and TS 130±7 mmHg). Furthermore, stanozolol significantly decreased developed tension and dT/dt (maximal and minimal) in U rats. However, the developed tension was completely restored by training. The Frank/Starling relationship was impaired in rats treated with stanozolol; however, again, training completely restored diastolic function. Taken together, the present data suggest that AAS treatment is able to decrease cardiac performance (systolic and diastolic functions). The combination of stanozolol and physical training improved cardiac performance, including diastolic and systolic functions, independent of changes in central hemodynamic parameters. Therefore, changes in ventricular myocyte calcium transients may play a cardioprotective role.

Angiogenic therapies for critical limb ischemia were tested in a mouse model. The mice were anesthetized and their femoral arteries were ligated. The animals were treated with bone marrow mononuclear cells (BMMCs) alone, BMMCs combined with plasmid vector encoding granulocyte macrophage colony-stimulating factor (GM-CSF), received no treatment, or no intervention (controls). The degree of ischemia was monitored for 4 weeks using a visual scale. Muscle atrophy and strength were assessed at 4 weeks postoperatively; the mice were then killed. In treated animals, total necrosis of the limb was not found, the weight of the gastrocnemius and quadriceps muscles was significantly higher, functional ability and tissue regeneration were significantly increased, and muscle impairment and adipocyte presence were significantly reduced compared with untreated animals. At inducing angiogenesis, the BMMCs alone was more effective than BMMCs combined with plasmid vector encoding GM-CSF. Treated animals showed increased angiogenesis compared with ischemic untreated ones.

ObjectiveTo evaluate the effect of bupivacaine on muscle force and histology. We hypothesize that bupivacaine will worsen the muscle's physiological activity.SettingControlled laboratory experiment.MethodsBupivacaine (0.5 mL, 0.5%) was injected into the mid belly and distal portions of the right gastrocnemius in 32 Wistar male rats (the left gastrocnemius was used as a control). After 5, 14, 21, and 28 days, in groups of 4, muscle force was evaluated and the animals were euthanized by an overdose of anesthetic for histologic evaluation. One-way analysis of variance was used to analyze data from force and weight measurements. Only the values of P < .05 were considered to be statistically significant.ResultsBupivacaine causes a process of degeneration-regeneration of the muscle fibers and it also causes a reduction in muscle force, which is significant at 2 and 3 weeks and does not normalize at 4 weeks. The muscle injury is obvious after 5 days, and the degenerative process is predominant at 2 and 3 weeks. We found an increase in muscle mass in the acute phase and a decrease in muscle force.ConclusionAlthough our results do not allow a direct clinical application, we believe that caution should be warranted when intramuscular bupivacaine is used.

Background Vascular endothelial growth factor (VEGF) has mostly been tested to treat ischemic diseases, although the outcomes obtained are not satisfactory. Our hypothesis is that the local transient expression of VEGF and stem cell mobilizer granulocyte colony-stimulating factor (G-CSF) genes in ischemic limbs can complement their activities and be more efficient for limb recovery.Methods Limb ischemia was surgically induced in mice and 50 mu g of VEGF and/or G-CSF genes were locally transferred by electroporation. After 3-4 weeks, evidence of necrosis by visual inspection, capillary density, muscle mass, muscle force and hematopoietic cell mobilization were evaluated.Results After 4 weeks, 70% and 90% of the animals of the ischemic group (IG) and VEGF-treated group (VG), respectively, presented limb necrosis, in contrast to only 10% observed in the group of mice treated with both VEGF and G-CSF genes (VGG). Recovery of muscle mass and muscle force was higher than 60% in the VGG compared to the non-ischemic group. The mobilization of Sca1+ cells and neutrophils was also higher in the VGG, which may explain the lower level of necrosis observed in this group (22%, in contrast to 70% in the IG). Capillary density and degree of fibrosis were determined in weeks 3 and 4, and also showed a clear benefit as a result of the use of the G-CSF and VEGF genes together.Conclusions Gene therapy using VEGF and G-CSF demonstrated a synergistic effect promoting vessel and tissue repair in mouse hind limb ischemia. Copyright (C) 2010 John Wiley & Sons, Ltd.

Granulocyte‐colony‐stimulating factor (GM‐CSF) is a pleiotropic factor for hematopoiesis that stimulates myeloblasts, monoblasts and mobilization of bone marrow stem cells. Therefore, the GM‐CSF gene is a potential candidate for vessel formation and tissue remodeling in the treatment of ischemic diseases.

A novel, efficient transfection method, based on ultrasound and hydrodynamics, has been developed to transfect heart tissue with plasmid DNA. An ultrasound probe was aimed at the heart of anesthetized rats for 30 sec, at an intensity of 1 MHz and 2 W/cm2. The aorta was clamped and a phosphate-buffered saline (PBS) solution containing pSV-LacZ was quickly injected into the left ventricle. Each animal was maintained in this condition for 20 sec, and then the clamp was opened and the needle was removed. Electrocardiography, performed after 4 weeks, showed mild or no sign of ischemia in all groups. Visual evaluation of heart tissue samples from rats that received 100 microg of pSV-LacZ in 100 microl had only a few blue cells, indicating transfection, and those that received only PBS had no blue cells. However, all heart tissue samples from rats transfected with 100 to 500 microg of pSV-LacZ in 200 microl, or with 200 to 500 microg of pSV-LacZ in 100 micro had many blue cells. The base and epicardium of the heart tissue samples had many more blue cells than did the rest of the samples. Histological results, based on staining with hematoxylin and eosin, showed similar results between control and transfected groups. Therefore, we concluded that gene delivery by plasmid vector in association with ultrasound and hydrodynamics was highly effective in transfecting rat heart.

Introduction Peripheral arterial diseases may be caused by any occlusive leg artery lesion that interferes with blood flow. When these conditions occur, there is an increase in vessel resistance that can lead to a reduction in distal perfusion pressure and blood flow. This process forces collateral vessel remodeling and it is known as arteriogenesis. Inflammation is the main triggering factor of arteriogenesis and several molecular factors are involved. One way to prolong the arteriogenic activity is extending monocytes half-life by apoptosis inhibition with GM-CSF (granulocyte and macrophage colony stimulating factor). This factor is also responsible for proliferation and differentiation of inflammatory cells, which are important growth factors producers and chemoattractants for arteriogenesis. The main objective of this study was to assess the capacity of reverting surgery-induced acute limb ischemia in mice by in vivo transference of plasmid vector with GM-CSF gene and to compare with the VEGF gene transfer. Materials and Methods Two plasmid vectors were used in this study: uP-GM and uP- VEGF to express GM-CSF and VEGF, respectively. To induce acute limb ischemia in mice BALB/c, the femoral artery was excised from its origin of the external iliac artery to the distal point, where it bifurcates into saphenous and popliteal arteries, without damaging the vein and femoral nerve. All branches of femoral artery were ligated. Gene therapy was performed by injecting 100 μg of plasmid vector at the thigh soon after the ischemia surgery. Results and Discussion About 30 % of animals that underwent ischemic surgery lost their limbs after 1 week, meanwhile none of uP-GM treated animals lost their limbs, even after 4 weeks of the surgery. Functional tests of the GM-CSF gene treated muscles revealed about 70 % of recovery of force, meanwhile the VEGF gene treated animals and the non- treated animals recovered only 25 % and 5 % of force respectively after 4 weeks of the gene transfer. Weight of gastrocnemius reached 100 % of recovery with ischemic animals treated with uP-GM after 4 weeks in comparison to the non-ischemic animals, but the VEGF gene treated or non-treated animals showed recovery of only 60 %. Capillary density had a significant increase in the VEGF treated animals after 1 week of surgery, but those treated with GM-CSF did not change in comparison to the non-treated ischemic animals, and this profile continued for 4 weeks. Histological data from the animals treated with GM-CSF for 4 weeks revealed that the muscle organization returned to the normal form. Therefore, we conclude that GM-CSF gene transference therapy in the ischemic muscle promoted good recovery of physical and physiological parameters in the long term meanwhile the use of VEGF gene promoted rapid recovery but it was not sustainable for 4 weeks.

A atividade vasomotora simpática é um dos determinantes da pressão arterial (PA). Estabelecer quais são os mecanismos geradores dessa atividade é importante para o entendimento de como o sistema cardiovascular opera, tanto em situações fisiológicas como fisiopatológicas. Os principais grupos pré-motores do simpático estão confinados no núcleo paraventricular do hipotálamo (PVN) e região rostoventrolateral bulbar (RVLM). Em diversas situações fisiopatológicas há aumento na atividade vasomotora simpática, em parte conseqüente a maior atividade dos neurônios do PVN e RVLM. Nesta breve revisão, foram discutidos os principais mecanismos de ativação simpática em diferentes modelos experimentais: 1) hipertensão renovascular, 2) hipertensão por baixa massa renal, 3) insuficiência cardíaca, 4) hipertensão por bloqueio do óxido nítrico, 5) obesidade e 6) dimorfismo sexual. As ações de diferentes mediadores sobre o PVN e RVLM podem em longo prazo determinar novos patamares de atividade simpática, modificando os níveis tensionais e dessa forma, contribuir para a progressão da doença cardiovascular.

Several studies support the hypothesis that chronic diseases in adulthood might be triggered by events that occur during fetal development. This study examined the consequences of perinatal salt intake on blood pressure (BP) and carbohydrate and lipid metabolism in adult offspring of dams on high-salt [HSD; 8% (HSD2) or 4% (HSD1)], normal-salt (NSD; 1.3%), or low-salt (LSD; 0.15% NaCl) diet during pregnancy and lactation. At 12 wk of age, female Wistar rats were matched with adult male rats that were fed NSD. Weekly tail-cuff BP measurements were performed before, during, and after pregnancy. After weaning, the offspring received only NSD and were housed in metabolic cages for 24-h urine collection for sodium and potassium and nitrate and nitrite excretion measurements. At 12 wk of age, intra-arterial mean BP was measured, a euglycemic-hyperinsulinemic clamp was performed, and plasma lipids and nitrate and nitrite concentrations were determined. Tail-cuff BP was higher during pregnancy in HSD2 and HSD1 than in NSD and LSD dams. Mean BP (mm Hg) was also higher in the offspring of HSD2 (110 ± 5) and HSD1 (107 ± 5) compared with NSD (100 ± 2) and LSD (92 ± 2). Lower glucose uptake and higher plasma cholesterol and triacylglycerols were observed in male offspring from LSD dams (glucose uptake: HSD2 17 ± 4, HSD1 15 ± 3, NSD 11 ± 3, LSD 4 ± 1 mg · kg−1 · min−1; cholesterol: HSD2 62 ± 6, HSD1 82 ± 11, NSD 68 ± 10, LSD 98 ± 17 mg/dL; triacylglycerols: HSD2 47 ± 15, HSD1 49 ± 12, NSD 56 ± 19, LSD 83 ± 11 mg/dL). In conclusion, maternal salt intake during pregnancy and lactation has long-term influences on arterial pressure, insulin sensitivity, and plasma lipids of the adult offspring.