Jorge José de Carvalho

Maternal high-fat intake predisposes nonalcoholic fatty liver disease in C57BL/6 offspring

OBJECTIVE This work aimed to verify the hypothesis that maternal intake of high-fat diet in critical periods of pregnancy and/or suckling period predisposes nonalcoholic fatty liver disease in adult C57BL/6 mice offspring. STUDY DESIGN Male pups were divided into 5 groups: (1) SC, from standard chow-fed dams; (2) G, from high-fat chow (HF)-fed dams during the gestation (G) period; (3) L, from HF-fed dams during the lactation (L) period; (4) GL, from HF-fed dams during the gestation and lactation (GL) periods; and (5) GL/HF, from HF-fed dams during GL, maintaining an HF diet from postweaning to adulthood. We analyzed body mass, plasma blood, and liver structure. RESULTS The G offspring showed insulin resistance and lower glucose transporter-2 expression. Hepatic steatosis was present in the G, L, GL, and mainly in GL/HF offspring. Sterol regulatory element-binding protein-1c expression was higher in G, GL, and GL/HF offspring. CONCLUSION Programming by HF chow predisposes hepatic adverse remodeling in the liver of adult offspring.

A new description of the reproductive system of Schistosoma mansoni (Trematoda: Schistosomatidae) analyzed by confocal laser scanning microscopy

Classical schemes of the adult Schistosoma mansoni reproductive system have been described. In our study, whole adult worms derived from unisexual or mixed infections and stained with carmine chlorine were virtually and tomographically analyzed under confocal laser scanning microscopy. We found that: (1) there were morphological differences in the ovary, vitteline glands and testicular lobes between specimens derived from unisexual or mixed infections; (2) there was always a single lobed ovary (three or four lobes), presenting differentiation from the anterior to the posterior lobes, where the most mature oocytes were located; (3) the proximal segment of oviduct was connected to an ampullary dilatation, full of tailed spermatozoa, characterizing a seminal receptacle; (4) there was no long vitelline duct, but a short one that begins at the end of the proximal region of the vitelline gland; (5) long cells of Mehlis’ gland placed radially around the ootype were not observed. Otherwise, the ootype was only lined by thick cuboidal epithelial cells with plaited bases and nuclei with flabby chromatin, making a clear distinction from the uterine epithelium. This morphological feature suggests that each cell represents a gland. (6) In coupled males, the specimens located inside the gynaecophoric canal had smaller testicular lobes, suckers, and body length and width when compared to their partners. Our results show that the reproductive system does not follow a unique pattern within flatworms. Due to its better resolution, confocal laser scanning microscopy, using a reflected mode with tomographic sections, allows new interpretations, modifying the adopted and current descriptions of the internal morphological structures of S. mansoni adult worms.

Connective tissue accumulation in the muscle layer in normal and varicose saphenous veins.

Varicose veins alternate areas of phlebosclerosis and hypertrophy of the vein wall. In this study, samples of long saphenous veins obtained from patients submitted for aortocoronary saphenous vein graft or for surgical resection of varicose saphenous veins were examined. Histologic changes in the intima, muscle, and adventitial layers were quantified. Thicknesses of the venous wall layers were obtained by linear measurements, and the volumetric density of the connective tissue in the muscle layer was determined by point counting. The muscle layer thickness was 300 +/- 13 and 581 +/- 25 microns in normal and varicose veins, respectively. A more severe connective tissue accumulation within the muscle bundles was found in the varicose condition. The volumetric density of the connective tissue in the circular muscle layer (CmC/MmC) showed also a marked difference between varicose (0.67 +/- 0.08) and normal veins (0.43 +/- 0.02), P < 0.05. The authors suggest that the varicose condition is associated with a connective tissue uniform accumulation among muscle cells in the circular muscle layer.Varicose veins alternate areas of phlebosclerosis and hypertrophy of the vein wall. In this study, samples of long saphenous veins obtained from patients submitted for aortocoro nary saphenous vein graft or for surgical resection of varicose saphenous veins were examined. Histologic changes in the intima, muscle, and adventitial layers were quanti fied. Thicknesses of the venous wall layers were obtained by linear measurements, and the volumetric density of the connective tissue in the muscle layer was determined by point counting. The muscle layer thickness was 300 ±13 and 581 ±25 μm in normal and varicose veins, respectively A more severe connective tissue accumulation within the muscle bundles was found in the varicose condition. The volumetric density of the connective tissue in the circular muscle layer (CmC/MmC) showed also a marked differ ence between varicose (0.67 ±0.08) and normal veins (0.43 ±0.02), P < 0.05. The authors suggest that the varicose condition is associated with a connective tissue uniform accumulation among muscle cells in the circular muscle layer.

Exercise training enhances elastin, fibrillin and nitric oxide in the aorta wall of spontaneously hypertensive rats.

This work aimed to analyze the effect of low-intensity exercise training on ultrastructural and molecular aortic remodeling. Male Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) were allocated into four groups: sedentary WKY (SED-WKY), exercised WKY (EX-WKY, 1 h/day, 5 days/week treadmill exercise training), sedentary SHR (SED-SHR), and exercised SHR (EX-SHR). EX-SHR showed blood pressure reduction of 26% in comparison to SED-SHR after 1 month of exercise (P<0.05). At the 20th week, BP level was not different between EX-SHRs and WKYs. Circumferential wall tension (CWT) was higher by 77% in SED-SHRs than in SED-WKYs (P<0.001). Exercise training reduced CWT by 30% in EX- vs. SED-SHR (P<0.001). In SED-SHRs, endothelial cells showed large and numerous cytoplasmatic vacuoles, fragmented inner elastic lamina and scarce elastin and fibrillin, while exercise training ameliorated it in EX-SHR group. The highest eNOS immunodensity was observed in EX-SHR, which was 50% higher than EX-WKY (P<0.01) and 120% higher than SED-SHR (P<0.0001). In conclusion, present findings indicate beneficial effects of exercise training in hypertensive rats since it increased elastin, fibrillin and eNOS content in the aortic wall.

Hypertension and kidney alterations in rat offspring from low protein pregnancies.

Objective Low birth weight contributes to the early onset of end-stage renal disease. Therefore, this study was undertaken to investigate early and late glomerular structural alterations in both sexes of Wistar rat offspring from dams submitted to severe low protein intake during gestation. Methods Offspring from dams fed normal protein (19% of protein) or low protein (5% of protein) were studied at days 0, 10, 90 and 180 of age. Results Inner cortical structure showed immature (comma-shaped and S-shaped forms) and mature corpuscles in different proportions in low protein offspring (less maturity) and normal protein offspring (more maturity). At day 10 (end of the nephrogenesis period), immature corpuscles were observed only in low protein offspring. In adulthood, low protein offspring had higher blood pressure, and showed thicker glomerular basement membrane (GBM) with effacement of the pedicles, and slit diaphragm absent with some podocytes directly adhering to the basal membrane with pedicles absent. The number of renal corpuscles was lower in low protein offspring than in normal protein offspring of the same sex, all age groups (P < 0.001). No interaction was observed between sex and maternal nutrition for the same sex and all age groups. Conclusion Gestational low protein leads to glomerulogenesis retardation and consequently a lower nephron number with thick GBM and structural alterations in the pedicles of podocytes.

Is the infertility in hypothyroidism mainly due to ovarian or pituitary functional changes

The objective of the present study was to examine whether hypothyroidism affects the reproductive system of adult female rats by evaluating ovarian morphology, uterus weight and the changes in serum and pituitary concentrations of prolactin and gonadotropins. Three-month-old female rats were divided into three groups: control (N = 10), hypothyroid (N = 10), treated with 0.05% 6-propyl-2-thiouracil (PTU) in drinking water for 60 days, and T4-treated group (N = 10), receiving daily sc injections of L-thyroxine (0.8 microg/100 g body weight) during the last 10 days of the experiment. At the end of 50 days of hypothyroidism no hypothyroid animal showed a regular cycle, while 71% of controls as well as the T4-treated rats showed regular cycles. Corpora lutea, growing follicles and mature Graafian follicles were found in all ovaries studied. The corpora lutea were smaller in both the hypothyroid and T4-replaced rats. Graafian follicles were found in 72% of controls and only in 34% of hypothyroid and 43% of T4-treated animals. Serum LH, FSH, progesterone and estradiol concentrations did not differ among the three groups. Serum prolactin concentration and the pituitary content of the three hormones studied were higher in the hypothyroid animals compared to control. T4 treatment restored serum prolactin concentration to the level found in controls, but only partially normalized the pituitary content of gonadotropins and prolactin. In conclusion, the morphological changes caused by hypothyroidism can be a consequence of higher prolactin production that can block the secretion and action of gonadotropins, being the main cause of the changes observed.

Effect of oral ingestion of an extract of the herb Uncaria tomentosa on the biodistribution of sodium pertechnetate in rats

The aim of the present study was to determine the effect of the oral ingestion of an extract of the herb Uncaria tomentosa (cats claw) on the biodistribution of the radiobiocomplex sodium pertechnetate (Na99mTcO4) in rats. The animals (male Wistar rats, 2 months old, 180-220 g), were treated (1 mL) with an U. tomentosa extract (32 mg/mL, N = 5) or 0.9% NaCl solution (control, N = 5) for 7 days. After this period, Na99mTcO4 (3.7 MBq, 0.3 mL) was injected through the ocular plexus and after 10 min the rats were killed, the organs isolated and counted in a well-gamma counter. A significant (P < 0.05) alteration in Na99mTcO4 uptake i) from 0.57 +/- 0.008 to 0.39 +/- 0.06 %ATI/organ (P < 0.05) and from 0.57 +/- 0.17 to 0.39 +/- 0.14 %ATI/g (P < 0.05) was observed in the heart, ii) from 0.07 +/- 0.02 to 0.19 +/- 0.07 %ATI/g in the pancreas, and iii) from 0.07 +/- 0.01 to 0.18 +/- 0.07 %ATI/g (P < 0.05) in muscle after treatment with this extract. Although these results were obtained with animals, caution is advisable in the interpretation of the nuclear medicine examination when the patient is using this herb. This finding is probably an example of drug interaction with a radiopharmaceutical, a fact that could lead to misdiagnosis of the examination in clinical practice with unexpected consequences for the patient.

Bioavailability of the sodium pertechnetate and morphometry of organs isolated from rats: study of possible pharmacokinetic interactions of a ginkgo biloba extract

Many compounds affect the bioavailability of radiobiocomplexes as radiopharmaceuticals. Ginkgo Biloba extract (EGb) has several effects. The influence of an EGb on the bioavailability of the radiobiocomplex sodium pertechnetate (Na99mTcO4) and on the morphometry of the organs was evaluated. Rats were treated with EGb and Na99mTcO4 was injected. The animals were sacrificed; the radioactivity in the organs was counted. The results showed that EGb altered the Na99mTcO4 bioavailability in the kidneys, liver and duodenum. Morphometric analysis of the organs showed significant alterations (P<0.05), probably caused by metabolites generated by EGb and capable of altering the bioavailability of the Na99mTcO4.

Pleiotropic effects of rosuvastatin on the glucose metabolism and the subcutaneous and visceral adipose tissue behavior in C57Bl/6 mice.

The aim of this study was to evaluate whether rosuvastatin (HMG-CoA reductase inhibitor) modulates the carbohydrate and lipid metabolism, the development of non-alcoholic fatty liver disease (NAFLD), and the increase in body mass in a model of diet-induced obesity. Male C57Bl/6 mice (3-months-old) were fed a high-fat diet (HF, 60% lipids) or the standard chow (SC, 10% lipids) for 15 weeks. The animals were then treated with 10 mg/kg/day (HF-R10 group), 20 mg/kg/day (HF-R20), or 40 mg/kg/day (HF-R40) of rosuvastatin for five weeks. The HF diet led to glucose intolerance, insulin resistance, weight gain, increased visceral adiposity with adipocyte hypertrophy, and hepatic steatosis (micro and macrovesicular). The rosuvastatin treatment decreased the adiposity and the adipocyte size in the HF-R10 and HF-R20 groups. In addition, rosuvastatin changed the pattern of fat distribution in the HF-R40 group because more fat was stored subcutaneously than in visceral depots. This redistribution improved the fasting glucose and the glucose intolerance. Rosuvastatin also improved the liver morphology and ultrastructure in a dose-dependent manner. In conclusion, rosuvastatin exerts pleiotropic effects through a dose-dependent improvement of glucose intolerance, insulin sensitivity and NAFLD and changes the fat distribution from visceral to subcutaneous fat depots in a mouse model of diet-induced obesity.

Exercise training in doxorubicin-induced heart failure: effects on the L-arginine–NO pathway and vascular reactivity

Heart failure (HF) is the end-stage of cardiovascular disease and is associated with a high incidence of thrombotic events. Nitric oxide (NO) mediates vasodilation and prevents platelet activation, providing an important antithrombotic effect. The aim of this study was to investigate the effects of aerobic training on survival, platelet L-arginine-NO pathway, and vasodilator properties in doxorubicin (DOX)-induced HF. Sprague Dawley rats were randomly assigned to saline/sedentary (SAL/SED), saline/exercise (SAL/EX), DOX/sedentary (DOX/SED), and DOX/exercise (DOX/EX) groups. Four weeks after intraperitoneal DOX injection (1mg/kg(-1)/d(-1); 10 days), shortening fraction in DOX/SED and DOX/EX was significantly reduced. Treadmill exercise was performed during 6 weeks, 5 days/week(-1), 30minutes/day(-1), 50% to 60% of maximum velocity. Survival was higher in DOX/EX (67%) than DOX/SED (33%). No differences were observed in intraplatelet L-arginine transport assessed by incubation with L- [(3)H]-arginine, nor in NOS activity measured by the conversion of L- [(3)H]-arginine into L- [(3)H]-citrulline among the groups. Vasodilation response to acetylcholine was impaired in DOX/SED and DOX/EX; in nitroglycerine, it was limited to DOX/SED. Aerobic training reduced mortality in DOX-induced HF animals and restored vascular smooth muscle relaxation properties. However, it did not ameliorate intraplatelet NO bioavailability and endothelial function during the period studied.