Vitamin D restriction enhances periovarian adipose tissue inflammation in a model of menopause.

Background: Menopause and deficiency in vitamin D (VD) are two health problems usually associated with aging women.Objective: We aimed to study inflammation in visceral adipose tissue when bilateral ovariectomy is combined with dietary restriction in VD.Methods: We studied 60 female C57BL/6 mice 3 months of age. Half of the animals had bilateral ovariectomy (Ovx group, n = 30) and half a sham procedure (Control [C] group, n = 30), and half of each Ovx or C group were fed a standard diet containing VD or a diet restricted in VD (D-) for an additional 12 weeks. Therefore, four groups were formed (n = 15 each group): C, C(D-), Ovx, and Ovx(D-). After sacrifice, the periovarian adipose tissue (PAT) was investigated.Results: In PAT, we observed different levels of hypertrophied adipocytes, enhanced proinflammatory cytokines, activation of inflammatory markers, and components of the extracellular signal-regulated kinase. The most affected PAT was seen in the Ovx(D-) group, followed by the Ovx group, the C(D-) group, and the C group (the least altered).Conclusion: The results demonstrate that ovariectomy and dietary restriction of VD are inducers of adverse effects on mouse visceral adipose tissue. When combined, these insults might enhance PAT inflammation.

High fat diets modulate nitric oxide biosynthesis and antioxidant defence in red blood cells from C57BL/6 mice.

The consumption of a high fat (HF) diet is considered a risk factor for the development of obesity. On the other hand, a monounsaturated HF diet has beneficial cardiometabolic effects. Since nitric oxide (NO) modulates vascular homeostasis, we investigate whether HF diets that vary in fatty acid composition have a different effect on theL-arginine-NO pathway and oxidative stress in C57BL/6 mice red blood cells (RBC). The olive oil diet induced an activation of L-arginine transport compared to other diets. NO synthase (NOS) activity was increased in all unsaturated HF diets (olive, sunflower and canola oils). Moreover, the expression of endothelial NOS (eNOS) and inducible NOS (iNOS) was increased in the olive oil group. In contrast, NOS activity from the lard group was decreased associated with diminished l-arginine transport. Olive oil also induced superoxide dismutase activation. Inhibition of the L-arginine-NO pathway in the lard group could contribute to cardiovascular diseases, while unsaturated HF diets may have a protector effect via enhanced NO bioavailability.

Left ventricular hypertrophy induced by overnutrition early in life.

BACKGROUND AND AIMS: Human overnutrition has caused a rise in the prevalence of obesity in recent years. In addition to the deleterious effects of obesity during childhood, the long-term effects in adulthood have also been described. For instance, cardiovascular diseases, particularly left ventricular hypertrophy, and type 2 diabetes are among the diseases associated with a history of obesity. However, it remains uncertain during which period of life this association is established. Several authors have suggested that the early period of life is critical for the emergence of cardiac disorders. METHODS AND RESULTS: In this study, we examined the impact of overnutrition on the heart morphology and stereology of obese animals using an experimental model to induce an overweight phenotype. In these animals, overfeeding during lactation was able to induce a significant increase in body weight and visceral fat starting at the 10th day of life, and this increased weight persisted until 21 days of age. Impairments in triglyceride levels and cholesterol were also observed in these animals. Moreover, an increased heart weight/tibia length ratio was observed, indicating an enlarged heart. The overfed animals also had left ventricular hypertrophy with an increased area of cardiomyocytes and a decreased vessel density. CONCLUSIONS: Our findings showed that overnutrition during early life induced obesity and cardiac hypertrophy in the pups, perhaps due to a decrease in the intramyocardial vessel.

Oral isotretinoin in photoaging: clinical and histopathological evidence of efficacy of an off-label indication.

BACKGROUND: Despite evidences of the beneficial clinical effects of oral isotretinoin in the treatment of cutaneous photoaging, scientific evidences are still scarce, mainly supported by histopathological and morphometric studies. OBJECTIVES: To analyse possible clinical and morphological changes resulting from the treatment of photoaging with oral isotretinoin. METHODS: Thirty female patients, aged 40 to 55 years, phototypes II to IV, with moderate to severe photoaging were randomly assigned to two groups of 15 each. Group I (G I) patients were treated with 10 mg of isotretinoin and group II (G II) with 20 mg of oral isotretinoin thrice a week for 3 months. Skin biopsies were performed before and after the end of therapy, and the various sections were submitted to specific staining for collagen and elastic fibres. To analyse the changes, morphometric studies were performed, and the results obtained were analysed by Student's t-test (paired and non-paired). Clinical results of therapy regarding texture, colouring and aspect of the wrinkles were assessed by both physician and patient. RESULTS: The increase in the amount of collagen fibres was statistically significant with both dosage regimens (mean, 37.8%, increasing to 44.4%; P = 0.029 with the 10-mg dosage; and mean, 36.6%, increasing to 41.9%; P = 0.01 with the 20-mg dosage). A pattern pointing toward a decrease in the number of elastic fibres was found (mean, 15.3-12%; P = 0.014 with the 10-mg dosage; mean, 15.5-14%; P = 0.125 with the 20-mg dosage). Additionally, there was improvement in the general aspect of the skin, regarding texture, wrinkles depth and skin coloration. LIMITATIONS: Despite ethical considerations, a lack of a control group using placebo may render the results less accurate. CONCLUSION: Low dosages of oral isotretinoin seem to be an effective therapeutic option for cutaneous photoaging.

Beneficial effects of maternal swimming during pregnancy on offspring metabolism when the father is obese.

We aimed to evaluate the impact of maternal exercise training on the offspring metabolism and body size caused by father obesity. C57BL/6 male 4-week-old mice were fed a high-fat diet (HF father) or control diet (C father), while equal age female mice were fed only a C diet and were separated into two groups: trained (T mother) and non-trained (NT mother), and at 12 weeks of age mice were mated. A continuous swimming protocol was applied for 10 weeks (before and during gestation), and offspring were followed since weaning until sacrifice (at 12 weeks of age). HF father, compared to C father, showed obesity, elevated total cholesterol (TC) and triglycerides (TG), and glucose intolerance. Both sexes HF/NT offspring showed hyperglycemia, glucose intolerance and high levels of TC and TG, without obesity. However, HF/T offspring showed data close to C/NT, demonstrating the beneficial effect of maternal exercise in the offspring of obese fathers.

Parathyroid glands: combination of sestamibi-(99m)Tc scintigraphy and ultrasonography for demonstration of hyperplasic parathyroid glands.

OBJECTIVE: To determine the frequency of the types of parathyroid gland hyperplasia and the sensitivity of sestamibi-(99m)Tc (MIBI) scintigraphy and ultrasonography (US) of patients with secondary hyperparathyroidism (SHPT) due to chronic renal failure. MATERIAL AND METHODS: We studied 43 patients with SHPT (26 females and 17 males with age range of 27-75 years). Blood tests were performed to determine intact parathyroid hormone (PTH), calcium and phosphorus concentration and parathyroid MIBI scintigraphy and US examinations were done, to evaluate each glandular function and structure. Nineteen of the 43 patients underwent total parathyroidectomy and 69 abnormal glands were removed at operation. The 69 abnormal and 4 normal glands from patients that underwent total thyroidectomy were studied by light microscopy. The results were compared and correlation was calculated to: weight, MIBI uptake and US results. RESULTS: All 43 patients had elevated serum PTH ranged from 400 to 4,075 pg/ml (1,868.0 +/- 975.9 and normal range 10-75 pg/ml). Serum calcium and phosphorus concentration were 10.13 +/- 2.02 mg/dl and 5.28 +/- 2.07 mg/dl respectively. Fifty eight of 69 glands from surgical resection were MIBI positive and 11 were negative, but their cellular composition and presentation were similar. Hyperplasic glands had increased number of all cell types considered (chief, oxyphil and clear) compared to the normal gland. Chief cell hyperplasia was the most frequent type (81 %) followed by oxyphil (9 %), clear (6 %) and adenomatous type (4 %). False negative results of 10 % to US and 4.6 % to scintigraphy were found. The correlation of gland weight and MIBI uptake were not significant (p = 0.09). The sensitivity of MIBI scintigraphy was 84 % and US was 72.5 %. CONCLUSIONS: The MIBI scintigraphy is a very sensitive tool for pre-operative localization of hyperplasic parathyroid gland and should be used as the first imaging method. The association of MIBI and US is recommended because increases the sensitivity for preoperative hyperplasic parathyroid glands identification.

Liver metabolism in adult male mice offspring: consequences of a maternal, paternal or both maternal and paternal high-fructose diet.

The study aimed to evaluate the consequences of the consumption of a high-fructose diet (HFR; fructose was responsible for 45% of the energy from carbohydrates) by the mother, the father, or both on C57BL/6 adult male offspring. Non-consanguineous parents received the diet (HFR or control, C) from 8 weeks before mating until weaning (n=10 fathers and n=10 mothers on each diet). After weaning, only the C diet was offered to offspring. The groups were formed by one male randomly taken from each litter. The offspring groups were identified according to the mother's diet (the first letter), then the father's diet (the second letter), that is, C/C, C/HFR, HFR/C, HFR/HFR (n=10 per group). The parents exhibited the following characteristics: compared with those of the C group, the HFR parents had higher blood pressure (BP), enlarged liver, increased hepatic triacylglycerol content, hypercholesterolemia, hypertriglyceridemia, high plasma leptin and low adiponectin. The offspring exhibited the following characteristics: compared with the C/C group, the HFR/HFR group had high BP. The C/HFR, HFR/C and HFR/HFR showed elevated uric acid and leptin levels and diminished adiponectin. The HFR/HFR group showed liver inflammation (increased NFκB, SOCS3, JNK, TNF-α, IL1-ß and IL6 levels). Likewise, SREBP-1c and FAS were upregulated. In conclusion, the consumption of a HFR by the mother and/or father is associated with adverse effects on liver metabolism in adult male offspring. When both mother and father are fed a HFR, the adverse effects on the offspring are more severe.

Assessment of Spleen Filtrate Function in Renal Transplant Recipients Using Technetium-99m Stannous Colloid Liver-Spleen Scan.

BACKGROUND: Functional hyposplenism (FH) is indicated by an anatomically present spleen that fails to take up radiolabeled colloid. The occurrence of FH has been reported in a small group of renal transplant recipients based on hematologic parameters. The aim of this study was to replicate this association in a larger group of renal transplant recipients with the use of technetium-99m-stannous colloid liver-spleen scan to assess the spleen function. METHODS: This survey based on single samples enrolled 101 unselected adult patients with functional kidney grafts >180 days after transplantation. All patients underwent 99mTc-stannous colloid scan to assess spleen function as well as bone marrow uptake of radiocolloid along with an anatomic and blood flow study of the spleen and kidney with the use of Doppler sonography. RESULTS: The prevalence of hyposplenism was 32.7% (33/101) for the cohort, and increased uptake of radiocolloid by the bone marrow was seen in 9.9% (10/101). According to the multivariate analysis, the frequency of hyposplenism was significantly influenced by indirect bilirubin and hemoglobin, and direct bilirubin and neutrophil count remained as independent predictors of bone marrow uptake. CONCLUSIONS: This study demonstrated that a group of renal transplant recipients has FH. In addition, bone marrow uptake might be interpreted as liver dysfunction. In this situation, the small amount of contrast (spleen compared with liver) would leave hyposplenism undiagnosed. Further prospective and longitudinal clinical studies are needed to determine the clinical impact of this condition on the management of renal transplant recipients.

Different edible oil beneficial effects (canola oil, fish oil, palm oil, olive oil, and soybean oil) on spontaneously hypertensive rat glomerular enlargement and glomeruli number.

We have tested the different edible oil effects on the blood pressure (BP) control and the following glomerular protection. Six groups of 12-week-old male spontaneously hypertensive rats (SHR) (n = 5), have received different edible oils (fish, canola, palm, olive, and soybean) or a placebo by gavage for 13 weeks. Renal cortex was analyzed through light microscopy and stereology. Usual BP increase, glomerulosclerosis, glomerular enlargement, and glomeruli loss in SHR has been prevented (fish, canola and palm oils) or attenuated (olive and soybean oils) by these oil long-term administration. The most favorable effect has been seen in the fish oil administration (source of n-3 polyunsaturated fatty acids, PUFA, eicosapentaenoic and docosahexaenoic acids), followed by both canola and palm oils (source of n-3 PUFA plus n-9 monounsaturated, MUFA, and saturated fatty acid, respectively), and finally both olive and soybean oils (source of n-9 MUFA and n-6 PUFA, respectively).

Leprosy: contribution of mast cells to epineurial collagenization.

BACKGROUND: Leprosy, a disease caused by Mycobacterium leprae, is an important health problem worldwide. It is responsible for an irreversible nerve damage in which fibrosis plays an important role. The existence of an interaction between mast cells and different fibrotic conditions has long been observed. Tryptase, the most abundant protein product of human mast cells, has been shown to be mitogenic for fibroblasts and to increase type I collagen production. PATIENTS AND METHODS: In order to explore the possible relationship between tryptase-rich mast cells and nerve fibrosis in leprosy, we studied 24 sural nerve biopsies of patients with leprous neuropathy. Mast cells stained with mouse antihuman mast cell antitryptase clone AA1 as well as fibrosis, were quantitatively estimated in both epi- and endoneurial compartments. RESULTS: There was a remarkable association between collagen increase and tryptase-rich mast cell density in the epineurium but not in the endoneurium of leprous nerves. CONCLUSION: Since the epineurium in leprosy is type I collagen rich, the present findings support a tryptase-rich mast cell contribution to epineurial collagenization in leprosy through their tryptase secretion.

A multivariate analysis of cardiac growth in human embryos: endocardial cushions and ventricular myocardium.

STUDY OBJECTIVE: In view of the controversy about the morphological significance of the endocardial cushion tissue and ventricular myocardium during cardiac development, the aim was to carry out quantitative studies of these structures. DESIGN: Endocardial cushion tissue and ventricular myocardium were quantified by point count planimetry. The relative growth of the volume of these structures, and also the embryonic crown-rump length, was studied by multivariate allometry (principal components analysis) with the covariance matrix calculated from natural logarithms of the data. EXPERIMENTAL MATERIAL: 27 serially sectioned human embryos were studied, ranging from stage 15 to stage 23 (Paris collection). MEASUREMENTS AND MAIN RESULTS: The relative growth of endocardial cushion tissue, ventricular myocardium, and crown-rump length was discontinuous during the postsomitic period. The first component in principal components analysis measures overall size and, in the present analysis, accounts for 88.6% of the total variance. The growth vector isometry hypothesis was checked with the chi 2 test. This showed that differences in growth between cardiac structures and crown-rump length were allometric (p less than 0.01). Endocardial cushion tissue volume and crown-rump length grew with negative allometry during the second month of gestation, while ventricular myocardium volume grew with positive allometry. CONCLUSIONS: The results agree with those researchers who consider that endocardial cushion tissue functions only in causing initial cardiac fusion and partitioning, with little influence on the formation of definitive cardiac structures.

High-intensity interval training (swimming) significantly improves the adverse metabolism and comorbidities in diet-induced obese mice.

AIM: Controlling obesity and other comorbidities in the population is a challenge in modern society. High-intensity interval training (HIIT) combines short periods of high-intensity exercise with long recovery periods or a low-intensity exercise. The aim was to assess the impact of HIIT in the context of diet-induced obesity in the animal model. METHODS: C57BL/6 mice were fed one of the two diets: standard chow (Lean group - LE) or a high--fat diet (Obese group - OB). After twelve weeks, the animals were divided into non-trained groups (LE--NT and OB-NT) and trained groups (LE-T and OB--T), and began an exercise protocol. For biochemical analysis of inflammatory and lipid profile, we used a colorimetric enzymatic method and an automatic spectrophotometer. One-way ANOVA was used for statistical analysis of the experimental groups with Holm-Sidak pos hoc test. Two-way ANOVA analyzed the interactions between diet and HIIT protocol. RESULTS: HIIT leads to significant reductions in body mass, blood glucose, glucose tolerance and hepatic lipid profile in T-groups compared to NT-groups. HIIT was able to reduce plasma levels of inflammatory cytokines. Additionally, HIIT improves the insulin immunodensity in the islets, reduces the adiposity and the hepatic steatosis in the T-groups. HIIT improves beta--oxidation and peroxisome proliferator--activated receptor (PPAR)-alpha and reduces lipogenesis and PPAR--gamma levels in the liver. In skeletal muscle, HIIT improves PPAR--alpha and glucose transporter-4 and reduces PPAR--gamma levels. CONCLUSION: HIIT leads to attenuate the adverse effects caused by a chronic ingestion of a high-fat diet.

Blood pressure, ventricular volume and number of cardiomyocyte nuclei in rats fed for 12 months on diets differing in fat composition.

Blood pressure (BP), body mass (BM), ventricular volume (V[vent]) and the number of ventricular cardiomyocyte nuclei (N[vcn]) were analysed in rats fed different dietary fats. A total of 20 Wistar male rats were studied from 21 days old to 12 months of age and divided in the groups: soybean oil (S), canola oil (CA), lard and egg yolk (LE) and canola oil+lard and egg yolk (CA+LE). The diets had the same basal diet that included cornstarch, casein, maize, egg white and mineral and vitamin mixtures. At the moment of the sacrifice, the LE group had the greatest BP and V[vent] which was significantly higher than the other groups, and the S group had the greatest BM. The myocardial structure was apparently normal in the S and CA groups while it showed areas of diffuse interstitial fibrosis and hypertrophied cardiomyocytes in the LE group, and intramyocardial coronaries with thick tunica media and little interstitial fibrosis in CA+LE group. The N[vcn] was significantly higher in the CA group and it was lower in the LE group. These results suggest that the different dietary fats affect the myocardial structure, and the canola oil diet reduces the cardiomyocyte loss in the old rats.

Glomerular profile numerical density per area and mean glomerular volume in rats submitted to nitric oxide synthase blockade.

Rats submitted to chronic inhibition of nitric oxide synthase (NOS) have developed systemic hypertension and consequent renal injury. The present study aims to determine glomerular quantitative changes due to NOS inhibition in rats. Adults and normotensive Wistar rats were separated into control and L-NAME groups (each group n=10). The animals received water and food ad libitum, while L-NAME rats received NG-Nitro-L-Arginine methyl Ester hydrochloride to inhibit NOS (50mg/kg/day) in drinking water during 40 days. After that period the rats were sacrificed, the kidneys were removed, measured, and prepared for histological and stereological analyses. The glomerular density per area [NA(glom)] and the mean glomerular volume [v] were determined per animal in 15 random fields. In L-NAME rat the blood pressure was 76% higher than the respective control group with the same age. Glomeruli had global or segmental glomerular sclerosis; some glomeruli only presented an atrophic structure. The renal volume was not different between control and L-NAME rats (p>0.05). However, L-NAME rats had the NA(glom) 33% smaller than the control rats (p=0.0001) and, concomitantly, L-NAME rats had the v (glom) 33% higher than the control ones (p=0.004). These results demonstrate morphological renal alterations caused by NOS inhibition and hypertension.

Effect of telmisartan on preexistent cardiac and renal lesions in spontaneously hypertensive mature rats.

Fifteen adult male spontaneously hypertensive rats (one year old) (SHR) were separated into three groups (n=5 each) during 15 weeks as follows: initial control group (IC); final control group (FC); and telmisartan group (T) (1.2 mg/kg/day of telmisartan). Serum and urinary creatinine and proteinuria were not different comparing untreated and telmisartan-treated SHRs. FC rats showed a continuous BP increase during the study while T rats reached the 15th week with a significantly low BP. The LV mass index was significantly smaller in the T group than in the FC group, as was the glomerular hypertrophy. The cardiomyocyte nuclei density per area and the cardiomyocyte mean cross-sectional area were smaller in the T group than in both the IC and FC groups. Intramyocardial artery densities (per area and per volume) were greater in the T group than in untreated SHRs, but myocardial fibrosis was reduced. In conclusion, telmisartan monotherapy effects on BP and also on the hypertension target organs, heart and kidney, are favorable. Telmisartan is able to attenuate SHR cardiomyocyte and glomerular hypertrophies, and myocardial reactive fibrosis as well. It also is favorable to the intramyocardial microcirculation.

Effect of unilateral nephrectomy on renal function of diabetic rats.

Glomerular alterations of experimental diabetes mellitus are observed in animals submitted to a reduction in renal mass, suggesting that some mechanisms responsible for the progression of renal disease are common. The aim of this study was to investigate the effect of nephrectomy on the renal function and morphology of diabetic rats. Male Wistar rats were divided into 4 groups: control (C), n=8; diabetic (DM), n=8; non-diabetic nephrectomized (Nx), n=8; (DMNx), n=9. DM was induced by streptozotocin (65 mg/Kg), and animals were treated with insulin. After 12 weeks, the glomerular filtration rate (GFR), renal plasma flow (RPF) and mean arterial pressure (MAP) were evaluated in unanaesthetized animals. Glomerular volume (GV), glomerular sclerosis index (GSI), mesangial volume density (Vvmes) and glomerular capillary surface density (Svcap) were also evaluated. Results show that kidney weight increased in Nx groups, being higher in DMNx. GFR was higher in Nx groups as was RPF, being higher in DMNx. RVR was lower in Nx groups, especially in DMNx. MAP was not different among the groups. RPF and GFR showed a high correlation for the DMNx group (r=0.95, p=0.02). The DMNx group showed a correlation between RVR and GFR (r=-0.96, p=0.005). The GV increased in Nx groups, and the GSI was higher in DMNx. Vvmes and Svcap increased in DMNx group. In summary, Nx groups developed similar degrees of glomerular hypertrophy, but only DMNx showed an increased value for GSI. The present data suggest that the acceleration of glomerular lesions in DMNx animals was more closely associated to hemodynamic adaptations than to glomerular hypertrophy.

Influence of the chronic nitric oxide synthesis inhibition on cardiomyocytes number.

This work analyzes the relationship between the number of viable cells and alteration of the cardiomyocytes growth response capacity of the hypertensive rat myocardium. Hypertension was induced in Wistar rats by means of nitric oxide synthesis blockade using NG-nitro-L-arginine methyl ester (L-NAME). L-NAME (12 mg/kg per day) was given to animals in drinking water ad lib for 15 weeks. Proliferating cell nuclear antigen (PCNA) protein expression and the disector method were used to evaluate the proliferation capacity of the cardiomyocytes and its numerical density alteration (Nv[m]), respectively. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) and monoclonal antibody to single-stranded DNA were two methods that detected the process of the apoptotic cell death. The association of the p53 expression with the apoptosis was investigated using anti-p53 antibody. The heart weight, body weight, and heart weight/body weight ratio of the control rats increased 114%, 77%, and 22%, respectively, and the Nv[m] decreased 60% (P<0.0001) relative to the L-NAME rats. The cardiomyocytes did not present PCNA labeling, indicating the absence of cellular proliferation. The decline of the Nv[m] was also associated with apoptotic cell death in the myocardium of the hypertensive rats. A p53-dependent pathway seems to mediate the programmed cell death in this model of hypertension.

Numerical density of cardiac myocytes in aged rats fed a cholesterol-rich diet and a canola oil diet (n-3 fatty acid rich).

We studied the myocardium of 45 aged rats fed from 21 days after birth until 15 months of age with a standard rat diet a cholesterol-rich diet (CHO) or canola oil (O). We analysed the cardiac weight (CW) and, using unbiased stereological estimates, studied isotropic, uniform, random sections of the free left ventricular wall to determine the numerical density of the myocytes (NV[myocyte]). The CW was not statistically different between groups A and CHO: it was smallest in animals in group O (21.2% smaller in group O than in group A and 15.3% smaller in group O than in group CHO). NV[myocyte] was statistically different in all three groups and was greatest in animals in group O. By comparison with rats in group A, group CHO rats had an NV[myocyte] than was 51.3% smaller and group O, 33.3% greater. Aged rats fed with canola oil diet have a well-vascularized myocardium, which is probably associated with preservation of NV[myocyte] in the myocardium of these animals.

Volume-weighted mean nuclear volume and numerical nuclear density in the cardiomyocyte following enalapril and verapamil treatment.

The estimation of the volume-weighted mean nuclear volume, nu(nu), and the numerical nuclear density in the plane, N(A), was used to analyze cardiomyocyte nuclei in the condition of cardiac hypertrophy caused by nitric oxide (NO) synthesis blockade and simultaneous antihypertensive treatment for 40 days (four groups of ten rats each: control, L-NAME, L-NAME+enalapril, L-NAME+verapamil). The blood pressure (BP) increased 71% in the L-NAME group. In the L-NAME+enalapril and L-NAME+verapamil groups, the BP did not show any alteration when compared with the respective controls. In comparison with the control group, nu(nu) was 250% greater, and the N(A) was 25% smaller in the L-NAME animals, while no difference occurred in the other two groups. With respect to cardiomyocyte nuclear size, the present results suggest a beneficial effect of the angiotensin-converting enzyme inhibitor enalapril and the calcium channel blocker verapamil when NO synthesis is blockaded.

Stereology of the myocardium in hypertensive rats. Differences in relation to the time of inhibition of nitric oxide synthesis.

Structural changes in the myocardium following inhibition of nitric oxide synthesis were studied quantitatively within two different periods. Four groups of 10 rats were studied: control and L-NAME (NG-nitro-methyl-ester-L-arginine) groups for 25 and for 40 days. L-NAME was administered at 50 mg/kg per day in the drinking water. On the 26th and 41st days, the hearts were examined. Volume densities of myocytes (Vv[m]), cardiac interstitium (Vv[int], numerical density of myocytes (Nv[m]) and mean cross-sectional area of the myocytes (A[m]) were determined. Comparing the L-NAME animals with their respective controls showed the arterial pressure (AP) and the heart weight (HW) to be increased in the L-NAME animals. At 25 days, and more obviously at 40 days, the myocytes were hypertrophied with increase of myofibrils (A[m], greater in L-NAME rats). There were some areas with ischaemic lesions, inflammatory infiltrates and perivascular and interstitial fibrosis. The intramyocardial arteries had a thick tunica media and tunica intima. At 25 days the myocardium showed no stereological difference between L-NAME and controls, but by 40 days there was decreased Vv[m] and Nv[m] and increased Vv[int] in the exposed group. Inhibition of NO synthesis provoked a time progressive myocardial change, quantified by stereology.