Aerobic exercise improves microvascular dysfunction in fructose fed hamsters.

Fructose is a major diet component directly related to severe damages to the microcirculation and to diseases such as obesity, diabetes and hypertension to which physical activity is pointed out as an important non-pharmacological treatment since its positive effects precede anthropometric improvements. In this study we have investigated the effects of a light/moderate aerobic exercise training (AET) on microcirculatory dysfunction elicited by carbohydrate overload during a period of 5 months. Male hamsters (Mesocricetus auratus) whose drinking water was substituted (F) or not (C) by 10% fructose solution, during 20 weeks, associated or not to AET in the last 4 weeks (EC and EF subgroups) had their microcirculatory function evaluated on the cheek pouch preparation, glucose and insulin tolerance (GTT and ITT) tested. Arterial blood was collected for pO2, pCO2, HCO3(-), pH, total CO2, saturated O2 and lactate determinations. Liver fragments were observed using an electron microscope. Microcirculatory responses to acetylcholine [Ach, an endothelium-dependent vasodilator; 10(-8)M - *123.3±7.5% (C), 119.5±1.3% (EC), *98.1±3.2% (F) and 133.6±17.2% (EF); 10(-6)M - *133.0±4.1% (C), 135.6±4.3% (EC), *103.4±4.3% (F) and 134.1±5.9% (EF); 10(-4)M - *167.2±5.0% (C), 162.8±5.4% (EC), *123.8±6.3% (F) and 140.8±5.0% (EF)] and to sodium nitroprusside [SNP, an endothelium-independent vasodilator; 10(-8)M - 118.8±6.8% (C), 114.0±5.0% (EC), 100.2±2.9% (F), 104.9±4.4% (EF); 10(-6)M - 140.6±11.7% (C), 141.7±5.5% (EC), 125.0±4.7% (F), 138.3±2.8% (EF); 10(-4)M - 150.4±10.9% (C), 147.9±6.5% (EC), 139.2±7.3% (F), 155.9±4.7% (EF)] and macromolecular permeability increase induced by 30 min ischemia/reperfusion (I/R) procedure [14.4±3.5 (C), 30.0±1.9 (EC), *112.0±8.8 (F) and *22.4±0.9 leaks/cm(2) (EF)] have shown that endothelium-dependent vasodilatation was significantly reduced and I/R induced macromolecular permeability augmented in sedentary fructose (F) subgroup and both improved after AET. Electron microscopy analysis of the liver showed significant differences between exercised and sedentary subgroups with greater amount of glycogen in F subgroups compared to other ones. No significant changes on mean arterial pressure, heart rate or blood gase between subgroups could be detected. Our results point out that AET could normalize microcirculatory dysfunction elicited by long term substitution of drinking water by 10% fructose solution.

Evaluation of a multiresidue method for measuring fourteen chemical groups of pesticides in water by use of LC-MS-MS.

European Council Directive 98/83/EC on the quality of water intended for human consumption brought a new challenge for water-quality control routine laboratories, mainly on pesticides analysis. Under the guidelines of ISO/IEC 17025:2005, a multiresidue method was developed, validated, implemented in routine, and studied with real samples during a one-year period. The proposed method enables routine laboratories to handle a large number of samples, since 28 pesticides of 14 different chemical groups can be quantitated in a single procedure. The method comprises a solid-phase extraction step and subsequent analysis by liquid chromatography-mass spectrometry (LC-MS-MS). The accuracy was established on the basis of participation in interlaboratory proficiency tests, with encouraging results (majority |z-score| <2), and the precision was consistently analysed over one year. The limits of quantitation (below 0.050 microg L(-1)) are in agreement with the enforced threshold value for pesticides of 0.10 microg L(-1). Overall method performance is suitable for routine use according to accreditation rules, taking into account the data collected over one year.

Experimental model to assess possible medicinal herb interaction with a radiobiocomplex: qualitative and quantitative analysis of kidney, liver and duodenum isolated from treated rats.

Ginkgo biloba extract (EGb) has been used as a medicinal herb. Several biological properties have been associated with this extract, especially, in the increase of the blood flow, in the action as platelet activating factor antagonism and in the prevention of the membrane against the damage caused by free radicals. Radiobiocomplexes have been utilized in various nuclear medicine procedures helping in the diagnosis and/or treatment of human diseases. Many substances have been reported to affect the bioavailability of different radiobiocomplexes. The aim of this work was to evaluate the possible influence of an EGb on the bioavailability of the sodium pertechnetate (99mTcO4Na) and on the morphometry of some organs isolated from rats. These animals were treated with EGb and 99mTcO4Na was injected. The animals were sacrificed, the organs isolated, counted in a well counter and the percentage of radioactivity per gram of each organ was calculated. The results showed that EGb decreased the uptake of the 99mTcO4Na in the duodenum (P<0.05). Moreover, morphometric analysis has revealed significant modifications (P<0.05) on kidney, liver and duodenum due to the cited treatment. It is speculated that the substances present in the EGb could act directly or generate metabolites capable to promote changes in organs (kidney, liver and duodenum), however, only significant alteration in the uptake of the 99mTcO4Na in the duodenum.

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 (cat's 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.

Optimization of supercritical fluid extraction of pesticide residues in soil by means of central composite design and analysis by gas chromatography-tandem mass spectrometry.

An environmentally friendly methodology is proposed for the analysis of pesticides in soil samples based on supercritical fluid extraction (SFE) and analysis at high selectivity and sensitivity, by gas chromatography-tandem mass spectrometry (GC-MS-MS). The pesticides investigated are among the most commonly used in intensive horticulture activities comprising organochlorine and organophosphorous insecticides, triazine and acetanilide herbicides, amongst others. An experimental design approach was used for modelling SFE and optimised extraction conditions were derived for the total pesticides extraction or for specific sub-groups of interest. Pesticide residues could be detected in soils in the sub-ppb range (0.1-3.7microgkg(-1)), with quite good precision (4.2-15.7%) and extraction efficiency (80.4-106.5%). The analysis of soil samples from an intensive horticulture area in Póvoa de Varzim, north of Portugal, revealed the presence of persistent pesticides, parent compounds and degradation products among the following: endosulfan, endosulfan sulfate, dieldrin, 4,4'-DDE, 4,4'-DDD, atrazine, alachlor, metolachlor, chlorpyrifos, pendimethalin and lindane. The important features to point out are the easy interpretation of chromatograms and straightforward confirmation of analytes that greatly facilitates the analyst judgement on the contamination of the sample.

Blood pressure in four remote populations in the INTERSALT Study.

Four remote population samples (Yanomamo and Xingu Indians of Brazil and rural populations in Kenya and Papua New Guinea) had the lowest average blood pressures among all 52 populations studied in INTERSALT, an international cooperative investigation of electrolytes and blood pressure. Average systolic blood pressure was 103 versus 120 mm Hg in the remaining INTERSALT centers; diastolic blood pressure in these four population samples averaged 63 versus 74 mm Hg in the 48 other centers. There was little or no upward slope of blood pressure with age; hypertension was present in only 5% of the rural Kenyan sample and virtually absent in the other three centers. Also in marked contrast with the rest of the centers was level of daily salt intake, as estimated by 24-hour urinary sodium excretion. Median salt intake ranged from under 1 g to 3 g daily versus more than 9 g in the rest of INTERSALT populations. Average body weight was also low in these four centers, with no or low average alcohol intake, again unlike the other centers. The association within these four centers between the above variables and blood pressure was low, possibly reflecting their limited variability. While several other INTERSALT centers also had low average body weight or low prevalence of alcohol drinking, when this was accompanied by much higher salt intake (7-12 g salt or 120-210 mmol sodium daily), hypertension prevalence ranged from 8% to 19%. These findings confirm previous reports that in populations with a low salt intake, there is little or no hypertension or rise of blood pressure with age.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood pressure and electrolyte excretion in the Yanomamo Indians, an isolated population.

As part of the INTERSALT study, blood pressure, urinary electrolytes, age, and body mass index were assessed in a sample of 195 Yanomamo Indians from the Brazilian Amazon. Mean sodium excretion was 0.9 mmol/24 hr. Urinary sodium was lower than 1 mmol/24 hr in 84% of participants. Mean blood pressure was 96.0/60.6 mmHg (range 78/37 to 128/86). Systolic and diastolic blood pressure were not higher at older than at younger ages in men. In women, systolic pressure was lower at older ages. Mean body mass index was low (20.8 in women and 21.5 in men), and there was no alcohol intake reported. Mean 24 hour urinary potassium was 60 mmol in women and 67 in men. In the presence of the low sodium intake among the Yanomamo, body mass index and potassium, found to be related to blood pressure in the overall INTERSALT study, were not consistently related to pressure in this population. Results demonstrated that there was low average population blood pressure, no hypertension and no positive slope of blood pressure with age in a population with very low salt intake.

Genotoxic effects of stannous chloride (SnCl2) in K562 cell line.

The toxic effects of SnCl2 in K562 cells were analyzed in this study. This cell line is resistant to reactive oxygen species (ROS) making it suitable to evaluate the impact of SnCl2 in culture either through ROS or by direct toxicity using Trypan blue dye exclusion, comet and flow cytometry assays. An important loss of viability induced by SnCl2 in a dose-response manner was observed in cells treated in Tris-buffered saline (TBS). This necrotic cell death was further confirmed by flow cytometry. On the other hand, there was no loss of viability when cells were treated in rich medium (RPMI). DNA damage was visualized in SnCl2-treated K562 cells in both tested conditions. The data indicate that SnCl2 induces DNA damage and reduces K562 viability. Both actions seem to be correlated with ROS formation and direct linkage to DNA.

Autoradiographic thyroid evaluation in short-term experimental diabetes mellitus.

Previous studies have shown that in vitro thyroid peroxidase (TPO) iodide oxidation activity is decreased and thyroid T4-5'-deiodinase activity is increased 15 days after induction of experimental diabetes mellitus (DM). In the present study we used thyroid histoautoradiography, an indirect assay of in vivo TPO activity, to determine the possible parallelism between the in vitro and in vivo changes induced by experimental DM. DM was induced in male Wistar rats (about 250 g body weight) by a single i.p. streptozotocin injection (45 mg/kg), while control (C) animals received a single injection of the vehicle. Seven and 30 days after diabetes induction, each diabetic and control animal was given i.p. a tracer dose of 125I (2 microCi), 2.5 h before thyroid excision. The glands were counted, weighted, fixed in Bouin's solution, embedded in paraffin and cut. The sections were stained with HE and exposed to NTB-2 emulsion (Kodak). The autohistograms were developed and the quantitative distribution of silver grains was evaluated with a computerized image analyzer system. Thyroid radioiodine uptake was significantly decreased only after 30 days of DM (C: 0.38 +/- 0.05 vs DM: 0.20 +/- 0.04%/mg thyroid, P < 0.05) while in vivo TPO activity was significantly decreased 7 and 30 days after DM induction (C: 5.3 and 4.5 grains/100 micron 2 vs DM: 2.9 and 1.6 grains/100 micron 2, respectively, P < 0.05). These data suggest that insulin deficiency first reduces in vivo TPO activity during short-term experimental diabetes mellitus.

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.

In vitro radioautographic studies of the biodistribution of radiopharmaceuticals on blood elements.

In the present study we evaluated the binding of the radiopharmaceuticals sodium pertechnetate (Na 99mTcO4), methylenediphosphonic acid (99mTc-MDP) and glucoheptonate acid (99mTc-GHA) to blood elements using centrifugation and radioautographic techniques. Heparinized blood was incubated with the labelled compounds for 0, 1, 2, 3, 4, 6 and 24 h. Plasma (P) and blood cells (BC) were isolated and precipitated with 5% trichloroacetic acid (TCA), and soluble (SF) and insoluble fractions (IF) were separated. Blood samples were prepared (0 and 24 h) and coated with LM-1 radioautographic emulsions and percent radioactivity (%rad) in P and BC was determined. The binding of Na 99mTcO4 (%rad) to P was 61.2% (0 h) and 46.0% (24 h), and radioautography showed 63.7% (0 h) and 43.3% (24 h). The binding to BC was 38.8% (0 h) and 54.0% (24 h), and radioautography showed 36.3% (0 h) and 56.7% (24 h). 99mTc-MDP study presented 91.1% (0 h) to P and 87.2% (24 h), and radioautography showed 67.9% (0 h) and 67.4% (24 h). The binding to BC was 8.9% (0 h) and 12.8% (24 h), and radioautography showed 32.1% (0 h) and 32.6% (24 h). 99mTc-GHA study was 90.1% (0 h) to P and 79.9% (24 h), and radioautography showed 67.2% (0 h) and 60.1% (24 h). The binding to BC was 9.9% (0 h) and 20.1% (24 h), and radioautography showed 32.8% (0 h) and 39.9% (24 h). The comparison of the obtained results suggests that the binding to plasma and blood cells in the two techniques used (radioautography and centrifugation) is qualitatively in accordance.

Correlates of blood pressure in Yanomami Indians of northwestern Brazil.

We determined associations of measures of body habitus with blood pressure for 100 adult Yanomami Indians (61 men, 39 women) examined during February and March 1990. Measurements included body weight and height, four skinfolds (triceps, subscapular, suprailiac, abdomen), four circumferences (wrist, upper arm, abdomen, hip), systolic and diastolic blood pressures, pulse rate, and estimated age. Various indices of fat distribution were determined from the measurements of skinfolds, circumferences, weight, and height. Estimated age averaged 35.0 years in men and 33.4 years in women (range: 15 to 63 years). Mean systolic and diastolic blood pressures were low in both men (104.8/70.4 mm Hg) and women (94.8/63.5 mm Hg), as was body mass index (men: 20.7; women: 21.4 kg/m2). In Yanomami women, all four skinfolds, wrist circumference, and the indices of hip and abdominal fat were significant correlates of systolic blood pressure, while the abdominal skinfold and wrist and hip circumferences correlated significantly with diastolic blood pressure. Among men, there was a negative correlation between estimated age and systolic blood pressure and a positive correlation between BMI and upper arm and hip circumferences and systolic blood pressure. There was a significant positive correlation between wrist, upper arm, and hip circumferences and diastolic blood pressure among Yanomami men. We used stepwise regression to generate sex-specific predictive equations for blood pressure. For men, estimated age and hip circumference, and for women, abdominal skinfold measurement and age were included in the model for systolic blood pressure. Among men, wrist circumference and height, and among women, wrist circumference alone entered the model for diastolic blood pressure. On the basis of these results, we suggest that even in a low-blood pressure, low-body fat, no-salt setting, systolic blood pressure is associated with the amount and placement of adipose tissue. However, diastolic blood pressure is more closely correlated with skeletal size.

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.

[Bilateral abducens nerve palsy in nasopharyngeal carcinoma]. / Paralisia bilateral do abducente em carcinoma de nasofaringe.

We report a case of bilateral abducens palsy as initial finding in nasopharyngeal carcinoma. Possible mode of involvement is discussed. Only one case has been reported in the past.

[Blood pressure in 6 Yanomami villages]. / Pressão arterial em seis aldeias Yanomami.

PURPOSE: To investigate in Yanomami Indians that not add salt to food, the relationship between blood pressure (BP), biological variables (age, body weight, height and pulse) and urinary electrolytes (Na+, K+, Ca++ and Mg++). PATIENT AND METHODS: We studied 125 males and 129 females from six villages on Surucuru plateau and on Catrimani and Ajarani rivers region in the state of Roraima, north Brazil. Two BP measurements were made and the mean of them were used in data analysis. RESULTS: None hypertensive was found. Systolic BP decreased with age and correlated with body weight, pulse and urinary Na+. Diastolic BP only correlated with body weight. Height, urinary K+, Ca++ and Mg++ did not correlate with BP. CONCLUSION: There was no hypertension nor increase of BP with increasing age in these isolated Yanomami.

[The absence of risk factors for coronary disease in Yanomami Indians and the influence of acculturation on arterial pressure]. / Ausência de fatores de risco de doença coronária em indios Yanomami e influência da aculturação na pressão arterial.

PURPOSE: To investigate the hypothesis that hypertension and coronary heart disease are "civilization diseases", analyzing the distribution of their antecedents in an Indian population and observing the influence of acculturation on blood pressure. METHODS: Seven hundred and twenty-five Yanomami Indians of both sexes, ages above 14 years, inhabitants of eight villages in Roraima and Amazonas states were examined. Measures related to all classical coronary risk factors were carried out and urine samples were collected to measure electrolytes. RESULTS: None of the known coronary risk factors were found. The blood pressure levels were low and did not increase with increasing age. There was influence of acculturation on blood pressure and it was in part mediated by increase in body weight and sodium intake. CONCLUSION: The absence of hypertension and other coronary risk factors and the increase of blood pressure with acculturation, among an isolated population, strongly suggest that these diseases are "civilization diseases".

Carotid body remodelling in l-NAME-induced hypertension in the rat.

The carotid body (CB) is a chemoreceptor organ located at the bifurcation of the common carotid artery. It is made up of the carotid glomus, a structure containing type 1 cells surrounded by type 2 cells. The aim of this study was to evaluate the morphological changes of the CB and carotid glomus in the rat model of l-NAME-induced hypertension. Male Wistar rats were divided in two groups: control untreated rats (C) and rats receiving l-NAME 40 mg/kg/day (LN) for 6 weeks. At the end of the experiment, the systolic blood pressure was 63% higher in the LN group compared with the C group. Morphometric analysis showed that the area of the CB was 29% greater in the LN group compared with the C group. The density of nuclei in the CB was similar between groups, but it was 31% less in the carotid glomus of the LN group. Cells in the CB of the LN group displayed cytoplasmic vacuolation and expressed several biogenic amines. There were more elastic fibres, proteoglycans and collagen fibres in the LN group compared with the C group. Immunohistochemistry showed increased expression of nuclear factor kB, substance P, vascular endothelial growth factor and neuronal nitric oxide synthase in the LN group, while expression of the protein gene product 9.5 was decreased. l-NAME alters cell morphology and the expression of extracellular matrix molecules in the CB and carotid glomus in rats with l-NAME-induced hypertension.