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.

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.

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.

Natural killer cells and alterations in collagen density: signs of periradicular herpesvirus infection?

This study evaluated the presence and density of natural killer (NK) cells as well as collagen density in chronic apical periodontitis lesions and tried to find any correlations with concomitant herpesvirus infection or histopathological status of the lesion. Surgical specimens of chronic apical periodontitis lesions were surveyed for the presence and density of NK cells by immunohistochemical analysis. Collagen density in these lesions was quantified by means of histochemistry. All specimens were positive for the presence of CD57-positive cells. Topographically, CD57-positive cells were found singly or forming clusters in the granulomatous tissue, as well as subjacent and within the cystic epithelium. No significant differences in the density of CD57-positive cells were found between nonepithelialized and epithelialized lesions or between herpesvirus-positive and herpesvirus-negative lesions. Significant differences were found in volumetric density of collagen when comparing nonepithelialized and epithelialized lesions, with the latter demonstrating higher values. When no distinction of lesion type was made, there was no significant difference in collagen density between herpesvirus-positive and herpesvirus-negative lesions. When comparing the collagen density in herpesvirus-positive and herpesvirus-negative specimens from the same lesion type, a significant difference was found in nonepithelialized lesions, with herpesvirus-positive lesions showing lower values. The presence of CD57-positive cells in all chronic apical periodontitis specimens may indicate that activated NK cells play a role in the pathogenesis of this disease, possibly by participating in innate immunity events involved in the control of virus infection. Collagen density may vary in function of the type of lesion and presence of herpesvirus infection.

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.

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 percent 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 percentATI/organ (P < 0.05) and from 0.57 ± 0.17 to 0.39 ± 0.14 percentATI/g (P < 0.05) was observed in the heart, ii) from 0.07 ± 0.02 to 0.19 ± 0.07 percentATI/g in the pancreas, and iii) from 0.07 ± 0.01 to 0.18 ± 0.07 percentATI/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.

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.

Prevalence of genital human papillomavirus infections established using different diagnostic techniques among males attending a urological clinic.

OBJECTIVE: To compare the accuracy of different diagnostic methods and their use in estimating the prevalence of genital human papillomavirus (HPV) infections in males attending a urological clinic. MATERIAL AND METHODS: The study population was derived from a series of 1,153 consecutive males attending a urological clinic in São Paulo between January 1996 and November 1998. Of these 1,153 males, 334 had clinically suspected genital HPV infection and comprised the study cohort. The diagnostic methods used included peniscopy, directed biopsy and HPV detection by means of the Hybrid Capture 2 (HC) assay for both oncogenic and non-oncogenic HPV types. RESULTS: Peniscopy was performed for 297 males, positive results being reported in 237 cases (79.8%). Directed biopsy was performed in 188 males, and histology suggested HPV in 140 of these cases (74.5%). HC confirmed the presence of HPV in only 35.2% of the histologically HPV-suggestive cases. Peniscopy has good sensitivity for identifying male carriers of genital HPV. However, the technique has an inherent low specificity, limiting its usefulness to the correct identification of those who never present with HPV infection. Characteristic histological alterations are useful in suggesting HPV infection, but their correlation with HPV detection using HC is not particularly good. CONCLUSIONS: These data suggest that both histology and peniscopy have low specificity in detecting male genital HPV. Accurate diagnosis of HPV infection can be confirmed by molecular detection methods only. Histology, however, plays an important role in the differential diagnosis. An appropriate diagnostic protocol for male genital HPV infections in a urological clinic should include peniscopy, histology and molecular diagnostic tools (HC or polymerase chain reaction).

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.

Evaluation of the cholesterol influence in type II collagen-induced arthritis in DBA/1J mice: an autoradiographic study.

In order to verify the cholesterol influence in RA severity in DBA/1J mice, we quantified the cholesterol present in the knee joints of normal (N) and with collagen II induced arthritis (CIA). Forty male DBA/1J mice, were divided in normal (n=20) and CIA group (n=20). Mice in CIA group were injected with 100 microg of collagen II emulsified in Freund's complete adjuvant. Sixteen DBA/1J (8 N and 8 CIA) received an injection of 2.96 x 10(6) Bq of (3)H-cholesterol and were anesthetized and sacrificed. Semi-fine sections were covered with LM-1 emulsion, exposed for six weeks and developed. Collagen induced edema, erythema and dysfunction of knee joints in CIA group. Radioactive cholesterol was located more on the synovial membrane, where we found the greatest density of silver grains, significantly (P<0.0001) higher in group CIA vs. controls (61-/+2.3 X 18-/+0.7). We conclude that the cholesterol deposits on the synovial membrane is related to CIA severity.

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.

Dna damage in peripheral blood nuclear cells assessed by comet assay from individuals submitted to scintigraphic examinations.

Stannous chloride (SnCl2) is employed as a reducing agent to obtain Technetium-99m-labelled radiophamaceuticals in nuclear medicine kits, being injected endovenously in humans. Toxic effects of these kits were not studied, thus making it important to evaluate their impact in humans. In this study, the toxic effects were evaluated from peripheral blood nuclear cells (PBNC) from patients who received radiopharmaceuticals obtained using such kits. The analyses included results performed by comet assay. DNA damage was visualized in PBNC samples collected within a time up to 2 hr, and 24 hr after radiopharmaceutical injection in the patients. Initially we observed an increase of comet signals, which subsequently were reduced to zero after 24 hr. The diminishing of comet amounts probably is associated with DNA repair of damaged cells or with the elimination by apoptosis of cells whose DNA are not repaired.

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 percent 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 æg/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 percent 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 percent of controls and only in 34 percent of hypothyroid and 43 percent 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

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.

Primary headache care delivery by nonspecialists in Brazil. Brazilian Headache Care Cooperative Group.

Headaches are common disorders usually examined by nonneurologists. In order to assess how primary headache patients (IHS groups 1, 2, and 3) are generally managed by nonspecialists, 414 patients were asked about their previous headache care. Correct diagnosis had previously been made in only 44.9%, 6.7%, and 26.7% of the migraine, tension-type headache, and cluster headache patients, respectively. The patients underwent 501 investigative procedures motivated by the headache, averaging 1.21 examinations per patient, mostly EEGs. Preventive treatment was largely overlooked irrespective of the headache type. It is concluded that scientific improvements in headache care may be ineffective unless educational programs improve headache knowledge in general.

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.