Focalização isoelétrica na identificação das hemoglobinas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Reduction of enantioselectivity in the kinetic disposition and metabolism of verapamil in rats exposed to toluene

Toluene and verapamil are subject to extensive oxidative metabolism mediated by CYP enzymes, and their interaction can be stereoselective. In the present study we investigated the influence of toluene inhalation on the enantioselective kinetic disposition of verapamil and its metabolite, norverapamil, in rats. Male Wistar rats (n = 6 per group) received a single dose of racemic verapamil (10 mg/kg) orally at the fifth day of nose-only toluene or air (control group) inhalation for 6 h/day (25, 50, and 100 ppm). Serial blood samples were collected from the tail up to 6 h after verapamil administration. The plasma concentrations of verapamil and norverapamil enantiomers were analyzed by LC-MS/MS by using a Chiralpak AD column. Toluene inhalation did not influence the kinetic disposition of verapamil or norverapamil enantiomers (p > 0.05, Kruskal-Wallis test) in rats. The pharmacokinetics of verapamil was enantioselective in the control group, with a higher plasma proportion of the S-verapamil (AUC 250.8 versus 120.4 ng.h.mL(-1); p <= 0.05, Wilcoxon test) and S-norverapamil (AUC 72.3 versus 52.3 ng.h.mL(-1); p <= 0.05, Wilcoxon test). Nose-only exposure to toluene at 25, 50, or 100 ppm resulted in a lack of enantioselectivity for both verapamil and norverapamil. The study demonstrates the importance of the application of enantioselective methods in studies on the interaction between solvents and chiral drugs.

Effects of chronic stress on nicotine-induced locomotor activity and corticosterone release in adult and adolescent rats

We examined nicotine-induced locomotion and increase in corticosterone plasma levels in adolescent and adult animals exposed to chronic restraint stress. Adolescent [postnatal day (P) 28-37] and adult (P60-67) rats were restrained for 2 hours once daily for 7 days. Three days after the last exposure to stress, the animals were challenged with saline or nicotine (0.4 mg/kg subcutaneously). Nicotine-induced locomotion was recorded in an activity cage. Trunk blood samples were collected in a subset of adolescent and adult rats and plasma corticosterone levels were determined by radioimmunoassay. Exposure to stress did not affect the nicotine-induced locomotor- or corticosterone-activating effects in both ages.

Análise molecular dos éxons 8 a 11 do gene da p53 em amostras de câncer de colo de útero no Rio Grande do Norte

Mutations on TP53 gene are common in human cancer but not in cervical cancer where they are rarely found and the inactivation and degradation of p53 protein are attributed to the action of E6 viral oncogene from high risk human papillomavirus (HPV). Analysis of cervical cancer cell lines suggests that HPV negative samples shows mutation on TP53, but clinical approaches didn t confirmed this hypothesis. However, in most TP53 mutations studies on cervical cancer, only the exons 5 to 8 were analyzed. Approximately 90% of mutations described are on this region. Recent studies on several cancer suggests that mutation frequency in the other exons must be considered. The aim of this work was to verify whether mutations on coding and non-coding regions occur in cancer tissue from cervical cancer in patients from Rio Grande do Norte using Denaturing Gradient Gel Electrophoresis (DGGE) as screening tool. Exons 8 to 11 were analyzed including some introns from 80 tumor samples and 8 peripheral blood samples from healthy women. DNA were submitted to PCR using primers with GC clamp on the end of one of them. The results were observed for each region after DGGE and silver staining. It was observed no amplified fragment with different migration profile from those obtained from DNA of peripheral blood. These results agree with those from literature where TP53 mutations in cervical cancer have been described in a very low frequency

Caracterização genética de vacas leiteiras por meio de marcadores moleculares e suas implicações na composição e qualidade do leite

The objective of this study was to identify DNA polymorphisms at the genes leptin, β-lactoglobulin and pituitary-specific transcription factor in three genetic groups of Holstein x Guzerat dairy cows and investigate the relationship between their genotypes and the composition and quality of milk of dairy cows. Samples were collected in August 2009, being 113 blood samples from lactating crossbred cows and 58 milk samples. For analysis of DNA polymorphisms blood samples were collected, analyzed later in the Genetic Laboratory affiliated to the Zootechny Institute of São Paulo and individual milk samples were collected according to standards established by the laboratory of Management Program of Northeast Dairy Herds (PROGEN), at Federal Rural University of Pernambuco (UFRPE) for analysis of milk composition and quality. The characterization of genotypes was performed by PCR-RFLP, for which were designed specific primers for each studied gene and restriction enzymes Kpn2I, HaeIII and HinfI that cut the DNA of the following genes: leptin, β-lactoglobulin and a PIT, respectively. The leptin estimate genotypic frequence were CC 0.112, TT 0.225 and CT 0.661, for β-lactoglobulin were AA 0.136, AB 0.323 and BB 0.539, and for PIT were ++ 0.655, -- 0.311 and +- 0.032. The results show that the population is in Hardy-Weinberg disequilibrium for leptin, β-lactoglobulin and a PIT due to excess of heterozygotes in the population, however, as these genes are associated with the milk production it is considered that the animals have genetic potential for milk production in the Brazilian semi-arid conditions. Through the characterization of the studied herd there were not found implications of the polymorphism of leptin, β-lactoglobulin and PIT in the composition and quality of milk from cows in the different genetic groups 1/2, 3/4 and 7/8 Holstein x Guzerat. Key words: β-lactoglobulin, crossbred cows, leptin, PCR-RFLP, PIT1, semi-arid.

Resposta comportamental e hormonal de machos não reprodutores de sagüi, callithrix jacchus, a estímulos sensoriais de filhotes não aparentados

Parental care in mammals is influenced by somatosensory stimuli from infants, such as vocalization and sight and by changes in the hormone levels of caretakers. The aim of this study was to evaluate the behavioral and hormonal responses of twelve non reproductive adult male common marmosets (Callithrix jacchus) to infant cues, vocalization recordings, sight and physical contact with newborn. Six out of twelve males had previous experience in caretaking. In article 1, adult males were exposed to newborn vocalizations for 10 minutes. On control condition no sound was presented. In article 2, males were tested on two conditions: a) Control: an empty acrylic transparent box (test box) was placed in male s cage for 15 minutes, and b) Experimental: males were exposed to newborns into a closed text box for 15 minutes. The cage was kept closed to prevented from tactile, smell and acoustic stimulation by the infant on common marmoset males. In article 3, males were exposed to an open or closed text box, which allowed or not their access to and social interaction with the infants. After each observation sessions, blood samples were collected to evaluate the cortisol levels of males. In all studies, behavioral response of adult males was significantly modified by newborns sight, vocalization and physical contact. Males approached and spent more time near the sound source and showed an increase in locomotion during sound exposure. Furthermore, males approached, smelled and spent more time near the test box when the newborn was inside it. There was no difference in behavioral pattern between experienced and non-experienced males in articles 1 and 2. In article 3, behavioral pattern of males was influence by previous caretaking experience. Experienced males recovered quicker and carried the infants more than the inexperienced ones. However, inexperienced males showed a decrease in recovery latency and an increase in carrying time after successive exposure to infants. Cortisol levels changed after exposure to infant s vocalization, especially for experienced adult males. Male hormonal profile was not affected by the sight of infants neither by their previous experienced in caretaking. The occurrence of social interaction between the caretaker and infant did not modify the hormonal profile of common marmoset males; however, as much as experienced males carried the infants their cortisol levels decreased. Thus, members of a social group or potential caretakers common marmosets exposed to sensory cues from dependent infant such as vocalization, sight, smell and physical contact, changed their behavioral and hormonal responses that are physiological modulators of parental behavior in common marmoset

Perfil hormonal anual de machos e fêmeas adultos de saguis (callithrix jacchus)

Many behavioral and biological variables of animals are expressed in the form of biological rhytms, down by the Circadian Timing System, that synchronize them with the environment from external stimuli such as light. One of them is the secretion profile of most circulating hormones regulated by the hypothalamuspirtuitary axis, which controls functions essential for the survival and reproduction of organisms. The sagüi, Callithrix jacchus, one of the most studied species about their endocrine physiology, is an appropriate subject for evaluating the profile of plasma prolactin and cortisol of adult males and females born in captivity throughout the year. Three male and two adult femelas were housed individually and subjected to natural environmental conditions over two years. Blood samples were used to measure the circulating levels of both hormones by methods radioimmunoassay (RIA) and immunoassay (ELISA), respectively. The analysis during the year of the plasmatic values of both hormones test was performed by ANOVA for repeated measures, the correlation of Spearman, and the test of Friedman and Student's t-test. The levels of prolactin in plasma were higher during the months in which there is a greater incidence of births of baby in the colony, possibly serving for modulating the expression of the behavior of parental care in both sexes. The plasma cortisol showed a lift in anticipation of the station with the highest birth rate and may be associated with the preparation of individual participation in caring for the baby, and also with the establishment of emotional bond between reproductive partners. Thus, this study shows that, despite the variations observed in the environment in which the animals live, plasma levels of prolactin and cortisol vary little throughout the year.

Avaliação do cortisol, dehidroepiandrosterona sulfato e dos perfis lipídico e imunológico e suas correlações com a sensibilidade dolorosa, depressão e funcionalidade em mulheres com fibromialgia na pós-menopausa

Fibromyalgia (FM) is a non-inflammatory rheumatic syndrome characterized by widespread musculoskeletal pain with palpable tender points, muscle stiffness, fatigue, and sleep disturbances. Patients with FM have hormonal changes that are directly correlated with symptoms of the syndrome. The neuroendocrine regulation may be impaired, with abnormalities in the hypothalamus-pituitary-adrenal (HPA) axis with various hormones showing changes in their levels. In women in fertile period, various gonadal hormones are associated with symptoms of the syndrome, but studies focusing only a population of women in post-menopausal period who do not use hormone replacement are rare. We developed an analytical cross sectional study to assess the plasma levels of cortisol and dehidroepiandrosterona sulfate (DHEA-S) with quimioluminescence method in a group of 17 women with FM and 19 healthy women in post-menopause who do not use hormone replacement and observe the correlation with the symptoms of pain through algometry, depression and physical functional capacity measured from the Beck Depression Index (BDI) and the Fibromyalgia Impact Questionnaire (FIQ). Three blood samples were collected in the morning (between 8:00 9:30) with an interval of 24 hours for the measurements of hormonal levels and biochemical profile. There were no immunological or lipid changes in patients with FM. Comparing the two groups, there is no difference in levels of cortisol and a tangential effect for DHEA-S (p=0,094) with the lowest levels in the FM. DHEA-S also correlated with pain threshold (r=0,7) and tolerance (r=0,65) in group FM. We found the presence of depressive state and low physical functional capacity in FM. It was also evident that women in post-menopausal period, DHEA-S should influence the symptoms of increased sensitivity to pain, but not the presence of depressive status and low physical functional

Indicadores da resposta ao estresse agudo associados com a familiaridade ao ambiente e aos procedimentos de banho e tosa em cães (Canis familiaris) da raça poodle

The stress responses can be parameters used in order to identify the welfare of animals. Behavior parameters can also be regarded as means to identify their relation to stressing situations. In this study, adult male and female poodle dogs, accustomed (group 1) or not (group 2) to the environment of a bath and grooming salon were analyzed through the plasmatic concentration of cortisol, cell counting of the immune system (total leukocytes and percentage of neutrophils and lymphocytes) and through observation of changes in behavior before and after the bath and grooming service. After arriving at the kennel, the dogs were taken to the bath and grooming facility, where they had their blood samples taken and were observed for ten minutes. The subsequent procedures consisted of the bath and grooming services and the collection of another blood samples. The research results were analyzed through non-parametric statistic tests and p < 0.05. Accustomed and non-accustomed dogs presented different hormonal and behavior responses: accustomed dogs presented an increasing of the cortisol level at the second blood samples collection, thus indicating stress towards the aforementioned procedures and presenting behavior responses which can be described as moderated stress. Non-accustomed dogs presented the same cortisol levels in both first and second samples suggesting that they were already reacting to the new environment. Dogs also shown behavioral responses which can be described as acute stress. Both male and female dogs showed similar immunological changes as well as different endocrine and behavioral profiles. Concerning aggression, this study demonstrates that biting the owner can be a predicting behavior of aggression during the bath and grooming service, male dogs were more aggressive than the female, and that males were more reactive to unknown caretaker. The results of this research propose that the familiarization of the animal with the bath and grooming environment and caretaker is important to the welfare of poodle dogs, especially to that of male dogs

Prevention of renal ischemia/reperfusion injury in rats using acetylcysteine after anesthesia with isoflurane

OBJETIVO: Avaliar o efeito da N-acetilcisteína na proteção renal contra lesão de isquemia/reperfusão, quando administrada logo após a indução anestésica, em ratos anestesiados com isoflurano. MÉTODOS: Dezoito ratos Wistar machos pesando mais que 300g foram anestesiados com isoflurano. A jugular interna direita e a carótida esquerda foram dissecadas e canuladas. Os animais foram distribuídos aleatoriamente em GAcetil, recebendo N-acetilcisteína por via intravenosa, 300mg/kg, e GIsot, solução salina. Foi realizada nefrectomia direita e clampeamento da artéria renal esquerda por 45 min. Os animais foram sacrificados após 48h, sendo colhidas amostras sanguíneas após a indução anestésica e ao sacrifício dos mesmos para avaliar a creatinina sérica. Realizou-se histologia renal. RESULTADOS: A variação da creatinina foi 2,33mg/dL ± 2,21 no GAcetil e 4,38mg/dL ± 2,13 no GIsot (p=0,074). Dois animais apresentaram necrose tubular intensa no GAcetil, comparados a cinco no GIsot. Apenas GAcetil apresentou animais livres de necrose tubular (dois) e degeneração tubular (um). CONCLUSÃO: Após isquemia/reperfusão renais, os ratos aos quais se administrou N-acetilcisteína apresentaram menor variação na creatinina sérica e lesões renais mais leves que o grupo controle.

DNA damage in patients who underwent minimally invasive surgery under inhalation or intravenous anesthesia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Evaluation of DNA damage and lipoperoxidation of propofol in patients undergoing elective surgery

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Plasma concentrations and behavioral, antinociceptive, and physiologic effects of methadone after intravenous and oral transmucosal administration in cats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genotoxicity, cytotoxicity and gene expression in patients undergoing elective surgery under isoflurane anaesthesia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cytokine Profile in Patients Undergoing Minimally Invasive Surgery with Balanced Anesthesia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)