Ciências Fisiológicas

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Programa de Pós-Graduação em Ciências Fisiológicas

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Agentes infecciosos associados à diarréia aguda em crianças até três anos de idade : estudo em um hospital de referência no município de Vitória-ES
(Universidade Federal do Espírito Santo, 2005-10-14) Sadovsky, Ana Daniela Izoton de; Pereira, Fausto Edmundo Lima; Spano, Liliana Cruz; Leite, José Paulo Gagliardi; Morais, Mauro Batista de
Acute diarrhea is one of the main causes of infantile mortality worldwide (WHO), mainly in developing countries. In the present work, the prevalence of Rotavirus (RV), adenovirus (Ad), diarrheogenic E. coli (EPEC, ETEC, EIEC, EHEC, EAEC, DAEC), Salmonella, Shigella, Cryptosporidium spp., Entamoeba histolytica and Giardia lamblia was studied among children up to 3 years old with acute diarrhea. From February 2003 to June 2004, stools samples were obtained prospectly from 253 children with acute diarrhea and 78 without diarrhea attending to the emergency room in a pediatric hospital - Hospital Infantil Nossa Senhora da Glória (HINSG), in Vitória Espírito Santo state, Brazil. Bacterial detection was done in 241 stools samples (12 were excluded because were in use of antimicrobian drugs) and E. coli were isolated in 219 and 68 cases with and without diarrhea, respectively. These cases were submited to serology with policlonal anti-seros (EPEC e EIEC) and hybridization tests (Hybr) to detect virulence genes of EPEC, ETEC, EIEC, EHEC, EAEC e DAEC. RV were studied in 147 cases for immune enzymatic assay (EIARA) and in 230 cases by poliacrylamide gel electrophoresis (PAGE) and Ad, only in 147 cases for immune enzymatic assay (EIARA). Protozoan infection was studied in 88 cases for immune enzymatic assay (EIA). Children with diarrhea were divided in Group I (88 cases = all enteropathogens studied), Group II (147 cases = bacterias, RV e Ad - EIARA) and Group III (230 cases = bacterias and RV - PAGE) and children without diarrhea were Group IV (78 cases = bactérias and RV - PAGE). Enteropathogens were detected in more than 60% in children with acute diarrhea and bacterial infection was the most prevalent: DEC were detected in 41,1%; EPEC in 3,6% (serology) e 9,1% (Hybr); Typical EPEC (0,9%); Atypical EPEC (8,2%); EAEC (9,1%); DAEC (20,6%); EIEC (0,9%); ETEC (4,2%). In stools samples from children without diarrhea, we found Atypical EPEC (10,3%); EAEC (20,6%); DAEC (16,2%); ETEC (1,5%). EHEC was not detected in the studied population. Shigella and Salmonella were detected in 4,6% e 2,9%, respectively, only in children with acute diarrhea. RV were detected in 35,2% (GEPA) and 50% (EIARA); Ad, in 8,2% and E. histolytica, Cryptosporidium spp. and G. lamblia in 8%, 11,4% and 14,8% of cases with diarrhea, respectively. In conclusion, Typical EPEC, EIEC and ETEC were detected only or predominantly in children with acute diarrhea. Atypical EPEC, EAEC and DAEC were not causes of acute diarrhea, except for EAEC in children more than two years old (p = 0,026). RV was the most prevalent agent when the classic enteropathogen DEC (Tipical EPEC, ETEC, EIEC and Shigella e Salmonella) was considered in this study. RV was more frequent in children below 18 months of life and in a period of March, 2003 up September, 2003. Associations among enteropathogens were frequent in the studied population and protozoa were the most of them. Comparing all of protozoa detected, only G. lamblia suggesting being a cause of acute diarrhea, isolately.
Alteraçães ponderais, hemodinâmicas e da função vascular do leito arterial caudal em ratas sete dias após o infarto do miocárdio
(Universidade Federal do Espírito Santo, 2009-01-01) Baldo, Thais de Oliveira Faria; Pereira, Raquel Binda; Stefanon, Ivanita; Vassallo, Dalton Valentim; Pereira, Fausto Edmundo Lima
Infarct area (AI) is an important determinant to heart failure (IC) development. However, studies from our laboratory have been shown that IC nor always correlates with AI. The aim of this study was to evaluate the IC development at 7 days after myocardial infarction (IM), and its consequence on vascular reactivity in the rat tail bed. Female Wistar rats were divided in Control group (CT); fictitious surgery (SHAM); and a group that was submitted to myocardial infarction (INF). Later, the INF group was subdivided in those developed (INFIC) or not (INF) IC. Seven days later, animals were anesthetized and catheterized to assess blood pressure and left ventricular function. After that, the vascular tail bed was removed and perfused under constant flow (2.5 mL/min). Alterations in the mean perfusion pressure (PPM) were acquire after concentration-response curve to phenilephrine (FE, 0.0001-300µg), before and after CHAPS-induced endothelial damage. To evaluate basal nitric oxide (NO) release, L-NAME was used in the presence of FE. To evaluate the dependent or independent relaxation, crescent concentration of acetilcholine (ACh) and sodium nitroprusside (NPS) were perfused after contraction with KCl (65mM). Data are shown as mean ± the standard error of mean, and statistical significance set at P<0.05. Seven days after, animals from INF-IC group showed decreased body weight (PC) (CT: 14.5±2.73; SHAM: 8.14±2.24; INF: - 4.06±2.65; INF-IC: -23.3±4.96g; P<0.05), increased lung/PC ratio (CT: 5.5±0.64; SHAM: 5.5±0.22; INF: 8.44±0.62; INF-IC: 9.90±0.91mg/g; P<0.05) and right ventricle/PC ratio (CT: 0.45±0.03; SHAM: 0.57±0.06; INF: 0.68±0.04; INF-IC: 0.78±0.06mg/g; P<0.05) when compared to other groups. Moreover, increased values for left ventricular end-diastolic pressure (PDFVE) were evidenced in INF-IC group as compared to the others (CT: 1.72±0.8; SHAM: 1.6±0.5; INF: 4.48±0.5; INF-IC: 14.5±1.3mmHg; P<0.05). AI was similar between infarcted groups (INF: 35.8±1.2; INF-IC: 38.8±2.3%; P>0.05). Furthermore, variation in the PC during the seven days correlates with PDFVE (r= -0.592; P<0.05). In the vascular tail bed, INF-IC group showed reduced maximal response (Rmáx) to FE (330.4±13.4 mmHg) as compared to other groups (CT: 423.2±25.4; SHAM: 403.6±28; IM: 425.5±30.4; P<0.05). Basal NO release acquire in the INF-IC group (13.35 ± 2.31) was significantly better as compared to other groups (CT 7.40 ± 1.36; SHAM 7.23 ± 0.8; INF 3.77 ± 0.7; P< 0.05). The relaxation mediated by Ach was reduced in SHAM and INF groups (SHAM: 50.0±2.8; INF: 48.4±2.7 %) as compared to CT and INF-IC groups (CT: 69.1±4.0; INF-IC: 66.9±3.4%; P<0.05). We conclude that seven days after IM is possible to identify animals that, with the same AI, develop or not the IC. Moreover, these animals depict different patterns of vascular response due, at least in part, to a better endothelial NO bioavailability.
Análise da correlação do espessamento médio-intimal proximal e distal nas carótidas comuns
(Universidade Federal do Espírito Santo, 2011-12-21) Roelke, Leonard Hermann; Mill, Jose Geraldo; França, Luiz Cláudio; Lotufo, Paulo Andrade
Background. The intima-media thickness (IMT) of the carotid arteries has been widely used as a noninvasive method to assess the cardiovascular risk because of its association with ischemic cardiovascular events. In the clinic practice, the IMT has been used as atherosclerosis predictor and as an additional tool to assess the cardiovascular risk according to the Framingham score. The high resolution ultrasound is used to measure carotid IMT in view of superficial localization of the artery and low cost of the exam. Different regions of carotid arteries, however, have been used to assess IMT, going from the common to the internal carotid artery, carotid bulb, either bilaterally or unilaterally. Aims: To correlate the thickening of the media-intima complex in different regions of the common carotid arteries in order to orientate the use of this index in daily clinical practice. Methods: IMT values were measured in the proximal and distal region of both common carotid arteries of 789 participants (both sexes, age 35-74 years) that attended to the Investigation Center of the ELSA study in Espírito Santo State. Images were obtained in the supine position with a Toshiba Aplio (model SSA-790A, version XG) ultrassonography platform by using a large band linear transducer with central frequency of 7.5 MHz (5.0-11.0 MHz). IMT was measured by using the dedicated software of the same equipment. Data were considered as following a normal distribution e the Pearson’s correlation coefficient ( r ) was used to determine the association between the IMT recorded at different sites of right and left common carotid arteries. The analysis were initially performed by the all group and then for the sub-groups without (<0.90 mm, 49% of the sample) a with (≥0.90 mm) subintimal thickening at least at one measured site. Statistical significance of associations was set at p<0.05. Results: A progressive increase in IMT was observed with age in all measured sites in the whole group and associations ranged from 0.56 to 0.69. An important decrease of association ( r ranging from 0.20 to 0.40) was observed in the group IMT≥0.90 mm suggesting low reproducibility of IMT when the thickness of the media-intima complex is increased. Conclusion: Despite the recent recommendations to measure the intima thickening in the distal region of the carotid artery, our data suggest that this recommendation should be restricted to epidemiological studies. The low correlations observed in the ≥0.90 mm group indicate that sub-intimal thickening is a focal process. Therefore, in the clinical practice, the whole extension of both common carotid arteries should be investigated to determine presence of IMT increase
Análise das propriedades mecânicas do miocárdio através do plano de fase
(Universidade Federal do Espírito Santo, 2008-05-09) Camilo, Luciana Moisés; Aquino, Regina Maria de; Vassallo, Dalton Valentim; Stefanon, Ivanita; Tucci, Paulo José Ferreira
The phase plane (PP) is a plot of the ventricular pressure in the X-axis and its time derivative in the Y-axis. One of the PP properties is that the final portion is a straight line segment and its slope (Ө) is a time constant that might represent a viscoelastic constant of the muscle. We aimed to compare the behavior of Ө among normal, hypertensives and hypertrophied rat hearts and to evaluate the influences of inotropic interventions on Ө. According an ideal mathematical model of approach of the isovolumic pressure curve, is given that PPA = P x dP/dt. As know that the external cardiac work (Wext) is the time rate of change of work, PdV, the equation can be rewrite in the following way: Text = P. dV . dP dP dt Since in this experimental model the volume is constant, the PPA is proportional to the Wext. The objective of this work was to compare the behavior of Ө and PPA among normal, hypertensives and hypertrophied rats heart and to evaluate the influences of the inotropics interventions on these variables. We used isolated hearts perfused by the Langendorff technique (male Wistar and spontaneously hypertensive rats- SHR), weighing between 210 and 280g, divided in 4 groups: (C) Wistar Control (n=7); SHR (n=7); (ISO,) rats with Isoproterenol-induced hypertrophy (0.3mg/Kg/day, n=9); and (V) rats treated only with vehicle (soyan bean oil, n=6). The experimental protocol consisted of performing ventricular function curves, changes in stimulation frequency and external Ca+2 (0.5, 1.25 and 2.5 mM) and β-adrenergic stimulation (0.1mL; 10- 5M). Left ventricle isovolumic pressure curve and its first derivative (dP/dt) were registered for the construction of the PP. The measures of Ө and PPA were obtained by using the Autocad 2004 software. Results are expressed as mean ± SEM. Statistical analysis: ANOVA 1 and/or 2 way; Fisher post-hoc test, significant for p<0,05. Slopes obtained after stabilization at DP of 10mmHg and during the performance of ventricular function curves were larger in the SHR and ISO groups. 26 The increase of the stimulation frequency significantly increased θ in all groups, as well as the increase of [Ca+2 ex] and the administration of isoproterenol. PPA was higher in SHR group than others. Positive inotropics interventions had provoked proportional increase of PPA in all groups. The increase of the stimulation frequency and the lower [Ca+2 ex] resulted in reduction of the PPA. Myocardial left ventricular hypertrophy in SHR and the chronic treatment with ISO modify the speed of isovolumic pressure decay increasing the slope. Regarding the inotropic interventions the increase of the slope might be linked to the increase of speed of isovolumic pressure decay, due to inotropic action in the helical ventricular myocardial band (HVMB) segments, specifically the ascending segment that operates on diastolic phase. PPA behavior contextualized the situations where the hearts had been submitted, in respect to the Wext.
Análise proteômica de tecido cardíaco de ratos com diferente capacidade aeróbica intrínseca
(Universidade Federal do Espírito Santo, 2012-12-17) Ribeiro, Leonardo Perin; Lunz, Wellington; Figueiredo, Suely Gomes de; Mill, Jose Geraldo; Pires, Simone da Fonseca
Exercise capacity is a complex attribute that involves different physiological systems under the influence of both genetic and environmental factors. Related to genetic influence, results have shown that more than 70% of aerobic exercise capacity is intrinsically determined. In this work, a comparative proteomic approach, twodimensional gel electrophoresis (2D) combined with MALDI-TOF/TOF tandem mass spectrometry, was used to investigate possible molecular differences at the protein expression level between rats heart (left ventricle - LV) with distinct intrinsic exercise capacity. Low running performance (LRP) and high running performance (HRP) rats were categorized by a maximal exercise test protocol performed on a motor-driven treadmill, according to total distance performed (TDP). The running capacity of HRPs was 3.5 fold greater than LRPs. Protein expression profiling revealed 29 statistically significant (p<0,05) differences between HRP and LRP, and 15 of these proteins were identified by MALDI-TOF/TOF (MS and MS/MS). Robust alterations were detected in components involved in antioxidant and stress response, miofibrillar and cytoskeletal proteins. Contractile proteins were found to have special expression modification: α-myosin heavy chain-6, myosin light chain-1 and creatine kinase up regulation in LV of HRP rats on patterns in HCR. In contrast, LV of LRP rats exhibited increase in abundance of protein associated with stress response. These animals exhibited enhanced expression of the antioxidant enzyme (aldehyde dehydrogenase 2), and heat shock proteins (α-crystallin B chain, heat shock protein β-2). In addition, the cytoskeletal proteins, desmin and α-actin, were upregulated in LCRs. In conclusion, our results suggest that the increased contractile proteins levels in HCRs rats may explain, in part, the improved exercise capacity. The increased stress protein expression in LCR suggests that the LV proteome of these animals are exposed to greater stress
Análise temporal da função renal em camundongos hipercolesterolêmicos
(Universidade Federal do Espírito Santo, 2009-10-02) Balarini, Camille de Moura; Gava, Agata Lages; Meyrelles, Silvana dos Santos; Stefanon, Ivanita; Pereira, Thiago de Melo Costa
Aging is a physiological process with deleterious consequences for renal function, which could be exacerbated when concurrent with pathological situations, such as dyslipidemia. The aim of this study was to determine whether hypercholesterolemia and aging could affect the renal function in mice. Male hypercholesterolemic apolipoprotein E-deficient (ApoE, n=18) mice and their age-matched C57BL/6 (C57, n=18) control mice were studied at 2-, 4- and 8-month-old. At each time point, animals were placed in metabolic cages for 24 hours in order to analyze urine volume and urinary creatinine determination. Samples of blood were collected for serum cholesterol, urea and creatinine determination. Glomerular filtration rate (GFR) was estimated by the creatinine clearance. Urine samples were submitted to polyacrilamide gel electrophoresis to verify the presence of albuminuria and renal senescence was evaluated by senescence associated beta-galactosidase activity technique. Glomerular morphometric measurements were evaluated in 10 hematoxilin-eosin stained sections (10 µm-thickness) and mesangial expansion was evaluated by Periodic Acid Schiff staining. Expression of nNOS was measured by Western Blotting. For statistical analysis, 2-way ANOVA was used followed by Fisher’s post hoc. Differences were considered statistically significant when p<0.05. Total plasma cholesterol was increased about 5-fold in ApoE mice at both time points compared with C57 animals (C57 2 months: 94.0±5.1; ApoE 2 months: 606.0±91.3; C57 4 months: 97.1±7.2; ApoE 4 months: 493.7±44.0; C57 8 months: 116.0±10.0 and ApoE 8 months: 636.1±76.4). At 2-month-old, GFR was already markedly reduced in ApoE (187±28) when compared to C57 mice (358±92). Aging caused a significant reduction of GFR in C57 mice (4 months: 211±60 and 8 months: 81±14) although did not worse this parameter in ApoE mice (4 months: 128±42 and 8 months: 93±18). In addition, serum urea was significantly increased in ApoE animals already at 2-month-old compared with C57 mice and this difference is not time-dependent (C57 2 months: 39.8±5.2; ApoE 2 months: 64.5±7.6; C57 4 months: 41.1±7.8; ApoE 4 months: 73.5±17.9; C57 8 months: 49.1±3.5; ApoE 8 months: 77.3±7.5). Only ApoE mice at 8-montold presented albuminuria. Aging promoted markedly renal senescence in C57 animals and this occurred earlier in ApoE. No differences were found in glomeruli number nor glomerular tuft area. A significant mesangial expansion was already observed at 2-month-old ApoE mice (ApoE: 35.3±0.8 vs. C57: 29.8±0.9) and this condition was aggravated by aging in ApoE mice (4 months: 40.4±1.2 and 8 months: 41.5±2.7) and induced in age-matched C57 animals (4 months: 37.8±1.3 and 8 months: 37.4±0.7). No differences were found in nNOS expression due to neither aging nor hypercholesterolemia. These data show that hypercholesterolemia can enhance the age-related loss of renal function.
Análise temporal do número e genotoxicidade de células-tronco de medula óssea de camundongos ateroscleróticos
(Universidade Federal do Espírito Santo, 2008-08-04) Tonini, Clarissa Loureiro; Meyrelles, Silvana dos Santos; Arruda, Robéria Maria Mendes Pontes; Vasquez, Elisardo Corral; Mill, José Geraldo
Currently stem cells have been target of many studies, many of them using cellular therapy in the cardiovascular diseases. However, little is known about the mechanisms that modify function and mobilization of the stem cells when submitted to the senescence and atherosclerosis. The analysis of stem cells of the ApoE-KO mice disclosed that atherosclerosis increases the number of mesenchymal and hematopoietic cells on the bone marrow probably through the increase of the symmetrical divisions stimulated by the progression of the atherosclerotic plaque. Analysis of control group showed increased number of hematopoietic stem cells and diminished number of mesenchymal stem cells. Genotoxicity assay revealed high level of DNA fragmentation in the mononuclear cells of the bone marrow in the atherosclerotic group that occurs because of the production of free radicals in atherosclerosis, what also it was evidenced by the senescence of the analyzed aortic root. In contrast, the C57 group presented low level of DNA fragmentation and, therefore low vascular senescence. We also observed that aging increased the number of hematopoietic stem cell in this group, that is, this lineage of cells does not loss the proliferative capacity, and it does not happen with the mesenchymal lineage.

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