Desnaturação de oxihemoglobinas humanas A, A2 e S pelo metodo de tampão isopropanol

Human oxyhaemoglobin A and A2 from normal individuals and oxyhaemoglobin S from patients with sickle cell anaemia and sickle cell trait were studied using Isopropanol/buffer method at 37°C and 40°C. Hb S was less stable than Hb A, whereas Hb A2 was considerably more stable than either. Denaturation of Hb S was dependent on temperature and its concentration. Between the patients with sickle cell trait it was not possible to verify the influence of the concentration probably due to the small range used (from 38% to 44%).

Avaliacao do efeito do diclofenaco de sodio topico sobre a midriase e sintomas oculares apos dilatacao pupilar

The aim of this study was evaluate the effectiveness of the sodium diclophenac on the mydriasis induced by drugs and on the tearing, blepharospasm and photophobia in normal individuals that received mydriatic or cycloplegic eyedrops. We observed that there was not increase of the pupil diameter with the use of the diclophenac (f = 0,52). The photophobia symptom was mainly present in the patients with dark iris and that received the cycloplegic drops (p < 0,01). The authors concluded that there is not benefit in the use of the sodium dyclophenac eyedrops for the mydriasis increase or the decrease of the ocular symptoms in routine ophthalmologic examination.

Immune reconstitution in HIV-1 infected patients treated for two years with highly active antiretroviral therapy

The aim of this paper was to evaluate the immune reconstitution of HIV-1 patients subjected to highly active antiretroviral therapy (HAART) for two years or more according to CD 45RA and CD 45RO cell count; determination of IL-2, IFN-γ, IL-4, IL-10 and TNF-α serum levels; CD 4 + T and CD 8 + T lymphocyte count; and plasma viral load (VL) determination. For this purpose, a cross sectional study was carried out in the Tropical Diseases Area, Botucatu School of Medicine, São Paulo State University, UNESP, Botucatu, São Paulo, Brazil. Between June 2001 and April 2002, 37 HIV-1 infected patients were evaluated, 13 with treatment indication but untreated (G1), 9 subjected to HAART for 5-7 months (G2), and 15 treated for two years or more (G3); both treated groups used medication regularly and without failure. Forty-nine normal individuals were studied as controls (GC-1 and GC-2). There was a tendency (p<0.10) for the predominance of two nucleoside reverse transcriptase inhibitors (NRTI) associated with one non-nucleoside reverse transcriptase inhibitor (NNRTI) regimen in G2; and two NRTI associated with a protease inhibitor (PI) in G3. Statistical differences between groups were seen for CD 45RA (G1<[G3=GC-2]; p<0.05) and CD 45RO (G1[G2=G3]; p<0.001), TNF-α serum determination ([G1>G3; G2=intermediate]>GC-1; p<0.001), IL-2 (G1<[G2=G3=GC-1]; p<0.01), IFN-γ ([G1=GC-1]>[GC-2=G3]; p<0.001), IL-4 and IL-10 ([G1=G2=G3]>GC-1; p<0.001), serum cytokine profiles, with a higher proportion of subtype 2 in G1 and mature subtype 0 in G2 and G3 (p<0.005). There was no statistical difference for CD 8 + T lymphocyte counts (G1=G2=G3; p<0.50). Consistency was seen between positive correlations of profile 1 definer cytokines (IL-2 and IFN-γ), CD 45RA and CD 45RO cells, and CD 4 + T lymphocyte counts and between positive correlations of profile 2 definer cytokines (IL-4 and IL-10) with TNF-α, and VL. The negative correlations were also consistent as they expressed the inverse of the positives. The variables with the highest number of correlations were IL-2, IFN-γ, and VL, followed by CD 45RA and CD 45RO cells, and IL-10. The variables with the lowest number of correlations were CD 4 + T and CD 8 + T lymphocytes. The results express the partial but important immune reconstitution in HIV-1 infected individuals with the interference of HAART and the importance of cytokines especially IL-2 and IFN-γ, and CD 45RA and CD 45RO cells as surrogate markers of this reconstitution.

Maximal clenching effort influence on the electromyographic activity of the trapezius muscle in healthy subjects

Alteration of the occlusion and the position of the jaw can affect the muscles of the neck, due to a relationship between the masticatory and cervical systems. Thus, the objective of this study was to verify whether the bite in maximal clenching effort, in centric occlusion, in individuals with clinically normal occlusion, and without a history of dysfunction in the masticatory system, influences the electromyographic activity of the upper trapezius muscle. A total of 19 normal individuals participated in the study, 14 of which were women (average age of 25.4 ± 4.14 years), and 5 were men (average age of 24.11 ± 3.28 years). The root mean square (RMS) amplitude and median frequency (MF) of the upper trapezium muscle with 40% and 60% of maximal voluntary contraction were analyzed under pre- and post-maximal clenching effort conditions in centric occlusion. The electromyographic signal was collected with a sampling frequency of 2. kHz and the value in RMS was obtained by a moving window of 200. ms. The paired Student's t-test was used to compare RMS amplitude and MF under pre- and post-maximal clenching effort conditions. The level of significance for each comparison was set to p<0.05. This study concluded that in individuals without a history of dysfunction of the masticatory system, maximum clenching effort in centric occlusion does not alter the electromyographic signal of the upper trapezius. © 2009 Elsevier Ltd.

Avaliação da função glomerular de cães sadios e nefropatas sob estimulação dopaminérgica

Dopamine is an endogenous compound widely used in intensive care. It has a broad spectrum of action, on the cardiovascular system and urinary tract. Increased glomerular filtration rate, renal blood flow and fractional excretion of sodium and phosphorus are expected renal effects in normal individuals, but are poorly explored in veterinary medicine. This study was conducted to evaluate the glomerular function of dogs with renal disease submitted to continuous infusion of dopamine. Different doses of dopamine were administered in healthy and nephropathic dogs. Laboratory evaluations of creatinine clearance and urinary protein/creatinine ratio were performed during and after treatments. Creatinine clearance showed dose-dependent increase in healthy dogs. In dogs with renal disease, the dose of 1μg/kg/min GFR increased slightly, without changing the urine P/C and blood pressure, while the dose of 3μg/kg/min increased urinary protein excretion.

Análise dos parâmetros clínicos periodontais e expressão genética de interferons alfa, gama e genes relacionados em indivíduos portadores de Síndrome de Down com doença periodontal

Pós-graduação em Ciências Odontológicas - FOAR

Estudo Doppler-ecocardiográfico de pacientes com talassemia major em regime de hipertransfusão sanguínea

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

ß-2 microglobulina e citocinas séricas como indicadores de falha terapêutica aos anti-retrovirais

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Os efeitos da idade sobre o padrão de assimetrias táteis, visuais e hemiespaciais em destros e canhotos

Em testes de bisecção de linhas horizontais, os indivíduos normais tendem a partilhá-las à esquerda do verdadeiro ponto zero, fenômeno este denominado de pseudonegligência. As investigações clínicas revelaram que os mecanismos de controle sobre a alocação de atenção ao lado esquerdo do corpo foram profundamente afetados quando houve lesão localizada na parte inferior do lóbulo parietal no hemisfério direito do cérebro; e mais, a omissão de atenção à esquerda da linha média do corpo foi igualmente observada em certas modalidades motoras e táteis, não se limitando apenas ao sistema visual. Neste contexto, estudos evidenciam, ainda, que mudanças na preferência e proficiência manual ocorreram durante o processo de envelhecimento. Por exemplo, diversos estudos mostraram que, com o avanço da idade, a prevalência de canhotismo tende a diminuir; além do mais, em alguns estudos, a partir da idade de 55 anos, a direção de pseudonegligência foi distribuída à direita do ponto zero. Surgem, então, três hipóteses que buscam explicar a fonte das variações através das faixas etárias: A hipótese da degradação diferencial dos hemisférios cerebrais; a hipótese de perda bilateral; e a hipótese de assimetria invariante. Neste estudo, uma amostra de 61 indivíduos foi selecionada, sendo composta de destros e canhotos, de ambos os sexos, em três faixas etárias: de 18 a 30 anos, de 35 a 55 anos e 60 anos e acima. Os indivíduos foram submetidos ao Inventário de Preferência Manual, ao Teste Tátil de Biseccionar Linhas (TTBL) e ao Teste Visual de Biseccionar Linhas (TVBL), com o objetivo de averiguar os efeitos da idade sobre a assimetria manual e hemiespacial através do desempenho dos mesmos. Os resultados, no TTBL, mostram que houve poucas diferenças significativas no desvio e as tendências que emergiram foram ou inconsistentes ou orientadas em direções contrárias às predições de Bowers & Heilman. Não foi identificada nenhuma divergência no grau e na direção do desempenho entre destros e canhotos. O modelo interdependente de ativação, não foi sustentado por estes achados, concluindo-se que a presente versão do TTBL não constitui uma medida válida da lateralidade desta função. Em contraste, no TVBL a pseudonegligência evidenciou-se consistentemente, sob condições específicas em todos os grupos. O modelo interdependente foi apoiado pela interação significativa existente entre as mãos e os hemicampos, em particular quando a mão esquerda partilhou as linhas no campo esquerdo e central. Não foi encontrada nenhuma divergência entre os canhotos e os destros (nas três faixas etárias), no nível e direção de desempenho no TVBL. Contudo, não existiu evidência empírica indicando que a degeneração precoce do hemisfério direito afetou a atenção visuoespacial no grupo de idosos. Por conseguinte, o modelo de assimetria invariante parece mais plausível.

Avaliação do bandeamento cromossômico por digestão enzimática e tratamento com solução tampão citratado

Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB

Análise molecular de pacientes com holoprosencefalia

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Análise molecular de pacientes com holoprosencefalia

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Dendritic cells from X-linked hyper-IgM patients present impaired responses to Candida albicans and Paracoccidioides brasiliensis

Background: Patients with X-linked hyper-IgM syndrome (X-HIGM) due to CD40 ligand (CD40L) mutations are susceptible to fungal pathogens; however, the underlying susceptibility mechanisms remain poorly understood. Objective: To determine whether monocyte-derived dendritic cells (DCs) from patients with X-HIGM exhibit normal responses to fungal pathogens. Methods: DCs from patients and controls were evaluated for the expression of costimulatory (CD80 and CD86) and MHC class II molecules and for their ability to produce IL-12 and IL-10 in response to Candida albicans and Paracoccidioides brasiliensis. We also evaluated the ability of C albicans- and P brasiliensis-pulsed mature DCs to induce autologous T-cell proliferation, generation of T helper (T-H) 17 cells, and production of IFN-gamma, TGF-beta, IL-4, IL-5, and IL-17. Results: Immature DCs from patients with X-HIGM showed reduced expression of CD80, CD86, and HLA-DR, which could be reversed by exogenous trimeric soluble CD40L. Most important, mature DCs from patients with X-HIGM differentiated by coculturing DCs with fungi secreted minimal amounts of IL-12 but substantial amounts of IL-10 compared with mature DCs from normal individuals. Coculture of mature DCs from X-HIGM patients with autologous T cells led to low IFN-g production, whereas IL-4 and IL-5 production was increased. T-cell proliferation and IL-17 secretion were normal. Finally, in vitro incubation with soluble CD40L reversed the decreased IL-12 production and the skewed T-H(2) pattern response. Conclusion: Absence of CD40L during monocyte/DC differentiation leads to functional DC abnormalities, which may contribute to the susceptibility to fungal infections in patients with X-HIGM. (J Allergy Clin Immunol 2012; 129: 778-86.)

Large-Scale Population Analysis Challenges the Current Criteria for the Molecular Diagnosis of Fascioscapulohumeral Muscular Dystrophy

Facioscapulohumeral muscular dystrophy (FSHD) is a common hereditary myopathy causally linked to reduced numbers (<= 8) of 3.3 kilobase D4Z4 tandem repeats at 4q35. However, because individuals carrying D4Z4-reduced alleles and no FSHD and patients with FSHD and no short allele have been observed, additional markers have been proposed to support an FSHD molecular diagnosis. In particular a reduction in the number of D4Z4 elements combined with the 4A(159/161/168)PAS haplotype (which provides the possibility of expressing DUX4) is currently used as the genetic signature uniquely associated with FSHD. Here, we analyzed these DNA elements in more than 800 Italian and Brazilian samples of normal individuals unrelated to any FSHD patients. We find that 3% of healthy subjects carry alleles with a reduced number (4-8) of D4Z4 repeats on chromosome 4q and that one-third of these alleles, 1.3%, occur in combination with the 4A161PAS haplotype. We also systematically characterized the 4q35 haplotype in 253 unrelated FSHD patients. We find that only 127 of them (50.1%) carry alleles with 1-8 D4Z4 repeats associated with 4A161PAS, whereas the remaining FSHD probands carry different haplotypes or alleles with a greater number of D4Z4 repeats. The present study shows that the current genetic signature of FSHD is a common polymorphism and that only half of FSHD probands carry this molecular signature. Our results suggest that the genetic basis of FSHD, which is remarkably heterogeneous, should be revisited, because this has important implications for genetic counseling and prenatal diagnosis of at-risk families.

Differential expression of toll-like receptor mRNAs in recurrent aphthous ulceration

BACKGROUND: Toll-like receptors (TLR) are membrane proteins that recognize conserved molecules derived from bacterial, viral, fungal or host tissues. They are responsible for promoting the production of cytokines and chemokines, increasing the expression of costimulatory molecules and influencing the T Helper response (Th) toward either a Th1 or Th2 profile, thereby modulating the regulatory T cell response and controlling the integrity of the epithelial barrier. The key factors responsible for increased susceptibility to recurrent aphthous ulceration (RAU) are unclear, and because TLRs are involved in both immune regulation and control of the epithelial barrier, a deficiency in TLR activity is likely to cause increased susceptibility. METHODS: We investigated the gene expression of TLRs one through 10 in tissue samples and peripheral blood mononuclear cells (PBMC) of RAU patients in comparison to healthy controls using real-time quantitative reverse transcription PCR. RESULTS: The analysis of mRNA expression levels in oral lesion showed significant (P < 0.01) overexpression of the TLR2(similar to 6-fold) gene and decreased expression of the TLR3 (similar to 5-fold) and TLR5 (similar to 6-fold) genes in comparison with healthy oral mucosa. The analysis of mRNA expression in PBMC indicated a down-regulation of TLR5 gene expression in the cells from RAU patients (P < 0.05; similar to 2-fold). CONCLUSION: Our results support the hypothesis that a subset of RAU patients has fewer TLR expression that have been tentatively implicated in antiinflammatory effects. This derangement of TLR gene expression may cause an overlay exuberant inflammation reaction in situations where normal individuals are resistant. J Oral Pathol Med (2012) 41: 8085