Downregulation of VAPB expression in motor neurons derived from induced pluripotent stem cells of ALS8 patients

Amyotrophic lateral sclerosis (ALS) is an incurable neuromuscular disease that leads to a profound loss of life quality and premature death. Around 10% of the cases are inherited and ALS8 is an autosomal dominant form of familial ALS caused by mutations in the vamp-associated protein B/C (VAPB) gene. The VAPB protein is involved in many cellular processes and it likely contributes to the pathogenesis of other forms of ALS besides ALS8. A number of successful drug tests in ALS animal models could not be translated to humans underscoring the need for novel approaches. The induced pluripotent stem cells (iPSC) technology brings new hope, since it can be used to model and investigate diseases in vitro. Here we present an additional tool to study ALS based on ALS8-iPSC. Fibroblasts from ALS8 patients and their non-carrier siblings were successfully reprogrammed to a pluripotent state and differentiated into motor neurons. We show for the first time that VAPB protein levels are reduced in ALS8-derived motor neurons but, in contrast to over-expression systems, cytoplasmic aggregates could not be identified. Our results suggest that optimal levels of VAPB may play a central role in the pathogenesis of ALS8, in agreement with the observed reduction of VAPB in sporadic ALS.

False-positive results of a rapid K39-based strip test and Chagas disease

Background: The definitive diagnosis of visceral. leishmaniasis (VL) requires invasive procedures with demonstration of amastigotes in tissue or promastigotes in culture. Unfortunately, these approaches require laboratory materials not available in poor countries where the disease is endemic. The correct diagnosis of VL is important, and made more difficult by the fact that several common tropical diseases such as malaria, disseminated tuberculosis, and enteric fever share the same clinical presentation. Serological tests have been developed to replace parasitological diagnosis in the field. A commercially available K39-based strip test for VL has been developed for this purpose. The endemic area of leishmaniasis in Brazil overlaps the endemic area of Chagas disease, a disease that can cause false-positive serological test results. The aim of this study was to evaluate the incidence of false-positive exams using a rapid test for VL in patients with Chagas disease. Methods: A rapid test based on the recombinant K39 antigen of Leishmania was used in: (1) 30 patients with confirmed Chagas disease, (2) 30 patients with a serological diagnosis of Chagas disease by ELISA, indirect immunofluorescence, indirect hemagglutination, and chemiluminescence, (3) 30 healthy patients from a non-endemic area as the control group, (4) 30 patients with confirmed VL, and (5) 20 patients with proved cutaneous leishmaniasis. Results: The sensitivity and specificity of the rapid strip test were 100% when compared with healthy volunteers and those with confirmed Chagas disease. One false-positive result occurred in the group with Chagas disease diagnosed by serological tests (specificity of 96%). Conclusion: The rapid test based on recombinant K39 is a useful diagnostic assay, and a false-positive result rarely occurs in patients with a serological diagnosis of Chagas disease. (C) 2008 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Preliminary results of a floating subirrigation system (Aquaforest system) as an alternative to conventional tree nurseries

A new methodology for tree seedling production (Aquaforest system) based on a continuous floating subirrigation system was developed and tested using creeks with different nutrient levels. Eucalyptus grandis seedlings were produced in water from polluted and clean creeks, and compared to conventional tree nursery production. The growth variables analyzed in the seedling phase were: survival, height, diameter, shoot and root dry weight, leaf area, and root/shoot ratio. Water and substrate were analyzed, as well as leaf nutritional status. Plant survival was 100% in all treatments. Height and diameter were greater in the higher nutrient water treatment. Leaf area and dry weight of plants produced in the higher nutrient water treatment were similar to those of the control. The results showed that polluted water can represent a good nutrient source. The preliminary results show that the Aquaforest system can be a viable alternative for tree seedling production. (C) 2004 Elsevier B.V. All rights reserved.

The role of apoptosis in the antigen-specific T cell hyporesponsiveness of paracoccidioidomycosis patients

Paracoccidioidomycosis is a deep endemic mycosis associated with an antigen-specific immunodeficiency. To examine the role of apoptosis in this immunodeficiency, peripheral blood mononuclear cells (PBMC) of patients with paracoccidioidomycosis and controls were stimulated with the main antigen of Paracoccidioides brasiliensis (gp43) and an unrelated fungal antigen (from Candida albicans, CMA) and analyzed for annexin V and propidium iodide staining by flow cytometry. Control PBMC proliferated well with both antigens. Patients' PBMC proliferated only with CMA, but presented higher levels of apoptosis with gp43 and CMA than in their own unstimulated cultures. Moreover, gp43-triggered apoptosis in control PBMC was lower than in those of the patients. Thus, patient but not control gp43-stimulated T cells apparently remained anergized and subsequently underwent apoptosis. While CMA-induced apoptosis is likely triggered by activation-induced cell death, this is apparently not the case in gp43-induced apoptosis because of the lack of cell cycling and IL-2 in the gp43-stimulated cultures. However, higher IL-10 levels were found in gp43-stimulated patient PBMC cultures. Addition of a neutralizing anti-IL-10 antibody to the cultures resulted in increased apoptosis levels only in gp43-stimulated patient PBMC cultures. Our results suggest that apoptosis plays a role in the patients' antigen-specific hyporesponsiveness and that IL-10 may have an antiapoptotic role. (C) 2002 Elsevier B.V. (USA).

The adaptive compensations in endocrine pancreas from glucocorticoid-treated rats are reversible after the interruption of treatment

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

Avaliacão da satisfação do usuário e da qualidade dos tratamentos endodônticos em Centros de Especialidades Odontológicas da Grande Natal-RN

Objective: To evaluate the degree of users satisfaction and technical quality of endodontic treatment in specialized dental clinics (CEO) of Grande Natal / RN between 2006 and 2008. Methodology: evaluated 282 endodontically treated teeth in CEOs through clinical and radiographic examinations. A questionnaire about the clinical condition of the tooth, evaluation of care and satisfaction with treatment was applied. Data on pre-and trans-operative were noted by the patient's clinical record. Endodontically treated teeth were examined by a specialist in endodontics, which compared with previous radiographs and current ones. The collected datas were presented descriptively by absolute numbers, percentages, averages. To determine the association between the independent and dependent variables was carried out through the bivariate association test Chi-square and Fisher exact test. Results: 79.8% presented with radiographic normal and 84.4% without pain symptoms. 8.2% of the teeth were fractured and 3.2% extracted. The persistence of the periapical lesion was associated with initial periapical status (p <0.05). 91.5% of patients are satisfied with the outcome of treatment. Such satisfaction is associated with absence of pain and an adequate esthetic tooth position (p <0.05). Conclusion: endodontic treatment in specialized dental clinics have an adequate technical quality, resulting in the success of endodontic therapy performed in these centers and that users are satisfied with the treatment

Postoperative renal function evaluation, through RIFLE criteria, of elderly patients who underwent femur fracture surgery under spinal anesthesia

Introduction. The postoperative acute renal failure (ARF) incidence in different kinds of surgery has rarely been studied. Age, cardiac dysfunction, previous renal dysfunction, intraoperative hypoperfusion, and use of nephrotoxic medications are mentioned as risk factors for ARF at the postoperative period. The postoperative ARF definition was based on the creatinine increase by the RIFLE classification (R = risk, I = injury, F = failure, L = loss, E = end stage), which corresponds to a 1.5 creatinine increase, two to three times, respectively, above the basal value. This study aimed to evaluate the postoperative ARF incidence in elderly patients who underwent femur fracture surgery under subarachnoid anesthesia and stratify it by the RIFLE criteria. Methods. Ninety patients older than 65 years under spinal anesthesia with fixed dosage of 15 mg of 0.5% isobaric bupivacaine associated with morphine 50 g were studied. Immediate postoperative creatinine was considered basal and compared with maximal creatinine evaluated at 24, 48, and 72 postoperative hours. Results. The mean age of the patients was 80.27 years. ARF incidence was 24.44% and stratified this way: R = 21.11% and I = 3.33%. Conclusions. In conclusion, the postoperative ARF incidence after femur fracture surgery in patients over 65 years was 24.44%. By analyzing the stratification based on the RIFLE classification, the incidence was categorized as Risk (R) = 21.11% and Injury (I) = 3.33%.

Clinical and pathological assessment of 82 patients with cardiovascular diseases undergoing autopsy at the Hospital das Clínicas of the Faculdade de Medicina de Botucatu from 1988 to 1993

OBJECTIVE: To evaluated the clinical diagnostic, efficiency for basic death causes in patients dying of circulatory disease and de relative frequency of those diseases. METHODS: Analysis of medical record data of 82 patients, ages from 16 to 84 years old (68 over 40 years old), whose died of circulatory disease and had undergone necropsy in the period from 1988 to 1993 years in the University Hospital of Medicine Faculty of Botucatu-UNESP, Br. RESULTS: The functional class of patients were III or IV, in 78%, and 81.7% needed urgent hospitalization. By the clinical judgment the death were by ischemic heart disease in 32 (21 acute myocardial infarction), Chagas'disease in 12, valvopathy in 11, cardiomyopathy in 7, heart failure with no specification of cardiopathy in 11 and other causes in 9. At the necropsy the death cause was ischemic heart disease in 34 patients, valvopathy in 10, Chagas'disease in 10, cardiomyopathy in 5, and heart failure with no specification of cardiopathy in 2.The concordance taxes were in thhe same order: 94,6%, 90,0%, 83.3%, 71.4% and 28.5%. CONCLUSION: There was a great efficiency of clinical diagnosis for death cause in a general university hospital. The ischemic heart disease were the main causes of death.

Influence of educational level on myocardial hypertrophy of hemodialysis patients

Introduction: Chronic renal disease is associated with a high cardiovascular risk. Data from the general population associate cardiovascular diseases with low educational level, but no study has evaluated this association in patients on hemodialysis. Objective: This study aimed at evaluating the association between educational level, hypertension, and left ventricular hypertrophy in patients on chronic hemodialysis. Methods: A standard socioeconomic questionnaire was applied to 79 hemodialysis patients at the Hospital das Clínicas of Faculdade de Medicina de Botucatu, state of São Paulo. Clinical, laboratory and echocardiographic data were obtained from medical records. The patients were divided into two groups according to the median educational level, as follows: G1, patients with three or less years of schooling; G2, patients with more than three years of schooling. Results: Blood pressure, interdialytic weight gain, and variables statistically different in the two groups (p < 0.2) underwent multiple analysis. Independent associations were stated with p < 0.05 in multiple analysis. The mean age of patients was 57 ± 12.8 years, 46 were males (57%), and 53 white (67%). The variables selected for multiple analysis were: age (p = 0.004); educational level (p < 0.0001); body mass index (p = 0.124); left ventricular diameter (p = 0.048); and left ventricular mass index (p = 0.006). Antihypertensive drugs were similar in both groups. Systolic blood pressure (p = 0.006) and years of schooling (p = 0.047) had a significant and independent correlation with left ventricular mass index. Conclusion: In hemodialysis patients, left ventricular mass associated not only with blood pressure but also with educational level.

Evaluation of hospitalized patients in terms of their knowledge related to smoking

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

Nutritional, immunological and microbiological profiles of burn patients

A burn is a lesion on an organic tissue resultant from direct or indirect action of heat on the organism. The present study aimed to evaluate the nutritional, immunological and microbiological status of burn patients at the Bauru State Hospital, São Paulo state, Brazil, in 2007. Eight patients, aged more than 18 years and injured up to 24 hours, were evaluated at the moment of hospitalization and seven days later. All victims were males with a mean age of 38 years. on average, 17.5% of their body surfaces were burned and 50% of the patients were eutrophic. There were significant alterations in levels of erythrocytes, hemoglobin, hematocrit, total protein and albumin due to increased endothelial permeability, direct destruction of proteins in the heat-affected area and blood loss from lesions or debridement. At a second moment, cytokines IL-6 and TNF-alpha had augmented significantly, with IL-6 presenting elevated levels in relation to controls at the first moment. Microbiological analysis showed that 100% of the samples collected at hospital admission were negative and after one week Staphylococcus aureus was found in all cultures. Therefore, a burn patient may be considered immunosuppressed and these results indicate significant nutritional, immunological and microbiological alterations that can interfere in his recovery.

Plasma glutathione of HIV+ patients responded positively and differently to dietary supplementation with cysteine or glutamine

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

Complete denture hygiene and nocturnal wearing habits among patients attending the Prosthodontic Department in a Dental University in Brazil

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

Programa de extrações seriadas: variáveis relacionadas com a extração de pré-molares

OBJETIVO: verificar o percentual de pacientes que necessitaram extração de dentes permanentes, pré-molares, dentre aqueles tratados com extração de dentes decíduos para correção do apinhamento primário na dentição mista, bem como analisar as possíveis variáveis relacionadas. MÉTODOS: a amostra foi composta por documentações ortodônticas de 70 pacientes na dentição permanente, cujo tratamento iniciou-se na dentição mista com planejamento de um programa de extrações seriadas (PES). Todos os prontuários foram analisados por um único examinador, no intuito de verificar se o PES havia sido cumprido com a extração de dentes permanentes ou se havia sido realizada apenas extração de dentes decíduos. Verificou-se a associação entre a extração de dentes permanentes e as variáveis padrão facial; relação sagital entre as arcadas dentárias; IMPA; proporção tamanho do segundo molar permanente inferior/espaço retromolar; mecânica de controle de espaço e discrepância de modelo (teste exato de Fisher para as variáveis categóricas e modelo de regressão logística para as variáveis numéricas). Os resultados foram considerados para p<0,05. RESULTADOS: dos pacientes que haviam sido tratados com extração de dentes decíduos para a correção do apinhamento na dentição mista, 70% necessitaram de extração de dentes permanentes. A análise estatística não mostrou associação significativa entre as variáveis estudadas e a necessidade de extração de dentes permanentes, com exceção da variável discrepância de modelo. CONCLUSÃO: a discrepância de modelo representou a principal determinante de extração de pré-molares no PES.

High incidences of DNA ploidy abnormalities in tongue squamous cell carcinoma of young patients: an international collaborative study

Aims: This multi-centre analysis assessed the DNA content of TSCC in 37 young patients (<40 years) and 28 old patients (>50 years) and determined the correlation of DNA ploidy findings with clinicopathological data.Methods and results: Image cytometry was carried out using an automated cellular imaging system on Feulgen-stained histological sections to obtain high-fidelity DNA histograms. Among young patients, 37.8% were females compared to 18.7% in the older group (P = 0.002). In total, 48.6% patients were non-smokers and 40.5% were non-drinkers compared to 10.7% non-smokers and non-drinkers in the older group (P < 0.0001). TNM, clinical stage of disease and histological grade of differentiation did not differ between groups. Tumour aneuploidy was detected in 86.5% and tetraploidy in 24.3% young patients; this was significantly greater than in the older group where 64.3% were aneuploid (P < 0.0001) and 7.2% tetraploid (P < 0.0001). The mean values of DNA index (DI) and DNA heterogeneity index as well as the percentage of cells with DI exceeding 5N were higher in young patients (P < 0.0001).Conclusions: Young patients with TSCC represent a distinct clinical entity. The high incidence of DNA ploidy abnormalities suggest that they may have increased genomic instability and indicates underlying genetic differences between TSCC in young and older patients.