Insights on PRAME and osteosarcoma by means of gene expression profiling

Osteosarcoma (OS) is the most frequent bone tumor in children and adolescents. Tumor antigens are encoded by genes that are expressed in many types of solid tumors but are silent in normal tissues, with the exception of placenta and male germ-line cells. It has been proposed that antigen tumors are potential tumor markers. The premise of this study is that the identification of novel OS-associated transcripts will lead to a better understanding of the events involved in OS pathogenesis and biology. We analyzed the expression of a panel of seven tumor antigens in OS samples to identify possible tumor markers. After selecting the tumor antigen expressed in most samples of the panel, gene expression profiling was used to identify osteosarcoma-associated molecular alterations. A microarray was employed because of its ability to accurately produce comprehensive expression profiles. PRAME was identified as the tumor antigen expressed in most OS samples; it was detected in 68% of the cases. Microarray results showed differences in expression for genes functioning in cell signaling and adhesion as well as extracellular matrix-related genes, implying that such tumors could indeed differ in regard to distinct patterns of tumorigenesis. The hypothesis inferred in this study was gathered mostly from available data concerning other kinds of tumors. There is circumstantial evidence that PRAME expression might be related to distinct patterns of tumorigenesis. Further investigation is needed to validate the differential expression of genes belonging to tumorigenesis-related pathways in PRAME-positive and PRAME-negative tumors.

Immunoendocrinology of the Thymus in Chagas Disease

During immune response to infectious agents, the host develops an inflammatory response which could fail to eliminate the pathogen or may become dysregulated. In this case, the ongoing response acquires a new status and turns out to be detrimental. The same elements taking part in the establishment and regulation of the inflammatory response (cytokines, chemokines, regulatory T cells and counteracting compounds like glucocorticoids) may also mediate harmful effects. Thymic disturbances seen during Trypanosoma cruzi (T. cruzi) infection fit well with this conceptual framework. After infection, this organ suffers a severe atrophy due to apoptosis-induced thymocyte exhaustion, mainly affecting the immature double-positive (DP) CD4+CD8+ population. Thymus cellularity depletion, which occurs in the absence of main immunological mediators involved in anti-T. cruzi defense, seems to be linked to a systemic cytokine/hormonal imbalance, involving a dysregulated increase in Tumor Necrosis Factor alpha (TNF-alpha) and corticosterone hormone levels. Additionally, we have found an anomalous exit of potentially autoimmune DP cells to the periphery, in parallel to a shrinkage in the compartment of natural regulatory T cells. In this context, our data clearly point to the view that the thymus is a target organ of T. cruzi infection. Preserved thymus may be essential for the development of an effective immune response against T. cruzi, but this organ is severely affected by a dysregulated circuit of proinflammatory cytokines and glucocorticoids. Also, the alterations observed in the DP population might have potential implications for the autoimmune component of human Chagas disease. Copyright (C) 2011 S. Karger AG, Basel

Gene expression profile analysis of primary glioblastomas and non-neoplastic brain tissue: identification of potential target genes by oligonucleotide microarray and real-time quantitative PCR

The prognosis of glioblastomas is still extremely poor and the discovery of novel molecular therapeutic targets can be important to optimize treatment strategies. Gene expression analyses comparing normal and neoplastic tissues have been used to identify genes associated with tumorigenesis and potential therapeutic targets. We have used this approach to identify differentially expressed genes between primary glioblastomas and non-neoplastic brain tissues. We selected 20 overexpressed genes related to cell cycle, cellular movement and growth, proliferation and cell-to-cell signaling and analyzed their expression levels by real time quantitative PCR in cDNA obtained from microdissected fresh tumor tissue from 20 patients with primary glioblastomas and from 10 samples of non-neoplastic white matter tissue. The gene expression levels were significantly higher in glioblastomas than in non-neoplastic white matter in 18 out of 20 genes analyzed: P < 0.00001 for CDKN2C, CKS2, EEF1A1, EMP3, PDPN, BNIP2, CA12, CD34, CDC42EP4, PPIE, SNAI2, GDF15 and MMP23b; and NFIA (P: 0.0001), GPS1 (P: 0.0003), LAMA1 (P: 0.002), STIM1 (P: 0.006), and TASP1 (P: 0.01). Five of these genes are located in contiguous loci at 1p31-36 and 2 at 17q24-25 and 8 of them encode surface membrane proteins. PDPN and CD34 protein expression were evaluated by immunohistochemistry and they showed concordance with the PCR results. The present results indicate the presence of 18 overexpressed genes in human primary glioblastomas that may play a significant role in the pathogenesis of these tumors and that deserve further functional investigation as attractive candidates for new therapeutic targets.

Maternal embryonic leucine zipper kinase transcript abundance correlates with malignancy grade in human astrocytomas

We have performed cDNA microarray analyses to identify gene expression differences between highly invasive glioblastoma multiforme (GBM) and typically benign pilocytic astrocytomas (PA). Despite the significant clinical and pathological differences between the 2 tumor types, only 63 genes were found to exhibit 2-fold or greater overexpression in GBM as compared to PA. Forty percent of these genes are related to the regulation of the cell cycle and mitosis. QT-PCR validation of 6 overexpressed genes: MELK, AUKB, ASPM, PRC1, IL13RA2 and KIAA0101 confirmed at least a 5-fold increase in the average expression levels in GBM. Maternal embryonic leucine zipper kinase (MELK) exhibited the most statistically significant difference. A more detailed investigation of MELK expression was undertaken to study its oncogenic relevance. In the examination of more than 100 tumors of the central nervous system, we found progressively higher expression of MELK with astrocytoma grade and a noteworthy uniformity of high level expression in GBM. Similar level of overexpression was also observed in medulloblastoma. We found neither gene promoter hypomethylation nor amplification to be a factor in MELK expression, but were able to demonstrate that MELK knockdown in malignant astrocytoma cell lines caused a reduction in proliferation and anchorage-independent growth in in vitro assays. Our results indicate that GBM and PA differ by the expression of surprisingly few genes. Among them, MELK correlated with malignancy grade in astrocytomas and represents a therapeutic target for the management of the most frequent brain tumors in adult and children. (C) 2007 Wiley-Liss, Inc.

Effects of changing blood viscosity and heart rate on vena contracta width as evaluated by color Doppler flow mapping. An in vitro study with a pulsatile flow model

Background: There is only limited knowledge on how the quantification of valvular regurgitation by color Doppler is affected by changing blood viscosity. This study was designed to evaluate the effect of changing blood viscosity on the vena contracta width using an in vitro model of valvular insufficiency capable of providing ample variation in the rate and stroke volume. Methods: We constructed a pulsatile flow model filled with human blood at varying hematocrit (15%, 35%, and 55%) and corresponding blood viscosity (blood/water viscosity: 2.6, 4.8, 9.1) levels in which jets were driven through a known orifice (7 mm(2)) into a 110 mL compliant receiving chamber (compliance: 2.2 mL/mmHg) by a pulsatile pump. In addition, we used variable pump stroke volumes (5, 7.5, and 10 mL) and rates (40, 60, and 80 ppm). Vena contracta region was imaged using a 3.5 MHz transducer. Pressure and volume in the flow model were kept constant during each experimental condition, as well as ultrasound settings. Results: Blood viscosity variation in the experimental range did not induce significant changes in vena contracta dimensions. Also, vena contracta width did not change from normal to low hematocrit and viscosity levels. A very modest increase only in vena contracta dimension was observed at very high level of blood viscosity when hematocrit was set to 55% . Pump rate, in the evaluated range, did not influence vena contracta width. These results in controlled experimental settings suggest that the vena contracta is an accurate quantitative method for quantifying valvular regurgitation even when this condition is associated with anemia, a frequent finding in patients with valvular heart disease.

Bone Deposition, Bone Resorption, and Osteosarcoma

Bone deposition and bone resorption are ongoing dynamic processes, constituting bone remodeling. Some bone tumors, such as osteosarcoma (OS), stimulate focal bone deposition. OS is the most common primary bone tumor in children and young adults. A complex network of genes regulates bone remodeling and alterations in its expression levels can influence the genesis and progression of bone diseases, including OS. We hypothesized that the expression profiles of bone remodeling regulator genes would be correlated with OS biology and clinical features. We used real-time PCR to evaluate the mRNA levels of the tartrate-resistant acid phosphatase (ACP5), colony stimulating factor-1 (CSF1R), bone morphogenetic protein 7 (BMP7), collagen, type XI, alpha 2 (COL11A2), and protein tyrosine phosphatases zeta 1 (PTPRZ1) genes, in 30 OS tumor samples and correlated with clinical and histological data. All genes analyzed, except CSF1R, were differentially expressed when compared with normal bone expression profiles. In our results, OS patients with high levels of COL11A2 mRNA showed worse overall (p = 0.041) and event free survival (p = 0.037). Also, a trend for better overall survival was observed in patients with samples showing higher expression of BMP7 (p =0.067). COL11A2 overexpression and BMP7 underexpression could collaborate to OS tumor growth, through its central role in bone remodeling process. (C) 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:1142-1148, 2010

Distribution of Hemoglobin Phenotypes in Four Different Districts of Porto Velho, Rondonia, Brazil

Hemoglobin profile studies have been carried out in four samples from different districts of Porto Velho (Rondonia State) in the western Amazonian region of Brazil: Candelaria, Bate Estaca, Hemeron (at the State Blood Bank), and Sao Carlos. Samples from 337 unrelated individuals were collected during medical and paramedical team visits by professionals from the Instituto de Pesquisa em Patologia Tropical and the Centro de Pesquisa em Patologias Tropicais (both research institutes in tropical diseases). The aim of this study is to assess the frequency of alleles in the hemoglobin system, mainly alleles HB*A, *S, and *E. The overall phenotype frequencies were FIB A,S = 0.025, HB A,E = 0.006, and HB A,A = 0.969. Samples from the blood bank subjects and samples from the homogeneous areas of Silo Carlos and Candelaria plus Bate Estaca have a chi-square of heterogeneity of 6.383 (p = 0.041) and 8.406 (p = 0.015), respectively. The allele frequencies (HB*A = 0.984, HB*S = 0.012, and HB*E = 0.003) do not significantly differ from frequencies found in other Brazilian regions.

Influence of aging, tidal volume, and respiratory rate on Doppler echocardiographic indices of diastolic function in normal male subjects: A quantitative evaluation

Background: Although the influence of respiration on ventricular filling, as evaluated by Doppler technique, and the evaluation of diastolic velocities of mitral valve annulus (MVA), as measured by Doppler tissue imaging (DTI), can provide valuable information for the study of left ventricular (LV) diastolic function, the concomitant effects of aging, tidal volume (TV), and respiratory rate (RR) on these velocities have not been quantitatively investigated. Methods: We evaluated 12 normal male volunteers (Group I) aged 20-26 years (mean: 22.8) and 8 normal subjects aged 41 to 54 years old (mean: 45.9) (Group II). Using DTI we measured peak early (E-a) and late (A(a)) velocities of longitudinal axis expansion at lateral and medial MVA. Doppler mitral and tricuspid flow velocities were measured: peak early (E) and late (A) inflow velocity, early (E-i) and late (A(i)) flow integral, and deceleration time of peak early mitral flow velocity (DT). Respiratory cycles were simultaneously recorded at RR of 9, 12, 15, and 18 cycles/min and TV of 600 and 900 mL during respiration (RESP). Results and conclusions: (1) E, A, and A(i) in MV had negligible change during respiration, but E-i was significantly reduced during inspiration; (2) DT reduced slightly with inspiration, but the change was significant only with TV of 900 mL; (3) an important increase of E in right ventricular flow was observed during inspiration; (4) variations of RR and TV did not significantly influence right and left ventricular inflow in normal subjects, in the conditions of this investigation; (5) a significant increase of E-a at medial MVA was documented during inspiration only in young subjects; (6) a significant decrease of A(a) at medial MVA was observed during inspiration in both groups of volunteers; (7) RR and TV did not influence MVA velocities in young and adult subjects; (8) a consistent reduction in E-a and a significant increase in A(a) were observed with increasing age; (9) these changes were more conspicuous and consistent than those documented in ventricular filling when young and middle-age men are compared, suggesting that the DTI is more sensitive to detect changes in diastolic function; and (10) in addition, these data suggest that, for evaluation of diastolic function, in clinical context, it is not necessary to control rigorously RR or TV.

Cytokine Signatures of Innate and Adaptive Immunity in 17DD Yellow Fever Vaccinated Children and Its Association With the Level of Neutralizing Antibody

Background. The live attenuated yellow fever (YF) vaccines have been available for decades and are considered highly effective and one of the safest vaccines worldwide. Methods. The impact of YF-17DD-antigens recall on cytokine profiles of YF-17DD-vaccinated children were characterized using short-term cultures of whole blood samples and single-cell flow cytometry. This study enrolled seroconverters and nonseroconverters after primovaccination (PV-PRNT(+) and PV-PRNT(-)), seroconverters after revaccination (RV-PRNT(+)), and unvaccinated volunteers (UV-PRNT(-)). Results. The analysis demonstrated in the PV-PRNT(+) group a balanced involvement of pro-inflammatory/regulatory adaptive immunity with a prominent participation of innate immunity pro-inflammatory events (IL-12(+) and TNF-alpha(+) NEU and MON). Using the PV-PRNT(+) cytokine signature as a reference profile, PV-PRNT(+) presented a striking lack of innate immunity proinflammatory response along with an increased adaptive regulatory profile (IL-4(+) CD4(+) T cells and IL-10(+) and IL-5(+) CD8(+) T cells). Conversely, the RV-PRNT(+) shifted the overall cytokine signatures toward an innate immunity pro-inflammatory profile and restored the adaptive regulatory response. Conclusions. The data demonstrated that the overall cytokine signature was associated with the levels of PRNT antibodies with a balanced innate/adaptive immunity with proinflammatory/regulatory profile as the hallmark of PV-PRNT(MEDIUM+), whereas a polarized regulatory response was observed in PV-PRNT(-) and a prominent proinflammatory signature was the characteristic of PV-PRNT(HIGH+).

Oposi????es a pedidos de patente de medicamentos

A Lei da Propriedade Industrial, lei n?? 9.279 de 1996, estabeleceu crit??rios para a concess??o de patentes. Desde ent??o, o Brasil passou a conceder patentes para produtos farmac??uticos. Fica proibida, assim, a produ????o e comercializa????o dos produtos e processos patenteados sem autoriza????o do titular, o que inclui a produ????o e comercializa????o de medicamentos patenteados para o tratamento de HIV/Aids. Com isso, os valores praticados passam a ser insustent??veis e, em 2007, na tentativa de reduzir o pre??o do medicamento Efavirenz, o pa??s teve de lan??ar m??o do licenciamento compuls??rio1, o que trouxe muitas discuss??es pol??ticas, j?? que o Brasil poderia sofrer retalia????es em n??vel internacional. A iniciativa de apresenta????o de oposi????es a pedidos de patentes no Instituto Nacional da Propriedade Industrial (INPI) evita a concess??o da patente, minimiza desgastes pol??ticos, reduz o gasto do Minist??rio da Sa??de na compra de medicamentos e cria compet??ncia na tecnologia descrita no pedido de patente. Com a apresenta????o de oposi????o ao pedido do medicamento Tenofovir, estima-se uma redu????o no pre??o de 75%

A????es premiadas no 14?? Concurso Inova????o na Gest??o P??blica Federal 2009

Nesta publica????o, referente ao 14?? Concurso Inova????o na Gest??o P??blica Federal, o leitor encontrar?? iniciativas inovadoras nas seguintes ??reas: Arranjos institucionais para coordena????o e/ou implementa????o de pol??ticas p??blicas (intra e inter-governamental); avalia????o e monitoramento de pol??ticas p??blicas; gest??o da informa????o; gest??o e desenvolvimento de pessoas; melhoria de processos de trabalho; planejamento, gest??o e desempenho institucional

Gest??o da informa????o

Cada vez mais, a integra????o das partes componentes de um processo se imp??e como vital, associada a necessidade de que as informa????es operacionais e gerenciais estejam dispon??veis ?? tempo e ?? hora, para servirem de apoio ?? tomada de decis??es gerenciais, com a efic??cia e a velocidade que s??o exigidas nos dias de hoje. Neste trabalho, est??o exemplificados como, atrav??s da integra????o dos processos, apoiados por um Sistema de Gest??o da Informa????o, elimina-se trabalhos redundantes, facilita-se o preenchimento de documentos, aprimora-se o fluxo dos procedimentos, disponibilizando, em tempo real, informa????es sobre o andamento dos servi??os. Este enfoque se aplica a qualquer servi??o que envolva diversas ??reas atrav??s da integra????o dos processos distribu??dos ou implantando novos procedimentos setoriais apoiados por um suporte informatizado espec??fico. Obedecendo ??s caracter??sticas particulares de cada ??rea envolvida os Sistemas integrados ou setoriais promovem um perfeito controle dos processos distribu??dos. A tomada de decis??es ?? agilizada e tem perfeito embasamento nas informa????es que est??o sempre dispon??veis no momento em que se fizerem necess??rias

Programa????o do sistema de manuten????o

Este projeto refere-se ?? implanta????o de um sistema operacional criado para sanar as dificuldades encontradas na manuten????o da ??rea de 800 ha ocupada pela Funda????o Oswaldo Cruz. O campus ?? dividido em cinco ??reas. Cada uma delas ?? visitada diariamente por um programador respons??vel que alimenta o sistema informatizado em rede com as requisi????es de servi??o, desdobrando-as em a????es internas, que por sua vez s??o distribu??das aos departamentos executantes. O sistema foi divulgado para a institui????o por meio de um Manual de Manuten????o explicando os novos procedimentos. A implanta????o do sistema possibilitou mudan??as qualitativas e quantitativas no conjunto da organiza????o

7?? Encontro Nacional das Escolas de Governo

Foi apresentada a atual situa????o do Mapeamento da Oferta de Capacita????o nas Escolas de Governo que consiste na coleta, organiza????o e sistematiza????o das informa????es sobre a oferta de capacita????o e sobre cada escola, por meio de uma comunidade virtual na plataforma Moodle, com foco na Rede Nacional de Escolas de Governo. Foram apresentados tamb??m os resultados alcan??ados pela Rede de Escolas no ??ltimo ano. Com a Comunidade Virtual e os Cursos a Dist??ncia foi poss??vel reduzir o isolamento e ampliar as capacidades das escolas de governo federais, estaduais e municipais, por meio do compartilhamento de conhecimento e experi??ncias, al??m do incentivo ?? forma????o de parcerias. Destacou-se a import??ncia da coopera????o internacional estabelecida com a Canada School of Public Service (CSPS) e com o Centro de Educa????o a Dist??ncia para Desenvolvimento Econ??mico e Tecnol??gico (CEDDET), da Espanha, que resultaram em in??meros eventos de aprendizagem. Durante o encontro, foram desenhadas a????es comuns ??s institui????es, projetando-as para 2010, bem como escolhida a Escola Nacional de Sa??de P??blica (ENSP), da Funda????o Oswaldo Cruz, no Rio de Janeiro, como sede do 3?? Encontro Nacional de Educa????o a Dist??ncia.

O patrimônio arquitetônico da saúde : discussões sobre a história da arquitetura hospitalar do século XIX

O Patrimônio Cultural da Saúde consiste nos bens materiais e imateriais que expressam o processo da saúde individual e coletiva nas suas dimensões científica, histórica e cultural. Com a inserção do Brasil, através da COC-Fiocruz e do Ministério da Saúde, na Rede Latino-americana de Patrimônio Cultural da Saúde, iniciou-se o incentivo ao estudo da história da medicina e da arquitetura hospitalar, buscando também a proteção e a salvaguarda da memória das edificações hospitalares históricas. O século XIX foi marcado pela construção de várias edificações voltadas para o controle e reclusão dos pobres, essas instituições eram: a Casa de Correção, a Santa Casa da Misericórdia, o Hospício de Pedro II, o Asilo da Mendicidade e as Instituições de acolhimento de Menores. Dessas edificações destacam-se a Santa Casa da Misericórdia, o Hospício de Pedro II e o Asilo da Mendicidade que formam o Patrimônio Arquitetônico da Saúde tombado em nível federal. O Hospital da Santa Casa da Misericórdia foi construído em 1840-1852 sob os modernos preceitos da medicina do século XIX. A edificação até hoje mantém o uso hospitalar e apresenta um estado de conservação bom em seu exterior. Porém as condições internas foram consideradas ruins devido à falta de salubridade e higiene nos ambientes. O Hospital da Santa Casa é um Hospital de Referência, realiza atendimentos ambulatoriais, cirúrgicos e de internação. O Hospício de Pedro II foi criado para atender exclusivamente os alienados do Império. O estilo neoclássico e a monumentalidade da edificação o fizeram ser reconhecido como Palácio dos Loucos. O hospício funcionou até 1944 e quatro anos depois a edificação foi cedida à Universidade do Brasil, que adaptou sua arquitetura ao uso educacional. A edificação apresenta estado de conservação regular, com exceção da área central composta pela Capela que está ruim, devido ao incêndio de 2011. O Palácio dos Loucos tornou-se Palácio Universitário, modificando sua identidade através das mudanças que foram feitas em sua arquitetura. O Asilo da Mendicidade foi criado em 1876 para fechar o pentágono asilar. A edificação panóptica buscava a efetiva observação e controle dos internos. A edificação funcionou como Asilo para mendigos até 1920, quando transformou-se em Hospital de São Francisco de Assis. Posteriormente o hospital seria transferido para a Universidade do Brasil, que funcionou como hospital escola até 1978. O Hospital foi desativado e ficou sem uso por dez anos, quando enfim voltou a funcionar como um estabelecimento destinado aos mais pobres. O conjunto da edificação é o que apresenta o pior estado de conservação, considerado de ruim a péssimo. Comprovou-se com essa pesquisa que o mais importante para a preservação das características arquitetônicas e artísticas do bem é a manutenção do uso, seja ele qual for. Os novos usos devem ser adequados também às características e à capacidade da arquitetura em questão. Através de reformas e planos adequados, os hospitais oitocentistas, que hoje se apresentam como Patrimônio Arquitetônico da Saúde, podem manter um uso similar para o qual foi construído, como uma edificação voltada à promoção da saúde da população.