Cellular Polyamine Catalogues of the Five Classes of the PhylumProteobacteria: Distributions of Homospermidine within the ClassAlphaproteobacteria, Hydroxyputrescine within the ClassBetaproteobacteria, Norspermidine within the ClassGammaproteobacteria, and Spermine within the ClassesDeltaproteobacteria and Epsilonproteobacteria

Cellular polyamines extracted from reclassified or newly validated 47 alphaproteobacteria, 46 betaproteobacteria, 96 gammaproteobacteria, 12 deltaproteobacteria and 10 epsilonproteobacteria were analyzed by high-performance liquid chromatography. Homospermidine was widely distributed within the class Alphaproteobacteria, however, homospermidine-dominant type, spermidine-dominant type and homospermidine/spermidinedominant type were found and the three triamine profiles were genus-specific. The all genera belonging to the class Betaproteobacteria, ubiquitously contained putrescine and 2- hydroxyputrescine. Triamines were absent in almost betaproteobacteria. Many genera, including psychrophilic species, of the class Gammaproteobacteria, contained putrescine and spermidine as the major polyaminenes. Diaminopropane and norspermidine were selectively distributed in several genera of the class Gammaproteobacteria. Spermidine was the major polyamine in the classes Deltaproteobacteria and Epsilonproteobacteria. Spermine was found in some thermophiles within Betaproteobacteria, Deltaproteobacteria and Epsilonproteobacteria, suggesting that the occurrence of spermine correlate to their thermophily. Additional these polyamine catalogues serve for the classification of the phylum Proteobacteria, as a chemotaxonomic marker

Comparison of weighed food record procedures for the reference methods in two validation studies of food frequency questionnaires

Background: Although open-ended dietary assessment methods, such as weighed food records (WFRs), are generally considered to be comparable, differences between procedures may influence outcome when WFRs are conducted independently. In this paper, we assess the procedures of WFRs in two studies to describe their dietary assessment procedures and compare the subsequent outcomes. Methods: WFRs of 12 days (3 days for four seasons) were conducted as reference methods for intake data, in accordance with the study protocol, among a subsample of participants of two large cohort studies. We compared the WFR procedures descriptively. We also compared some dietary intake variables, such as the frequency of foods and dishes and contributing foods, to determine whether there were differences in the portion size distribution and intra- and inter-individual variation in nutrient intakes caused by the difference in procedures. Results: General procedures of the dietary records were conducted in accordance with the National Health and Nutrition Survey and were the same for both studies. Differences were seen in 1) selection of multiple days (non-consecutive days versus consecutive days); and 2) survey sheet recording method (individual versus family participation). However, the foods contributing to intake of energy and selected nutrients, the portion size distribution, and intra- and inter-individual variation in nutrient intakes were similar between the two studies. Conclusion: Our comparison of WFR procedures in two independent studies revealed several differences. Notwithstanding these procedural differences, however, the subsequent outcomes were similar

A radioprotective agonist for p53 transactivation

Inhibiting p53-dependent apoptosis by inhibitors of p53 is an effective strategy for preventing radiation-induced damage in hematopoietic lineages, while p53 and p21 also play radioprotective roles in the gastrointestinal epithelium. We previously identified some zinc(II) chelators, including 8-quinolinol derivatives that suppress apoptosis in attempts to discover compounds that target the zinc-binding site in p53. We found that 5-chloro-8-quinolinol (5CHQ) has a unique p53-modulating activity that shifts its transactivation from proapoptotic to protective responses including enhancing p21 induction and suppressing PUMA induction. This p53-modulating activity also influenced p53 and p53-target gene expression in unirradiated cells without inducing DNA damage. The specificity of 5CHQ for p53 and p21 was demonstrated by silencing the expression of each protein. These effects seems to be attributable to the sequence-specific alteration of p53 DNA-binding, as evaluated by chromatin immunoprecipitation and electrophoretic mobility shift assays. In addition, 5-chloro-8-methoxyquinoline itself had no antiapoptotic activity, indicating that the hydroxyl group at the 8-position is required for its antiapoptotic activity. We applied this remarkable agonistic activity to protecting the hematopoietic and gastrointestinal system in mouse irradiation models. The dose-reduction factors of 5CHQ in total-body and abdominally irradiated mice were about 1.2 and 1.3, respectively. 5CHQ effectively protected mouse epithelial stem cells from a lethal dose of abdominal irradiation. Furthermore, the specificity of 5CHQ for p53 in reducing the lethality induced by abdominal irradiation was revealed in Trp53-KO mice. These results indicate that the pharmacological upregulation of radioprotective p53-target genes is an effective strategy for addressing the gastrointestinal syndrome

Epstein-Barr virus (EBV) detection, FOXP3 expression and evaluation of EBV viral load as prognostic markers in Aids-related lymphomas

Introdução: Pacientes com infecção pelo HIV têm risco aumentado para o desenvolvimento de linfomas não-Hodgkin de células B comparado à população geral. Dentre os mecanismos que podem estar relacionados a esta patologia, encontra-se a reativação do vírus de Epstein-Barr secundária a imunossupressão. O papel do sistema imune para desenvolvimento de tumores é citado há longa data, e seu equilíbrio é mantido pelos linfócitos T regulatórios, cujo principal regulador e marcador é o fator de transcrição FOXP3. Neste estudo, avaliamos a presença de EBER e FOXP3 em amostras diagnósticas, além da carga viral para o vírus de Epstein-Barr em pacientes com linfomas relacionados à Aids a fim de avaliar e correlacionar os resultados como marcadores prognósticos nesta população. Métodos: Análise prospectiva da carga viral para Epstein-Barr no plasma e em células mononucleares do sangue periférico em 15 pacientes com linfomas relacionados à Aids acompanhados no Serviço de Hematologia do Instituto de Infectologia Emílio Ribas e do Hospital das Clínicas/Instituto do Câncer do Estado de São Paulo da Faculdade de Medicina da Universidade de São Paulo. As mensurações foram realizadas para cada paciente por reação da cadeia de polimerase em tempo real ao diagnóstico, término do tratamento e três meses após o término do tratamento. Dois grupos controles constituídos de 26 pacientes infectados pelo HIV em uso de anti-retroviral e sem diagnóstico de linfoma ou infecção oportunista e 30 indivíduos saudáveis também foram analisados para comparação da carga viral para o vírus de Epstein-Barr. Amostras coletadas por biópsia para o diagnóstico de linfoma foram submetidas a análise imuno-histoquímica para FOXP3 e para EBER por hibridização in situ. Resultados: 13 pacientes eram do sexo masculino e dois do sexo feminino, dos quais 14 foram tratados com quimioterapia e um com radioterapia de sistema nervoso central. Nove de 15 pacientes (60%) completaram o tratamento proposto e obtiveram remissão completa. A mediana da carga viral para o vírus de Epstein-Barr antes do tratamento foi 13 cópias/106 nas células mononucleares do sangue periférico (1-1472 cópias/106) e 70 cópias/mL (0-24900 cópias/mL) no plasma. Após o tratamento foi de 0,5/106 (0-109,5) e indetectável no plasma, com diminuição significativa da carga viral em células mononucleares do sangue periférico (p=0,022) e no plasma (p=0,003) ao término do tratamento em comparação ao diagnóstico. Nos pacientes em remissão completa, a carga viral para o vírus de Epstein-Barr diminuiu tanto no plasma como em células mononucleares do sangue periférico na maioria dos casos. A hibridização in situ para EBER resultou positiva em 7/15 (46,7%) casos, sendo significativamente superior no grupo de pacientes com linfomas relacionados a AIDS com mais de um sítio extralinfonodal comprometido (p=0,041) e com linfócitos T CD4 <100 células/L (p=0,026). A expressão de FOXP3 foi negativa em 15/15 (100%) dos pacientes com ARL. Conclusões: A expressão de EBER foi positiva em 7/15 (46,7%) dos pacientes com linfomas relacionados à Aids e superior de forma significativa nos pacientes com estádio mais avançado do linfoma e maior grau de imunossupressão. Observou-se diminuição estatisticamente significativa da mediana de carga viral para o vírus de Epstein-Barr em células mononucleares do sangue periférico (p=0,022) e plasma (p=0,003) após o tratamento do linfoma em comparação aos valores do diagnóstico em pacientes que atingiram remissão completa, o que poderia ser considerado um marcador prognóstico de resposta a terapiaIntroduction: Patients with HIV infection have increased risk for development of non-Hodgkins lymphoma compared to general population. Among mechanisms that could be related to this disease is the reactivation of Epstein-Barr virus infection secondary to immunosuppression. The role of immune system in development of tumors was reported a long time ago, and balance of this system is maintained by regulatory T cells; FOXP3 transcription factor is the main regulator and marker of these cells. In this study we evaluated the presence of EBER and FOXP3 in diagnostic samples, and also viral load of Epstein-Barr virus in patients with Aids-related lymphoma to evaluate and correlate the results as prognostic markers in this population. Methods: Prospective analysis of viral load of Epstein-Barr virus in plasma and peripheral blood mononuclear cells from 15 patients with Aids-related lymphoma treated at Instituto de Infectologia Emílio Ribas and Hospital das Clínicas/Instituto do Câncer do Estado de São Paulo da Faculdade de Medicina da Universidade de São Paulo. Viral load measures were performed by real time polymerase chain reaction at diagnosis of lymphoma, completion of treatment and three months afterwards. Two control groups composed by 26 HIV-positive patients in use of HAART and without diagnosis of lymphoma or opportunistic infection and 30 healthy persons were also analyzed for viral load comparison. Biopsy samples performed to lymphoma diagnosis were submitted to immunohistochemistry for FOXP3 and in situ hybridization to EBER. Results: 13 patients were male and two females, 14 were treated with chemotherapy and one with radiotherapy of central nervous system. Nine of 15 patients (60%) completed treatment achieving complete remission. Median viral load of Epstein-Barr virus before treatment was 13 copies/106 in peripheral blood mononuclear cells (1-1472 copies/106) and 70 copies/mL (0-24900 copies/mL) in plasma. After treatment it was 0,5/106 (0-109,5) and not detectable in plasma, with significant decrease of viral load in peripheral blood mononuclear cells (p=0,022) and in plasma (p=0,003) after treatment compared to diagnosis. In patients with complete remission, viral load decreased in the majority of cases. In situ hybridization for EBER was positive in 7/15 (46,7%), and significant higher in the group of patients with Aids-related lymphoma with more than one extra nodal site (p=0,041) and CD4 T-cells <100 cells/L (p=0,026). FOXP3 expression was negative in 15/15 (100%) of patients with ARL. Conclusions: EBER expression was positive in 7/15 (46,7%) of patients with Aids-related lymphoma and significantly higher in patients with advanced stages of lymphoma and higher degree of immunosuppression. Significant decrease in median viral load of Epstein-Barr virus was observed in peripheral blood mononuclear cells (p=0,022) and plasma (p=0,003) after lymphoma treatment compared to diagnosis in patients that achieved complete remission, what could be considered a prognostic marker of response to therap