Magnetic carbon nanotubes modified with proteins and hydrophilic monomers: Cytocompatibility, in-vitro toxicity assays and permeation across biological interfaces.

Carbon nanotubes are promising materials for biomedical applications like delivery systems and tissue scaffolds. In this paper, magnetic carbon nanotubes (M-CNTs) covered with bovine serum albumin (M-CNTs-BSA) or functionalized with hydrophilic monomers (M-CNTs-HL) were synthesized, characterized, and evaluated concerning their interaction with Caco-2 cells. There is no comparison between these two types of functionalization, and this study aimed to verify their influence on the material's interaction with the cells. Different concentrations of the nanotubes were applied to investigate cytotoxicity, cell metabolism, oxidative stress, apoptosis, and capability to cross biomimetic barriers. The materials showed cytocompatibility up to 100 µg mL-1 and a hemolysis rate below 2 %. Nanotubes' suspensions were allowed to permeate Caco-2 monolayers for up to 8 h under the effect of the magnetic field. Magnetic nanoparticles associated with the nanotubes allowed estimation of permeation through the monolayers, with values ranging from 0.50 to 7.19 and 0.27 to 9.30 × 10-3 µg (equivalent to 0.43 to 6.22 and 0.23 to 9.54 × 10-2 % of the initially estimated mass of magnetic nanoparticles) for cells exposed and non-exposed to the magnets, respectively. Together, these results support that the developed materials are promising for applications in biomedical and biotechnological fields.

Espiritualidade na concepção do paciente oncológico em tratamento antineoplásico / Conception of spirituality of cancer patients undergoing antineoplastic treatment / Espiritualidad en la concepción del paciente oncológico en tratamiento antineoplásico

Resumo Além de ser uma modalidade de enfrentamento, ressignificação do sofrimento e concepção de vida, a espiritualidade é fonte de esperança e confiança para pacientes oncológicos. Com base nessas premissas, objetivou-se desvelar a concepção de espiritualidade de pacientes oncológicos em tratamento antineoplásico a fim de compreender sua influência no enfrentamento da doença. Trata-se de estudo descritivo, de delineamento qualitativo, realizado no ambulatório de quimioterapia de um hospital público brasileiro. A amostra foi composta por 18 pacientes oncológicos submetidos a sessões de quimioterapia, em tratamento ambulatorial, maiores de 18 anos, de ambos os sexos. Para confecção dos resultados, utilizou-se a análise de conteúdo temática, que revelou duas categorias: 1) espiritualidade como fonte de esperança e confiança; e 2) espiritualidade como fonte de ressignificação do sofrimento e de uma renovada concepção de vida.

Abstract In addition to being a way of coping with and attributing new meanings to suffering and conceiving life, spirituality is a source of hope and confidence for cancer patients. From these premises, the aim of this study was to present the perception of spirituality of cancer patients undergoing antineoplastic treatment in order to understand its influence on how patients cope with the disease. This is a descriptive and qualitative study carried out in the outpatient chemotherapy clinic of a Brazilian public hospital. The study sample consisted of 18 cancer patients undergoing outpatient chemotherapy, over 18 years of age, of both genders. Thematic content analysis was used, which revealed two thematic categories: 1) spirituality as a source of hope and confidence; and 2) spirituality as a source of new meanings to suffering and of a renewed view of life.

Resumen Además de ser una modalidad de afrontamiento, resignificación del sufrimiento y concepción de la vida, la espiritualidad es una fuente de esperanza y confianza para los pacientes oncológicos. Partiendo de estas premisas, el objetivo de este trabajo fue desvelar la concepción de la espiritualidad de los pacientes con cáncer sometidos a tratamiento antineoplásico para comprender su influencia en el afrontamiento de la enfermedad. Se trata de un estudio descriptivo de diseño cualitativo, realizado en el ambulatorio de quimioterapia de un hospital público brasileño. La muestra se compuso de 18 pacientes oncológicos sometidos a sesiones de quimioterapia, en tratamiento ambulatorio, mayores de 18 años, de ambos sexos. Para confeccionar los resultados, se utilizó el análisis de contenido temático, que reveló dos categorías: 1) la espiritualidad como fuente de esperanza y confianza; y 2) la espiritualidad como fuente de resignificación del sufrimiento y de una concepción renovada de la vida.

Characterization of levan produced by a Paenibacillus sp. isolated from Brazilian crude oil.

A levan-type fructooligosaccharide was produced by a Paenibacillus strain isolated from Brazilian crude oil, the purity of which was 98.5% after precipitation with ethanol and dialysis. Characterization by FTIR, NMR spectroscopy, GC-FID and ESI-MS revealed that it is a mixture of linear ß(2 â†’ 6) fructosyl polymers with average degree of polymerization (DP) of 18 and branching ratio of 20. Morphological structure and physicochemical properties were investigated to assess levan microstructure, degradation temperature and thermomechanical features. Thermal Gravimetric Analysis highlighted degradation temperature of 218 °C, Differential Scanning Calorimetry (DSC) glass transition at 81.47 °C, and Dynamic Mechanical Analysis three frequency-dependent transition peaks. These peaks, corresponding to a first thermomechanical transition event at 86.60 °C related to the DSC endothermic event, a second at 170.9 °C and a third at 185.2 °C, were attributed to different glass transition temperatures of oligo and polyfructans with different DP. Levan showed high morphological versatility and technological potential for the food, nutraceutical, and pharmaceutical industries.

Characterization of phospholipid vesicles containing lauric acid: physicochemical basis for process and product development.

Lauric acid (LAH) strongly inhibits the growth of acne-causing bacteria. LAH is essentially water-insoluble and the solubility of laurate (LA) salts are medium and temperature dependent. Hence, LAH/LA preparations are difficult to formulate. Here we fully characterized phospholipid vesicles containing up to 50 mol% LAH. Vesicles of dipalmitoylphosphatidylcholine (DPPC) containing LAH, at pHs 7.4 and 5.0, were characterized measuring size, charge, bilayer phase transition temperature (Tm) and permeability of water-soluble probes. Small angle X-ray scattering and cryotransmission electron microscopy showed multilamellar vesicles at low LAH %. Increasing LAH % had a negligible effect on particle size. An internal aqueous compartment in all vesicle's preparations, even at equimolar DPPC: LAH fractions, was demonstrated using water-soluble probes. At pH 5.0, the interaction between DPPC and LAH increased the Tm and phase transition cooperativity showing a single lipid phase formed by hydrogen-bonded DPPC: LAH complexes. At pH 7.4, vesicles containing 50 mol% LAH exhibited distinct phases, ascribed to complex formation between LAH and LA or LAH and DPPC. LAH incorporated in the vesicles minimally permeated a skin preparation at both pHs, indicating that the primary sites of LAH solubilization were the skin layers. These results provide the foundations for developing processes and products containing DPPC: LAH.

Collaborative Brazilian pediatric renal transplant registry (CoBrazPed-RTx): A report from 2004 to 2018.

The Brazilian collaborative registry for pediatric renal transplantation began in 2004 as a multicenter initiative aimed at analyzing, reporting, and disseminating the results of pediatric renal transplantation in Brazil. Data from all pediatric renal transplants performed from January 2004 to May 2018 at the 13 participating centers were analyzed. A total of 2744 pediatric renal transplants were performed in the thirteen participating centers. The median age at transplantation was 12.2 years, with the majority being male recipients (56%). The main underlying diseases were CAKUT (40.5%) and glomerulopathy (28%). 1981 (72%) of the grafts were from deceased donors (DD). Graft survival at one year (censored by death) was 94% in the live donor group (LD) and 91% in the DD group (log-rank test P < 0.01). The patient's survival at one and 5 years was 97% and 95% for the LD group and 96% and 93% for the DD group (log-rank test P = 0.02). The graft loss rate was 19% (n = 517), more frequently caused by vascular thrombosis (n = 102) and chronic graft nephropathy (n = 90). DD recipients had 1.6 (1.0-2.2) times greater chance of death and 1.5 (1.2-1.8) times greater chance of graft loss compared to LD recipients. The mortality rate was 5.4% (n = 148), mainly due to infection (n = 69) and cardiovascular disease (n = 28). The results of this collaborative pediatric renal transplant record are comparable to other international registries, although we still have a high infection rate as a cause of death.

Forensic characterization of Brazilian regional populations through massive parallel sequencing of 124 SNPs included in HID ion Ampliseq Identity Panel.

Use of Massive Parallel Sequencing (MPS) techniques has been investigated by forensic community aiming introduction of such methods in routine forensic casework analyses. Interesting features presented by MPS include high-throughput, ability to simultaneous genotyping of significant number of samples and forensic markers, workflow automation, among others. Emergence of single nucleotide polymorphism (SNP) as forensic relevant markers was facilitated in this process, since concurrent typing of larger marker sets is necessary for obtaining same levels of individual discrimination provided by other marker categories. In this context, HID Ion Ampliseq Identity Panel is a commercial solution with forensic purposes comprising simultaneous analysis of 90 highly informative autosomal SNPs and 34 Y -chromosome superior clade SNPs for male lineage haplotyping. SNP typing can be obtained with smaller amplicons, and this panel was designed for efficient processing of critical or challenging forensic samples. In this work, a sample of 432 individuals from all five Brazilian geopolitical regions was evaluated with this panel, in order to access feasibility of this panel use in a national basis. Results obtained for all five regions, including forensic parameters, show that this marker set can be efficiently employed for Brazilian nationals in human identification or kinship determination applications, due to high levels of genetic discriminative information content displayed by Brazilians. Interpopulation comparison studies were executed among Brazilian regional populations and 26 worldwide populations, in order to access genetic stratification occurrence. Some levels of population structure were identified, and impact on database design was discussed. Y-chromosome haplotyping of Brazilian samples revealed high levels of European ancestry in Brazilian male lineages, and utility of haplotyping in real forensic casework is addressed. Finally, genotyping and sequencing efficiency with this panel were addressed, as an effort to appraise the adequacy of this panel use in Brazilian national forensic demands.

Proportional weight loss in six months as a risk factor for mortality in stage IV non-small cell lung cancer / Perda de peso proporcional em seis meses como fator de risco para mortalidade no câncer de pulmão de células não pequenas estádio IV

ABSTRACT Objective: To evaluate different weight loss (WL) cut-off points as prognostic markers of 3-month survival after diagnosis of stage IV non-small cell lung cancer (NSCLC). Methods: This was a prospective study involving 104 patients with metastatic (stage IV) NSCLC who were admitted to a cancer treatment center in southern Brazil between January of 2014 and November of 2016. We evaluated total WL and WL per month, as well as WL and WL per month in the 6 months preceding the diagnosis. The patients were followed for 3 months after diagnosis. A Cox proportional hazards regression model and Kaplan-Meier curves were used in order to evaluate 3-month survival. Results: The median WL in the 6 months preceding the diagnosis was 6% (interquartile range, 0.0-12.9%). Patients with WL ≥ 5% had a median survival of 78 days, compared with 85 days for those with WL < 5% (p = 0.047). Survival at 3 months was 72% for the patients with WL ≥ 5% (p = 0.047), 61% for those with WL ≥ 10% (p < 0.001), and 45% for those with WL ≥ 15% (p < 0.001). In the multivariate analysis, the hazard ratio for risk of death was 4.51 (95% CI: 1.32-15.39) for the patients with WL ≥ 5%, 6.34 (95% CI: 2.31-17.40) for those with WL ≥ 10%, and 14.17 (95% CI: 5.06-39.65) for those with WL ≥ 15%. Conclusions: WL in the 6 months preceding the diagnosis of NSCLC is a relevant prognostic factor and appears to be directly proportional to the rate of survival at 3 months.

RESUMO Objetivo: Avaliar diferentes pontos de corte da perda de peso (PP) como marcadores prognósticos de sobrevida em 3 meses após o diagnóstico de câncer de pulmão de células não pequenas estádio IV (CPCNP). Métodos: Estudo prospectivo envolvendo 104 pacientes com CPCNP metastático (estádio IV) que foram internados em um centro de tratamento de câncer no sul do Brasil entre janeiro de 2014 e novembro de 2016. Avaliamos a PP total e PP por mês, bem como PP e PP por mês nos 6 meses anteriores ao diagnóstico. Os pacientes foram acompanhados por 3 meses após o diagnóstico. Um modelo de regressão de riscos proporcionais de Cox e curvas de Kaplan-Meier foram utilizados para avaliar a sobrevida em 3 meses. Resultados: A mediana da PP nos 6 meses anteriores ao diagnóstico foi de 6% (intervalo interquartil, 0,0-12,9%). Pacientes com PP ≥ 5% tiveram uma sobrevida mediana de 78 dias, comparados a 85 dias para aqueles com PP < 5% (p = 0,047). A sobrevida em 3 meses foi de 72% para os pacientes com PP ≥ 5% (p = 0,047), 61% para aqueles com PP ≥ 10% (p < 0,001) e 45% para aqueles com PP ≥ 15% (p < 0,001). Na análise multivariada, a taxa de risco para óbito foi de 4,51 (IC95%: 1,32-15,39) para os pacientes com PP ≥ 5%, 6,34 (IC95%: 2,31-17,40) para aqueles com PP ≥ 10%, e 14,17 (IC95%: 5,06-39,65) para aqueles com PP ≥ 15%. Conclusões: A PP nos 6 meses anteriores ao diagnóstico de CPCNP é um fator prognóstico relevante e parece ser diretamente proporcional à taxa de sobrevida em 3 meses.

Can NO-indomethacin counteract the topical gastric toxicity induced by indomethacin interactions with phospholipid bilayers?

Nitric oxide (NO)-releasing nonsteroidal anti-inflammatory drugs (NSAIDs) have been developed to overcome the gastrointestinal and cardiovascular toxicity of NSAIDs, by chemically associating a NO-releasing moiety with commercial NSAIDs. Since increasing evidence supports that NSAIDs toxicity is related to their topical actions in membrane lipids, this work aims to evaluate the impact of adding a NO-releasing moiety to parent NSAIDs regarding their effect on lipid bilayers. Thus, the interactions of NO-indomethacin and indomethacin (parent drug) with 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) bilayers were described herein at pH 3.0 and 7.4. Diverse experimental techniques were combined to characterize the partitioning and location of drugs in DMPC bilayers, and to analyze their effect on the lipid phase transition and the bilayer structure and dynamics. The partitioning of NO-indomethacin into DMPC bilayers was similar to that of charged indomethacin and smaller than that of neutral indomethacin. Both drugs were found to insert the DMPC bilayer and the membrane location of indomethacin was pH-dependent. NO-indomethacin and indomethacin induced a decrease of the main phase transition temperature of DMPC. The effect of these drugs on the bilayer structure and dynamics was dependent on diverse factors, namely drug ionization state, drug:lipid molar ratio, temperature and lipid phase. It is noteworthy that NO-indomethacin induced more pronounced alterations in the biophysical properties of DMPC bilayers than indomethacin, considering equivalent membrane concentrations. Such modifications may have in vivo implications, particularly in the gastric mucosa, where NO-NSAIDs-induced changes in the protective properties of phospholipid layers may contribute to the occurrence of adverse effects.

Proportional weight loss in six months as a risk factor for mortality in stage IV non-small cell lung cancer.

OBJECTIVE: To evaluate different weight loss (WL) cut-off points as prognostic markers of 3-month survival after diagnosis of stage IV non-small cell lung cancer (NSCLC). METHODS: This was a prospective study involving 104 patients with metastatic (stage IV) NSCLC who were admitted to a cancer treatment center in southern Brazil between January of 2014 and November of 2016. We evaluated total WL and WL per month, as well as WL and WL per month in the 6 months preceding the diagnosis. The patients were followed for 3 months after diagnosis. A Cox proportional hazards regression model and Kaplan-Meier curves were used in order to evaluate 3-month survival. RESULTS: The median WL in the 6 months preceding the diagnosis was 6% (interquartile range, 0.0-12.9%). Patients with WL ≥ 5% had a median survival of 78 days, compared with 85 days for those with WL < 5% (p = 0.047). Survival at 3 months was 72% for the patients with WL ≥ 5% (p = 0.047), 61% for those with WL ≥ 10% (p < 0.001), and 45% for those with WL ≥ 15% (p < 0.001). In the multivariate analysis, the hazard ratio for risk of death was 4.51 (95% CI: 1.32-15.39) for the patients with WL ≥ 5%, 6.34 (95% CI: 2.31-17.40) for those with WL ≥ 10%, and 14.17 (95% CI: 5.06-39.65) for those with WL ≥ 15%. CONCLUSIONS: WL in the 6 months preceding the diagnosis of NSCLC is a relevant prognostic factor and appears to be directly proportional to the rate of survival at 3 months.

Composition of pectic polysaccharides in a Portuguese apple (Malus domestica Borkh. cv Bravo de Esmolfe)

Malus domestica Borkh. cv Bravo de Esmolfe is a typical Portuguese apple cultivar classified as Protected Designation and Origin (PDO). It is a traditional product produced under strict conditions and labelled with a specific law protected designation. This cultivar presents quite good sweetness and flavor. The monosaccharide composition of the pectic polysaccharides from this traditional apple is herein reported for the first time. Based on the molar ratios obtained from the sugar composition, the presumable pectin structure could be inferred. The cell-wall polysaccharides present in the alcohol-insoluble residue (AIR) of unpeeled BE apple were sequentially fractionated. In addition, pectic material was also extracted by citric acid treatment prior to heat extraction at acidic pH. The water soluble pectin, imidazole soluble pectin and sodium carbonate soluble pectin account for 44, 16 and 40 % of the AIR, respectively. The pectic polysaccharides extracted in the presence of citric acid had lower galacturonic acid content and higher neutral sugars content. The homogalacturonan (HG) and less-substituted rhamnogalacturonan (RG) domains are extracted first. Pectin treated with citric acid has been shown to contain more substituted polymers, especially RG-I. In addition, the relatively higher Xylose/Galacturonic acid ratio found in the citric acid extract demonstrates that the xylogalacturonan (XG) domain presumably is present in the pectic material of the unpeeled BE apple.

Composition of pectic polysaccharides in a Portuguese apple (Malus domestica Borkh. cv Bravo de Esmolfe)

Malus domestica Borkh. cv Bravo de Esmolfe is a typical Portuguese apple cultivar classified as Protected Designation and Origin (PDO). It is a traditional product produced under strict conditions and labelled with a specific law protected designation. This cultivar presents quite good sweetness and flavor. The monosaccharide composition of the pectic polysaccharides from this traditional apple is herein reported for the first time. Based on the molar ratios obtained from the sugar composition, the presumable pectin structure could be inferred. The cell-wall polysaccharides present in the alcohol-insoluble residue (AIR) of unpeeled BE apple were sequentially fractionated. In addition, pectic material was also extracted by citric acid treatment prior to heat extraction at acidic pH. The water soluble pectin, imidazole soluble pectin and sodium carbonate soluble pectin account for 44, 16 and 40 % of the AIR, respectively. The pectic polysaccharides extracted in the presence of citric acid had lower galacturonic acid content and higher neutral sugars content. The homogalacturonan (HG) and less-substituted rhamnogalacturonan (RG) domains are extracted first. Pectin treated with citric acid has been shown to contain more substituted polymers, especially RG-I. In addition, the relatively higher Xylose/Galacturonic acid ratio found in the citric acid extract demonstrates that the xylogalacturonan (XG) domain presumably is present in the pectic material of the unpeeled BE apple.(AU)

Alterations in dendritic cell function in aged mice: potential implications for immunotherapy design.

It is known that immune system functions decrease with age, and that adaptive immune responses, especially CD4+ T cell function, seem to be the main affected point in immunity with aging. Dendritic cells (DC) are the major antigen presenting cell (APC), and at least part of the defects observed in adaptive immunity of aged individuals could be due to diminished potential of bone marrow to generate new DC, or defects in DC function. In this study, we investigated if the ability of aged bone marrow (BM) to generate new DC in vitro, as well as aged BM-derived DC responses to lypopolysaccharide (LPS). Because DC are important tools in newly developing anti-tumor therapies, we also studied the ability of aged DC to phagocytose and present antigen from necrotic tumor cells. We found that aged BM generated fewer DC in vitro compared to young BM. While LPS-induced DC maturation is reduced in DC of aged mice, a high TNF-alpha production is observed in aged DC even without LPS stimulation. While phagocytosis of tumor cells is not affected by age, and DC derived from aged BM show a higher TNF-alpha production in response to phagocytosis, presentation of tumor antigens was decreased in aged DC. Because class II upregulation in response to phagocytosis was similar between aged and young DC, this could indicate an age associated processing defect in the exogenous pathway. These findings suggest that age of BM used to generate DC does not impair their phagocytic ability or TNF-alpha production, however leads to a decreased yield in mature DC, reduced response to LPS, and diminished antigen processing/presentation potential. Our results are relevant to optimization DC-based vaccine design for aged populations.

Leishmaniose visceral: o que a população de três lagoas, ms, brasil, sabe sobre esta enfermidade?

RESUMO A leishmaniose visceral (LV) é um importante problema de saúde pública e as medidas de prevenção preconizadas nem sempre são conhecidas pela população. Esta pesquisa avaliou, pela aplicação de um questionário, o conhecimento que a população de Três Lagoas, MS, tem sobre esta zoonose. Dos 384 entrevistados, 100% afirmaram que já tinham conhecimento prévio da LV, 64,5% sabiam que é transmitida por inseto vetor e 65,4% sabiam que a prevenção se dá evitando o criadouro deste. Observaram-se 93,5% de respostas para manutenção do quintal limpo como medida preventiva conhecida. Pelo menos um método de prevenção era utilizado no animal por 50,5% dos entrevistados e observou-se associação estatisticamente significante entre o grupo de proprietários cujos cães nunca apresentaram leishmaniose visceral canina e o grupo que fazia uso de alguma prevenção no animal (p=0,0006). Observou-se ainda que o conhecimento não difere entre as classes econômicas, mas a adoção de medidas preventivas nos cães depende do poder aquisitivo da população. Palavras-chave: leishmaniose visceral; epidemiologia; prevenção; controle; conhecimento. Visceral leishmaniasis - what the population from the city of Três Lagoas, MS, Brazil, knows about this disease? ABSTRACT Visceral leishmaniasis (VL) is an important public health problem and the measures to prevent it are not always known by the popul

LEISHMANIOSE VISCERAL: O QUE A POPULAÇÃO DE TRÊS LAGOAS, MS, BRASIL, SABE SOBRE ESTA ENFERMIDADE?

RESUMO A leishmaniose visceral (LV) é um importante problema de saúde pública e as medidas de prevenção preconizadas nem sempre são conhecidas pela população. Esta pesquisa avaliou, pela aplicação de um questionário, o conhecimento que a população de Três Lagoas, MS, tem sobre esta zoonose. Dos 384 entrevistados, 100% afirmaram que já tinham conhecimento prévio da LV, 64,5% sabiam que é transmitida por inseto vetor e 65,4% sabiam que a prevenção se dá evitando o criadouro deste. Observaram-se 93,5% de respostas para manutenção do quintal limpo como medida preventiva conhecida. Pelo menos um método de prevenção era utilizado no animal por 50,5% dos entrevistados e observou-se associação estatisticamente significante entre o grupo de proprietários cujos cães nunca apresentaram leishmaniose visceral canina e o grupo que fazia uso de alguma prevenção no animal (p=0,0006). Observou-se ainda que o conhecimento não difere entre as classes econômicas, mas a adoção de medidas preventivas nos cães depende do poder aquisitivo da população. Palavras-chave: leishmaniose visceral; epidemiologia; prevenção; controle; conhecimento. Visceral leishmaniasis - what the population from the city of Três Lagoas, MS, Brazil, knows about this disease? ABSTRACT Visceral leishmaniasis (VL) is an important public health problem and the measures to prevent it are not always known by the popul

Dehydroepiandrosterone sulphate enhances IgG and Interferon-gamma production during immunization to tuberculosis in young but not aged mice.

Ageing of the endocrine system (endocrinosenescence) has been closely related to immunosenescence. Dehydroepiandrosterone sulphate (DHEAS), a steroid hormone produced by the adrenals with reported enhancing immunomodulatory properties, consistently decline during ageing in parallel to detrimental increase in peripheral glucocorticoids. We investigated here the adjuvant effects of DHEAS during intraperitoneal immunization to Mycobacterium tuberculosis heat shock protein 70 (mycHSP70) in old (24 months) as well as young (3 months) BALB/c mice. Both young and old mice had significantly higher Immunoglobulin G (IgG) levels following immunization. Young mice co-immunized with mycHSP70-DHEAS presented an early increase in specific IgG levels and showed increased Interferon-gamma production compared to old mice. Also, T cells of immunized young animals were consistently more resistant to the immunosuppressive effects of glucocorticoids and to DHEAS. DHEAS was not effective in modulating antigen-specific T-cell proliferation, Interleukin-2 production or percentage of recent activated T-cell subsets (CD4 + CD69 + and CD8 + CD69 +). Our data further indicate mycHSP70 as a putative good antigen in vaccine to tuberculosis. Our data also suggest that DHEAS produced adjuvant effects upon humoral and some cellular immune responses of young, but not old mice and indicate that immunization with DHEAS is capable of changing T-cell responses to steroids.

Molecular discrimination of pouched four-eyed opossums from the Mamirauá Reserve in the Brazilian Amazon

Previous cytochrome B (CytB) mtDNA studies have suggested four species for the opossum genus Philander (four-eyed opossums), three (P. mcilhennyi, P. andersoni and P. opossum) from the Amazon and one (P. frenata) from the Brazilian Atlantic forest. During a faunal survey nine specimens of Philander sp. and four of Didelphis marsupialis were collected in the Mamirauá Sustainable Reserve, Amazonas State, Brazil. Preliminary analyses based on morphology and geographical distributions were not conclusive, suggesting that Philander specimens could belong to either P. andersoni or P. opossum. In order to elucidate the relationship of this taxon to the remaining Amazonian taxa, seven Philander and two Didelphis specimens animals were sequenced for the cytB mtDNA gene and compared to other previously studied taxa. The maximum likelihood (ML), neighbor-Joining (NJ) and maximum parsimony (MP) consensus bootstrap trees depicted six groups: Didelphis., P. frenata, P andersoni, P. mcilhennyi, P.o. opossum and Philander sp. and Philander canus in a common assemblage supported by significant bootstrap values, suggesting that the Philander sp. from Mamiraua in fact belongs to the species Philander canus.

14-3-3 gene characterization and description of a second 14-3-3 isoform in both Echinococcus granulosus and E. multilocularis.

Members of the 14-3-3 protein family have been identified as regulatory molecules in intracellular signaling pathways and cell cycle control. Previously, the first Echinococcus 14-3-3 isoform (E14-3-3.1) was isolated from E. granulosus and E. multilocularis metacestode stages. Hyperexpression of this isoform was claimed to be associated with non-restricted tumor-like growth of the E. multilocularis metacestode. In this report, we describe the characterization of a 14-3-3 cDNA from E. granulosus and E. multilocularis corresponding to a second isoform of this family, E14-3-3.2. The characterized 14-3-3 gene was interrupted by two introns whose sequence and positions were conserved in both Echinococcus species. The deduced amino acid sequence of E14-3-3.2 showed 88% identity to the E14-3-3.1 isoform and 52% identity to a third Echinococcus isoform (E14-3-3.3) described by other authors. These findings, coupled to Southern blot analysis, suggest the presence of more than one 14-3-3 gene in Echinococcus. Phylogenenetically, the Echinococcus 14-3-3.1 and 14-3-3.2 isoforms appeared to cluster with zeta-type ("pro-tumorigenic") 14-3-3 isoforms from closely related organisms, whereas the E14-3-3.3 isoform grouped with 14-3-3 epsilon isoforms. The presence of more than one 14-3-3 isoform might indicate isoform-specific roles in the different parasite stages of Echinococcus.

Diffusely adherent Escherichia coli as a cause of acute diarrhea in young children in Northeast Brazil: a case-control study.

In a prospective study carried out in two urban centers in northeastern Brazil, 195 HEp-2-adherent Escherichia coli strains were isolated; 110 were identified as the only pathogen in stools of children with diarrhea, and 85 were from controls. Enteropathogenic E. coli isolates were identified in 21 children with diarrhea (8.9%) and 7 children without diarrhea (3.0%), and they were significantly associated with diarrhea (P < 0.01). Enteroaggregative E. coli strains were isolated from 40 children with diarrhea (16.9%) and 38 children without diarrhea (16.4%) and showed no correlation with diarrhea (P > 0.5). In 49 children with diarrhea (20.7%) and 40 children without diarrhea (17.3%), diffusely adherent E. coli (DAEC) isolates were detected and were not found to be associated with diarrhea (P = 0.41). However, after stratification, for children older than 12 months of age a significant correlation between DAEC infection and diarrhea was detected (P = 0.01). These results suggest that DAEC isolates should be considered potential pathogens in northeastern Brazil and also confirm the association of DAEC with age-dependent diarrhea.

Diffuse and enteroaggregative patterns of adherence of escherichia coli isolated from stools of children in northeastern Brazil

Childhood diarrheal diseases remain highly endemic in northeastern Brazil. The attributable fraction of all diarrheal diseases among children less than 2 years of age due to Escherichia coli was examined in a 2 year prospective study in two large urban centers of Brazil. Between May 1997 and June 1999, fecal E.coli isolates from 237 children with diarrhea (217 acute and 20 persistent cases) and 231 children without diarrhea (controls) attending two hospitals in Northeast Brazil were tested for their pattern of adherence to HEp-2 cells and for colony hybridization with DNA probes specific for the six pathotypes of diarrheagenic E.coli. Enteroinvasive E.coli, enterotoxigenic E.coli and enterohemorrhagic E.coli were not isolated from any children. Diffusely adherent E.coli (DAEC) and enteroaggregative E.coli (EAEC) were the most frequent isolates with similar frequencies from children with or without diarrhea. Atypical EPEC (EAF-negative) strains were isolated with similar frequency from both cases 5.5 per cente and controls 5.6 per cente. Enteropathogenic E.coli (typical EPEC) strains, characterized by localized adherence pattern of adherence, hybridization with the EAF probe, and belonging to the classical O serogroups, were significantly associated with diarrhea (P=0.03). These E.coli strains associated with diarrhea accounted for 9 per cente of all children with diarrhea. Collectively, in Northeast Brazil, E.coli strains comprise a small proportion of severe diarrhea prevalence in children.

Diffuse and enteroaggregative patterns of adherence of Escherichia coli isolated from stools of children in northeastern Brazil

Childhood diarrheal diseases remain highly endemic in northeastern Brazil. The attributable fraction of all diarrheal diseases among children less than 2 years of age due to Escherichia coli was examined in a 2-year prospective study in two large urban centers of Brazil. Between May 1997 and June 1999, fecal E. coli isolates from 237 children with diarrhea (217 acute and 20 persistent cases) and 231 children without diarrhea (controls) attending two hospitals in Northeast Brazil were tested for their pattern of adherence to HEp-2 cells and for colony hybridization with DNA probes specific for the six pathotypes of diarrheagenic E. coli. Enteroinvasive E. coli, enterotoxigenic E. coli and enterohemorrhagic E. coli were not isolated from any children. Diffusely adherent E. coli (DAEC) and enteroaggregative E. coli (EAEC) were the most frequent isolates with similar frequencies from children with or without diarrhea. Atypical EPEC (EAF-negative) strains were isolated with similiar frequency from both cases (5.5%) and controls (5.6%). Enteropathogenic E. coli (typical EPEC) strains, characterized by localized adherence pattern of adherence, hybridization with the EAF probe, and belonging to the classical O serogroups, were significantly associated with diarrhea (P = 0.03). These E. coli strains associated with diarrhea accounted for 9% of all children with diarrhea. Collectively, in Northeast Brazil, E. coli strains comprise a small proportion of severe diarrhea prevalence in children.

Na região nordeste do Brasil a doença diarréica na infância continua sendo altamente endêmica. Com o objetivo de determinar a prevalência das diferentes categorias de E. coli diarreiogênica foi realizado um estudo prospectivo de dois anos em dois grandes centros urbanos dessa região. Entre maio de 1997 a junho de 1999, foram examinadas amostras de Escherichia coli isoladas de 237 fezes de crianças com diarréia (217 aguda e 20 persistente casos) e 231 de crianças sem diarréia (controles) atendidas em dois hospitais na região nordeste do Brasil quanto a adesão a células HEp-2 e hibridização com sondas genéticas especîficas para as seis categorias de E. coli diarreiogênica. E. coli que adere difusamente (DAEC) e E. coli enteroagregativa (EAEC) foram as categorias mais frequentemente isoladas tanto em casos como em controles. E. coli enteropatogênica atípica (EPEC) foi isolada tanto em casos (5.5%) como em controles (5.6%). Amostras de E. coli enteropatogênica (EPEC típica), caracterizadas pelo padrão de adesão localizada, hibridização positiva com a sonda EAF e pertencentes aos sorogrupos clássicos foram mais frequentes em casos do que em controles, significantemente relacionadas com diarréia (P = 0.03). Essas amostras constituíram o terceiro patógeno, depois de rotavirus e Shigella spp, com predomínio significantemente maior entre os casos. Concluindo, na região nordeste do Brasil as EPEC respondem por 9% dos casos de diarréia grave em crianças menores de dois anos.