RESUMO
Defective apoptosis might be involved in the pathogenesis of multiple sclerosis (MS). We evaluated apoptosis-related molecules in MS patients before and after autologous haematopoietic stem cell transplantation (AHSCT) using BCNU, Etoposide, AraC and Melphalan (BEAM) or cyclophosphamide (CY)-based conditioning regimens. Patients were followed for clinical and immunological parameters for 2 years after AHSCT. At baseline, MS patients had decreased proapoptotic BAD, BAX and FASL and increased A1 gene expression when compared with healthy counterparts. In the BEAM group, BAK, BIK, BIMEL , FAS, FASL, A1, BCL2, BCLXL , CFLIPL and CIAP2 genes were up-regulated after AHSCT. With the exception of BIK, BIMEL and A1, all genes reached levels similar to controls at day + 720 post-transplantation. Furthermore, in these patients, we observed increased CD8+ Fas+ T cell frequencies after AHSCT when compared to baseline. In the CY group, we observed increased BAX, BCLW, CFLIPL and CIAP1 and decreased BIK and BID gene expressions after transplantation. At day + 720 post-AHSCT, the expression of BAX, FAS, FASL, BCL2, BCLXL and CIAP1 was similar to that of controls. Protein analyses showed increased Bcl-2 expression before transplantation. At 1 year post-AHSCT, expression of Bak, Bim, Bcl-2, Bcl-xL and cFlip-L was decreased when compared to baseline values. In summary, our findings suggest that normalization of apoptosis-related molecules is associated with the early therapeutic effects of AHSCT in MS patients. These mechanisms may be involved in the re-establishment of immune tolerance during the first 2 years post-transplantation.
Assuntos
Apoptose/genética , Proteína 5 Relacionada à Autofagia/genética , Esclerose Múltipla/genética , Adulto , Linfócitos T CD8-Positivos/efeitos dos fármacos , Ciclofosfamida/uso terapêutico , Feminino , Expressão Gênica/genética , Transplante de Células-Tronco Hematopoéticas/métodos , Humanos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/tratamento farmacológico , Condicionamento Pré-Transplante/métodos , Transplante Autólogo/métodos , Adulto JovemRESUMO
Despite all the knowledge, the cellular and molecular mechanisms involved in myeloproliferative neoplasm (MPN) pathophysiology remain unclear. Authors have shown galectin-1 (Gal-1) and 3 playing roles in tumour angiogenesis and fibrosis, which were correlated with poor prognosis in patients with MPN. In the present study LGALS1 and LGALS3 were differently expressed between polycythemia vera, essential thrombocythemia (ET) and primary myelofibrosis (PMF) diseases. Increased LGALS3 expression was associated with a negative JAK2 V617F status mutation in leucocytes from PMF but not in patients with ET without this mutation. However, a positive Janus kinase 2 (JAK2) V617F cell line established from patients with ET (SET-2 cells) when treated with JAK inhibitor presented high levels of LGALS3. Additionally, high LGALS1 expression was found in CD34(+) cells but not in leucocytes from patients with PMF, in absence of JAK2 V617F mutation, and also in SET-2 cells treated with JAK inhibitor. Thus, our findings indicate that differential expression of LGALS1 and/or LGALS3 in patients with MPN is linked with JAK2 V617F status mutation in these diseases and state of cell differentiation.
Assuntos
Galectina 1/genética , Galectina 3/genética , Janus Quinase 2/genética , Policitemia Vera/genética , Mielofibrose Primária/genética , Trombocitemia Essencial/genética , Adulto , Substituição de Aminoácidos , Antígenos CD34/genética , Proteínas Sanguíneas , Medula Óssea/patologia , Neoplasias da Medula Óssea/diagnóstico , Neoplasias da Medula Óssea/genética , Neoplasias da Medula Óssea/metabolismo , Linhagem Celular , Galectina 1/metabolismo , Galectina 3/metabolismo , Galectinas , Regulação Neoplásica da Expressão Gênica , Humanos , Janus Quinase 2/metabolismo , Masculino , Pessoa de Meia-Idade , Mutação , Transtornos Mieloproliferativos/diagnóstico , Transtornos Mieloproliferativos/genética , Transtornos Mieloproliferativos/metabolismo , Policitemia Vera/diagnóstico , Policitemia Vera/metabolismo , Mielofibrose Primária/diagnóstico , Mielofibrose Primária/metabolismo , Trombocitemia Essencial/diagnóstico , Trombocitemia Essencial/metabolismoRESUMO
Este estudo foi conduzido com o objetivo de determinar o melhor teor de substituição da proteína do farelo de algodão pela proteína da torta de girassol em dietas para cordeiros Santa Inês, por meio da avaliação do consumo, ganho de peso, conversão alimentar e medidas morfométricas. Foram confinados, por 60 dias, 30 cordeiros da raça Santa Inês, machos inteiros, com idade e peso médio no início do experimento de 80 dias e 21,45±2,16kg, respectivamente. O delineamento experimental foi completamente casualizado, sendo os animais divididos em 5 tratamentos, com 6 repetições por tratamento, de acordo com a quantidade de proteína do farelo de algodão substituída pela proteína da torta de girassol (0, 20, 40, 60 e 80% de substituição). As variáveis peso final, consumo de matéria seca e conversão alimentar não foram afetadas pelas dietas experimentais, embora o ganho médio diário tenha sido inferior para as dietas com maior teor de torta de girassol. Para os parâmetros de consumo, verificou-se diferença apenas no consumo de extrato etéreo em relação ao peso metabólico (g/kg PV0,75), em que cordeiros recebendo dieta com maior teor de torta de girassol ingeriram maiores quantidades de extrato etéreo. Em relação às medidas morfométricas, houve decréscimo linear da altura de dorso e largura de garupa com a inclusão da torta de girassol na dieta. A substituição da proteína do farelo de algodão pela proteína da torta de girassol afetou negativamente o ganho de peso de cordeiros Santa Inês em confinamento. Porém, teve pouca ou nenhuma influência nos parâmetros de consumo e nas medidas morfométricas in vivo.(AU)
This study was conducted with the aim to determine the optimal level of protein replacement of cottonseed meal protein by sunflower cake in diets for Santa Inês lambs, through the evaluation of consumption, weight gain, feed conversion and morphometric measurements. Thirty Santa Inês lambs, bulls, with age and weight at the beginning of the experiment of 80 days and 21.45±2.16kg, respectively, were confined for 60 days. A completely randomized experimental design was used, where the animals were divided into 5 treatments with 6 replicates per treatment. For the treatments, the protein from the cottonseed meal was replaced by the protein of the sunflower cake (0, 20, 40, 60 and 80% of substitutions). For morphometric determinations, the following measurements were taken: pre-slaughter body length, leg length, leg perimeter, height of the dorsum; hip height; chest girth, hip width and chest width. Final weight, dry matter intake and feed conversion were not affected by the experimental diets, although the average daily gain was lower for diets with higher content of sunflower cake. For consumption parameters, there was a difference only in the consumption of ether extract in relation to metabolic weight (g/kg PV 0, 75), where lambs receiving diets with higher levels of sunflower cake ingested larger amounts of ether extract. Regarding the morphometric measurements, there was a linear decrease of the height of the dorsum and hip width with the inclusion of sunflower cake in the diet. The replacement of cottonseed meal protein by sunflower cake protein negatively affected the weight gain of Santa Inês lambs. However, it affected little or nothing the parameters of consumption and in vivo morphometric measurements.(AU)
Assuntos
Animais , Ovinos/metabolismo , Ração Animal/análise , Proteínas/análise , Aumento de Peso , Pesos e Medidas Corporais/veterinária , Helianthus , Dieta/veterinária , Fenômenos Fisiológicos da Nutrição AnimalRESUMO
Este estudo foi conduzido com o objetivo de determinar o melhor teor de substituição da proteína do farelo de algodão pela proteína da torta de girassol em dietas para cordeiros Santa Inês, por meio da avaliação do consumo, ganho de peso, conversão alimentar e medidas morfométricas. Foram confinados, por 60 dias, 30 cordeiros da raça Santa Inês, machos inteiros, com idade e peso médio no início do experimento de 80 dias e 21,45±2,16kg, respectivamente. O delineamento experimental foi completamente casualizado, sendo os animais divididos em 5 tratamentos, com 6 repetições por tratamento, de acordo com a quantidade de proteína do farelo de algodão substituída pela proteína da torta de girassol (0, 20, 40, 60 e 80% de substituição). As variáveis peso final, consumo de matéria seca e conversão alimentar não foram afetadas pelas dietas experimentais, embora o ganho médio diário tenha sido inferior para as dietas com maior teor de torta de girassol. Para os parâmetros de consumo, verificou-se diferença apenas no consumo de extrato etéreo em relação ao peso metabólico (g/kg PV0,75), em que cordeiros recebendo dieta com maior teor de torta de girassol ingeriram maiores quantidades de extrato etéreo. Em relação às medidas morfométricas, houve decréscimo linear da altura de dorso e largura de garupa com a inclusão da torta de girassol na dieta. A substituição da proteína do farelo de algodão pela proteína da torta de girassol afetou negativamente o ganho de peso de cordeiros Santa Inês em confinamento. Porém, teve pouca ou nenhuma influência nos parâmetros de consumo e nas medidas morfométricas in vivo.(AU)
This study was conducted with the aim to determine the optimal level of protein replacement of cottonseed meal protein by sunflower cake in diets for Santa Inês lambs, through the evaluation of consumption, weight gain, feed conversion and morphometric measurements. Thirty Santa Inês lambs, bulls, with age and weight at the beginning of the experiment of 80 days and 21.45±2.16kg, respectively, were confined for 60 days. A completely randomized experimental design was used, where the animals were divided into 5 treatments with 6 replicates per treatment. For the treatments, the protein from the cottonseed meal was replaced by the protein of the sunflower cake (0, 20, 40, 60 and 80% of substitutions). For morphometric determinations, the following measurements were taken: pre-slaughter body length, leg length, leg perimeter, height of the dorsum; hip height; chest girth, hip width and chest width. Final weight, dry matter intake and feed conversion were not affected by the experimental diets, although the average daily gain was lower for diets with higher content of sunflower cake. For consumption parameters, there was a difference only in the consumption of ether extract in relation to metabolic weight (g/kg PV 0, 75), where lambs receiving diets with higher levels of sunflower cake ingested larger amounts of ether extract. Regarding the morphometric measurements, there was a linear decrease of the height of the dorsum and hip width with the inclusion of sunflower cake in the diet. The replacement of cottonseed meal protein by sunflower cake protein negatively affected the weight gain of Santa Inês lambs. However, it affected little or nothing the parameters of consumption and in vivo morphometric measurements.(AU)
Assuntos
Animais , Ovinos/crescimento & desenvolvimento , Aumento de Peso , Helianthus , Ração Animal , BiocombustíveisRESUMO
Primary myelofibrosis (PM) is a Philadelphia-negative clonal hematopoietic stem cell disorder characterized by intense reactive changes of bone marrow stroma with collagen fibrosis, osteosclerosis and angiogenesis. PM usually affects elderly people, and approximately half of the patients present JAK2V617F mutation. PM clinical course varies from 1 to 30 years, evolving from asymptomatic into progressive bone marrow failure, symptomatic splenomegaly or acute leukemia in 10-20 % of cases. PM risk stratification is based on parameters predicting survival, and several attempts have been made to identify clinical and laboratory features that could predict PM patient survival. This study applied five prognostic scores: Dupriez, Cervantes, Mayo, IPSS and DIPSS-Plus in 62 Brazilians patients from three centers, and compared their relevance and clinical usefulness considering the scores' parameters, fibrosis, JAK2V617F mutation, splenomegaly, hepatomegaly and treatment. According to the Cervantes, Dupriez and Mayo scores, most patients were stratified into low-risk group. However, when IPSS and DIPSS-Plus were applied, most patients were classified into an intermediate range, being low risk in only 11 and 13 % of patients, respectively. Overall survival at 4 years was 84 %. The Cervantes score was the only one that remained significantly associated with survival in a multivariate analysis. In conclusion, the Cervantes score remains important to the prognostication of PM.
Assuntos
Mielofibrose Primária/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Brasil , Feminino , Humanos , Janus Quinase 2/genética , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Mielofibrose Primária/classificação , Mielofibrose Primária/enzimologia , Mielofibrose Primária/genética , Prognóstico , Análise de SobrevidaRESUMO
Type 1 diabetes (T1D) is a chronic autoimmune disease characterized by T cell-mediated destruction of pancreatic ß cells, resulting in insulin deficiency and hyperglycaemia. Recent studies have described that apoptosis impairment during central and peripheral tolerance is involved in T1D pathogenesis. In this study, the apoptosis-related gene expression in T1D patients was evaluated before and after treatment with high-dose immunosuppression followed by autologous haematopoietic stem cell transplantation (HDI-AHSCT). We also correlated gene expression results with clinical response to HDI-AHSCT. We observed a decreased expression of bad, bax and fasL pro-apoptotic genes and an increased expression of a1, bcl-x(L) and cIAP-2 anti-apoptotic genes in patients' peripheral blood mononuclear cells (PBMCs) compared to controls. After HDI-AHSCT, we found an up-regulation of fas and fasL and a down-regulation of anti-apoptotic bcl-x(L) genes expression in post-HDI-AHSCT periods compared to pre-transplantation. Additionally, the levels of bad, bax, bok, fasL, bcl-x(L) and cIAP-1 genes expression were found similar to controls 2 years after HDI-AHSCT. Furthermore, over-expression of pro-apoptotic noxa at 540 days post-HDI-AHSCT correlated positively with insulin-free patients and conversely with glutamic acid decarboxylase autoantibodies (GAD65) autoantibody levels. Taken together, the results suggest that apoptosis-related genes deregulation in patients' PBMCs might be involved in breakdown of immune tolerance and consequently contribute to T1D pathogenesis. Furthermore, HDI-AHSCT modulated the expression of some apoptotic genes towards the levels similar to controls. Possibly, the expression of these apoptotic molecules could be applied as biomarkers of clinical remission of T1D patients treated with HDI-AHSCT therapy.
Assuntos
Diabetes Mellitus Tipo 1/imunologia , Diabetes Mellitus Tipo 1/cirurgia , Proteína Ligante Fas/genética , Expressão Gênica , Transplante de Células-Tronco Hematopoéticas , Tolerância Imunológica/genética , Leucócitos Mononucleares/efeitos dos fármacos , Receptor fas/genética , Adolescente , Adulto , Apoptose/genética , Autoanticorpos/metabolismo , Regulação para Baixo , Feminino , Seguimentos , Glutamato Descarboxilase/imunologia , Humanos , Imunossupressores/administração & dosagem , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/patologia , Masculino , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/imunologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Transplante Autólogo , Regulação para Cima , Adulto Jovem , Proteína bcl-X/genética , Proteína bcl-X/imunologia , Proteína bcl-X/metabolismoRESUMO
Tumor necrosis factor-related apoptosis-inducing ligand-TNFSF10 (TRAIL), a member of the TNF-α family and a death receptor ligand, was shown to selectively kill tumor cells. Not surprisingly, TRAIL is downregulated in a variety of tumor cells, including BCR-ABL-positive leukemia. Although we know much about the molecular basis of TRAIL-mediated cell killing, the mechanism responsible for TRAIL inhibition in tumors remains elusive because (a) TRAIL can be regulated by retinoic acid (RA); (b) the tumor antigen preferentially expressed antigen of melanoma (PRAME) was shown to inhibit transcription of RA receptor target genes through the polycomb protein, enhancer of zeste homolog 2 (EZH2); and (c) we have found that TRAIL is inversely correlated with BCR-ABL in chronic myeloid leukemia (CML) patients. Thus, we decided to investigate the association of PRAME, EZH2 and TRAIL in BCR-ABL-positive leukemia. Here, we demonstrate that PRAME, but not EZH2, is upregulated in BCR-ABL cells and is associated with the progression of disease in CML patients. There is a positive correlation between PRAME and BCR-ABL and an inverse correlation between PRAME and TRAIL in these patients. Importantly, knocking down PRAME or EZH2 by RNA interference in a BCR-ABL-positive cell line restores TRAIL expression. Moreover, there is an enrichment of EZH2 binding on the promoter region of TRAIL in a CML cell line. This binding is lost after PRAME knockdown. Finally, knocking down PRAME or EZH2, and consequently induction of TRAIL expression, enhances Imatinib sensibility. Taken together, our data reveal a novel regulatory mechanism responsible for lowering TRAIL expression and provide the basis of alternative targets for combined therapeutic strategies for CML.
Assuntos
Antígenos de Neoplasias/metabolismo , Proteínas de Ligação a DNA/metabolismo , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Ligante Indutor de Apoptose Relacionado a TNF/metabolismo , Fatores de Transcrição/metabolismo , Adenocarcinoma/tratamento farmacológico , Adenocarcinoma/genética , Adenocarcinoma/metabolismo , Antígenos de Neoplasias/análise , Antineoplásicos/uso terapêutico , Benzamidas , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Carcinoma/tratamento farmacológico , Carcinoma/genética , Carcinoma/metabolismo , Linhagem Celular , Proteínas de Ligação a DNA/análise , Progressão da Doença , Proteína Potenciadora do Homólogo 2 de Zeste , Proteínas de Fusão bcr-abl/análise , Proteínas de Fusão bcr-abl/metabolismo , Humanos , Mesilato de Imatinib , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Melanoma/tratamento farmacológico , Melanoma/genética , Melanoma/metabolismo , Piperazinas/uso terapêutico , Complexo Repressor Polycomb 2 , Regiões Promotoras Genéticas , Pirimidinas/uso terapêutico , Interferência de RNA , Ligante Indutor de Apoptose Relacionado a TNF/análise , Ligante Indutor de Apoptose Relacionado a TNF/genética , Fatores de Transcrição/análise , Células Tumorais Cultivadas , Regulação para CimaRESUMO
Iron and oxidative stress have a regulatory interplay. During the oxidative burst, phagocytic cells produce free radicals such as hypochlorous acid (HOCl). Nevertheless, scarce studies evaluated the effect of either iron deficiency anemia (IDA) or anemia of chronic disease (ACD) on phagocyte function in the elderly. The aim of the present study was to determine the oxidative burst, phagocytosis, and nitric oxide (*NO) and HOCl, reactive species produced by monocytes and neutrophils in elderly with ACD or IDA. Soluble transferrin receptor, serum ferritin, and soluble transferrin receptor/log ferritin (TfR-F) index determined the iron status. The study was constituted of 39 patients aged over 60 (28 women and 11 men) recruited from the Brazilian Public Health System. Oxidative burst fluorescence intensity per neutrophil in IDA group and HOCl generation in both ACD and IDA groups were found to be lower (p < 0.05). The percentages of neutrophils and monocytes expressing phagocytosis in ACD group were found to be higher (p < 0.05). There was an overproduction of *NO from monocytes, whereas the fundamental generation of HOCl appeared to be lower. Phagocytosis, oxidative burst, and *NO and HOCl production are involved in iron metabolism regulation in elderly patients with ACD and IDA.
Assuntos
Anemia/metabolismo , Fagocitose , Espécies Reativas de Oxigênio/metabolismo , Explosão Respiratória , Idoso , Idoso de 80 Anos ou mais , Anemia Ferropriva/metabolismo , Células Cultivadas , Doença Crônica , Feminino , Sequestradores de Radicais Livres/metabolismo , Humanos , Ácido Hipocloroso/metabolismo , Masculino , Pessoa de Meia-Idade , Monócitos/metabolismo , Neutrófilos/metabolismo , Óxido Nítrico/metabolismoRESUMO
INTRODUÇÃO: Em nossa sociedade, temos observado uma oferta cada vez maior de modelos, cores, estilos, altura e diversos tipos de salto. OBJETIVO: Avaliar se o uso de calçados de salto alto influencia nas alterações posturais com base em um conjunto de variáveis mensuradas por meio da fotogrametria computadorizada. MÉTODOS: Vinte indivíduos que utilizam salto alto com freqüência (grupo 1) e 20 indivíduos que utilizam salto alto esporadicamente (grupo 2) foram fotografados no plano frontal anterior e sagital em três momentos: a) sem utilização de calçado, b) utilizando salto agulha e c) utilizando salto plataforma, sendo estas fotografias aleatorizadas e analisadas por um experimentador cego por meio da fotogrametria. A análise estatística foi realizada a partir da análise de variância em esquema fatorial 2x3, ou seja, comparando-se a freqüência do uso de salto com o tipo de calçado, com 5 por cento de significância. RESULTADOS: Apenas o ângulo protrusão da cabeça apresentou diferença quando comparados grupo 1 e 2 (p<0,01). O efeito do tipo de calçado ocorreu na variável alinhamento do joelho direito, sendo que houve diferença apenas entre o sapato agulha e os pés descalços (p=0,03); também para a variável ângulo tibiotársico, o efeito esteve presente em todos os tipos de calçado. Os demais ângulos avaliados não apresentaram diferenças entre a freqüência no uso de salto e os outros tipos de sapato. CONCLUSÕES: A freqüência do uso de salto e o tipo de salto praticamente não modificam a postura estática avaliada pela fotogrametria.
INTRODUCTION: In our society, it is observed an increasing number of models, colors, styles, heights and types of high heels. OBJECTIVE: To evaluate whether the use of high heel shoes results in postural changes, based on a set of variables measured through computerized photogrammetry. METHODS: Twenty individuals who often used high heels (group 1) and 20 individuals who only used high heels sporadically (group 2) were photographed in the frontal and sagittal planes at three conditions: a) without using footwear; b) using stiletto heels; and c) using high platform heels. These photographs were randomized and analyzed by a blinded examiner, by means of photogrammetry. Statistical analysis was performed, using a 2x3 factorial analysis of variance to compare the frequency of high heel use with the type of shoe, at the 5 percent significance level. RESULTS: Only the head protrusion angle showed a difference between groups 1 and 2 (p<0.01). The effect of the type of shoe observed in the alignment of the right knee, which only showed a difference between stiletto heels and barefoot (p=0.03). For the tibiotarsal angle variable, the effect was also observed for all types of footwear. The other angles evaluated did not present any differences regarding the frequency of high heel use and the types of shoe. CONCLUSIONS: The frequency and type of high heel practically did not change the static posture evaluated by photogrammetry.
RESUMO
CD95 (Fas/Apo-1)-mediated apoptosis was shown to occur through two distinct pathways. One involves a direct activation of caspase-3 by large amounts of caspase-8 generated at the DISC (Type I cells). The other is related to the cleavage of Bid by low concentration of caspase-8, leading to the release of cytochrome c from mitochondria and the activation of caspase-3 by the cytochrome c/APAF-1/caspase-9 apoptosome (Type II cells). It is also known that the protein synthesis inhibitor cycloheximide (CHX) sensitizes Type I cells to CD95-mediated apoptosis, but it remains contradictory whether this effect also occurs in Type II cells. Here, we show that sub-lethal doses of CHX render both Type I and Type II cells sensitive to the apoptogenic effect of anti-CD95 antibodies but not to chemotherapeutic drugs. Moreover, Bcl-2-positive Type II cells become strongly sensitive to CD95-mediated apoptosis by the addition of CHX to the cell culture. This is not the result of a restraint of the anti-apoptotic effect of Bcl-2 at the mitochondrial level since CHX-treated Type II cells still retain their resistance to chemotherapeutic drugs. Therefore, CHX treatment is granting the CD95-mediated pathway the ability to bypass the mitochondria requirement to apoptosis, much alike to what is observed in Type I cells.
Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Apoptose/fisiologia , Mitocôndrias/metabolismo , Transdução de Sinais/fisiologia , Receptor fas/metabolismo , Anticorpos/farmacologia , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/antagonistas & inibidores , Proteína Agonista de Morte Celular de Domínio Interatuante com BH3/efeitos dos fármacos , Proteína Agonista de Morte Celular de Domínio Interatuante com BH3/metabolismo , Caspases/efeitos dos fármacos , Caspases/metabolismo , Cicloeximida/farmacologia , Citocromos c/efeitos dos fármacos , Citocromos c/metabolismo , Relação Dose-Resposta a Droga , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/fisiologia , Sinergismo Farmacológico , Células HL-60 , Humanos , Mitocôndrias/efeitos dos fármacos , Inibidores da Síntese de Proteínas/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Transdução de Sinais/efeitos dos fármacos , Receptor fas/antagonistas & inibidoresRESUMO
Using S. cerevisiae as a eukaryotic cell model we have analyzed the involvement of both glutathione transferase isoforms, Gtt1 and Gtt2, in constitutive resistance and adaptive response to menadione, a quinone which can exert its toxicity as redox cycling and/or electrophiles. The detoxification properties, of these enzymes, have also been analyzed by the appearance of S-conjugates in the media. Direct exposure to menadione (20 mM/60 min) showed to be lethal for cells deficient on both Gtt1 and Gtt2 isoforms. However, after pre-treatment with a low menadione concentration, cells deficient in Gtt2 displayed reduced ability to acquire tolerance when compared with the control and the Gtt1 deficient strains. Analyzing the toxic effects of menadione we observed that the gtt2 mutant showed no reduction in lipid peroxidation levels. Moreover, measuring the levels of intracellular oxidation during menadione stress we have shown that the increase of this oxidative stress parameter was due to the capacity menadione possesses in generating reactive oxygen species (ROS) and that both GSH and Gtt2 isoform were required to enhance ROS production. Furthermore, the efflux of the menadione-GSH conjugate, which is related with detoxification of xenobiotic pathways, was not detected in the gtt2 mutant. Taken together, these results suggest that acquisition of tolerance against stress generated by menadione and the process of detoxification through S-conjugates are dependent upon Gtt2 activity. This assessment was corroborated by the increase of GTT2 expression, and not of GTT1, after menadione treatment.
Assuntos
Glutationa Transferase/deficiência , Saccharomyces cerevisiae/genética , Vitamina K 3/farmacologia , Sequência de Bases , Primers do DNA , Genótipo , Glutationa/metabolismo , Glutationa Transferase/genética , Peroxidação de Lipídeos , Mutação , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/enzimologiaRESUMO
Propor avaliacao quantitativa das assimetrias posturais e verificar a confiabilidade intra e interexaminadores e a repetibilidade do metodo. Metodos: participaram 21 voluntarios (24+-1,9 anos) que tiveram fotografados a face e o corpo todo nos planos frontal anterior, posterior e sagital. Os angulos estudados form tracados nas fotos digitais, a partir de marcadores fixos a pele em varios pontos anatomicos, que sao referencias frequentes na avaliacao postural tradicional. A analise da confiabilidade foi efetuada a partir das medidas angulares de uma mesma foto, obtidas por um unico examinador em duas ocasioes (intra-examinadores) intervaladas por um mes e de uma so medida realizada por um terceiro examinador (interexaminadores). Arepetibilidade do metodo foi analisada pelas medidas angulares tomadas por um examinador em 2 fotos diferentes do mesmo sujeito, realizadas com intervalos de uma semana. O coeficiente de correlacao intraclasse (ICC) foi aplicado com nivel de significancia de 5 por cento. Resultados: O metodo proposto apresenta significativa confiabilidade intra e interexaminadores para a maioria dos angulos estudados. No entanto, a cifose toracica apresentou niveis nao aceitaveis de confiabilidade para a analise inter (ICC=0,603) e intra-examinador (ICC=0,031). Na repetibilidade do metodo, a maioria dos angulos estudados apresentou baixo nivel de confiabildade, exceto pelos angulos de inclinacao do pe direito (ICC=0,863) e triangulos de Talles (esquerdo ICC=0,922 e direito = 0,867). Conclusao: nessas condicoes experimentais a maioria dos valores angulares obtidos pelo metodo proposto apresentou confiabilidade intra e interexaminadores. No entanto, sua repetibilidade e baixa, sugerindo que o metodo e pouco indicado para o acompanhamento de mudancas posturais
Assuntos
Face , Fotogrametria , PosturaRESUMO
Allogeneic bone marrow transplantation (alloBMT) is the only curative therapy for chronic myelogenous leukemia (CML). This success is explained by the delivery of high doses of antineoplastic agents followed by the rescue of marrow function and the induction of graft-versus-leukemia reaction mediated by allogeneic lymphocytes against host tumor cells. This reaction can also be induced by donor lymphocyte infusion (DLI) producing remission in most patients with CML who relapse after alloBMT. The immunological mechanisms involved in DLI therapy are poorly understood. We studied five CML patients in the chronic phase, who received DLI after relapsing from an HLA-identical BMT. Using flow cytometry we evaluated cellular activation and apoptosis, NK cytotoxicity, lymphocytes producing cytokines (IL-2, IL-4 and IFN-gamma), and unstimulated (in vivo) lymphocyte proliferation. In three CML patients who achieved hematological and/or cytogenetic remission after DLI we observed an increase of the percent of activation markers on T and NK cells (CD3/DR, CD3/CD25 and CD56/DR), of lymphocytes producing IL-2 and IFN-gamma, of NK activity, and of in vivo lymphocyte proliferation. These changes were not observed consistently in two of the five patients who did not achieve complete remission with DLI. The percent of apoptotic markers (Fas, FasL and Bcl-2) on lymphocytes and CD34-positive cells did not change after DLI throughout the different study periods. Taken together, these preliminary results suggest that the therapeutic effect of DLI in the chronic phase of CML is mediated by classic cytotoxic and proliferative events involving T and NK cells but not by the Fas pathway of apoptosis.
Assuntos
Transplante de Medula Óssea/efeitos adversos , Leucemia Mielogênica Crônica BCR-ABL Positiva/cirurgia , Transfusão de Linfócitos , Adulto , Transplante de Medula Óssea/imunologia , Feminino , Seguimentos , Doença Enxerto-Hospedeiro/etiologia , Doença Enxerto-Hospedeiro/imunologia , Doença Enxerto-Hospedeiro/prevenção & controle , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/imunologia , Masculino , Recidiva Local de Neoplasia/imunologia , Recidiva , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Quimeras de Transplante/imunologia , Resultado do TratamentoRESUMO
Allogeneic bone marrow transplantation (alloBMT) is the only curative therapy for chronic myelogenous leukemia (CML). This success is explained by the delivery of high doses of antineoplastic agents followed by the rescue of marrow function and the induction of graft-versus-leukemia reaction mediated by allogeneic lymphocytes against host tumor cells. This reaction can also be induced by donor lymphocyte infusion (DLI) producing remission in most patients with CML who relapse after alloBMT. The immunological mechanisms involved in DLI therapy are poorly understood. We studied five CML patients in the chronic phase, who received DLI after relapsing from an HLA-identical BMT. Using flow cytometry we evaluated cellular activation and apoptosis, NK cytotoxicity, lymphocytes producing cytokines (IL-2, IL-4 and IFN-gamma), and unstimulated (in vivo) lymphocyte proliferation. In three CML patients who achieved hematological and/or cytogenetic remission after DLI we observed an increase of the percent of activation markers on T and NK cells (CD3/DR, CD3/CD25 and CD56/DR), of lymphocytes producing IL-2 and IFN-gamma, of NK activity, and of in vivo lymphocyte proliferation. These changes were not observed consistently in two of the five patients who did not achieve complete remission with DLI. The percent of apoptotic markers (Fas, FasL and Bcl-2) on lymphocytes and CD34-positive cells did not change after DLI throughout the different study periods. Taken together, these preliminary results suggest that the therapeutic effect of DLI in the chronic phase of CML is mediated by classic cytotoxic and proliferative events involving T and NK cells but not by the Fas pathway of apoptosis.
Assuntos
Humanos , Masculino , Feminino , Adulto , Transplante de Medula Óssea , Leucemia Mielogênica Crônica BCR-ABL Positiva , Transfusão de Linfócitos , Transplante de Medula Óssea , Seguimentos , Doença Enxerto-Hospedeiro , Leucemia Mielogênica Crônica BCR-ABL Positiva , Recidiva Local de Neoplasia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Quimeras de Transplante , Resultado do TratamentoRESUMO
Natural killer (NK) cells are CD3- CD56+ and/or CD16+ cytotoxic lymphocytes that mediate first-line defense against various types of target cells without prior immunization. To assess the effect of the menstrual cycle and gender on NK activity we evaluated 30 healthy women (mean age 28.1 years, range 21-39) in follicular and luteal phases, 29 postmenopausal women (mean age 58.8 years, range 42-72) and 48 healthy men (mean age 31.6 years, range 21-40). In a flow cytometric test of NK activity, peripheral blood mononuclear effector cells were mixed with K562 targets cells labeled with DiO (3,3'-dioctadecyloxacarbocyanine perchlorate) at effector:target cell ratios of 40, 20, 10 and 5:1. Dead cells were stained with propidium iodide and results were expressed as lytic units per 10(7) cells. In addition, progesterone levels were determined in the luteal phase of the menstrual cycle of healthy women by a chemiluminescence assay. Our results showed that (1) NK cytotoxicity was higher in the follicular than in the luteal phase of the menstrual cycle (P < 0.0001); (2) postmenopausal women and men showed NK activity similar to women in the follicular phase but higher than women in the luteal phase of the menstrual cycle (P < 0.05); and (3) there was no correlation between NK activity and levels of progesterone. The data suggest that progesterone does not influence NK activity directly and that other factors may explain the reduction of NK activity in the luteal phase of the menstrual cycle.
Assuntos
Células Matadoras Naturais/imunologia , Ciclo Menstrual/imunologia , Adulto , Idoso , Citotoxicidade Imunológica , Feminino , Fase Folicular/imunologia , Humanos , Técnicas In Vitro , Células K562 , Fase Luteal/sangue , Fase Luteal/imunologia , Masculino , Menopausa/imunologia , Pessoa de Meia-Idade , Progesterona/sangue , Caracteres SexuaisRESUMO
OBJECTIVE: To evaluate the frequency of diabetes mellitus and glucose intolerance in patients with cystic fibrosis treated at the Pediatric Gastroenterology Service of HC-FMRP-USP. METHODS: A cross-sectional analytical study was conducted on a group of 25 patients with mucoviscidosis who were followed up at HC-FMRP-USP. Oral glucose tolerance tests (OGTT) were performed, with simultaneous determination of glycemia and insulinemia. Areas under the curve were obtained for glycemia (G) and insulinemia (I) and the I/G ratio was calculated and correlated with the duration of clinical manifestation and pancreatic exocrine function. RESULTS: Five patients presented alterations: one was diabetic and four had glucose intolerance and/or hyperinsulinemia. There was a direct correlation between the area under the curve for insulinemia and the duration of mucoviscidosis. A significant inverse correlation was also observed between the area under the curve for insulinemia and I/G ratio, and number of enzyme capsules/kg/day. CONCLUSIONS: The frequency of alterations in glucose homeostasis observed in patients with mucoviscidosis was higher than in the population at large (20% of the total sample and 33% of the group of patients with glycemia and insulinemia on OGTT). Therefore, it is important that glucose tolerance tests be performed systematically in patients with mucoviscidosis so that metabolic abnormalities can be early detected, and proper treatment can be initiated.
RESUMO
A polymorphism in the coagulation factor XIII gene (FXIII Val34Leu) has been recently described to confer protection for arterial and venous thrombosis and to predispose to intracerebral hemorrhage. At present it is known that FXIII Val34Leu is prevalent in Caucasians, but information upon its distribution in different ethnic groups is scarce. We investigated the prevalence of FXIIIVal34Leu in 450 unrelated subjects of four ethnic groups: 97 Caucasians (Brazilians of European descent and Portuguese), 149 Blacks (Brazilians, and Africans from Cameroon, Zaire and Angola), 40 Asians (Japanese descendents) and 164 Amerindians from South America. PCR amplification of exon 2 of FXIII gene followed by MseI restriction-digestion was employed to define the genotypes. FXIIIVal34Leu was detected in 44.3% of the Caucasians, in 28.9% of the Blacks, in 2.5% of the Asians and in 51.2% of the Amerindians. These data confirm that FXIII Val34Leu is highly prevalent in Caucasians and indicate that it is rarer in populations of African origin. The very high frequency among Amerindians indicates that FXIII Val34Leu is not absent among Asians, and since it has a very low prevalence in Japanese, a heterogeneity in its distribution in Asia may be inferred. Taken together, our data showed that FXIII Val34Leu exhibits a significant ethnic heterogeneity, a finding that is relevant for studies relating this polymorphism with thrombotic and bleeding disorders.
Assuntos
Fator XIII/genética , Polimorfismo Genético , Grupos Raciais , Frequência do Gene , Humanos , Mutação Puntual , PrevalênciaRESUMO
The antiarthritic effect of a hydroalcohol extract of Pterodon pubescens (HEPp) seeds was tested using collagen-induced arthritis (CIA) in DBA1/J mice treated with daily oral doses of HEPp in different schedules. The preventive treatment significantly reduced both the arthritic index (AI) and the CIA incidence. Using a therapeutic protocol, only the lower dose of HEPp induced a decrease in both parameters. These results provide a scientific foundation for the popular use of Pp seed infusions in rheumatoid arthritis (RA) treatment.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Artrite Reumatoide/tratamento farmacológico , Colágeno/toxicidade , Extratos Vegetais/uso terapêutico , Plantas Medicinais/química , Animais , Modelos Animais de Doenças , Masculino , Camundongos , Camundongos Endogâmicos DBA , Sementes/químicaRESUMO
Searching for the natural vector of Plasmodium juxtanucleare in an enzootic locality: Granjas Calábria (33% of the chickens infected), Jacarepaguá, in Rio de Janeiro, Brazil, 13 comparative captures of mosquitoes were carried out, simultaneously on man (out-doors) and on chicken (in a poultry-yard), between 6 and 9 p.m., from September 1988 to March 1989. Culex saltanensis was the most frequent species in captures on chicken, accounting for 41.7% of the mosquitoes collected on this bait, showing to be highly ornithophilic (90% captured on chicken versus 10% on man). Seven specimens of Cx. saltanensis were found naturally infected in Granjas Calábria: five with mature pedunculate oocysts and two with sporozoites (one in the haemocoele and one in the salivary glands). These sporozoites produced an infection by P. juxtanucleare in a chick, which had parasitemia on day 41 after inoculation. One Cx. coronator was found with mature pedunculate oocysts. Culex saltanensis was regarded as primary vector of P. juxtanucleare in Rio de Janeiro for being highly ornithophilic and in enough density to maintain the transmission, having been found with infective sporozoites in its salivary glands, and being susceptible to the parasite and able to transmit experimentally it by the bite.
Assuntos
Culex/parasitologia , Insetos Vetores/parasitologia , Plasmodium/isolamento & purificação , Animais , Brasil , Galinhas , Comportamento Alimentar/fisiologia , Feminino , Malária Aviária/etiologia , Masculino , Plasmodium/fisiologiaRESUMO
Os autores estudaram prospectivamente 160 pacientes portadores de ITU. Instituiram o tratamento convencional e seguiram os pacientes em protocolo especial. Quarenta e nove pacientes (30,62%) de ITU apresentaram fatores predisponentes no trato urinario, sendo que 34(21,25%) apresentaram ITU alta e 18 pacientes (11,25%) refluxo vesicoureteral. Apos o tratamento preconizado, 39 pacientes (24,37%) dos que apresentaram fatores predisponentes no trato urinario foram curados, salientando a cura de todos os casos de refluxo vesicoureteral. Concluiram os autores que nos casos de ITU com fatores predisponentes locais removiveis consegue-se a cura