RESUMO
Cowpea mild mottle virus (CPMMV) is a flexuous filamentous virus that belongs to the genus Carlavirus (family Betaflexiviridae). The CPMMV genome contains six open reading frames (ORFs), among which the triple gene block (TGB), encoded by ORFs 2 to 4, has been reported to encode movement proteins for different viruses. The subcellular localization of the TGB proteins of CPMMV isolate CPMMV:BR:MG:09:2 was analysed by transient expression of each protein fused to a fluorophore. Overall, the accumulation pattern and interactions among CPMMV TGB proteins (TGBp) were similar to those of their counterparts from the potex-like group. Considering these similarities, we evaluated the potential interactions between the TGB proteins of CPMMV and of potato virus X, which could complement cell-to-cell movement. The TGBp2 and TGBp3 of PVX had an effect on CPMMV TGBp1, directing it to the plasmodesmata, but the reverse was not true.
Assuntos
Carlavirus , Flexiviridae , Potexvirus , Nicotiana , Proteínas Virais/genética , Proteínas Virais/metabolismo , Carlavirus/genética , Potexvirus/genética , Flexiviridae/genéticaRESUMO
Abstract The Brazilian Cerrado has several botanical species for medicinal purposes used by traditional communities and many of these plants are not included in the list of Medicinal Plants Research Program of Central Medicines. The purpose of this study was the phytochemical screening and toxicology bioassay front of Brine shrimp L. of ethanolic extracts of eight species of plants used in folk medicine of Campo Grande, Mato Grosso do Sul, Brazil. The ethanolic extracts were submitted to phytochemical screening, determination of phenolic compounds and flavonoids. Toxicological assays were carried front of Brine shrimp according to standard methodology. The analysis TLC and HPLC-DAD confirmed the predominance of phenolic compounds and derivatives, specifically for the quercetin. The most toxic was the C. affinis DC. with death at all concentrations, the A. humile A. St. Hil. showed no mortality and the other species showed intermediate toxicity. The extracts of the investigated species are rich in phenolic compounds and derivatives, specifically quercetin, and feature toxicity between moderate to high, a fact that requires attention, since much of the traditional communities rely on herbal drugs in its raw form with preventive and curative purpose, meeting the basic health care.
Resumo O Cerrado brasileiro possui diversas espécies botânicas com fins medicinais utilizadas por comunidades tradicionais e muitas dessas plantas não estão incluídas na lista do Programa de Pesquisa de Plantas Medicinais da Central de Medicamentos. O objetivo deste estudo foi a triagem fitoquímica e o bioensaio toxicológico frente ao Brine shrimp L. de extratos etanólicos de oito espécies de plantas utilizadas na medicina popular de Campo Grande, Mato Grosso do Sul. Os extratos etanólicos foram submetidos à triagem fitoquímica, determinação de compostos fenólicos e flavonoides. Os ensaios toxicológicos foram realizados frente ao Brine shrimp de acordo com a metodologia padrão. A análise de cromatografia de camada delgada e HPLC-DAD confirmou a predominância de compostos fenólicos e derivados, especificamente para a quercetina. O mais tóxico foi o C. affinis DC. com a morte em todas as concentrações, o A. humile A. St. Hil. não apresentou mortalidade e as demais espécies apresentaram toxicidade intermediária. Os extratos das espécies investigadas são ricos em compostos fenólicos e derivados, especificamente a quercetina e apresentam toxicidade entre moderada a alta, fato que requer atenção, pois grande parte das comunidades tradicionais conta com fitoterápicos em sua forma bruta com preventivo e finalidade curativa, atendendo aos cuidados básicos de saúde.
Assuntos
Plantas Medicinais , Medicina Tradicional , Brasil , Extratos Vegetais , Compostos FitoquímicosRESUMO
O objetivo deste trabalho foi avaliar a influência da inclusão da rama de mandioca na dieta de cordeiros confinados e semiconfinados, em substituição parcial de alimentos concentrados, como forma de redução de custos de produção. O experimento foi conduzido no Centro de Tecnologia de Ovinos (CTO), município de Campo Grande, MS. Foram utilizados 56 cordeiros do grupo genético Pantaneiro, sendo 28 machos e 28 fêmeas, com pesos médios de 18,84kg±2,02kg. As dietas recebidas foram: tratamento controle com ração comercial e tratamentos contendo a parte aérea de mandioca (PAM) em substituição de 10% dessa ração. Os machos confinados obtiveram ganho de peso médio diário de 210g e os semiconfinados, de 178g. No semiconfinamento, as fêmeas controle tiveram GMD de 120g e menor desempenho entre os grupos experimentais. A inclusão PAM acarretou uma redução de 31,29% para a mesma categoria animal. Entre os sistemas de produção, o semiconfinamento apresentou maior lucratividade média, 6,4%, e uma rentabilidade de 7,99% em relação ao sistema de confinamento, que obteve 0,04% de lucratividade e 0,87% de rentabilidade. Conclui-se que a inclusão da parte aérea da mandioca aumenta a eficiência econômica dos sistemas produtivos sem afetar o desenvolvimento dos animais em terminação.(AU)
The scope of this study was to evaluate the inclusion of raw cassava aerial parts in the diet of lambs in partial substitution of rations, in order to reduce production costs. The experiment was conducted at Sheep Technology Centre (CTO) in Campo Grande City, MS. A sample of 56 lambs from Pantaneiro genetic group was used, 28 males and 28 females, average weight of 18.84kg±2.02kg. The inclusion of PAM decreased the costs by 31.29% for the same category of animal. Among the production systems the semi-feedlot, showed the highest average profitability of 6.4% and a return of 7.99%, compared to the feedlot system, which obtained 0.04% 0.87% profitability and return. In conclusion, the inclusion of the aerial part of cassava increases the economic efficiency of production systems without affecting the development of finishing animals.(AU)
Assuntos
Animais , Ovinos/metabolismo , Manihot/economia , Ração Animal/economia , EficiênciaRESUMO
The objective of this study was to analyze the factors that influence the performance of lambs in pasture confinement, by substituting a concentrate with different levels of the aerial part of cassava in relation to hematocrit indices. The experiment was conducted at the Sheep Technology Center using 56 Pantaneira sheep, of which 28 were male and 28 were female. For female lambs, the globular volume showed a decreasing quadratic effect, with a minimum inclusion of 10.24% PAM in the diet for a hematocrit index of 32.28. A linear effect was observed for the mean crude protein intake in the male lambs based on the diet provided and was estimated to be equivalent to 87.4% of the lambs' final weight. In conclusion, mean crude protein consumption is most strongly correlated with the final weight of male lambs, and NDF is most strongly correlated with the final weight of female lambs, with the level of cyanide consumption influencing the average daily weight gain in female lambs. The hematocrit level of the male lambs decreased with an increase in the inclusion of the aerial part of cassava in the ration of the animals.(AU)
O objetivo do presente trabalho foi analisar os fatores de influência no desempenho de cordeiros em confinamento a pasto, com níveis de substituição de concentrado por parte aérea de mandioca, e sua relação com os índices de hematócritos. O experimento foi conduzido no Centro de Tecnologia de Ovinos (CTO), utilizando-se 56 ovinos da raça Pantaneira, sendo 28 machos e 28 fêmeas. No volume globular do grupo de fêmeas, observou-se efeito quadrático decrescente, com o ponto de mínimo para uma inclusão de 10,24% de PAM na ração com índice de níveis de hematócritos de 32,28. O efeito linear observado para o consumo médio de PB no modelo para cordeiros machos, conforme a dieta fornecida, em função do peso final, foi estimada o equivalente a 87,4% da formação do peso final dos cordeiros. Conclui-se que o consumo médio de proteína bruta é o fator com maior correlação com o peso final dos cordeiros machos e para as fêmeas o fator com maior correlação foi FDN, os níveis de consumo de cianeto influenciaram no ganho de peso médio diário das fêmeas. O nível de hematócritos dos cordeiros machos foi reduzido com o aumento da inclusão da parte aérea da mandioca na ração dos animais.(AU)
Assuntos
Animais , Pastagens/análise , Ovinos/crescimento & desenvolvimento , Hematócrito/veterinária , Ovinos/sangue , Ração AnimalRESUMO
Abstract The Brazilian Cerrado has several botanical species for medicinal purposes used by traditional communities and many of these plants are not included in the list of Medicinal Plants Research Program of Central Medicines. The purpose of this study was the phytochemical screening and toxicology bioassay front of Brine shrimp L. of ethanolic extracts of eight species of plants used in folk medicine of Campo Grande, Mato Grosso do Sul, Brazil. The ethanolic extracts were submitted to phytochemical screening, determination of phenolic compounds and flavonoids. Toxicological assays were carried front of Brine shrimp according to standard methodology. The analysis TLC and HPLC-DAD confirmed the predominance of phenolic compounds and derivatives, specifically for the quercetin. The most toxic was the C. affinis DC. with death at all concentrations, the A. humile A. St. Hil. showed no mortality and the other species showed intermediate toxicity. The extracts of the investigated species are rich in phenolic compounds and derivatives, specifically quercetin, and feature toxicity between moderate to high, a fact that requires attention, since much of the traditional communities rely on herbal drugs in its raw form with preventive and curative purpose, meeting the basic health care.
Resumo O Cerrado brasileiro possui diversas espécies botânicas com fins medicinais utilizadas por comunidades tradicionais e muitas dessas plantas não estão incluídas na lista do Programa de Pesquisa de Plantas Medicinais da Central de Medicamentos. O objetivo deste estudo foi a triagem fitoquímica e o bioensaio toxicológico frente ao Brine shrimp L. de extratos etanólicos de oito espécies de plantas utilizadas na medicina popular de Campo Grande, Mato Grosso do Sul. Os extratos etanólicos foram submetidos à triagem fitoquímica, determinação de compostos fenólicos e flavonoides. Os ensaios toxicológicos foram realizados frente ao Brine shrimp de acordo com a metodologia padrão. A análise de cromatografia de camada delgada e HPLC-DAD confirmou a predominância de compostos fenólicos e derivados, especificamente para a quercetina. O mais tóxico foi o C. affinis DC. com a morte em todas as concentrações, o A. humile A. St. Hil. não apresentou mortalidade e as demais espécies apresentaram toxicidade intermediária. Os extratos das espécies investigadas são ricos em compostos fenólicos e derivados, especificamente a quercetina e apresentam toxicidade entre moderada a alta, fato que requer atenção, pois grande parte das comunidades tradicionais conta com fitoterápicos em sua forma bruta com preventivo e finalidade curativa, atendendo aos cuidados básicos de saúde.
RESUMO
Psychotic disorders affect ~3% of the general population and are among the most severe forms of mental diseases. In early stages of psychosis, clinical aspects may be difficult to distinguish from one another. Undifferentiated psychopathology at the first-episode of psychosis (FEP) highlights the need for biomarkers that can improve and refine differential diagnosis. We investigated gene expression differences between patients with FEP-schizophrenia spectrum (SCZ; N=53) or FEP-Mania (BD; N=16) and healthy controls (N=73). We also verified whether gene expression was correlated to severity of psychotic, manic, depressive symptoms and/or functional impairment. All participants were antipsychotic-naive. After the psychiatric interview, blood samples were collected and the expression of 12 psychotic-disorder-related genes was evaluated by quantitative PCR. AKT1 and DICER1 expression levels were higher in BD patients compared with that in SCZ patients and healthy controls, suggesting that expression of these genes is associated more specifically to manic features. Furthermore, MBP and NDEL1 expression levels were higher in SCZ and BD patients than in healthy controls, indicating that these genes are psychosis related (independent of diagnosis). No correlation was found between gene expression and severity of symptoms or functional impairment. Our findings suggest that genes related to neurodevelopment are altered in psychotic disorders, and some might support the differential diagnosis between schizophrenia and bipolar disorder, with a potential impact on the treatment of these disorders.
Assuntos
Transtorno Bipolar/genética , Regulação da Expressão Gênica/genética , Transtornos Psicóticos/genética , Esquizofrenia/genética , Psicologia do Esquizofrênico , Adulto , Proteínas de Transporte/genética , Estudos de Casos e Controles , RNA Helicases DEAD-box/genética , Diagnóstico Diferencial , Feminino , Humanos , Masculino , Proteína Básica da Mielina/genética , Proteínas Proto-Oncogênicas c-akt/genética , Escalas de Graduação Psiquiátrica/estatística & dados numéricos , Psicometria , Transtornos Psicóticos/diagnóstico , Transtornos Psicóticos/psicologia , Valores de Referência , Ribonuclease III/genética , Esquizofrenia/diagnóstico , Estatística como Assunto , Adulto JovemRESUMO
RESUMO O fato de possuir espécies com potencial econômico ilimitado, faz com que o ambiente amazônico se torne alvo constante da biopirataria e da extração predatória dos recursos. Muitas espécies deste ambiente sequer possuem catalogação e, mesmo assim, encontram risco eminente de desaparecimento. Neste âmbito, trabalhos que reúnam dados sobre estas espécies possuem grande valor científico, cultural e econômico, sendo este o estímulo que desencadeou a produção desta revisão. Aqui a espécie Calycophyllum spruceanum (Benth.) Hook. f. ex K. Schum., uma Rubiaceae amazônica, é contemplada por meio de características botânicas, indicações etnofarmacológicas e propriedades químicas. Popularmente conhecida como mulateiro, a espécie é frequentemente receitada na etnomedicina como cicatrizante e rejuvenescedor, além de ser usada no controle de manchas de pele. Como peculiaridade botânica, mulateiro apresenta tronco retilíneo com epiderme fina esverdeada que evolui para uma periderme castanho-escuro, que é anualmente renovada. Em termos químicos, há destaque para a presença de alcalóides, taninos e, sobretudo, secoiridóides (7-metoxididerrosideo, 6´-acetil-β-D-glucopiranosildiderrosideo e 8-0-tigloildiderrosideo são peculiares à espécie). Mesmo com propriedades fotoprotetoras comprovadas, C. spruceanum ainda carece de pesquisas, sobretudo àquelas voltadas para a produção ex situ da planta e àquelas que demonstrem a relação entre a ecologia da planta e a produção de metabólitos funcionais para a indústria.
ABSTRACT The Amazonian environment is a constant target for biopiracy and predatory extraction of resources due to the species with vast economic potential. Many of these species have not even been cataloged, and yet are already at imminent risk of extinction. In this paper, Calycophyllum spruceanum (Benth.) Hook. f. ex k. Schum., an Amazonian Rubiaceae, is studied with information on its botanical characteristics, ethnopharmacological uses, and chemical properties. Popularly known as mulateiro, the species is often prescribed in ethnomedicine for healing and vitality, and is also used to control skin patches. As a botanical peculiarity, it has a thin green epidermis that evolves into a dark brown periderm, which is renewed annually. In terms of chemistry, there is emphasis on the presence of alkaloids, tannins, and, especially, secoiridoids (7-methoxydiderroside, 6′-acetyl-β-D-glucopyranosyldiderroside, and 8-O-tigloyldiderroside are peculiar to the species). Even with proven photoprotective properties, research on C. spruceanum is still lacking, in particular studies aimed at the ex situ production of the plant and those that show the relationship between the plant’s ecology and the production of functional metabolites for the industry.
Assuntos
Botânica/instrumentação , Exploração de Recursos Naturais/análise , Rubiaceae/classificação , Etnofarmacologia/instrumentaçãoRESUMO
Changes in the expression of the protein disulfide isomerase genes PDIA3 and PDIA6 may increase endoplasmic reticulum stress, leading to cellular instability and neoplasia. We evaluated the expression of PDIA3 and PDIA6 in invasive ductal carcinomas. Using reverse transcription-quantitative polymerase chain reaction, we compared the mRNA expression level in 45 samples of invasive ductal carcinoma with that in normal breast samples. Increased expression of the PDIA3 gene in carcinomas (P = 0.0009) was observed. In addition, PDIA3 expression was increased in tumors with lymph node metastasis (P = 0.009) and with grade III (P < 0.02). The PDIA6 gene showed higher expression levels in the presence of lymph node metastasis (U = 99.00, P = 0.0476) and lower expression for negative hormone receptors status (P = 0.0351). Our results suggest that alterations in PDIA3/6 expression levels may be involved in the breast carcinogenic process and should be further investigated as a marker of aggressiveness.
Assuntos
Biomarcadores Tumorais/genética , Neoplasias da Mama/genética , Carcinoma Ductal de Mama/genética , Regulação Neoplásica da Expressão Gênica , Isomerases de Dissulfetos de Proteínas/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Carcinoma Ductal de Mama/metabolismo , Carcinoma Ductal de Mama/patologia , Feminino , Humanos , Metástase Linfática , Pessoa de Meia-Idade , Gradação de Tumores , Invasividade Neoplásica , Isomerases de Dissulfetos de Proteínas/metabolismo , Receptores de Estrogênio/genética , Receptores de Estrogênio/metabolismo , Receptores de Progesterona/genética , Receptores de Progesterona/metabolismoRESUMO
BACKGROUND: Wheezing is one of the most frequent causes of visit to emergency rooms among children. However, data on wheezing burden are mostly provided at healthcare setting, and particularly only for infants. AIMS: We sought to estimate the prevalence of wheezing in children under 4 years and to assess potential risk factors in the community. DESIGN: This was a cross-sectional analysis of a population-based cohort study. METHODS: The sample comprised children aged <4 years living in Salvador, Brazil. Data were collected via home visits when the parents/guardians were interviewed. Data were recorded on standardized forms. RESULTS: Of 1534 children, mean age was 21 ± 14 months (minimum 3 days; maximum 47 months; 6% <2 months); 780 (51%) were males and 501 [33%; 95% confidence interval (95% CI): 30-35%] reported wheezing in the last 12 months. Among wheezers, 321 (64%) had occasional wheezing. Overall, 180 (12%; 95% CI: 10-14%) had recurrent wheezing and 157 (10%; 95% CI: 9-12%) had asthma. For children in the first, second, third and fourth year of life wheezing was reported in 23, 41, 34 and 37%, respectively. Mother atopic-related disease was independently associated with recurrent wheezing (AdjPR[95% CI]: 1.54 [1.12-2.11]) and asthma (AdjPR[95% CI]: 1.54 [1.10-2.16]). Smoker at home (AdjPR[95% CI]: 1.34 [1.07-1.67]) and low birth weight (AdjPR[95%CI]: 1.38 [1.05-1.81]) were independently associated with occasional wheezing. CONCLUSIONS: One-third of under 4 years reported wheezing; history of mother's atopic-related disease was an independent risk factor for recurrent wheezing and asthma; smoker at home and low birth weight were independent risk factors for occasional wheezing.
Assuntos
Sons Respiratórios/etiologia , Distribuição por Idade , Asma/epidemiologia , Brasil/epidemiologia , Pré-Escolar , Estudos Transversais , Saúde da Família , Feminino , Humanos , Lactente , Recém-Nascido de Baixo Peso/fisiologia , Recém-Nascido , Masculino , Prevalência , Recidiva , Fatores de Risco , Poluição por Fumaça de Tabaco/estatística & dados numéricosRESUMO
OBJECTIVE: Stem cells have the ability to renew themselves and differentiate into various cell types. For this reason, numerous research groups have been studying these cells for their therapeutic potential. Some of the therapies, however, are not producing the expected results because of contamination by other cell types, especially by fibroblasts. In the cosmetic industry, stem cells are used to test the efficacy of anti-ageing and rejuvenation products. The purpose of this work was to gain a better understanding of the differences in phenotype, in gene expression associated with stem cells, in the pattern of cell surface proteins and in the differentiation capacity of adipose-derived stem cells, of skin-derived stem cells and of commercially available fibroblasts. METHODS: In this study, we compared fibroblasts with mesenchymal stem cells derived from bone marrow, skin (dermis) and adipose tissue, to assess the differentiation potential of fibroblasts. Dermal and adipose stem cells were isolated from aesthetic surgery patients, and fibroblasts were obtained from a commercial source. The following parameters were used in this study: immunophenotypic profile (positive: CD29, CD73, CD90 and CD105; negative: CD14, CD45 and HLA-DR); differentiation into osteoblastic, chondrogenic and adipogenic cell types; and PCR array to analyse the gene expression of cells isolated from different culture passages. RESULTS: Fibroblasts express the same cell immunophenotypic markers, as well as the genes that are known to be expressed in stem cells, and were shown to be expressed also in adipose and dermis stem cells. Fibroblasts are also able to differentiate into the three cell lineages mentioned above, that is, adipocytes, osteocytes and chondrocytes. CONCLUSION: Human dermal fibroblasts have a potential to adhere to plastic surfaces and differentiate into other cell types. However, for stem cells intended to be used in cosmetics, experiments conducted with contaminated fibroblasts may produce poor or even falsely negative results for the efficacy of the active ingredient or formulation and thus conceal their promising effects as anti-ageing and skin rejuvenation products.
Assuntos
Tecido Adiposo/citologia , Fibroblastos/citologia , Células-Tronco Mesenquimais/citologia , Pele/citologia , Tecido Adiposo/fisiologia , Diferenciação Celular/fisiologia , Feminino , Fibroblastos/fisiologia , Citometria de Fluxo , Humanos , Imunofenotipagem , Células-Tronco Mesenquimais/fisiologia , RNA/química , RNA/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Specific reversed-phase high-performance liquid chromatography conditions are reported for the analysis of recombinant and native human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) preparations. Heterodimeric hLH, hCG and their alpha- and beta-subunits migrated with significantly different retention times (t(R)) in the following order of increasing hydrophobicity: alpha-hCGAssuntos
Gonadotropina Coriônica/análise
, Cromatografia Líquida de Alta Pressão/métodos
, Cromatografia de Fase Reversa/métodos
, Hormônio Luteinizante/análise
, Preparações Farmacêuticas/análise
, Gonadotropina Coriônica Humana Subunidade beta/análise
, Subunidade alfa de Hormônios Glicoproteicos/análise
, Humanos
, Proteínas Recombinantes/análise
RESUMO
Complete dissociation into subunits was attained by incubating Chinese hamster ovary (CHO)-derived or native human thyrotropin, follitropin and lutropin overnight at 37 degrees C in acetic acid. The alpha-and beta-subunits of the pituitary glycoprotein hormones were rapidly and quantitatively isolated by reversed-phase high-performance liquid chromatography (RP-HPLC). A dissociation efficiency of > 98% was obtained on the basis of mass determinations of the heterodimers and subunits carried out via mass spectrometry. CHO-derived or native subunits were isolated on a C4 column (80-90% total recovery) and characterized comparatively for purity, hydrophobicity, molecular mass and charge distribution by HPLC, mass spectrometry, sodium dodecylsulfate-polyacrylamide gel electrophoresis and isoelectric focusing. Thyrotropin was used as a model for showing that, after subunit reassociation, the in vivo bioactivity of the hormone was completely restored. The method described is mild, practical, flexible, and can be adapted to dissociate microgram amounts of native or recombinant glycoprotein hormones, allowing characterization of each subunit.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Subunidade alfa de Hormônios Glicoproteicos/isolamento & purificação , Hormônios Adeno-Hipofisários/isolamento & purificação , Subunidades Proteicas/isolamento & purificação , Proteínas Recombinantes/isolamento & purificação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Animais , Células CHO , Cricetinae , Cricetulus , Subunidade alfa de Hormônios Glicoproteicos/metabolismo , Humanos , Interações Hidrofóbicas e Hidrofílicas , Focalização Isoelétrica , Hormônios Adeno-Hipofisários/metabolismo , Subunidades Proteicas/metabolismo , Proteínas Recombinantes/metabolismoRESUMO
The present study was conducted to clarify the interference of selenomethionine (SeMet) on methylmercury (MeHg) toxicity through the evaluation of changes in biomarkers of exposure and effect in rats exposed to MeHg and co-exposed to MeHg and SeMet. Male Wistar rats received two intraperitoneally (i.p.) administrations, either MeHg (1.5mg/kg body weight), SeMet alone (1mg/kg body weight) or combined MeHg and SeMet, followed by 3 weeks of rat urine collection and neurobehavioural assays. The effects of different administrations were investigated by the quantification of total mercury in kidney and brain, analysis of urinary porphyrins, determination of hepatic GSH and evaluation of motor activity functions (rearing and ambulation). MeHg exposure resulted in a significant increase of urinary porphyrins during the 3 weeks of rat urine collection, where as it caused a significant decrease in motor activity only at the first day after cessation of rat exposure. Additionally, SeMet co-exposure was able to normalize the porphyrins excretion, and a tendency to restore rat motor activity was observed, on the first day after cessation of exposure. Brain and kidney mercury levels increased significantly in rats exposed to MeHg; however, in co-exposed rats to SeMet no significant changes in Hg levels were found as compared to rats exposed to MeHg alone. Hence, the present study shows that urinary porphyrins are sensitive and persistent indicators of MeHg toxicity and demonstrates for the first time that SeMet reduces its formation. Finally, these results confirm that the mechanism of interaction between SeMet and MeHg cannot be explained by the reduction of Hg levels in target organs and suggestions are made to clarify the interference of SeMet on MeHg toxicity.
Assuntos
Intoxicação do Sistema Nervoso por Mercúrio/tratamento farmacológico , Intoxicação do Sistema Nervoso por Mercúrio/metabolismo , Compostos de Metilmercúrio/toxicidade , Selenometionina/farmacologia , Animais , Comportamento Animal/efeitos dos fármacos , Biomarcadores/metabolismo , Biomarcadores/urina , Interações Medicamentosas , Glutationa/metabolismo , Rim/metabolismo , Fígado/metabolismo , Masculino , Intoxicação do Sistema Nervoso por Mercúrio/urina , Compostos de Metilmercúrio/antagonistas & inibidores , Compostos de Metilmercúrio/farmacocinética , Atividade Motora/efeitos dos fármacos , Atividade Motora/fisiologia , Porfirinas/urina , Ratos , Ratos WistarRESUMO
Diabetic retinopathy is a leading cause of blindness among adults in the western countries. It has been reported that neurodegeneration may occur in diabetic retinas, but the mechanisms underlying retinal cell death are poorly understood. We found that high glucose increased the number of cells with condensed nuclei and the number of TUNEL-positive cells, and caused an increase in the translocation of phosphatidylserine to the outer leaflet of the plasma membrane, indicating that high glucose induces apoptosis in cultured retinal neural cells. The activity of caspases did not increase in high glucose-treated cells, but apoptosis-inducing factor (AIF) levels decreased in the mitochondria and increased in the nucleus, indicating a translocation to the nucleus where it may cause DNA fragmentation. These results demonstrate that elevated glucose induces apoptosis in cultured retinal neural cells. The increase in apoptosis is not dependent on caspase activation, but is mediated through AIF release from the mitochondria.
Assuntos
Apoptose/efeitos dos fármacos , Glucose/farmacologia , Neurônios/citologia , Neurônios/efeitos dos fármacos , Retina/citologia , Trifosfato de Adenosina/metabolismo , Animais , Antropologia Cultural , Apoptose/fisiologia , Fator de Indução de Apoptose/metabolismo , Caspases/metabolismo , Núcleo Celular/metabolismo , Retinopatia Diabética/patologia , Mitocôndrias/metabolismo , Neurônios/metabolismo , Ratos , Ratos WistarRESUMO
Several technologically sophisticated high-throughput techniques have been recently developed for the study of human single nucleotide polymorphisms and the diagnosis of point mutations in human diseases. However, there is also a need for simple and inexpensive techniques suitable for clinical services and small research laboratories. Minisequencing meets the latter requirements. It is simple, non-radioactive and can be easily multiplexed by adding oligonucleotide tails of increasing size to the sequencing oligonucleotide primers. To optimize the minisequencing protocol, we designed a test multiplex system capable of typing simultaneously 12 different human autosomal single nucleotide polymorphisms. We discovered that the quality of minisequencing primers and the careful selection of the tail sequences were especially critical for success. This optimized protocol permits rapid genotyping at low cost and can serve as a blueprint for the creation of multiplex minisequencing systems suitable to virtually any typing application in population studies and medical genetics.
Assuntos
Humanos , Análise de Sequência de DNA/métodos , Mutação Puntual/genética , Polimorfismo de Nucleotídeo Único/genética , Primers do DNA/genética , Reação em Cadeia da Polimerase/métodos , GenótipoRESUMO
Microorganisms with large genomes are commonly the subjects of single-round partial sequencing of cDNA, generating expressed sequence tags (ESTs). Usually there is a great distance between gene discovery by EST projects and submission of amino acid sequences to public databases. We analyzed the relationship between available ESTs and protein sequences and used the sequences available in the secondary database, clusters of orthologous groups (COG), to investigate ESTs from eight microorganisms of medical and/or economic relevance, selecting for candidate ESTs that may be further pursued for protein characterization. The organisms chosen were Paracoccidioides brasiliensis, Dictyostelium discoideum, Fusarium graminearum, Plasmodium yoelii, Magnaporthe grisea, Emericella nidulans, Chlamydomonas reinhardtii and Eimeria tenella, which have more than 10,000 ESTs available in dbEST. A total of 77,114 protein sequences from COG were used, corresponding to 3,201 distinct genes. At least 212 of these were capable of identifying candidate ESTs for further studies (E. tenella). This number was extended to over 700 candidate ESTs (C. reinhardtii, F. graminearum). Remarkably, even the organism that presents the highest number of ESTs corresponding to known proteins, P. yoelii, showed a considerable number of candidate ESTs for protein characterization (477). For some organisms, such as P. brasiliensis, M. grisea and F. graminearum, bioinformatics has allowed for automatic annotation of up to about 20 of the ESTs that did not correspond to proteins already characterized in the organism. In conclusion, 4093 ESTs from these eight organisms that are homologous to COG genes were selected as candidates for protein characterization
Assuntos
Animais , Bases de Dados de Proteínas , Etiquetas de Sequências Expressas , Análise de Sequência de Proteína , Chlamydomonas reinhardtii/genética , Dictyostelium/genética , Eimeria tenella/genética , Emericella/genética , Fusarium/genética , Genoma , Magnaporthe/genética , Paracoccidioides/genética , Plasmodium yoelii/genética , Proteínas/genética , Homologia de Sequência de AminoácidosRESUMO
Mutation of p53 is rare in localized prostate carcinoma. The oncoprotein MDM2, whose gene has a response element for p53, promotes the degradation of p53 protein and inhibits its transcriptional activation of genes related to cell cycle arrest and apoptosis, constituting a negative feedback control. We studied p53 and MDM2 expression by immunohistochemistry and looked for mutations in p53 exons 5 to 8 by polymerase chain reaction-single strand conformational polymorphism in 118 patients submitted to radical prostatectomy for localized prostate cancer. In 28 cases, we studied cell proliferation by immunohistochemistry, using antibody for Ki-67, and apoptosis by the deoxynucleotidyl transferase mediated dUTP biotin nick end labeling technique. Although no p53 mutations were found, p53 protein was detected in 31.4% of the cases, and these cases had higher Gleason scores (P = .03) and more advanced tumor stages (P = .02). MDM2 was overexpressed in 40.7% of the cases, and these cases had greater tumor volumes (P = .001). Tumors that were positive for both p53 and MDM2 were larger (P = .003) and of more advanced stage (P = .03). Within the 28-case subset, the proliferative index was higher among MDM2-positive tumors (P = .046), and the apoptotic index was lower among p53-positive tumors (P = .01). We conclude that, although p53 mutation is a rare event in prostate carcinogenesis, the detection of p53 protein by immunohistochemistry is common and is associated with decreased apoptosis and increased histologic grade and tumor stage. We also conclude that the overexpression of MDM2 has a role in prostate carcinogenesis, being frequently detected and associated with increased cell proliferation and tumor volume. Finally, we propose that the MDM2-positive/p53-positive phenotype identifies prostate cancers with aggressive behavior.
Assuntos
Adenocarcinoma/metabolismo , Proteínas de Neoplasias/metabolismo , Proteínas Nucleares , Neoplasias da Próstata/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Adenocarcinoma/genética , Adenocarcinoma/patologia , Adenocarcinoma/cirurgia , Idoso , Apoptose , Divisão Celular , DNA de Neoplasias/análise , Genes p53/genética , Humanos , Marcação In Situ das Extremidades Cortadas , Antígeno Ki-67/metabolismo , Masculino , Pessoa de Meia-Idade , Mutação , Proteínas de Neoplasias/genética , Reação em Cadeia da Polimerase , Polimorfismo Conformacional de Fita Simples , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Neoplasias da Próstata/cirurgia , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas c-mdm2 , Proteína Supressora de Tumor p53/metabolismoRESUMO
OBJECTIVES: Lymphoma is the most frequent testicular malignancy in men over 60 years of age. Even though patients present initially with localized disease, the high incidence of bilateral involvement, synchronous or not, and early systemic dissemination are characteristic of these neoplasms. Sometimes the interval between tumor involvement of both testes is long. The question is raised whether either the patient has a predisposition to present new clones of transformed lymphocytes, or the same disease using the same pathway from a systemic reservoir infiltrates the contralateral testis. METHOD: Polymerase chain reaction and DNA sequencing were used to detect immunoglobulin heavy chain (IgH) rearrangement in paraffin-embedded specimens from asynchronous tumors affecting the right and left testis of a 85-year-old man with an interval period of 13 months. RESULTS: Both tumors showed the same IgH rearrangement. CONCLUSIONS: The lymphoma affecting the left and right testis derived from the same clone. It makes a strong case that lymphoma of the testis is the first manifestation of a systemic disease and should be treated aggressively early at the beginning of the disease.
Assuntos
Linfoma Difuso de Grandes Células B/genética , Neoplasias Testiculares/genética , Idoso , Idoso de 80 Anos ou mais , DNA de Neoplasias/química , Humanos , Cadeias Pesadas de Imunoglobulinas/química , Linfoma Difuso de Grandes Células B/patologia , Masculino , Reação em Cadeia da Polimerase , Neoplasias Testiculares/patologia , Testículo/patologia , Fatores de TempoRESUMO
CONTEXT: The presence of fetal fibronectin in the cervix or vagina has been investigated as a possible marker for the risk of preterm birth. Fetal fibronectin in cervical fluid can provide direct evidence of pathologic changes at the interface of fetal and maternal tissues. OBJECTIVE: To evaluate the presence of fetal fibronectin as a predictor of premature delivery in twin pregnancies in relation to gestational age. DESIGN: Accuracy study. SETTING: University referral unit. PARTICIPANTS: 52 pregnant women with twin pregnancies and gestational age of between 24 and 34 weeks. MAIN MEASUREMENTS: Sensitivity, specificity, predictive values and relative risk ratios of the correlation between fetal fibronectin and preterm birth before 34 and 37 weeks using an immediate-reading membrane test on cervicovaginal secretions obtained from participants. RESULT: The sensitivity varied between 66.7% and 85.7%, whereas the specificity was from 58.3% to 81.8% according to gestational age at the time of sampling. The relative risk of spontaneous preterm birth after a positive fetal fibronectin test, as compared with a negative fetal fibronectin test, rose from 2.8 at 24-26 weeks to 4.1 at 27-30 weeks. Analyses of the risk of delivery before 34 weeks were not statistically significant. CONCLUSION: Fetal fibronectin in the cervicovaginal secretions of patients with twin pregnancies is a useful tool for the early identification of twin pregnancies likely to deliver before 37 weeks. However, the clinical value of the fibronectin test is limited because of low indices for prediction of delivery before 34 weeks. The best period for performing the fetal fibronectin test in twin pregnancies to predict preterm delivery is between 27 and 30 weeks.
Assuntos
Fibronectinas/análise , Idade Gestacional , Trabalho de Parto Prematuro/diagnóstico , Gêmeos , Biomarcadores/análise , Feminino , Humanos , Recém-Nascido , Razão de Chances , Valor Preditivo dos Testes , Gravidez , Estudos Prospectivos , Sensibilidade e Especificidade , Esfregaço VaginalRESUMO
The changes in the intracellular free Ca2+ concentration, [Ca2+]i, mediated by glutamate and D-aspartate into rat hippocampal synaptosomes was studied. Glutamate increased the [Ca2+]i in a dose-dependent manner with an EC50 of 1.87 microM and a maximal increase of 31.5 +/- 0.9 nM. We also observed that stimulation of the synaptosomes with 100 microM alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), 100 microM kainate, or 100 microM D-aspartate increased the synaptosomal [Ca2+]i. The effect of either of these non-NMDA receptor agonists and of D-aspartate was additive, suggesting the activation of two different components (the ionotropic non-NMDA receptors or the glutamate transporters). Stimulation of synaptosomes with 100 microM glutamate increased the [Ca2+]i and prevented the effect of either non-NMDA receptor agonists and the effect of D-aspartate. We also observed that incubation of the synaptosomes with D-aspartate induced the Ca(2+)-independent release of glutamate, possibly through the reversal of the glutamate carrier. The aim of incubating the synaptosomes with D-aspartate was to avoid undesirable secondary activation of glutamate receptors. After incubating the synaptosomes with 100 microM D-aspartate (10 min at 37 degrees C), the subsequent stimulation with D-aspartate increased the [Ca2+]i due to glutamate transport. This increase in [Ca2+]i induced by 100 microM D-aspartate was insensitive to 1 microM nitrendipine, but was inhibited by about 50% by the presence of both 500 nM omega-CgTx GVIA and 100 nM omega-Aga IVA or by 500 nM omega-CgTx MVIIC. We clearly identified two different processes by which glutamate increased the [Ca2+]i in rat hippocampal synaptosomes: activation of non-NMDA receptors and activation of the glutamate transporters. We also characterized the voltage sensitive Ca2+ channels (VSCC) activated as a consequence of the glutamate transport, and determined that class B (N-type) and class A (P or Q-type) Ca2+ channels were responsible for about 50% of the signal.