RESUMO
MAIN CONCLUSION: Host-derived suppression of nematode essential genes decreases reproduction of Meloidogyne incognita in cotton. Root-knot nematodes (RKN) represent one of the most damaging plant-parasitic nematode genera worldwide. RNAi-mediated suppression of essential nematode genes provides a novel biotechnological strategy for the development of sustainable pest-control methods. Here, we used a Host Induced Gene Silencing (HIGS) approach by stacking dsRNA sequences into a T-DNA construct to target three essential RKN genes: cysteine protease (Mi-cpl), isocitrate lyase (Mi-icl), and splicing factor (Mi-sf), called dsMinc1, driven by the pUceS8.3 constitutive soybean promoter. Transgenic dsMinc1-T4 plants infected with Meloidogyne incognita showed a significant reduction in gall formation (57-64%) and egg masses production (58-67%), as well as in the estimated reproduction factor (60-78%), compared with the susceptible non-transgenic cultivar. Galls of the RNAi lines are smaller than the wild-type (WT) plants, whose root systems exhibited multiple well-developed root swellings. Transcript levels of the three RKN-targeted genes decreased 13- to 40-fold in nematodes from transgenic cotton galls, compared with those from control WT galls. Finally, the development of non-feeding males in transgenic plants was 2-6 times higher than in WT plants, indicating a stressful environment for nematode development after RKN gene silencing. Data strongly support that HIGS of essential RKN genes is an effective strategy to improve cotton plant tolerance. This study presents the first application of dsRNA sequences to target multiple genes to promote M. incognita tolerance in cotton without phenotypic penalty in transgenic plants.
Assuntos
Gossypium , Tylenchoidea , Animais , Gossypium/genética , Doenças das Plantas/genética , Plantas Geneticamente Modificadas/genética , RNA de Cadeia Dupla , Tylenchoidea/genéticaRESUMO
The Coffea arabica HB12 gene (CaHB12), which encodes a transcription factor belonging to the HD-Zip I subfamily, is upregulated under drought, and its constitutive overexpression (35S:CaHB12OX) improves the Arabidopsis thaliana tolerance to drought and salinity stresses. Herein, we generated transgenic cotton events constitutively overexpressing the CaHB12 gene, characterized these events based on their increased tolerance to water deficit, and exploited the gene expression level from the CaHB12 network. The segregating events Ev8.29.1, Ev8.90.1, and Ev23.36.1 showed higher photosynthetic yield and higher water use efficiency under severe water deficit and permanent wilting point conditions compared to wild-type plants. Under well-irrigated conditions, these three promising transformed events showed an equivalent level of Abscisic acid (ABA) and decreased Indole-3-acetic acid (IAA) accumulation, and a higher putrescine/(spermidine + spermine) ratio in leaf tissues was found in the progenies of at least two transgenic cotton events compared to non-transgenic plants. In addition, genes that are considered as modulated in the A. thaliana 35S:CaHB12OX line were also shown to be modulated in several transgenic cotton events maintained under field capacity conditions. The upregulation of GhPP2C and GhSnRK2 in transgenic cotton events maintained under permanent wilting point conditions suggested that CaHB12 might act enhancing the ABA-dependent pathway. All these data confirmed that CaHB12 overexpression improved the tolerance to water deficit, and the transcriptional modulation of genes related to the ABA signaling pathway or downstream genes might enhance the defense responses to drought. The observed decrease in IAA levels indicates that CaHB12 overexpression can prevent leaf abscission in plants under or after stress. Thus, our findings provide new insights on CaHB12 gene and identify several promising cotton events for conducting field trials on water deficit tolerance and agronomic performance.
Assuntos
Secas , Gossypium , Regulação da Expressão Gênica de Plantas , Gossypium/genética , Gossypium/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Estresse Fisiológico/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
BACKGROUND: Drought is one of the most harmful abiotic stresses for plants, leading to reduced productivity of several economically important crops and, consequently, considerable losses in the agricultural sector. When plants are exposed to stressful conditions, such as drought and high salinity, they modulate the expression of genes that lead to developmental, biochemical, and physiological changes, which help to overcome the deleterious effects of adverse circumstances. Thus, the search for new specific gene promoter sequences has proved to be a powerful biotechnological strategy to control the expression of key genes involved in water deprivation or multiple stress responses. RESULTS: This study aimed to identify and characterize the GmRD26 promoter (pGmRD26), which is involved in the regulation of plant responses to drought stress. The expression profile of the GmRD26 gene was investigated by qRT-PCR under normal and stress conditions in Williams 82, BR16 and Embrapa48 soybean-cultivars. Our data confirm that GmRD26 is induced under water deficit with different induction folds between analyzed cultivars, which display different genetic background and physiological behaviour under drought. The characterization of the GmRD26 promoter was performed under simulated stress conditions with abscisic acid (ABA), polyethylene glycol (PEG) and drought (air dry) on A. thaliana plants containing the complete construct of pGmRD26::GUS (2.054 bp) and two promoter modules, pGmRD26A::GUS (909 pb) and pGmRD26B::GUS (435 bp), controlling the expression of the ß-glucuronidase (uidA) gene. Analysis of GUS activity has demonstrated that pGmRD26 and pGmRD26A induce strong reporter gene expression, as the pAtRD29 positive control promoter under ABA and PEG treatment. CONCLUSIONS: The full-length promoter pGmRD26 and the pGmRD26A module provides an improved uidA transcription capacity when compared with the other promoter module, especially in response to polyethylene glycol and drought treatments. These data indicate that pGmRD26A may become a promising biotechnological asset with potential use in the development of modified drought-tolerant plants or other plants designed for stress responses.
Assuntos
Ácido Abscísico/farmacologia , Glycine max/genética , Biotecnologia/métodos , Secas , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/genética , Regiões Promotoras Genéticas/efeitos dos fármacos , Regiões Promotoras Genéticas/genética , Glycine max/efeitos dos fármacos , Estresse Fisiológico/genética , Estresse Fisiológico/fisiologiaRESUMO
Genetically modified (GM) cotton plants that effectively control cotton boll weevil (CBW), which is the most destructive cotton insect pest in South America, are reported here for the first time. This work presents the successful development of a new GM cotton with high resistance to CBW conferred by Cry10Aa toxin, a protein encoded by entomopathogenic Bacillus thuringiensis (Bt) gene. The plant transformation vector harbouring cry10Aa gene driven by the cotton ubiquitination-related promoter uceA1.7 was introduced into a Brazilian cotton cultivar by biolistic transformation. Quantitative PCR (qPCR) assays revealed high transcription levels of cry10Aa in both T0 GM cotton leaf and flower bud tissues. Southern blot and qPCR-based 2-ΔΔCt analyses revealed that T0 GM plants had either one or two transgene copies. Quantitative and qualitative analyses of Cry10Aa protein expression showed variable protein expression levels in both flower buds and leaves tissues of T0 GM cotton plants, ranging from approximately 3.0 to 14.0 µg g-1 fresh tissue. CBW susceptibility bioassays, performed by feeding adults and larvae with T0 GM cotton leaves and flower buds, respectively, demonstrated a significant entomotoxic effect and a high level of CBW mortality (up to 100%). Molecular analysis revealed that transgene stability and entomotoxic effect to CBW were maintained in T1 generation as the Cry10Aa toxin expression levels remained high in both tissues, ranging from 4.05 to 19.57 µg g-1 fresh tissue, and the CBW mortality rate remained around 100%. In conclusion, these Cry10Aa GM cotton plants represent a great advance in the control of the devastating CBW insect pest and can substantially impact cotton agribusiness.
Assuntos
Proteínas de Bactérias/metabolismo , Endotoxinas/metabolismo , Gossypium/metabolismo , Gossypium/parasitologia , Proteínas Hemolisinas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/parasitologia , Gorgulhos/patogenicidade , Animais , Toxinas de Bacillus thuringiensis , Proteínas de Bactérias/genética , Endotoxinas/genética , Gossypium/genética , Proteínas Hemolisinas/genética , Imunidade Vegetal/genética , Imunidade Vegetal/fisiologia , Plantas Geneticamente Modificadas/genética , Reação em Cadeia da PolimeraseRESUMO
Hepatitis C virus (HCV) quasispecies constitute a dynamic population in a continuous process of variation and selection. To investigate effect of the immune system on the genetic variability of HCV, we compared the hypervariable region 1 (HVR1) of immunosuppressed patients with chronic renal failure (CRF group) to immunocompetent patients with HCV chronic infection (control group). The HVR1 from ten samples of each group was amplified, cloned and sequenced. The HCV quasispecies from the control group had a higher frequency of variable sites in HVR1 (83.9 % vs 59.3 %, p < 0.05), as well as a greater diversity within (intra-patient) and between samples, compared to the CRF group. The clustering of the majority of the quasispecies of the CRF group in the phylogenetic tree also showed the limited diversity of the quasispecies in immunosuppressed patients. Moreover, a higher variability of amino acids at positions 384, 386, 391, 394, 397, 398, 400, 405 and 410 was observed in the control group than in the CRF group, which showed a greater variability only at position 388 (p < 0.05). These data corroborates the hypothesis that the major selective pressure factor is the immune system, which promotes a high degree of diversity in the viral progeny and contributes to a constant evolution of HCV.
Assuntos
Variação Genética , Hepacivirus/genética , Hepatite C Crônica/virologia , Falência Renal Crônica/etiologia , Adulto , Idoso , Brasil/epidemiologia , Feminino , Hepacivirus/classificação , Hepacivirus/isolamento & purificação , Hepacivirus/fisiologia , Hepatite C Crônica/complicações , Humanos , Falência Renal Crônica/virologia , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Filogenia , Adulto JovemRESUMO
BACKGROUND: Root-knot nematodes (RKN- Meloidogyne genus) present extensive challenges to soybean crop. The soybean line (PI 595099) is known to be resistant against specific strains and races of nematode species, thus its differential gene expression analysis can lead to a comprehensive gene expression profiling in the incompatible soybean-RKN interaction. Even though many disease resistance genes have been studied, little has been reported about phytohormone crosstalk on modulation of ROS signaling during soybean-RKN interaction. RESULTS: Using 454 technology to explore the common aspects of resistance reaction during both parasitism and resistance phases it was verified that hormone, carbohydrate metabolism and stress related genes were consistently expressed at high levels in infected roots as compared to mock control. Most noteworthy genes include those encoding glycosyltransferases, peroxidases, auxin-responsive proteins and gibberellin-regulated genes. Our data analysis suggests the key role of glycosyltransferases, auxins and components of gibberellin signal transduction, biosynthesis and deactivation pathways in the resistance reaction and their participation in jasmonate signaling and redox homeostasis in mediating aspects of plant growth and responses to biotic stress. CONCLUSIONS: Based on this study we suggest a reasonable model regarding to the complex mechanisms of crosstalk between plant hormones, mainly gibberellins and auxins, which can be crucial to modulate the levels of ROS in the resistance reaction to nematode invasion. The model also includes recent findings concerning to the participation of DELLA-like proteins and ROS signaling controlling plant immune or stress responses. Furthermore, this study provides a dataset of potential candidate genes involved in both nematode parasitism and resistance, which can be tested further for their role in this biological process using functional genomics approaches.
Assuntos
Resistência à Doença/genética , Perfilação da Expressão Gênica , Glycine max/parasitologia , Interações Hospedeiro-Parasita , Doenças das Plantas/parasitologia , Reguladores de Crescimento de Plantas/metabolismo , Tylenchoidea/fisiologia , Animais , Giberelinas/metabolismo , Ácidos Indolacéticos/metabolismo , Reguladores de Crescimento de Plantas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Análise de Sequência , Glycine max/genética , Glycine max/imunologia , Glycine max/metabolismo , Estresse Fisiológico/genética , Transcrição GênicaRESUMO
PURPOSE: The functional vision assessment (AVIF-2 to 6 years) for low vision children with ages from 2 to 6 years old was analyzed. The inter-rater, test-retest and the internal consistency were verified for reliability. METHODS: The test, composed by seven domains (visual fixation, visual following, visual field confrontation, eye-hand coordination and surrounding locomotion, contrast vision and color vision discrimination), was applied to 40 children, 20 with low vision and 20 without low vision. The inter-rater and test-retest reliability were analyzed with 12 children and the internal consistency with 40 children. RESULTS: Concerning test-retest reliability, six domains exhibited intra-class correlation coefficients ranging from 0.5361 to 1.000. For the movement within surroundings category, this coefficient was 0.37. For inter-rater reliability, only the category of confrontation visual field was below expectations (0.3901). The weighted kappa coefficient exhibited values ranged from -0.087 to 1.000. Cronbach's alfa ranged from 0.584 (movement within surroundings) to 0.973 (contrast sensitivity). CONCLUSION: AVIF-2 to 6 years presented good reliability indexes, however a few items within the domains need minor adjustments in order to improve scores' consistency of some items.
Assuntos
Baixa Visão , Testes de Campo Visual/instrumentação , Criança , Pré-Escolar , Humanos , Reprodutibilidade dos Testes , Campos VisuaisRESUMO
OBJETIVOS: Avaliar a confiabilidade interexaminadores, fazer o teste-reteste e verificar a consistência interna da avaliação da visão funcional para crianças com baixa visão de dois a seis anos de idade (AVIF-2 a 6 anos). MÉTODOS: Foram testados sete domínios: fixação visual, seguimento visual, campo visual de confrontação, coordenação olho-mão, visão de contraste, deslocamento no ambiente e visão de cores, em 40 crianças, 20 com baixa visão e 20 sem alterações visuais. A consistência interna foi examinada para o teste de todas as crianças e a confiabilidade interexaminadores e teste-reteste para 12 delas. RESULTADOS: Seis domínios apresentaram ao teste-reteste coeficiente de correlação intraclasse com valores de 0,5361 a 1,000. Para o domínio deslocamento no ambiente esse coeficiente foi de 0,37. Na confiabilidade interexaminadores, somente o campo visual de confrontação apresentou valor abaixo do esperado (0,3901). O coeficiente kappa ponderado obteve resultados que variaram de -0,087 a 1,000. O valor do alfa de Cronbach variou de 0,584 (no deslocamento no ambiente) a 0,973 (na visão de contraste). CONCLUSÃO: A AVIF-2 a 6 anos apresentou bons índices de confiabilidade, mas a confiabilidade dos domínios, quando analisados isoladamente, precisa ser aprimorada.
PURPOSE: The functional vision assessment (AVIF-2 to 6 years) for low vision children with ages from 2 to 6 years old was analyzed. The inter-rater, test-retest and the internal consistency were verified for reliability. METHODS: The test, composed by seven domains (visual fixation, visual following, visual field confrontation, eye-hand coordination and surrounding locomotion, contrast vision and color vision discrimination), was applied to 40 children, 20 with low vision and 20 without low vision. The inter-rater and test-retest reliability were analyzed with 12 children and the internal consistency with 40 children. RESULTS: Concerning test-retest reliability, six domains exhibited intra-class correlation coefficients ranging from 0.5361 to 1.000. For the movement within surroundings category, this coefficient was 0.37. For inter-rater reliability, only the category of confrontation visual field was bellow expectations (0.3901). The weighted kappa coefficient exhibited values ranged from -0.087 to 1.000. Cronbach's alfa ranged from 0.584 (movement within surroundings) to 0.973 (contrast sensitivity). CONCLUSION: AVIF-2 to 6 years presented good reliability indexes, however a few items within the domains need minor adjustments in order to improve scores' consistency of some items.
Assuntos
Criança , Pré-Escolar , Humanos , Baixa Visão , Testes de Campo Visual/instrumentação , Reprodutibilidade dos Testes , Campos VisuaisRESUMO
PURPOSE: The Functional Vision Assessment (AVIF-2 to 6 years)'s capacity to differentiate visual ability levels in children with low vision was analyzed. The AVIF-2 to 6 years was created at the Infantile Low Vision Sector from São Geraldo Hospital, Brazil. METHODS: After a debate among professionals from different areas regarding the appropriateness of the test items, AVIF-2 to 6 years was applied to 40 children aged from 2 to 6 years; twenty children with low vision (Group 1) and twenty without it (Group 2) were assessed. Group 1 was recruited from the Infantile Low Vision Sector from São Geraldo Hospital. Group 2 comprised children from two different public day care centers and children were matched by age, gender and social level with group 1. Seven domains were studied: visual fixation, visual following, visual field confrontation, eye-hand coordination and surrounding locomotion, contrast vision and color vision discrimination. Group 1 children were submitted to a complete ophthalmologic exam and group 2 to ophthalmologic screening. Children with low vision and neurologic disease were excluded. Comparative analyzes were performed for both groups and for distinct subgroups classified by age (24 to 35 months, 36 to 59 months and 60 to 78 months) and by visual acuity subgroups (<1.0 logMAR and ≥1.0 logMAR). RESULTS: The scores at total AVIF-2 to 6 years and its domains were statistically significant (p<0.05), except for the contrast vision and color vision discrimination domains. The total AVIF- 2 to 6 years median was lower for group 1 at the three interval ages. CONCLUSION: AVIF- 2 to 6 years can discriminate different levels of functional vision of low vision children, however the authors emphasize that although the results are encouraging, further studies shall be done until the test is ready for clinical use.
Assuntos
Baixa Visão/fisiopatologia , Acuidade Visual/fisiologia , Testes de Campo Visual/métodos , Campos Visuais/fisiologia , Estudos de Casos e Controles , Criança , Pré-Escolar , Estudos Transversais , Feminino , Humanos , Lactente , Masculino , Sensibilidade e Especificidade , Fatores SocioeconômicosRESUMO
OBJETIVO: O objetivo deste trabalho foi apresentar a capacidade da AVIF-2 a 6 anos para discriminar os diferentes níveis de resposta visual de crianças com baixa visão. A AVIF-2 a 6 anos foi criada no Setor de Baixa Visão Infantil do Hospital São Geraldo, Brasil. MÉTODOS: Após a discussão da adequação dos itens do teste, com especialistas de diversas áreas, a AVIF-2 a 6 anos foi aplicada em 40 crianças de dois a seis anos de idade, 20 das quais com baixa visão (Grupo 1) e 20, sem baixa visão (Grupo 2). O grupo 1 foi recrutado do Setor de Baixa Visão Infantil do Hospital São Geraldo. As crianças do grupo 2 foram selecionadas em duas creches públicas. Sete domínios foram estudados: fixação visual, seguimento visual, campo visual de confrontação, coordenação olho-mão, visão de contraste, deslocamento no ambiente e visão de cores. As crianças do grupo 1 foram submetidas a exame oftalmológico completo e as do grupo 2 a triagem oftalmológica. Crianças com baixa visão e alterações neurológicas foram excluidas. Os dois grupos foram emparelhados por idade, sexo e nível socioeconômico, e os dados comparados entre as crianças do grupo geral e dos subgrupos de idade (24 a 35 meses, 36 a 59 meses e 60 a 78 meses) e de acuidade visual (< que 1,0 logMAR e >1,0 logMAR). RESULTADOS: No total da AVIF-2 a 6 anos e dos domínios fixação visual, seguimento visual, campo visual de confrontação, coordenação olho-mão e deslocamento no ambiente foram constatadas diferenças com significância estatística (p<0,05) entre os dois grupos. As medianas da pontuação da AVIF-2 a 6 anos foram inferiores para as crianças com baixa visão nas três faixas etárias. CONCLUSÃO: A AVIF-2 a 6 anos demonstrou ter potencial para discriminar diferentes níveis de visão funcional, entretanto, serão necessários ainda novos estudos para que o teste possa ser disponibilizado para uso clínico. (ETIC 684/07).
PURPOSE: The Functional Vision Assessment (AVIF-2 to 6 years)'s capacity to differentiate visual ability levels in children with low vision was analyzed. The AVIF-2 to 6 years was created at the Infantile Low Vision Sector from São Geraldo Hospital, Brazil. METHODS: After a debate among professionals from different areas regarding the appropriateness of the test items, AVIF-2 to 6 years was applied to 40 children aged from 2 to 6 years; twenty children with low vision (Group 1) and twenty without it (Group 2) were assessed. Group 1 was recruited from the Infantile Low Vision Sector from São Geraldo Hospital. Group 2 comprised children from two different public day care centers and children were matched by age, gender and social level with group 1. Seven domains were studied: visual fixation, visual following, visual field confrontation, eye-hand coordination and surrounding locomotion, contrast vision and color vision discrimination. Group 1 children were submitted to a complete ophthalmologic exam and group 2 to ophthalmologic screening. Children with low vision and neurologic disease were excluded. Comparative analyzes were performed for both groups and for distinct subgroups classified by age (24 to 35 months, 36 to 59 months and 60 to 78 months) and by visual acuity subgroups (<1.0 logMAR and >1.0 logMAR). RESULTS: The scores at total AVIF-2 to 6 years and its domains were statistically significant (p<0.05), except for the contrast vision and color vision discrimination domains. The total AVIF- 2 to 6 years median was lower for group 1 at the three interval ages. CONCLUSION: AVIF- 2 to 6 years can discriminate different levels of functional vision of low vision children, however the authors emphasize that although the results are encouraging, further studies shall be done until the test is ready for clinical use.
Assuntos
Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Baixa Visão/fisiopatologia , Acuidade Visual/fisiologia , Testes de Campo Visual/métodos , Campos Visuais/fisiologia , Estudos de Casos e Controles , Estudos Transversais , Sensibilidade e Especificidade , Fatores SocioeconômicosRESUMO
OBJECTIVE: Juvenile Neuronal Ceroid-Lipofuscinosis (JNCL, CLN 3, Batten Disease) (OMIM #204200) belongs to the most common group of neurodegenerative disorders of childhood. We report the clinical data and molecular analysis of a large Brazilian family. METHOD: Family composed of two consanguineous couples and thirty-two children. Clinical data of ten JNCL patients and molecular analyses on 13 participants were obtained. RESULTS: The large 1.02 kb deletion was detected. The most severe phenotype, with autistic behavior, tics and parkinsonism was seen in a 12-year-old female and a milder phenotype in a 14-year-old male. Nyctalopia was the first symptom in one deceased child. The visual loss of six patients has been first observed in the school and not at home. CONCLUSION: The report highlights the phenotypical intrafamily variation in 10 affected children of this family. The molecular investigation of this large family in our metabolic center turned possible the diagnosis, right approach and genetic counseling.
Assuntos
Consanguinidade , Deleção de Genes , Lipofuscinoses Ceroides Neuronais/genética , Adolescente , Brasil , Causas de Morte , Criança , Eletroforese em Gel de Ágar , Éxons/genética , Feminino , Humanos , Masculino , Cegueira Noturna/genética , Linhagem , Fenótipo , Transtornos da Visão/genéticaRESUMO
OBJECTIVE: Juvenile Neuronal Ceroid-Lipofuscinosis (JNCL, CLN 3, Batten Disease) (OMIM #204200) belongs to the most common group of neurodegenerative disorders of childhood. We report the clinical data and molecular analysis of a large Brazilian family. METHOD: Family composed of two consanguineous couples and thirty-two children. Clinical data of ten JNCL patients and molecular analyses on 13 participants were obtained. RESULTS: The large 1.02 kb deletion was detected. The most severe phenotype, with autistic behavior, tics and parkinsonism was seen in a 12-year-old female and a milder phenotype in a 14-year-old male. Nyctalopia was the first symptom in one deceased child. The visual loss of six patients has been first observed in the school and not at home. CONCLUSION: The report highlights the phenotypical intrafamily variation in 10 affected children of this family. The molecular investigation of this large family in our metabolic center turned possible the diagnosis, right approach and genetic counseling.
OBJETIVO: Lipofuscinose Ceróide Neuronal Juvenil (JNCL, CLN 3, Doença de Batten) (OMIM # 204200) pertence ao grupo mais comum de doenças neurodegenerativas na infância. É causada por mutações no gene CLN3, com padrão de herança recessiva. A deleção de 1,02 kb é a mutação mais comum. Relatamos os dados clínicos e análise molecular de uma família consanguínea numerosa. MÉTODO: Família composta por dois casais consanguíneos e trinta e duas crianças. Foram obtidos dados clínicos de dez pacientes e análises moleculares de 13 participantes. RESULTADOS: Foi detectada deleção de 1,02 kb. O fenótipo mais grave, com comportamento autista, tiques e parkinsonismo foi visto em uma paciente do sexo feminino de 12 anos e o fenótipo mais leve em um paciente do sexo masculino de 14 anos. Nictalopia foi o primeiro sintoma de uma criança falecida. A perda visual de seis pacientes foi observada pela primeira vez na escola e não em casa. CONCLUSÃO: Destaca-se a variação fenotípica intrafamiliar em 10 pacientes. A investigação molecular desta família numerosa tornou possível o diagnóstico, a abordagem correta e aconselhamento genético.
Assuntos
Adolescente , Criança , Feminino , Humanos , Masculino , Consanguinidade , Deleção de Genes , Lipofuscinoses Ceroides Neuronais/genética , Brasil , Causas de Morte , Eletroforese em Gel de Ágar , Éxons/genética , Cegueira Noturna/genética , Linhagem , Fenótipo , Transtornos da Visão/genéticaRESUMO
Early promoter activity of HPV-58 molecular variants isolated from high-grade cervical lesions in Brazil was compared. Luciferase reporter assays were conducted in C33 cells transfected with the complete long control region of 3 molecular variants of HPV-58 as well as HPV-58, -18 or -16 prototypes. The HPV-58 prototype and Bsb-329 and Bsb-367 variants showed a promoter activity similar to that of HPV-16, but lower than that of Bsb-295 and HPV-18. The introduction of the Bsb-295 7788 mutation into the HPV-58 prototype resulted in the enhancement of transcription closer to Bsb-295 and HPV-18. These results could impact the expression of E6 and E7 viral oncogenes.
Assuntos
Tipagem Molecular , Papillomaviridae/classificação , Papillomaviridae/genética , Transcrição Gênica , Fusão Gênica Artificial , Brasil , Linhagem Celular , Colo do Útero/patologia , Colo do Útero/virologia , Feminino , Genes Reporter , Genótipo , Humanos , Luciferases/genética , Luciferases/metabolismo , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/virologia , Regiões Promotoras GenéticasRESUMO
Crianças nascidas prematuras, especialmente aquelas com peso <1.500g, apresentam frequentemtente complicações neonatais e riscos para problemas de desenvolvimento. Nesse estudo, 20 crianças com inteligência e exame neurológico normal, aos sete anos, nascidas <33 semanas de gestação e peso <1.500g, foram comparadas com 20 crianças nascidas a termo. Avaliou-se a inteligência pela WISC-III e o comportamento pelo Questionário de Conners. O Exame Neurológico Evolutivo-Lefèvre foi aplicado nas prematuras. As crianças prematuras apresentaram nível intelectual inferior ao das nascidas a termo. No Conners não houve diferença entre os grupos. Apenas 20 por cento das prematuras completaram todas as provas do Exame Evolutivo previstas para a idade. Os dados alertam para a necessidade de avaliações específicas, na idade escolar, em crianças nascidas prematuras, mesmo que apresentem exame neurológico normal.(AU)
Preterm children - especially those with birth weight <1,500g - often present neonatal complications and higher risk of suffering from developmental disorders. Twenty children aged seven, with average intelligence and normal neurological exams, born with gestational age <33 weeks and weight <1,500g, were enrolled in a study and compared with 20 children born at full term. Intelligence was measured by WISC-III and behavior by the Conners Questionnaire. Lefèvre Evolutive Neurological Examination was applied in the preterm children. Children born preterm had lower IQ than those born at full term. In the Conners, there was no difference between groups. Only 20 percent of preterm children performed all tests from the Evolutive Exam for their age. This study highlights the importance of specific cognitive evaluations in preterm children during school age, even if they have normal neurological exams.(AU)
Assuntos
Humanos , Masculino , Feminino , Criança , Recém-Nascido de Baixo Peso/psicologia , Recém-Nascido Prematuro/psicologia , Neuropsicologia , Cognição , Estudos Transversais , Desenvolvimento Infantil , Comportamento Infantil/psicologiaRESUMO
Crianças nascidas prematuras, especialmente aquelas com peso <1.500g, apresentam frequentemtente complicações neonatais e riscos para problemas de desenvolvimento. Nesse estudo, 20 crianças com inteligência e exame neurológico normal, aos sete anos, nascidas <33 semanas de gestação e peso <1.500g, foram comparadas com 20 crianças nascidas a termo. Avaliou-se a inteligência pela WISC-III e o comportamento pelo Questionário de Conners. O Exame Neurológico Evolutivo-Lefèvre foi aplicado nas prematuras. As crianças prematuras apresentaram nível intelectual inferior ao das nascidas a termo. No Conners não houve diferença entre os grupos. Apenas 20 por cento das prematuras completaram todas as provas do Exame Evolutivo previstas para a idade. Os dados alertam para a necessidade de avaliações específicas, na idade escolar, em crianças nascidas prematuras, mesmo que apresentem exame neurológico normal.
Preterm children - especially those with birth weight <1,500g - often present neonatal complications and higher risk of suffering from developmental disorders. Twenty children aged seven, with average intelligence and normal neurological exams, born with gestational age <33 weeks and weight <1,500g, were enrolled in a study and compared with 20 children born at full term. Intelligence was measured by WISC-III and behavior by the Conners Questionnaire. Lefèvre Evolutive Neurological Examination was applied in the preterm children. Children born preterm had lower IQ than those born at full term. In the Conners, there was no difference between groups. Only 20 percent of preterm children performed all tests from the Evolutive Exam for their age. This study highlights the importance of specific cognitive evaluations in preterm children during school age, even if they have normal neurological exams.
Assuntos
Humanos , Masculino , Feminino , Criança , Cognição , Neuropsicologia , Recém-Nascido Prematuro/psicologia , Recém-Nascido de Baixo Peso/psicologia , Desenvolvimento Infantil , Estudos Transversais , Comportamento Infantil/psicologiaRESUMO
Hepatitis C virus (HCV) genotypes and subtypes were determined in hemodialysis patients in the Federal District, Brazil, by sequencing of the 5' noncoding (NC) and nonstructural 5B (NS5B) regions. From 761 patients, 66 anti-HCV-positive samples were tested for HCV RNA. All 51 HCV RNA-positive samples by PCR of the 5' NC region were genotyped as genotypes 1 (90.2%) and 3 (9.8%). Subtype 1a (82.3%) was the most prevalent, followed by subtypes 3a (9.8%), 1b (5.9%) and 1a/1b (2.0%). Forty-two samples could be amplified and genotyped in the NS5B region: 38 (90.5%) as genotype 1, subtypes 1a, and 8 (9.5%) as genotype 3, subtype 3a. For the 42 samples sequenced in both regions, the genotypes and subtypes determined were concordant in 100% and 95.2% of cases, respectively. Two samples presented discrepant results, with the 5' NC region not distinguishing correctly the subtypes 1a and 1b. These findings indicate that the HCV genotype 1, subtype 1a, is the most prevalent among hemodialysis patients in the Federal District, Brazil.
Assuntos
Regiões 5' não Traduzidas/genética , Hepacivirus/genética , Hepatite C/virologia , RNA Viral/análise , Proteínas não Estruturais Virais/genética , Sequência de Bases , Genótipo , Hepacivirus/classificação , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Diálise RenalRESUMO
Hepatitis C virus (HCV) genotypes and subtypes were determined in hemodialysis patients in the Federal District, Brazil, by sequencing of the 5' noncoding (NC) and nonstructural 5B (NS5B) regions. From 761 patients, 66 anti-HCV-positive samples were tested for HCV RNA. All 51 HCV RNA-positive samples by PCR of the 5' NC region were genotyped as genotypes 1 (90.2 percent) and 3 (9.8 percent). Subtype 1a (82.3 percent) was the most prevalent, followed by subtypes 3a (9.8 percent), 1b (5.9 percent) and 1a/1b (2.0 percent). Forty-two samples could be amplified and genotyped in the NS5B region: 38 (90.5 percent) as genotype 1, subtypes 1a, and 8 (9.5 percent) as genotype 3, subtype 3a. For the 42 samples sequenced in both regions, the genotypes and subtypes determined were concordant in 100 percent and 95.2 percent of cases, respectively. Two samples presented discrepant results, with the 5' NC region not distinguishing correctly the subtypes 1a and 1b. These findings indicate that the HCV genotype 1, subtype 1a, is the most prevalent among hemodialysis patients in the Federal District, Brazil.
Os genótipos e subtipos do vírus da hepatite C (HCV) foram determinados em pacientes em hemodiálise no Distrito Federal, Brasil, pelo sequenciamento das regiões 5' não codificante (NC) e não estrutural 5B (NS5B). De 761 pacientes, 66 amostras anti-HCV positivas foram testadas para RNA-HCV. Todas as 51 amostras RNA-HCV positivas por PCR para a região 5' NC foram genotipadas como dos genótipos 1 (90,2 por cento) e 3 (9,8 por cento). O subtipo 1a (82,3 por cento) foi o mais prevalente, seguido pelos subtipos 3a (9,8 por cento), 1b (5,9 por cento) e 1a/1b (2,0 por cento). Quarenta e duas amostras puderam ser amplificadas e genotipadas na região NS5B: 38 (90,5 por cento) como genótipo 1, subtipo 1a, e 8 (9,5 por cento) como genótipo 3, subtipo 3a. Para as 42 amostras sequenciadas nas duas regiões, os genótipos e subtipos determinados foram concordantes em 100 por cento e 95,2 por cento dos casos, respectivamente. Duas amostras apresentaram resultados discrepantes, sendo que a região 5' NC não diferenciou corretamente os subtipos 1a e 1b. Estes achados indicam que o genótipo 1, subtipo 1a, do HCV é o mais prevalente em pacientes em hemodiálise no Distrito Federal, Brasil.
Assuntos
Humanos , /genética , Hepacivirus/genética , Hepatite C/virologia , RNA Viral/análise , Proteínas não Estruturais Virais/genética , Sequência de Bases , Genótipo , Hepacivirus/classificação , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Diálise RenalRESUMO
More than 100 HPV types have been described, 13 of which are classified as high-risk due to their association with the development of cervical cancer. The intratype genomic diversity of HPV-16 and -18 has been studied extensively, while little data have been generated for other less common high-risk types. The present study explores the nucleotide variability and phylogeny of the high-risk HPV-31, -33, -35, -52, and -58, in samples from Central Brazil. For this purpose, the LCR and the E6 and L1 genes were sequenced. Several variants of these HPV types were detected, some of which have been detected in other parts of the world. Furthermore, new variants of all types examined were characterized in a total of 13 new variants. Based on the E6 and L1 sequences, variants were described comprising conservative and non-conservative amino acid changes. For phylogenetic tree construction, samples characterized in this study were compared to others described and submitted to GenBank previously. The phylogenetic analysis of HPV-31, -33, -35, and -58 isolates did not reveal ethnic or geographical clustering as observed previously for HPV-16 and -18. HPV-35 analysis showed a dichotomic branching characteristic of viral subtypes. Interestingly, four clusters relative to the analysis of HPV-52 isolates were identified, two of which could be classified as Asian and European branches. The genomic characterization of HPV variants is crucial for understanding the intrinsic geographical relatedness and biological differences of these viruses and contributes further to studies on their infectivity and pathogenicity.
Assuntos
Variação Genética , Papillomaviridae , Infecções por Papillomavirus/epidemiologia , Filogenia , Brasil/epidemiologia , DNA Viral/análise , DNA Viral/genética , DNA Viral/isolamento & purificação , Feminino , Humanos , Dados de Sequência Molecular , Proteínas Oncogênicas Virais/genética , Papillomaviridae/classificação , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/virologia , Risco , Análise de Sequência de DNARESUMO
BACKGROUND AND OBJECTIVES: The severity of liver disease among hepatitis C patients on hemodialysis is controversial. The aim of this study was to compare the clinical, biochemical, and liver histologic characteristics of hepatitis C virus (HCV) in hemodialysis patients and in those with normal renal function. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: A case-control study was carried out with 36 HCV patients on hemodialysis and 37 HCV patients with normal renal function matched for gender, age at infection, and estimated time of infection. RESULTS: HCV patients on hemodialysis had lower levels of alanine aminotransferase and lower viral load. Hepatic fibrosis was significantly higher in the patients with normal renal function (73%) than in hemodialysis patients (47.2%, P < 0.025); the same was observed for inflammatory activity (control group 59.5% versus hemodialysis patients 27.7%, P = 0.003). In addition, the risk of tissue inflammation was four times lower in hemodialysis patients (odds ratio = 0.23, P < 0.004), and severe inflammatory activity on biopsy was the only independent risk factor for fibrosis (P < 0.001). CONCLUSIONS: The lower biochemical and inflammatory activities observed in hemodialysis patients suggest that hemodialysis and uremia may have a protective role against progression of the disease caused by HCV.
Assuntos
Hepatite C Crônica/diagnóstico , Falência Renal Crônica/terapia , Cirrose Hepática/virologia , Fígado/patologia , Diálise Renal , Uremia/terapia , Adulto , Idoso , Alanina Transaminase/sangue , Estudos de Casos e Controles , Progressão da Doença , Feminino , Hepatite C Crônica/complicações , Hepatite C Crônica/metabolismo , Hepatite C Crônica/patologia , Humanos , Mediadores da Inflamação/metabolismo , Falência Renal Crônica/complicações , Falência Renal Crônica/metabolismo , Falência Renal Crônica/patologia , Fígado/enzimologia , Cirrose Hepática/patologia , Masculino , Pessoa de Meia-Idade , Razão de Chances , Medição de Risco , Fatores de Risco , Índice de Gravidade de Doença , Uremia/etiologia , Carga ViralRESUMO
Este trabalho é parte de uma pesquisa em Educação e Saúde, desenvolvida pela autora, no mestrado em Ciências da Saúde da criança e do Adolescente, da Faculdade de Medicina - UFMG e foi realizado na Clínica-escola de Fonoaudiologia do Centro Universitário Metodista Izabela Hendrix, BH-MG. Aborda a construção e a análise do Discurso do Sujeito Coletivo de familiares de crianças com alterações no desenvolvimento da linguagem, que aguardam vaga para atendimento especializado. O discurso analisado revela que as crenças e as concepções que os familiares têm a respeito das dificuldades de seus filhos interferem no modo como conduzem suas ações e no enfrentamento de suas dificuldades. As alterações de linguagem podem criar problemas nas relações familiares e comprometer o desenvolvimento e a aprendizagem da criança, sendo, portanto, um tema de saúde coletiva