Evidence for a nuclear compartment of transcription and splicing located at chromosome domain boundaries

The nuclear topography of splicing snRNPs, mRNA transcripts and chromosome domains in various mammalian cell types are described. The visualization of splicing snRNPs, defined by the Sm antigen, and coiled bodies, revealed distinctly different distribution patterns in these cell types. Heat shock experiments confirmed that the distribution patterns also depend on physiological parameters. Using a combination of fluorescencein situ hybridization and immunodetection protocols, individual chromosome domains were visualized simultaneously with the Sm antigen or the transcript of an integrated human papilloma virus genome. Three-dimensional analysis of fluorescence-stained target regions was performed by confocal laser scanning microscopy. RNA transcripts and components of the splicing machinery were found to be generally excluded from the interior of the territories occupied by the individual chromosomes. Based on these findings we present a model for the functional compartmentalization of the cell nucleus. According to this model the space between chromosome domains, including the surface areas of these domains, defines a three-dimensional network-like compartment, termed the interchromosome domain (ICD) compartment, in which transcription and splicing of mRNA occurs

Experience of taking care of children exposed to HIV: a trajectory of expectations

OBJETIVO: conhecer a experiência do cuidador/mãe em relação ao cuidado para com a criança exposta ao vírus da imunodeficiência humana por transmissão vertical, na trajetória pós-natal. MÉTODO: pesquisa qualitativa, que utilizou o Interacionismo Simbólico como referencial teórico. Foram realizadas entrevistas com 39 cuidadores de crianças expostas ao vírus da imunodeficiência humana e em seguimento em um serviço especializado. Os dados foram analisados pela análise de conteúdo indutiva. RESULTADOS: apreenderam-se quatro categorias que reportam à experiência solitária de manuseio da terapia antirretroviral da criança, no que se refere principalmente à ausência ou incompletude de informações recebidas; estar atento aos cuidados, que incluem o uso da profilaxia para pneumonia, vacinas e outros restritos à interação mãe/criança; querer omitir a presença do vírus da imunodeficiência humana pelo medo do preconceito e olhar o futuro e temer a doença. CONCLUSÃO: a presença do vírus da imunodeficiência humana e a ameaça dessa infecção na criança são capazes de gerar apreensão e tantos outros sentimentos como medo, culpa e ansiedade no cuidador. Os profissionais de saúde precisam trabalhar conjuntamente com a mãe para o enfrentamento das demandas e sofrimentos. Assim, o tratamento para evitar a transmissão vertical será eficiente e a mãe, juntamente à criança, viverá essa trajetória com apoio, apesar da apreensão pelo resultado.OBJETIVO: conocer la experiencia del cuidador/madre en relación al cuidado de niño expuesto al VIH por transmisión vertical en la trayectoria posnatal. MÉTODO: investigación cualitativa, que utilizó el Interaccionismo Simbólico como referencial teórico. Fueron realizadas entrevistas con 39 cuidadores de niños expuestos al VIH y que eran acompañados en un servicio especializado. Los datos fueron analizados por el análisis de contenido inductivo. RESULTADOS: se encontraron cuatro categorías que informan sobre la experiencia solitaria de la administración de la terapia antirretroviral del niño, en lo que se refiere principalmente a: la ausencia o a informaciones incompletas recibidas; al estar atento a los cuidados, que incluyen el uso de la profilaxis para neumonía, vacunas y otros restringidos a la interacción madre-niño; al querer omitir la presencia del VIH por miedo al prejuicio; y al mirar al futuro y temer a la enfermedad. CONCLUSIÓN: la presencia del VIH y la amenaza de infectar al niño son capaces de generar preocupación y varios otros sentimientos en el cuidador, como miedo, culpa y ansiedad. Los profesionales de la salud precisan trabajar conjuntamente con la madre para el enfrentamiento de las demandas y sufrimientos. Así, el tratamiento para evitar la transmisión vertical será eficiente y la madre con el niño vivirán esa trayectoria con apoyo, a pesar de la preocupación por el resultado.'OBJECTIVE: to learn about the experience of caregivers/mothers providing care to infants exposed to HIV through vertical transmission. METHODS: this qualitative study used Symbolic Interactionism as the theoretical framework. A total of 39 caregivers of children exposed to HIV in follow-up at a specialized service were interviewed. Data were analyzed through inductive content analysis. RESULTS: four categories were identified that report on the lonely experience of handling the child's antiretroviral therapy, mainly due to a lack of information or incomplete information; being attentive to required care, such as the use of prophylaxis for pneumonia, vaccines, and other practices restricted to the mother-child interaction; the desire to omit the HIV out of fear of prejudice and fear of the disease, considering future prospects. CONCLUSION: the HIV and the threat this infection may affect the child cause apprehension and feelings such as fear, guilt and anxiety in the caregivers. Healthcare workers need to work together with mothers so they are able to cope with demands and distress. Only then will the treatment to avoid vertical transmission be efficient and will mother and child be supported during the process, despite apprehension with the outcome

Cardiac Alpha-Myosin (MYH6) Is the Predominant Sarcomeric Disease Gene for Familial Atrial Septal Defects

Secundum-type atrial septal defects (ASDII) account for approximately 10% of all congenital heart defects (CHD) and are associated with a familial risk. Mutations in transcription factors represent a genetic source for ASDII. Yet, little is known about the role of mutations in sarcomeric genes in ASDII etiology. To assess the role of sarcomeric genes in patients with inherited ASDII, we analyzed 13 sarcomeric genes (MYH7, MYBPC3, TNNT2, TCAP, TNNI3, MYH6, TPM1, MYL2, CSRP3, ACTC1, MYL3, TNNC1, and TTN kinase region) in 31 patients with familial ASDII using array-based resequencing. Genotyping of family relatives and control subjects as well as structural and homology analyses were used to evaluate the pathogenic impact of novel non-synonymous gene variants. Three novel missense mutations were found in the MYH6 gene encoding alpha-myosin heavy chain (R17H, C539R, and K543R). These mutations co-segregated with CHD in the families and were absent in 370 control alleles. Interestingly, all three MYH6 mutations are located in a highly conserved region of the alpha-myosin motor domain, which is involved in myosin-actin interaction. In addition, the cardiomyopathy related MYH6-A1004S and the MYBPC3-A833T mutations were also found in one and two unrelated subjects with ASDII, respectively. No mutations were found in the 11 other sarcomeric genes analyzed. The study indicates that sarcomeric gene mutations may represent a so far underestimated genetic source for familial recurrence of ASDII. In particular, perturbations in the MYH6 head domain seem to play a major role in the genetic origin of familial ASDII

Genes from Chagas Susceptibility Loci That Are Differentially Expressed in T. cruzi-Resistant Mice Are Candidates Accounting for Impaired Immunity

Variation between inbred mice of susceptibility to experimental Trypanosoma cruzi infection has frequently been described, but the immunogenetic background is poorly understood. The outcross of the susceptible parental mouse strains C57BL/6 (B6) and DBA/2 (D2), B6D2F1 (F1) mice, is highly resistant to this parasite. In the present study we show by quantitative PCR that the increase of tissue parasitism during the early phase of infection is comparable up to day 11 between susceptible B6 and resistant F1 mice. A reduction of splenic parasite burdens occurs thereafter in both strains but is comparatively retarded in susceptible mice. Splenic microarchitecture is progressively disrupted with loss of follicles and B lymphocytes in B6 mice, but not in F1 mice. By genotyping of additional backcross offspring we corroborate our earlier findings that susceptibility maps to three loci on Chromosomes 5, 13 and 17. Analysis of gene expression of spleen cells from infected B6 and F1 mice with microarrays identifies about 0.3% of transcripts that are differentially expressed. Assuming that differential susceptibility is mediated by altered gene expression, we propose that the following differentially expressed transcripts from these loci are strong candidates for the observed phenotypic variation: H2-Eα, H2-D1, Ng23, Msh5 and Tubb5 from Chromosome 17; and Cxcl11, Bmp2k and Spp1 from Chromosome 5. Our results indicate that innate mechanisms are not of primary relevance to resistance of F1 mice to T. cruzi infection, and that differential susceptibility to experimental infection with this protozoan pathogen is not paralleled by extensive variation of the transcriptome