Dados preliminares sobre partículas semelhantes a virus em células de triatomíneos

Viral particles distributed within cytolysomes or other cytoplasmic globule types, giving rise to membrane enclosed paracrystalline arrays, and between membranes of contiguous cells, were found in the Malpighian tubes of Triatoma infestans Klug and Panstrongylus megistus Burmeister. Other tissues of the same insects also display paracrystalline arrays of these viruses. A RNP composition is assumed for the viruses based on their response to cytochemical methods at the EM level. Drastic morphological changes were not found in the injected tissues. This infection assumed to be promoted by ingestion of a bird infected blood meal.Nos tubos de Malpighi de Triatoma infestans Klug e de Panstrongylus megistus Burmeister foram encontradas estruturas víricas distribuídas no citoplasma, no interior de citolissomos ou outros tipos de glóbulos, formando às vezes arranjos para cristalinos envolvidos por membrana, e entre plasmalemas de células contíguas. Outros tecidos dos mesmos insetos apresentam arranjos paracristalinos desses virus. A análise citoquímica ao nível da microscopia eletrônica indica serem estes virus compostos de RNP. Não são encontradas alterações morfológicas drásticas nos tecidos infectados. Admite-se que tais virus tenham sido ingeridos durante uma alimentação de sangue infectado de ave

Changes In Liver Cell Dna Methylation Status In Diabetic Mice Affect Its Ft-ir Characteristics.

Lower levels of cytosine methylation have been found in the liver cell DNA from non-obese diabetic (NOD) mice under hyperglycemic conditions. Because the Fourier transform-infrared (FT-IR) profiles of dry DNA samples are differently affected by DNA base composition, single-stranded form and histone binding, it is expected that the methylation status in the DNA could also affect its FT-IR profile. The DNA FT-IR signatures obtained from the liver cell nuclei of hyperglycemic and normoglycemic NOD mice of the same age were compared. Dried DNA samples were examined in an IR microspectroscope equipped with an all-reflecting objective (ARO) and adequate software. Changes in DNA cytosine methylation levels induced by hyperglycemia in mouse liver cells produced changes in the respective DNA FT-IR profiles, revealing modifications to the vibrational intensities and frequencies of several chemical markers, including νas -CH3 stretching vibrations in the 5-methylcytosine methyl group. A smaller band area reflecting lower energy absorbed in the DNA was found in the hyperglycemic mice and assumed to be related to the lower levels of -CH3 groups. Other spectral differences were found at 1700-1500 cm(-1) and in the fingerprint region, and a slight change in the DNA conformation at the lower DNA methylation levels was suggested for the hyperglycemic mice. The changes that affect cytosine methylation levels certainly affect the DNA-protein interactions and, consequently, gene expression in liver cells from the hyperglycemic NOD mice.9e10229

Spatial distribution of heterochromatin bodies in the nuclei of Triatoma infestans (Klug)

Constitutive heterochromatin typically exhibits low gene density and is commonly found adjacent or close to the nuclear periphery, in contrast to transcriptionally active genes concentrated in the innermost nuclear region. In Triatoma infestans cells, conspicuous constitutive heterochromatin forms deeply stained structures named chromocenters. However, to the best of our knowledge, no information exists regarding whether these chromocenters acquire a precise topology in the cell nuclei or whether their 18S rDNA, which is important for ribosome function, faces the nuclear center preferentially. In this work, the spatial distribution of fluorescent Feulgen-stained chromocenters and the distribution of their 18S rDNA was analyzed in Malpighian tubule cells of T. infestans using confocal microscopy. The chromocenters were shown to be spatially positioned relatively close to the nuclear periphery, though not adjacent to it. The variable distance between the chromocenters and the nuclear periphery suggests mobility of these bodies within the cell nuclei. The distribution of 18S rDNA at the edge of the chromocenters was not found to face the nuclear interior exclusively. Because the genome regions containing 18S rDNA in the chromocenters also face the nuclear periphery, the proximity of the chromocenters to this nuclear region is not assumed to be associated with overall gene silencing.133CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP304668/2014-1; 304668/2014-1Não tem2015/10356-2This research was supported by the São Paulo State Research Foundation (FAPESP, Brazil; grant no. 2015/10356-2) and the Brazilian National Council for Research and Development (CNPq, grant no. 304668/2014-1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. C.H.L.I. received a fellowship from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brazil – Finance Code 001). M.L.S.M. received a fellowship from CNPq (grant no. 304668/2014-1)

Nuclear phenotype changes after heat shock in Panstrongylus megistus (Burmeister)

The nuclear phenotypes of Malpighian tubule epithelial cells of male nymphs of the blood-sucking insect, Panstrongylus megistus, subjected to short- and long-duration heat shocks at 40ºC were analyzed immediately after the shock and 10 and 30 days later. Normal nuclei with a usual heterochromatic body as well as phenotypes indicative of survival (unravelled heterochromatin, giants) and death (apoptosis, necrosis) responses were observed in control and treated specimens. However, all nuclear phenotypes, except the normal ones, were more frequent in shocked specimens. Similarly altered phenotypes have also been reported in Triatoma infestans following heat shock, although at different frequencies. The frequency of the various nuclear phenotypes observed in this study suggests that the forms of cell survival observed were not sufficient or efficient enough to protect all of the Malpighian tubule cells from the deleterious effects of stress. In agreement with studies on P. megistus survival following heat shock, only long-duration shock produced strongly deleterious effects.27127

Genomic profiling of type-1 adult diabetic and aged normoglycemic mouse liver

Background: Hyperglycemia induces chromatin remodeling with consequences on differential gene expression in mouse hepatocytes, similar to what occurs during aging. The liver is the central organ for the regulation of glucose homeostasis and xenobiotic and lipid metabolism and is affected by insulin signaling. The precise transcriptional profiling of the type-1 diabetic liver and its comparison to aging have not been elucidated yet. Methods: Here, we studied the differential genomic expression of mouse liver cells under adult hyperglycemic and aged normoglycemic conditions using expression arrays. Results: Differential gene expression involved in an increase in glucose and impaired lipid metabolism were detected in the type-1 diabetic liver. In this regard, Ppargc1a presents an increased expression and is a key gene that might be regulating both processes. The differential gene expression observed may also be associated with hepatic steatosis in diabetic mouse liver, as a secondary disease. Similarly, middle-aged mice presented differential expression of genes involved in glucose, lipid and xenobiotic metabolism. These genes could be associated with an increase in polyploidy, but the consequences of differential expression were not as drastic as those observed in diabetic animals. Conclusions: Taken together, these findings provide new insights into gene expression profile changes in type-1 diabetic liver. Ppargc1a was found to be the key-gene that increases glucose metabolism and impairs lipid metabolism impairment. The novel results reported here open new areas of investigation in diabetic research and facilitate the development of new strategies for gene therapy

Sobrevivência e ocorrência de muda em Triatoma infestans Klug (Hemiptera, Reduviidae) após choque de temperatura

Survival and molting occurrence were studied in specimens of Triatoma infestans over 30 days after temperature shocks. Hyperthermal and hypotermal shocks could be found to affect both survival and molting incidence as a function of temperature and period of the developement phase and sex of the specimens. Considering the various test conditions, the shock at 0°C for 12 h was found to elicit the most deleterious effect, whereas shocks at 40°C and 0°C even for 1 h are interpreted as affecting the hormonal balance which controls molting. Cases of a rise in post-shock survival are suggested to have been favored by heat-shock protein action.A sobrevivência e a ocorrência de mudas em espécimes de Triatoma infestans foram estudados num período de 30 dias após choques de temperatura. Foi demonstrado que choques hipertérmico e hipotérmico interferem nesses processos, na dependência da temperatura do choque, tempo de sua duração, fase de desenvolvimento e sexo dos espécimes. Dentre as situações experimentais utilizadas, o choque a 0°C por 12 h pareceu produzir a ação mais deletéria, porém choques a 40°C e 0°C mesmo por uma hora são admitidos como afetando o desenvolvimento hormonal que controla a muda. Casos de aumento de sobrevivência pós-choque são sugeridos como tendo sido favorecidos por ação de proteínas de choque térmico

Valproic acid influences the expression of genes implicated with hyperglycaemia-induced complement and coagulation pathways

Because the liver plays a major role in metabolic homeostasis and secretion of clotting factors and inflammatory innate immune proteins, there is interest in understanding the mechanisms of hepatic cell activation under hyperglycaemia and whether this can be attenuated pharmacologically. We have previously shown that hyperglycaemia stimulates major changes in chromatin organization and metabolism in hepatocytes, and that the histone deacetylase inhibitor valproic acid (VPA) is able to reverse some of these metabolic changes. In this study, we have used RNA-sequencing (RNA-seq) to investigate how VPA influences gene expression in hepatocytes. Interesting, we observed that VPA attenuates hyperglycaemia-induced activation of complement and coagulation cascade genes. We also observe that many of the gene activation events coincide with changes to histone acetylation at the promoter of these genes indicating that epigenetic regulation is involved in VPA action11CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP304668/2014-12010/50015-6; 2012/03238-5; 2014/10198-5; 2015/10356-2NHMRC; International Joint Program. Professor Sam El-Osta is a National Health and Medical Research Council; Senior Research Fello