Mitochondrial mutation in adult patient with Hypertrophic Cardiomyopathy and renal failure

Background: Mitochondrial diseases (MDs) (1:5000-10000) represents a wide group of human disorders associated with mitochondrial DNA (mtDNA) variations causing defect of oxidative phosphorylation system, whereas nuclear genome mutations are somewhat rare. The extremely heterogeneous clinical phenotype, extending from oligosymptomatic condition to complex syndromes involving neurological, ophtalmological, gastroenterological and endocrine features, depends to the involved tissue well as to the specific mtDNA mutations and their heteroplasmic level. Diabetes and deafness are common features of mitochondrial diseases, while renal alterations are rarely reported, especially in adults, probably because of lack of association to mitochondrial conventional phenotypes. Case Presentation: We investigated a 62 years old male affected by hypertrophic cardiomyopathy (HCM) and renal failure that caused already a bilateral transplantation. Pathological anamnesis revealed also diabetes, deafness and Crohn disease. Family history of cardiomyopathy showed a strong mitochondrial involvement. The proband's mother, three brothers (one of which died of renal failure at 26 years), the sister and her child were affected. Materials and Methods: Genomic DNA from peripheral blood and buccal cells was extracted with the Kit-Nucleon-BACC2 (Illustra DNA-Extraction Kit-BACC2-GE Healthcare, UK) and the whole mitochondrial genome was amplified by two pair of primers designed in our laboratory to generate two overlapping fragments. The PCR products were then sequenced and compared to mitochondrial reference sequence (rCRS NC_012920). Results and Discussion: In both biological samples the mtDNA analysis showed the heteroplasmic A3243G mutation in the tRNALeu (UUR), frequently associated with MDs. A cardiological involvement leading to hypertrophic remodelling, caused to mitochondria intermyofibrillar proliferation, occurs up to 40% of patients with mtDNA disease. Molecular backgrounds of mitochondrial cardiomyopathy of adult age are still quite poorly known and the A3243G mutation in tRNA Leu(UUR) of mtDNA has been reported in 40-60% of patients with HCM. The interesting finding presented here support the knowledge that mitochondrial gene altertation represents a possible etiology in cardiological patients with unexplained renal failure. This is particularly true, as in this case, when other associated symtoms linked with dysfunctional oxidative phosphorylation are present. The case presented in this report further suggests that a differential diagnosis in presence of HCM should be solved by a multidisciplinary approach together with mutation analysis of mitochondrial DNA

Investigation on genetically modified soybean (RoundUp Ready) in goat nutrition: DNA detection in suckling kids

The presence of plant DNA fragments in blood, kidney, hearth, liver, spleen and muscle tissue from suckling kids was investigated by using PCR approach. Fragments of high copy number chloroplast and low copy soybean lectin genes were found in several samples of kids whose mother were fed diet containing conventional (control) or transgenic soybean (treated). Only in treated group, fragments of 35S and CP4 epsps soybean genes were found in several samples

Comparative analysis of the SOL plasma in DEMO using EDGE2D/EIRENE and TECXY codes

In this contribution a benchmark of the 2D edge codes TECXY and EDGE2D-EIRENE is presented. A conventional DEMO scenario is considered by assuming a simplified geometry, with the target plates perpendicular to the separatrix, and a pure Deuterium plasma. Despite the different models adopted in the two codes, mainly related to the description of the neutral dynamics and to the different boundary conditions, the results show a good match both in terms of power load profiles on the outer target and predicted trends for global quantities. A scan in density and in diffusion coefficients is performed in order to identify the characteristic conditions and the different regimes of the SOL. Comparable values and similar dependency of the global quantities as a function of the power decay length is also observed. Keywords: EDGE2D, EIRENE, TECXY, DEMO, Diverto

In vitro fermentation of ten cultivars of barley silage.

The fermentation characteristics of whole-crop barley silages from ten different cultivars were evaluated by the in vitro gas production technique. The organic matter degradability of barley silage (62.9% in average) was comparable to those reported in our previous trials for oat (59.7%) and sorghum silages (65.5%); while the maximum gas production rate (5.38 ml/h in average) was slightly lower respect to oat (6.71 ml/h) and sorghum silage (6.74 ml/h). The mean nutritive value (4.00 MJ/kg DM) calculated on the basis of both chemical composition and in vitro fermentation data was comparable to that (4.16 MJ/kg DM) obtained in our previous research performed on corn silage, from crop sowed in the same area

1.6 Radiofrequency catheter ablation in paediatric age: For nodal re-entry tachycardia and atrio-ventricular re-entry tachycardia

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Detection of plant species-specific dna (barley and soybean) in blood, muscle tissue, organs and gastrointestinal contents of rabbit

The aim of this study was to detect plant DNA sequences from low copy number genes of barley grain and soybean meal, the latter being subjected to solvent extraction process, in blood, liver, kidney, spleen, muscle tissue and digesta (duodenum, caecum and faeces from rectal ampulla) of rabbits. For fattening, Hyla rabbits (20 males and 20 females) were fed a diet including barley grain (15%) and soybean meal (12%). Animals were slaughtered at 74 d of age (2 ± 0.2 kg live weight) and samples collected from each animal. The quality of each DNA sample was verified using the UNIV P/Q primers used to amplify a mammalian specifi c portion of mtDNA 16S rRNA gene. The presence of plant DNA was subsequently ascertained on the same DNA samples, as well as on barley and soybean (control). Two classes of plant DNA sequences were monitored via real-time PCR, using SYBR(R) Green I Dye: a high copy number chloroplast gene (trnl) and a low copy number specific for barley (metal-dependent hydrolase-like protein) and soybean (lectin) genes. Melting curve analysis was used to identify the PCR products. The chloroplast fragment detection frequency was higher (P<0.01) in muscle (90%), liver (80%), kidney (80%) and spleen (80%) than in blood (40%) and digesta samples. In the latter, chloroplast DNA was found in 40 and 30% of duodenum and caecum contents respectively, and in 30% of faeces. The specificity of the amplicons obtained was checked by sequencing and annotation. In the samples positive for chloroplast fragments, the frequency of detection of barley specific sequence was higher (P<0.01) in liver (62.5%), kidney (62.5%), spleen (62.5%) and digesta (100%) than in blood (25%) and muscle (22.2%) samples. The soybean lectin gene was not detected in animal samples, although it was seen in plant samples. Results confirm that, except for gastrointestinal tract (GIT), plant single copy genes are more difficult to identify in animal samples.Tudisco, R.; Calabrò, S.; Bovera, F.; Cutrignelli, M.; Nizza, A.; Piccolo, V.; Infascelli, F. (2010). Detection of plant species-specific dna (barley and soybean) in blood, muscle tissue, organs and gastrointestinal contents of rabbit. World Rabbit Science. 18(2). doi:10.4995/WRS.2010.18.1118

LCAT cholesterol esterification is associated with the increase of ApoE/ApoA-I ratio during atherosclerosis progression in rabbit

Apolipoprotein A-I and Apolipoprotein E promote different steps of reverse cholesterol transport, including lecithin-cholesterol acyltransferase stimulation. Our aim was to study the changes in the levels of Apolipoprotein A-I, Apolipoprotein E, and lecithin-cholesterol acyltransferase activity during atherosclerosis progression in rabbits. Quantitative echocardiographic parameters were analyzed in order to evaluate, for the first time, whether atherosclerosis progression in rabbit is associated to apolipoproteins changes and alteration of indices of cardiac function, such as systolic strain and strain rate of the left ventricle. Atherosclerosis was induced by feeding rabbits for 8 weeks with 2 % cholesterol diet. The HDL levels of cholesterol and cholesteryl esters were measured by HPLC. The lecithin-cholesterol acyltransferase activity was evaluated both ex vivo, as cholesteryl esters/cholesterol molar ratio, and in vitro. Apolipoproteins levels were analyzed by ELISA. The HDL levels of cholesterol and cholesteryl esters increased, during treatment, up to 3.7- and 2.5-fold, respectively, compared to control animals. The lecithin-cholesterol acyltransferase activity in vitro was halved after 4 weeks. During cholesterol treatment, Apolipoprotein A-I level significantly decreased, whereas Apolipoprotein E concentration markedly increased. The molar ratio Apolipoprotein E/Apolipoprotein A-I was negatively correlated with the enzyme activity, and positively correlated with both increases in the intima-media thickness of common carotid wall and cardiac dysfunction signs, such as systolic strain and strain rate of the left ventricle. © 2012 University of Navarra

LCAT cholesterol esterification is associated with the increase of ApoE/ApoA-I ratio during atherosclerosis progression in rabbit

Apolipoprotein A-I and Apolipoprotein E promote different steps of reverse cholesterol transport, including lecithin-cholesterol acyltransferase stimulation. Our aim was to study the changes in the levels of Apolipoprotein A-I, Apolipoprotein E, and lecithin-cholesterol acyltransferase activity during atherosclerosis progression in rabbits. Quantitative echocardiographic parameters were analyzed in order to evaluate, for the first time, whether atherosclerosis progression in rabbit is associated to apolipoproteins changes and alteration of indices of cardiac function, such as systolic strain and strain rate of the left ventricle. Atherosclerosis was induced by feeding rabbits for 8 weeks with 2 % cholesterol diet. The HDL levels of cholesterol and cholesteryl esters were measured by HPLC. The lecithin-cholesterol acyltransferase activity was evaluated both ex vivo, as cholesteryl esters/cholesterol molar ratio, and in vitro. Apolipoproteins levels were analyzed by ELISA. The HDL levels of cholesterol and cholesteryl esters increased, during treatment, up to 3.7- and 2.5-fold, respectively, compared to control animals. The lecithin-cholesterol acyltransferase activity in vitro was halved after 4 weeks. During cholesterol treatment, Apolipoprotein A-I level significantly decreased, whereas Apolipoprotein E concentration markedly increased. The molar ratio Apolipoprotein E/Apolipoprotein A-I was negatively correlated with the enzyme activity, and positively correlated with both increases in the intima-media thickness of common carotid wall and cardiac dysfunction signs, such as systolic strain and strain rate of the left ventricle. © 2012 University of Navarra