Evidence for multi-copy Mega-NUMTs in the human genome

The maternal mode of mitochondrial DNA (mtDNA) inheritance is central to human genetics. Recently, evidence for bi-parental inheritance of mtDNA was claimed for individuals of three pedigrees that suffered mitochondrial disorders. We sequenced mtDNA using both direct Sanger and Massively Parallel Sequencing in several tissues of eleven maternally related and other affiliated healthy individuals of a family pedigree and observed mixed mitotypes in eight individuals. Cells without nuclear DNA, i.e. thrombocytes and hair shafts, only showed the mitotype of haplogroup (hg) V. Skin biopsies were prepared to generate ρ° cells void of mtDNA, sequencing of which resulted in a hg U4c1 mitotype. The position of the Mega-NUMT sequence was determined by fluorescence in situ hybridization and two different quantitative PCR assays were used to determine the number of contributing mtDNA copies. Thus, evidence for the presence of repetitive, full mitogenome Mega-NUMTs matching haplogroup U4c1 in various tissues of eight maternally related individuals was provided. Multi-copy Mega-NUMTs mimic mixtures of mtDNA that cannot be experimentally avoided and thus may appear in diverse fields of mtDNA research and diagnostics. We demonstrate that hair shaft mtDNA sequencing provides a simple but reliable approach to exclude NUMTs as source of misleading results.     

Feed consumption and performance of dairy cows with alternate feedings of grass silage and maize silage

A feeding experiment was conducted with 10 dairy cows of the Fleckvieh breed and the cross Red Holstein Friesian × Fleckvieh, to study whether feeding with grass silage at the morning meal and maize silage at the evening meal (treatment B: alternating forage allocation) affects forage intake and milk production, in comparison with combined feeding with these two silages at each meal (treatment A). In order to prevent a selective forage consumption in treatment A, the two silages were given as a homogeneous mixture of nearly equal portions (51.6% maize silage, 48.4% grass silage) of dry matter (DM). The experiment was of switch-back design, with the treatment sequences ABA and BAB, and three experimental periods of 6 weeks.The daily forage consumption averaged 12.3 kg DM when the silages were given as a mixture and was significantly higher than the total forage consumption of 11.8 kg DM (P < 0.05) during the alternating allocation of the silages. In treatment B, daily intake of maize silage (7.10 kg DM) was greater than that of grass silage (4.70 kg DM/day). Furthermore, variation between cows in forage intake was significantly higher in this treatment than in treatment A. Average daily milk yield for treatment A was 18.75 kg with 3.84% fat and 3.70% protein, and 18.10 kg with 3.76% fat and 3.68% protein for treatment B. Production was significantly higher (P < 0.05), by 0.65 kg milk or 0.90 kg FCM, for treatment A.     

Human genes involved in lipolysis of plasma lipoproteins: mapping of loci for lipoprotein lipase to 8p22 and hepatic lipase to 15q21

We have used cDNA probes for lipoprotein lipase and hepatic lipase to determine the chromosomal and subchromosomal locations of the human genes for these lipolytic enzymes. Southern blot analysis of genomic DNA from 17 independent mouse-human somatic cell hybrids demonstrated the presence of the gene for human lipoprotein lipase on chromosome 8, whereas the gene for hepatic lipase was on chromosome 15. Regional mapping of the genes by in situ hybridization to human chromosomes indicated that the lipoprotein lipase gene (LPL) resides in the p22 region of chromosome 8, while hepatic lipase gene (HL) resides in the q21 region of chromosome 15. We previously reported, on the basis of nucleotide and amino acid homologies, that these genes are members of a gene family of lipases, and, thus, the present findings indicate that the members of this family are dispersed. The results are also of significance with respect to disorders involving deficiencies of the enzymes. In particular, they suggest that certain rare combined deficiencies of both enzymes do not involve mutations of the structural loci.     

Effects of different diets and levels of feeding on retention and efficiency of utilization of energy and protein by carp (Cyprinus carpio L.)

In two experiments the effects of wide range of feeding intensities on energy and protein retention and on efficiency of utilization of energy and protein by carp ( Cyprinus carpio L.) were studied. The feeding period varied between an initial live weight (LW) of 160g to a final LW of 520g. The utilization of ME for gain (k_g) was high with values of 0.6–0.8 at an adequate feeding level but decreased significantly to 0.3–0.1 at the lower levels of energy supplied. Deposition of 1 g protein required 44 kJ and 1g fat required 52 kj ME.