Remarks on the differentiation of lysosomes from cultured human fibroblasts by silica gradient centrifugation

Two bands of lysosomal marker enzyme activity from cultured skin fibroblasts can be obtained by silica gradient centrifugation (Rome, L H, Garvin, L H, Allietta, M M & Neufeld, E F, Cell 17 (1979) 143) [3]. It is shown that the distribution of lysosomes into the bands in the upper and lower part of the density gradient depends on the starting density and the shape of the self-forming gradient. A plot of marker enzyme activity vs density reveals that the two bands stem from a lysosomal population with unimodal density distribution.     

Lipolytic activity of porcine adipose tissue after preincubation

1. The stimulated lipolytic rate in porcine adipose tissue slices was markedly increased after preincubation with or without isoproterenol. 2. Preincubation with and without insulin, dibutyryl-cAMP or theophylline suggested that the activation of the stimulated rate resulted from inhibition or inactivation of cAMP-phosphodiesterase activity during preincubation. 3. Preincubation with isoproterenol also caused activation of the basal (no exogenous hormone) lipolytic activity. 4. However, much of this activation appeared to result from carry-over of isoproterenol into the incubation medium because it could be lowered by additional washing of the tissue, by lower isoproterenol concentration or by propranolol.     

Simultaneous fine mapping of closely linked epistatic quantitative trait loci using combined linkage disequilibrium and linkage with a general pedigree

Causal mutations and their intra- and inter-locus interactions play a critical role in complex trait variation. It is often not easy to detect epistatic quantitative trait loci (QTL) due to complicated population structure requirements for detecting epistatic effects in linkage analysis studies and due to main effects often being hidden by interaction effects. Mapping their positions is even harder when they are closely linked. The data structure requirement may be overcome when information on linkage disequilibrium is used. We present an approach using a mixed linear model nested in an empirical Bayesian approach, which simultaneously takes into account additive, dominance and epistatic effects due to multiple QTL. The covariance structure used in the mixed linear model is based on combined linkage disequilibrium and linkage information. In a simulation study where there are complex epistatic interactions between QTL, it is possible to simultaneously map interacting QTL into a small region using the proposed approach. The estimated variance components are accurate and less biased with the proposed approach compared with traditional models.     

The effect of insulin on porcine adipose tissue lipogenesis

1. This laboratory and others have not been able to demonstrate consistent insulin stimulation of glucose incorporation into lipid by porcine adipose tissue in vitro. 2. A multiplicity of tissue handling procedures, additions to the incubation medium, and pig size (age) did not allow the expression of a consistent and substantial insulin stimulation. 3. It is suggested that the twofold or greater stimulation of glucose metabolism observed occasionally in this laboratory results from pig genetics, husbandry, or seasonal effects.