Bidimensional electrophoresis of factor VIII antigen on cellulose acetate

It is described two-dimensional immunoelectrophoresis using cellulose acetate as supporting medium compared with agarose gel method. The results show that the determination of the FVIIIR: Ag on cellulose acetate is a technique more simple and rapid than agarose gel method and many free from the technical failure.     

Histological localization of citrus infectious variegation virus (CVV) inPhaseolus vulgaris

Summary Ultrathin sections ofPhaseolus vulgaris leaves were studied in the electron microscope. The leaves were taken from plants, both healthy and experimentally infected with CVV. The sieve tubes and companion cells of all samples contained a slime-like substance, more or less organized into compact systems. In the mature sieve elements of virus-infected plants systems of parallel membranes were seen along which spherical particles, of about 30 m diameter, were aligned in simple rows.Pubbl No. 117 of Gruppo di Ricerca per le Virosi, del C. N. R.     

A procedure for selecting mammalian cells with an impairment in oxidative phosphorylation.

The mitochondrial encephalomyopathies in man are characterized by heterogeneous defects leading to an impairment in the pathway of aerobic energy production. As a means of investigating the molecular and genetic mechanisms underlying these disorders we have developed a procedure for selecting mammalian cell lines with features resembling the human pathological phenotypes. The principle of the selection is the use of a fluorescent amphiphilic dye, 2,4-(dimethylamino)-1-styrylmethylpyridiniumiodine, a cation showing two main features. Firstly, it is accumulated by mitochondria to an extent correlated with the magnitude of the electrochemical gradient of protons across the mitochondrial inner membrane. Secondly, upon irradiation with UV light, it gives rise to formation of free radicals, which inflict damage to the cell. Mutant cells with an impairment in oxidative phosphorylation will have more chance to survive than wild type cells. The selection procedure was applied to a stock of mutagenized Chinese hamster ovary cells. After subcloning of the cells which survived the selection procedure, twenty-six independent clones were isolated. Eighteen of the clones had a partial deficiency of cytochrome c oxidase ranging from 30 to 60% of the activity in control cells. The properties of two of the clones are described. One clone has been cultured under non-selective conditions for at least 12 months with retention of the partial deficiency of cytochrome c oxidase.     

Subcellular localization of cholesterol ester hydrolase in the human intestine

Immunocytochemistry and subcellular fractionation were used to localize the cholesterol ester hydrolase in the human small intestine. A positive immunoreaction, when using antibodies directed against pancreatic cholesterol ester hydrolase, was mainly found in endocytotic vesicles. Moreover, a label by gold particles was observed in intercellular spaces where lymphatic tissue merges. No specific immunoreactivity was obtained with the mucosa when sera directed against human pancreatic chymotrypsinogen and human pancreatic lipase were used. Conventional subcellular fractionation was performed after extensive washing of enterocytes to rule out any possible contamination by pancreatic enzymes. In these conditions a bile salt-dependent cholesterol ester hydrolase activity was detected in the soluble fraction of cells. Data agree with the concept that the intestinal cholesterol ester hydrolase may have a pancreatic origin. The absorption, if any, of this enzyme by enterocytes seems specific since other pancreatic (pro)enzymes tested (lipase, chymotrypsinogen) are not detected in these cells.     

RNA synthesis in neuronal and glial cell nuclei from rat cerebral hemispheres during early postnatal development

RNA synthesis in rat cerebral hemispheres at 1, 5, and 10 days of age and the relative contribution brought by neuronal and glial nuclei to RNA synthesis was investigated. The experiments were carried out both in vivo (by i.p. injection of [³H]uridine) and in vitro (either by incubation of tissue slices with [³H]uridine or by determination of RNA polymerase activities). The labeling of RNA decreases from 1 to 10 days of age both in vivo and in vitro; the decrease is of the same extent in neuronal and glial nuclei. RNA polymerase activity Mg²⁺-dependent does not change significantly from 1 to 10 days of age either in total, in neuronal, or in glial nuclei, whereas the Mn²⁺-dependent activity increases significantly over the same developmental period studied. The significance of RNA polymerase assay as an index of in vivo RNA synthesis is discussed.     

Local breeding experience and the reproductive performance of Tree Swallows

ABSTRACT The potential advantages of repeated breeding at a particular location should improve reproductive performance in long‐lived species of birds. However, for short‐lived species, natural selection should favor individuals that most quickly develop competency in reproduction. Therefore, we hypothesized that local breeding experience beyond the first breeding attempt at a particular location would have little effect on subsequent reproductive performance of Tree Swallows (Tachycineta bicolor), a species where about 50% of adults breed only once in their lives. We tested this hypothesis using data collected from Tree Swallows in Michigan from 1993 to 2002. Because we were specifically interested in examining the effects of local breeding experience on reproductive performance, we restricted our analyses to after‐second‐year (ASY) females and their mates that we first encountered as breeders. Consistent with our hypothesis, we found no relationship between repeated local breeding experience and the reproductive performance of ASY female Tree Swallows and their mates as measured by clutch size and number of fledged young. However, pairs with more combined total local experience tended to lay eggs earlier in the season. These results suggest that Tree Swallows may benefit from breeding site fidelity, not because repeated local experience improves reproductive performance as measured by the production of fledglings, but because returning individuals acquire nest cavities earlier and are able to begin breeding earlier, providing time to renest in case of early nest failure.     

Human sperm motility: a molecular study of mitochondrial DNA,mitochondrial transcription factor A gene and DNA fragmentation

Molecular Biology Reports - Alterations affecting the mitochondrial genome and chromatin integrity of spermatozoa impair male reproductive potential. This study aimed to evaluate the impact of...