Search results for "Density gradient"

A rapid and simple method for the preparation of yeast mitochondrial DNA

1990

Isolation and Characterization of Epidermal DNA and RNA from Guinea Pig Skin

1971

DNA and RNA were isolated from mammalian epidermis in a relatively small scale procedure. The high purity and native state of the DNA isolated is reflected by its molar absorptivity E (P), its thermal hyperchromicity and its hyperchromicity upon DNase treatment and by its sedimentation profile as well as by its profile in a cesium chloride density gradient. The very low content of protein and RNA, as well as the data of DNA determination, indicate that this method permits the isolation of a highly purified product. This is further substantiated by the determination of UV absorption spectra and by analysis of the base composition.The mammalian skin DNA showed the following properties: Mol. w…

Univacuolar refractile hemocytes from the tunicate Ciona intestinalis are cytotoxic for mammalian erythrocytes in vitro

1996

A discontinuous, Percoll density gradient was used to separate hemocyte populations from the hemolymph of Ciona intestinalis. Hemocytes from each band were examined for their frequency, morphology, and cytotoxic activity against rabbit and sheep erythrocytes; results were expressed as a percentage of hemolysis. Statistical analysis revealed that only the "univacuolar" granulocytes from Band 5, which contain a vacuole of refractile material, were cytotoxic. Cytotoxic activity was inhibited by sphingomyelin. For the first time in tunicates, lytic activity against erythrocytes was assessed by an assay based on plaque-forming cells. Plaques of lysis were revealed against rabbit erythrocytes but…

Centrifugation does not alter spatial distribution of `BEP4' mRNA in paracentrotus lividus EGG

1997

AbstractParacentrotus lividus unfertilized eggs were centrifuged in a sucrose gradient, so to split each into two parts: a nucleated light fragment and an anucleated heavy fragment. Northern blot analyses utilizing a bep4 probe as animal marker and H2A histone gene and 12S-mit RNA as controls indicate that the eggs are elongated along the animal-vegetal axis during centrifugation and thereafter split into an animal and a vegetal half. Treatment of the eggs with colchicine before centrifugation abolishes the animal localization of bep4 mRNA.

Highly protective alkalinization by ammonia vapor diffusion in viscosimetric DNA damage assessment.

1988

A method for the measurement of viscosities correlated to DNA alterations in alkaline homogenate suspensions is described. The alkaline pH shift to afford cell lysis, DNA unfolding, and denaturation is attained by gaseous ammonia diffusion, thus avoiding shear stress from mechanical mixing. At the same time a stabilizing density gradient is established. This solution is run through a plastic measuring tube that is wide enough to minimize the influence of uneven swelling of the lysing DNA-containing components. Flow times under a carefully controlled water head are registered, and their ratios to control solutions are evaluated. The relative viscosities show a strong and irreversible depende…

Isolation of Cholinergic Synaptic Vesicles from the Myenteric Plexus of Guinea-Pig Small Intestine

1980

The acetylcholine-rich myenteric plexus-longitudinal muscle preparation of the guinea-pig small intestine has been subjected to subcellular fractionation using modifications of both classical methods and that originally devised for bulk isolation of cholinergic synaptic vesicles from the electromotor nerve terminals of Torpedo marmorata by means of density gradient centrifugation in a zonal rotor. The latter method gave a vesicle fraction with the highest acetylcholine content so far recorded for a mammalian particulate fraction, 30.9 +/- S.E.M. 1.8 (5) nmol of acetylcholine . mg of protein-1. Electron-microscopical examination showed that it consisted of a homogeneous preparation of vesicl…

Tissue evacuated during joint replacement procedure as a source of mononuclear cells.

2017

Background Different cell populations from bone marrow were used in various clinical trials for cardiac diseases during last decade. Four clinical studies are ongoing in our institution and enroll patients with cardiac diseases, coronary disease, type 2 diabetes, and osteoarthritis. The density gradient is used to separate bone marrow mononuclear cells. Joint replacement procedures were associated with significant loss of tissue. Usually, excess tissue as bone marrow, peripheral blood and fat are removed to clean operation site. The aim of this study is to prove whether removed tissue during joint replacement procedure can be considered as a significant source of mononuclear cells. Methods …

Involvement of microsomal vesicles in part of the sensitivity of carnitine palmitoyltransferase I to malonyl-CoA inhibition in mitochondrial fraction…

1994

Liver mitochondrial fractions as normally isolated contain only 10-20% of total mitochondria and may not be representative of the whole mitochondrial population. This study was designed to evaluate the dependence of the sensitivity of carnitine palmitoyl-transferase I (CPT I) to malonyl-CoA inhibition in mitochondrial fractions that are not normally studied. Four fractions prepared from rat liver were found to be contaminated to different extents by microsome vesicles, on the basis of marker-enzyme activities and micrographic data. Purification of mitochondrial fractions on a Percoll gradient decreased to some extent the microsomal contamination, which was due in part to the existence of cl…

Steroid hormone receptors in human melanoma.

1981

Human melanomas were investigated for the presence of high-affinity estrogen-, gestagen-, and glucocorticoid-binding proteins. A statistically significant difference was found for mean estrogen receptor (ER) concentrations in melanomas of male versus female origin: female origin 37.6 (0-107) fmol/mg protein, male origin 3.9 (0-8.3) fmol/mg protein. No significant difference between sexes was found for gestragen receptors: 41.5 (0-194) fmol/mg protein for melanomas of female origin versus 99 (0-362) fmol/mg protein for male. Sucrose density gradient analyses revealed specific binding for both receptor types in the 4-5 S region as well as in the 8 S region. The binding affinities were in the …

The Measurement of Enzyme Activities in the Resting Human Polymorphonuclear Leukocyte — Critical Estimate of a Method

1993

As a system for study, the isolated human polymorphonuclear leukocyte combines the advantages of a quasi-non-invasive preparation with a nearly complete complement of enzymes of carbohydrate and energy metabolism. However, small sample volumes and, in some cases, very low enzyme activities make high demands on sample processing, storage, and performance of continuous measurements, if the enzyme activities are to be measured with acceptable reproducibility. In the presented study several aspects of homogenization, storage, and continuous measurement were scrutinized, to identify critical steps and consider ways of optimizing the method. Polymorphonuclear leukocytes were separated from the bl…