Search results for " centri"

A new method for the in situ determination of phospholipids after thin-layer separation

1973

Abstract A very sensitive method has been devised for the in situ determination of phospholipids after thin-layer chromatographic separation, which enabled us to investigate the phospholipid content of erythrocytes and ATPase preparations. The phospholipid compositions of the ATPase preparations and of the erythrocytes are different, and the relative phospholipid compositions of the Na,K-ATPase preparations are also different, which indicates that Na,K- and Ca-ATPase seem to be different with regard to their phospholipid composition. An increase in temperature during the preparation procedure yields a Ca-ATPase preparation (II), which exhibits different kinetic properties and a different ph…

Purification of Large Cytosolic Proteases for In Vitro Assays: 20S and 26S Proteasomes

2012

Proteasomes are the main cytosolic proteases responsible for generating peptides for antigen processing and presentation in the MHC (major histocompatibility complex) class-I pathway. Purified 20S and 26S proteasomes have been widely used to study both specificity and efficiency of antigen processing. Here, we describe the purification of active human 20S and 26S proteasomes from human erythrocytes by DEAE-ion exchange chromatography, ammonium sulfate precipitation, glycerol density gradient centrifugation, and Superose-6 size exclusion chromatography and their characterization using fluorogenic substrates and specific inhibitors.

Isolation and Functional Analysis of Organelles

2006

Fusarium proliferatum: induction and intracellular location of a lipoxygenase

1998

The fungus Fusarium proliferatum was grown in a soya oil- and glucose-supplemented medium. Induction of lipoxygenase activity (13-fold) in an extract of Fusarium biomass has been observed at day four on soya oil culture medium. A band at 232 kDa was detected using a specific lipoxygenase stain combined with native PAGE. The method of fungal homogenate obtained has been checked via subcellular marker enzymes activities: succinate dehydrogenase (SDH) for mitochondrial fraction and NADH-cytochrome C reductase (NADH-cyt. C) for the microsomal fraction. Protoplasts production and disintegration followed by subcellular fractionation using differential centrifugation was the best method to recover…

Chitin synthetase activity in Candida albicans: subcellular distribution in yeast cells and protoplasts

1991

Chitin synthetase activity was detected in a partly zymogenic form in cell-free homogenates obtained from C. albicans yeast cells and protoplasts of the same type of cells. By isopycnic centrifugation on sucrose gradients of the cell-free homogenates two fractions with chitin synthetase activity were obtained: one was associated with the plasma membrane (labelled with [ 3 H]concanavalin A (con A) and buoyant density 1·195 g ml −1 ) in a partly active state, and the second was in the cytoplasm, where the enzyme was in a particulate fully zymogenic form, lacking affinity to con A. The buoyant density of the enzyme found in this location depended on the method of cell breakage. Lysis of partly…

Numerical analysis of density gradient centrifugation profiles from eukaryotic DNA

1990

A numerical method for the deconvolution of superimposed Gaussian distributions with a unique solution has been proposed by Medgyessy [10]. We have tested the usefulness of this method for the analysis of density gradient centrifugation profiles from eukaryotic DNA, which are normally composed from overlapping Gaussian distributed profiles of several subcomponents with different mean buoyant densities. From the analysis of human DNA and from model calculations we conclude that major subcomponents can be identified by this method, if they differ in their buoyant density by approximatly 0.005 g/ml. Minor components can only be identified if the total DNA has been fractionated according to buo…

Nuclear DNA fractions with grossly different base ratios in the genome of the marine sponge Geodia cydonium

2008

The DNA of the marine sponge Geodia cydonium (G.c.), a member of the phylogenetically old phylum Porifera, was characterized by density gradient centrifugation and by determining its genetic complexity by reassociation kinetics. At least five subcomponents were identified by curve-fit analyses of analytical density gradient centrifugation profiles of total G.c.-DNA. Four of these subcomponents were isolated from total G.c.-DNA by preparative density gradient centrifugation. The GC-contents of the subcomponents were determined to be 36.4%, 44.0%, 58.7%, and 66.1%, respectively. To our knowledge, such an extreme heterogeneity of DNA composition has never before been observed for any organism.…

Demonstration and partial characterization of an intermediate HBcAG (Dane particle) population.

1981

Abstract Hepatitis-B core antigen (HBcAg) was released from Dane particles previously separated from anti-HBc by repeated pelleting through sucrose gradients separated into three HBcAg populations when analysed by cesium chloride density gradient centrifugation. Heavy HBcAg particles banded at a density of 1.355 gm/ml, intermediate HBcAg particles at a density of 1.33 gm/ml, and light mediate HBcAg particles at a density of 1.30 gm/ml. Like heavy HBcAg particles, intermediate HBcAg particles contained DNA polymerase activity, but the ratio of HBcAg to DNA polymerase activity was significantly different in both populations. Intermediate HBcAg particles could not be separated from heavy HBcAg…

Steroid hormone receptors in the female urogenital tract

1994

The increasing clinical use of estrogens in the treatment of stress-incontinent women caused us to examine receptor patterns in the female bladder and urethral tissue, and to characterize possible receptors by means of Scatchard plot analysis, competition experiments and density gradient centrifugation.

L'intervento nei centri storici: l'ANCSA e il caso Erice

2014

Il Convegno “Salvaguardia e risanamento dei centri storico-artistici”, svoltosi a Gubbio nel Settembre del 1960, offre l’occasione per fare un bilancio sulla situazione dei centri storici, sull’insufficienza normativa dei piani regolatori, sulla tecnica urbanistica e confrontare contemporaneamente le esperienze concrete di alcuni piani e il relativo approccio al tema dell’intervento nei centri storici. Il comitato promotore del Convegno di Gubbio, noto per avere prodotto la celebre Dichiarazione finale poi denominata “Carta di Gubbio”, si trasformerà l’anno successivo in Comitato permanente fondando a Gubbio l’Associazione nazionale centri storico-artistici (ANCSA). Appare significativo che…