Identification of centromere proteins in different mammalian cells

The characterization of centromeric proteins is facilitated using anti-centromere antibodies present in the sera of patients with the CREST variant of scleroderma. We have employed these sera to determine whether or not those proteins are present in different mammalian species, as well as to study their tissue distribution. Here, we describe the immunofluorescent pattern and the proteins recognized by CREST sera in dividing and resting cells from mouse, rat, swine, hamster, rabbit, and man. In nuclear preparations from cultured cells, thymocytes and spermatozoa from these species, the antigens recognized by CREST sera are proteins of 18 to 20 kDa in all species tested, except in rat. Additionally, two peptides of 80 and 140 kDa were observed in human preparations. In contrast, a 50 kDa peptide is the primary protein detected by the sera in rat nuclei.     

Subcellular Distribution of UDP-Galactose:Ceramide Galactosyltransferase in Rat Brain Oligodendroglia

Oligodendrocytes isolated from 18-19-day-old rat brain were homogenized in 0.32 M sucrose. The homogenate was centrifuged at 100,000 g for 50 min in a gradient containing 0.8, 1.05, and 1.3 M sucrose. Three discrete bands were obtained at the interfaces 0.32-0.8 (F1), 0.8-1.05 (F2), and 1.05-1.3 M (F3). The distribution of UDP-galactose:ceramide galactosyltransferase (CgalT) activity in each fraction was measured using liposomes containing normal fatty acid-containing ceramides (NFA-CgalT activity) or 2-hydroxy fatty acid-containing ceramides (HFA-CgalT activity). Although detection of both CgalT activities was possible in all fractions, HFA-CgalT activity was enriched in F1 and F2 fractions, which also showed an enrichment of Golgi and endoplasmic reticulum markers, respectively. It is interesting that NFA-CgalT activity was significantly enriched in the F2 fraction. These results suggest that hydroxylated and nonhydroxylated galactocerebrosides may be synthesized at different intracellular locations.     

Study on the mutagenicity of nifurtimox and eight derivatives with the L-arabinose resistance test of Salmonella typhimurium

The mutagenicity of nifurtimox (nfx) and 8 nfx analogues has been investigated with the L-arabinose forward-mutation assay of Salmonella typhimurium. The nfx analogues tested were obtained by replacing the 3-methyl-4-yl-tetrahydro-1,4-thiazine-1,1-dioxide group of the parent compound with the following other groups: indazol-1-yl (1); pyrazol-1-yl (2); benzimidazol-1-yl (3); 1,2,4-triazol-4-yl (4); 1-methyl-3-methylthio-1,2,4-triazol-4-yl-5-thione (5); 3,5-bis(methylthio)-1,2,4-triazol-4-yl (6); 1-adamantyl (7); 4,6-diphenylpyridin-1-yl-2-one (8). The mutagenic activity of each chemical was determined by the standard plate-incorporation test, in the presence or absence of the S9 activation mixture. The 9 compounds were mutagenic and exhibited linear dose-mutagenic response relationships. They were direct-acting mutagens and showed a nearly 1000-fold range in mutagenic potency from chemical 1 to nfx. In most cases, the addition of S9 mixture to the test plates decreased the mutagenicity of compounds. This effect was particularly noticeable in the case of chemicals 1-3, 5 and 7 where a more than 70% decrease in mutagenic activity was observed in the presence of the S9 mixture. The mutagenic potency of compounds in the Ara test showed a negative linear correlation with previously reported antitrypanosomal activity. Thus, chemicals 6 and 8 with in vitro activities against Trypanosoma cruzi clearly superior to that of nfx showed 2 of the lowest mutagenic potencies in the Ara test and these were only somewhat higher than the mutagenicity of the reference drug.     

The prophylactic effect of diphenylhidantoin in the kindling effect on reptiles (Gallotia galloti)

1. Lizards Gallotia galloti received daily 3 mg/kg body wt of diphenylhidantoin (DPH) over a period of 15 days and at the same time the animals were kindled. 2. The progression of the kindling effect was evaluated by counting the number of spontaneous epileptiform potentials, the duration of afterdischarges and the duration of electroencephalographic spontaneous seizures. 3. The diphenylhidantoin treated group, relative to controls presented: (a) significant reduction in the duration of afterdischarges and spontaneous electroencephalographic seizures; and (b) increased frequency of the spontaneous epileptiform potentials.     

Serotonin uptake in the central nervous system of rats fed a corn-diet

1. Endogenous serotonin (5-HT), 5-hydroxyindol acetic acid (5-HIAA) content and exogenous 5-HT uptake (Km and Vmax) were measured in different brain regions (cerebellum, diencephalon, brain stem and telencephalon) of rats fed with a corn diet and restricted protein (8%) diet during 6 weeks. 2. A reduction of 5-HT levels was found in all regions studied of animals fed a corn diet, whereas, 5-HIAA was only decreased in brain stem and diencephalon. 3. An important increase in Km and Vmax were registered in brain stem and diencephalon of protein restricted animals, whereas, an increase of 5-HT uptake affinity in cerebellum, brain stem and telencephalon (35, 42 and 33% respectively) was observed. Simultaneously, under corn diet conditions, the Vmax decreased 40, 30 and 34% respectively in those regions. 4. It is suggested that the brain stem was the more sensitive area under nutritional restricted conditions and the development of some possible compensatory mechanisms of the 5-HTergic system is discussed.     

Structure and floristic composition of the lowland rain forest of Los Tuxtlas,Mexico

Physiognomy, structure and floristic composition of one hectare of lowland tropical rain forest was studied in detail at Los Tuxtlas, Mexico. Physiognomically, the Los Tuxtlas forest should be classified as lowland tropical high evergreen rain forest. The forest showed a closed canopy at 30–35 m. Of all woody, non-climbing species with a DBH1.0 cm 89.4% (94.5% of all individuals) were evergreen, 25.4% (59.5% of the individuals) had compound leaves, and over 80% of species (and individuals) had leaves in the notophyll and mesophyll size classes. The forest structure was characterized by a low density (2976 individuals with a DBH1.0 cm, 346 individuals with a DBH10.0 cm, per ha, excluding vines) with an average basal area (38.1 m², DBH1.0 cm, 34.9 m², DBH10.0 cm, per ha, excluding vines). This was attributed to the relative maturity of the forest on the study plot. The study plot contained 234 species (11 208 individuals with a height 0.5 m), of which 55.1% (34.8% of individuals) were trees, 9.4% (6.8%) shrubs, 3.4% (44.3%) palms, 20.1% (5.2%) vines, 6.8% (8.7%) herbs and 5.1% (0.3%) of unknown lifeform. Furthermore, 58 species of epiphytes and hemi-epiphytes were found. Diversity of trees, shrubs and palms with a DBH1.0 cm was calculated as Shannon-Wiener index (4.65), Equitability index (0.65), and Simpson index (0.10). The dominance-diversity curve showed a lognormal form, characteristic for tropical rain forest. The community structure was characterized by a relative dominance of Astrocaryum mexicanum in the understorey, Pseudolmedia oxyphyllaria in the middle storeys, and Nectandra ambigens in the canopy. Species population structures of 31 species showed three characteristic patterns, differentiated by recruitment: continuously high, discontinuously high, and continuously low recruitment. Height/diameter and crown cover/diameter diagrams suggested a very gradual shift from height growth to crown growth during tree development. Forest turnover was calculated as 138 years. Compared to other tropical rain forests the Los Tuxtlas forest had 1. similar leaf physiognomical characteristics, 2. a lower diversity, 3. a lower density, 4. an average basal area, and 5. a slow canopy turnover.     

The distribution of A1 adenosine receptor and 5′-nucleotidase in pig brain cortex subcellular fractions

Pig brain cerebral cortex was subfractionated by isopycnic centrifugation in sucrose gradients. In each subfraction the content of the agonist [³H]R-PIA binding, the activity of adenosine metabolizing enzymes (5-nucleotidase and adenosine deaminase) and the activity of membrane marker enzymes were determined. The fractions were also examined by electron microscope. In general, the results suggest a widespread distribution of A₁ adenosine receptors in membranes from different origins. Marker enzyme profile characterization indicated an enrichment of A₁ adenosine receptor in pre-synaptic membranes isolated from the crude synaptosomal fraction (P2B subfraction) as well as in membranes of glial origin such as myelin. The receptor is also present in the endoplasmic reticulum and in membranes isolated from the microsomal fraction that seem to have a post-synaptic origin (P3B). In subfractions having a high content of adenosine receptor the equilibrium binding paramters were obtained as well as the proportion of high- to low-affinity sites. From the values of the equilibrium constants it was not possible to find differences between the receptor in the different subfractions. Analysis of the affinity state distribution showed a diminished percentage of high-affinity sites in fraction P3A, which can be accounted by the existence of myelin membranes; in contrast the percentage of high-affinity states was higher in P2 and P3B, indicating that in these fractions the receptor is present in synaptosomal membranes. The close correlation shown between the enzyme 5-nucleotidase specific activity and the specific ligand binding distributions led us to postulate an important role for the enzyme in the regulation of adenosine action in pig brain cortex.     

Physiological and biochemical studies on Fusarium oxysporum f. sp. lycopersici race 2 for its biocontrol by nonpathogenic fungi

Abstract The self-degradation of the phytopathogenic fungus Fusarium oxysporum f. sp. lycopersici race 2 ( F. oxysporum l. 2), which reached an autolysis degree of 72% after 60 days of incubation in stationary culture, occurred principally during the first 14 days of incubation, when considerable β-(1,3)-glucanase, pectinase, xylanase and chitinase activities were detected in the culture fluids. The levels of β-(1,3)-glucanase, pectinase, cellulase, chitinase and xylanase activities increased in the culture fluids of this fungus, when the culture medium was supplemented with different inducers. The vegetable juice (V8) that contained tomato juice, was the best inducer for most of these activities. Chitosan, glucosamine oligomers and Mucor rouxii mycelium extract were found to have an inhibitory effect on F. oxysporum l. 2 growth. When incubating cell walls from young mycelia of F. oxysporum l. 2 with enzymic precipitates obtained from autolyzed cultures of Mucor rouxii, Aspergillus nidulans, Penicillin oxalicum and Penicillium purpurogenum , degradations of 45%, 22%, 21% and 12%, respectively, were detected.     

Metabolism of oxygen radicals in peroxisomes and cellular implications.

Peroxisomes are subcellular respiratory organelles which contain catalase and H2O2-producing flavin oxidases as basic enzymatic constituents. These organelles have an essentially oxidative type of metabolism and have the potential to carry out different important metabolic pathways. In recent years the presence of different types of superoxide dismutase (SOD) have been demonstrated in peroxisomes from several plant species, and more recently the occurrence of SOD has been extended to peroxisomes from human and transformed yeast cells. A copper,zinc-containing SOD from plant peroxisomes has been purified and partially characterized. The production of hydroxyl and superoxide radicals has been studied in peroxisomes. There are two sites of O2- production in peroxisomes: (1) in the matrix, the generating system being xanthine oxidase; and (2) in peroxisomal membranes, dependent on reduced nicotinamide adenine dinucleotide (NADH), and the electron transport components of the peroxisomal membrane are possibly responsible. The generation of oxygen radicals in peroxisomes could have important effects on cellular metabolism. Diverse cellular implications of oxyradical metabolism in peroxisomes are discussed in relation to phenomena such as cell injury, peroxisomal genetic diseases, peroxisome proliferation and oxidative stress, metal and salt stress, catabolism of nucleic acids, senescence, and plant pathogenic processes.