Studies on plating efficiency and estimation of viability of suspensions of Paracoccidioides brasiliensis yeast cells

Mild sonication was used to obtain single cell suspensions of Paracoccidioides brasiliensis. These cells were intact by microscopic criteria. Direct cell counts in a given inoculum and colony formation on various media were used to determine plating efficiency. Sonicated and nonsonicated cell suspensions were used to study plating efficiency and to estimate viability by means of vital dyes. Methylene blue, Erythrosin B, and Janus green were unreliable when used with P. brasiliensis, but vital dyes were accurate when tested with Candida albicans.Acridine orange gave more meaningful results of viability. Estimates of viability, however, changed significantly as a result of relatively minor alterations in the composition of the suspending medium.In initial experiments, the plating efficiency of P. brasiliensis was dismally low. It descended abruptly with increasing dilution of inoculum. Efficiency was much improved if horse serum was added to brain heart infusion plates or if glucose glycine yeast extract (GGY) plates were incubated at room temperature and mycelial colonies were counted. With the technique we report, current plating efficiency of sonicated suspensions is of the order of 25 %. Our results and procedures have an important bearing upon those studies concerned with in vitro killing of P. brasiliensis in suspensions or with isolating this fungus from clinical or environmental specimens.     

A SURVEY FOR 1,3,6,7-TETRAHYDROXY-C-GLYCOSYLXANTHONES EMPHASIZING THE “PRIMITIVE” LEPTOSPORANGIATE FERNS AND THEIR ALLIES

A survey for 1,3,6,7-tetrahydroxy-C-glycosylxanthones of representative species within the primitive vascular plants, emphasizing the leptosporangiate ferns, has indicated a limited distribution of these compounds within three leptosporangiate families: Hymenophyllaceae, Aspleniaceae and Marsileaceae. In the Hymenophyllaceae the distribution of these compounds appears to be a useful criterion for segregating species of Mecodium from other species of Hymenophyllum (sensu lato) and suggests that the tubulate vs. the valvate indusial condition may not be an ideal character for separating all species of Hymenophyllum (s.l.) from those of Trichomanes (s.l.). These compounds appear useful for delimiting several species of Elaphoglossum section Pachyglossa and support a relationship among the Aspleniaceae, Athyriaceae, and Elaphoglossaceae. Their presence in Marsilea also raises questions as to the origin of this group of plants.     

Effect of streptozotocin diabetes on some urea cycle enzymes

Streptozotocin induced diabetes in rats increased the activities of the three mitochondrial enzymes, carbamylphosphate synthetase, ornithine transcarbamylase and N-acetylglutamate synthetase, but not of the cytosolic N-acetylglutamate deacylase. Levels of both N-acetylglutamate and arginine, which are activators of carbamylphosphate synthetase and N-acetylglutamate synthetase respectively, increased in diabetes. These results serve to explain the increase both of mitochondrial citrulline and urea formation in hepatocytes and the increased urea excretion in diabetes.     

Plasma determination of the novel anticonvulsant d,l-3-hydroxy-3-ethyl-3-phenylpropionamide and preliminary pharmacokinetic studies in the rat

A sensitive gas chromatographic method with flame ionization detection was developed for the analysis in plasma of the novel anticonvulsant d,l-3-hydroxy-3-ethyl-3-phenylpropionamide (HEPP), using d,l-2-hydroxy-2-ethyl-2-phenylacetamide as the internal standard. HEPP was extracted from alkalinized plasma into dichloromethane and quantified after derivatization with bis(trimethylsilyl)-trifluoroacetamide. Standard curves were linear from 0.5 to 50 and from 2 to 100 μg/ml of plasma, using 1.5 and 5 μg of the internal standard, respectively. The lower limit of detection at a signal-to-noise ratio of 3 standard deviations was 0.33 μg/ml of sample. The sensitivity, accuracy and reproducibility of the method were shown to be satisfactory for pharmacokinetic studies of HEPP. After intraperitoneal administration of 50 mg/kg to Wistar rats, the principal kinetic parameters were: absorption half-life = 0.04 h; volume of distribution = 1.32 l/kg; clearance = 4.40 ml/min; peak concentration = 50 μg/ml; peak time = 0.25 h; mean residence time = 4.55 h.     

Boron,copper, iron,manganese and zinc contents in leaves of flowering sunflower plants (Helianthus annuus,L.), grown with different boron supplies

Summary Boron, copper, iron, manganese and zinc concentrations were measured in sunflower leaves from plants grown hydroponically in a closed continuous flow system and with boron concentrations ranging from deficient to toxic. Leaves were analyzed at the stage of flower development. There was a highly significant inverse correlation between B concentration in solution and Mn concentration in leaves. Cu, Fe and Zn concentrations in the leaves were not changed by the different B levels, although the B concentration in leaves increased with an increasing concentration in solution, showing a close correlation each other.     

Effect of aeration on minimal medium recovery of heated Salmonella typhimurium.

The effect of presence or absence of air on minimal medium recovery of heated Salmonella typhimurium was investigated. It was determined that the expression of minimal medium recovery is not only dependent on heat and a nutritionally complex medium but also on air. Unlike in the presence of air, in the presence of nitrogen, cells were able to recover their ability to grow on Trypticase soy agar enriched with 0.5% yeast extract (TSY) when incubated in TSY broth. It was established that in the presence of nitrogen the number of heat-TSY- induced, single-straneded breaks in deoxyribonucleic acid (DNA) were less than in the presence of air. Furthermore, the DNA breaks in nitrogen were repaired, whereas DNA breaks in air were not. The ability of cells to grow on TSY agar corresponded well with their ability to repair damage to DNA.     

Pericarditis,pleural effusion,and pneumonitis with transient mitochondrial antibodies.

Four women with fever, arthromyalgias, pericarditis, pleural effusion, high erythrocyte sedimentation rates, and lymphopenia had mitochondrial antibodies in the serum in the absence of antinuclear antibody. Their illness lasted 5-12 weeks and the antibody test results became negative on remission. Absence of specific bacteriological findings, normal antistreptolysin O titres, resistance to antibiotics, and good response to steroids suggested that these cases represented a milder and less persistent form of the syndrome resembling systemic lupus erythematosus described by German authors.     

Patients with Alzheimer's disease show an increased content of 15 Kdalton somatostatin precursor and a lowered level of tetradecapeptide in their cerebrospinal fluid

The relative proportions of both somatostatin-14 and its precursors somatostatin-28 and the 15 Kdalton prosomatostatin were evaluated by radioimmunoassay in the cerebrospinal fluid of patients with Alzheimer's disease. It was observed that the patients have a lowered content in the tetradecapeptide somatostatin while they exhibit a significant increase in unprocessed 15 Kda precursor. These results indicate that these patients possess impaired processing mechanisms which may be responsible for the lowered content in mature somatostatin-14. These observations may provide a valuable test for the ante-mortem diagnosis of the disease. They are discussed in connection with others suggesting that Alzheimer's patients may be selectively altered in their somatostatinergic neurones of their cerebral cortex (Morrison et al. (1985) Nature 314, 90-92. Roberts et al. (1985) Nature 314, 92-94).     

Multiscale change in reef coral species diversity and composition in the Tropical Eastern Pacific

Coral Reefs - Both natural and anthropogenic factors are changing coral-reef structure and function worldwide. Long-term monitoring has revealed declines in the local composition and species...