The biosynthesis of nicotinamide adenine dinucleotide during early stages of frog embryonic development of haploid and diploid embryos.

1. Concentration of NAD during embryonic development of haploid and diploid embryos of frog was followed. NAD content in haploid embryonic forms is twice that in diploid embryos. 2. The variation of the NMN adenylyltransferase activity in the oocytes and during the first states of embryonic development as surveyed in the nuclear soluble fraction and the nuclear insoluble fraction (chromatin). 3. The enzyme activity in the soluble fraction is low during embryonic development and shows higher values in haploid embryos. 4. In the nonfertilized mature oocytes, the NMN adenylyltransferase activity is sixfold higher in the insoluble chromatin fraction than in the soluble fraction. 5. The evolution of the NMN adenylyltransferase in the insoluble chromatin fraction also shows higher values in haploid embryos, as compared with diploid forms.     

All-reagent test tablets and method for rapid and selective alpha-amylase iodometric determination

The new type of test tablets (Iodocrom) for alpha-amylase assay contain crosslinked (CL)-amylose or CL-starch (specific substrates for alpha-amylase only) and the reagent (KIO3/KI) generating iodine (in acidic medium, when the reaction is stopped). The method--amyloclastic in nature--is based on selective action of alpha-amylase on the CL-substrate liberating soluble polysaccharide chains large enough and in a conformation suitable to allow the formation of iodine inclusion complexes. Unlike the classical iodometric methods, the reaction is followed by an increase in iodine complex blue color. The method has some common points with the well-known chromogenic (e.g., Phadebas) methods. Both use insoluble substrates which are not susceptible to attack by exoamylases and in both cases the enzymatic reaction is followed by the release of soluble products. The amounts of these released chains and the absorbances of their inclusion complexes with iodine are in a linear dependence with the enzyme concentration (activity).     

Sensitive and fast simultaneous photometric determination of oxygen consumption and NAD(P)H formation or disappearance using a continuous flow cuvette

The photometric determination of the oxygen consumption and NAD(P)H formation or disappearance on the same sample were performed by alternative recordings at 334 nm (in the vicinity of an isosbestic point of HbO₂-Hb spectra) and at 436 nm. The optical homogeneity of the medium during the reaction was ensured by continuous stirring, and the automatic emptying and filling of the cuvettes were achieved with glass cuvettes specially built for this purpose. This method permits rapid and accurate estimations on 20–100 μg of microsomal protein, for hydroxylation reactions with rates which are comparable to, or at least one-tenth of the “free” oxidation rate of NADPH. On the other hand, using the same amount of mitochondrial protein, informations concerning oxidative phosphorylation could be obtained within a time interval of 3–4 min/sample.     

Effects of Long-Term exposure to Heavy Metals upon Rhizosphere Bacteria from Baia Mare Area (Maramureş County,Romania)

Abstract

The aim of this study was to investigate the extent of heavy metal (HM) pollution and its effect on microorganisms from rhizosphere soil in Baia Mare area (Maramure? County, Romania). Two sites with different contamination degrees were included in the study: one with a long history of mining activities and one within a drinking water safeguard zone. Rhizosphere soil samples were characterized with respect to physico-chemical parameters and the Cd, Cu, Pb and Zn contents. Native bacteria were investigated for HM tolerance and biofilm formation under toxic exposure by the microdilution assay. The most resistant strains were identified and the minimum inhibitory concentrations for HMs were determined. Cd, Cu, Pb and Zn exceeded the intervention threshold in Bozânta tailings site, while Pb content exceeded the intervention level within the area of the drinking water treatment plant. Cd showed a very high potential ecological risk in Bozânta area. The long-term exposure to HMs contributed to the selection of HM-tolerant and weakly adherent strains. Biofouling was significantly reduced under the influence of copper ions. Arthrobacter, Rhodococcus and Acidovorax strains with exceptional resistant profiles were isolated from the tailings site, indicating the important role of native microorganisms in rhizosphere ecosystems of contaminated sites.