Julijan Kandrač

NITROGEN AND PROTEIN METABOLISM IN YOUNG PEA PLANTS AS AFFECTED BY DIFFERENT CONCENTRATIONS OF NICKEL, CADMIUM, LEAD, AND MOLYBDENUM

A comparative study was carried out on the effect of molybdenum (Mo), nickel (Ni), cadmium (Cd), and lead (Pb) using concentrations of 1×10− 7, 1×10− 5, and 1×10− 3 mol dm− 3, on the metabolism of nitrogen and proteins in young pea plants (Pisum sativum L. ‘NS Lim’). The highest concentrations of the investigated metals were noted to suppress the development of the aboveground parts and roots. The smallest inhibiting effect was observed in the presence of Ni, which at lowest concentration produced a stimulating effect; Pb and Cd had a similar effect. The most pronounced inhibition was caused by Mo, which decreased the growth of the roots and aboveground parts by 50% and 35%, respectively. An increase in concentration of heavy metals in the medium resulted in their increased accumulation in both the plant roots and aboveground parts. The largest accumulation in the aboveground parts was observed for Ni and Mo, followed by Cd, and least for Pb. Nitrate and protein metabolism decreased according to the order of Cd > Pb > Ni > Mo. However, when expressed per amount of heavy metal accumulated in particular plant parts, toxicity was in the order of Pb > Cd > Ni > Mo.

Effect of heavy metals on nitrate and protein metabolism in sugar beet

Nitrate content, activities of nitrate reductase and glutamine synthetase, soluble protein content, and proportion of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) protein in total proteins were measured in leaves of Beta vulgaris L. plants affected by nickel, cadmium, and molybdenum in concentrations of 10-4, 10-2, and 1 mM. The most harmful effect on the above mentioned parameters had Cd, less harmful Ni, whereas Mo stimulated the investigated parameters. The proportion of RuBPCO protein showed a high tolerance to heavy metals.

Dynamics of cadmium distribution in the intercellular space and inside cells in soybean roots, stems and leaves

Soybean (Glycine max L.) plants grown in nutrient solution were exposed to 1 mM Cd(NO3)2 for 24 h. Dynamics of distribution of cadmium among its different forms (water soluble, Ca-exchangeable and complexed) in the intercellular space and the ratio of the intercellular and intracellular cadmium in roots, stems and leaves were studied. In roots, in the beginning of treatment the largest portion of Cd was found in the intercellular space and 1 h later Cd content started to decrease, so that between 13- and 24-h treatment an equilibrium was reached in which about 70 % of Cd was found inside cells. In stems, already after 1-h treatment, the Cd concentrations in the cells and intercellular space were similar, the equilibrium being disturbed after 13 h, so that after 24-h treatment 80 % of Cd was found inside cells. In leaves, up to the 13 h Cd distribution showed fluctuation, after that equilibrium was reached, with 70 % of intracellular Cd. The highest contents of all Cd forms in the intercellular space was observed in roots.

Isolation and determination of bile acids

SummaryIn this article, the methods of isolation and determination of bile acids are reviewed. Methods for separation of bile acids from cattle and pig bile are given in detail. Isolation of a mixture of cholic acid and deoxycholic acids from cattle bile and their subsequent purification are described. The isolation and purification of hyodeoxycholic acid and other components of pig bile are also included. Methods for the determination of bile acids in various biological samples are reviewed, including enzyme assays, radioimmunoassay, enzyme immunoassay and chromatographic methods. Among chromatographic methods, separation and determination of bile acids by thin-layer chromatography, gas chromatography and high performance liquid chromatography are reviewed. Particular attention is given to the use of high performance liquid chromatography since this has recently been the most commonly applied method for the separation and determination of bile acids.

The role of sulphur in detoxication of cadmium in young sugar beet plants

In young sugar beet plants cadmium suppressed the activity of nitrate reductase, glutamine synthetase and glutamate dehydrogenase, whereas sulphur exhibited a protective role towards activity of these enzymes, except of glutamine synthetase. Protein synthesis was suppressed in the absence of S in nutrient medium; the lowest level was at 10-3 M Cd2+. Chloroplast pigment contents were increased by S while Cd2+, even in the lowest concentration, (10−5 M) showed a repressive effect. The highest concentrations of Cd2+ (10−3 M) caused a decrease in dry mass, whereas S induced its increase. Nitrate content was increased in the presence of Cd2+ and decreased by increased concentration of S.

Biosynthesis of bile acids in mammalian liver

SummaryThe biosynthesis of bile acids in mammalian liver and its regulation, together with the physiological role of bile acids, are reviewed in this article. Bile acids are biosynthesized from cholesterol in hepatocytes. Several steps are involved including epimerisation of the 3β-hydroxyl group, reduction of the Δ4 double bond to the 5β-H structural arrangement, introduction of α-hydroxyl groups at C7 or C7 and C12 and, finally, oxidative degradation of the side chain by three carbon atoms. This gives the primary bile acids, cholic and chenodeoxycholic acids. Cholesterol-7α-hydroxylation is the rate determining step in the biosynthesis of cholic and chenodeoxycholic acids. Feedback regulation of cholesterol biosynthesis occurs by various mechanisms including termination of the synthesis of specific cytochromes P-450, modulation of specific cytosol proteins, short-term changes in the process of phosphorylation-dephosphorylation and changes in the capacity of the cholesterol pool as a substrate. Prior to being exported from the liver, bile acids are conjugated with glycine and taurine to produce the bile salts. After excretion into the intestinal tract, primary bile acids are partly converted to secondary bile acids, deoxycholic and lithocholic acids, by intestinal microorganisms. The majority of bile acids is absorbed from the intestinal tract and returned to the liver via the portal blood, so that only a small fraction is excreted in the feces. Bile acids returned to the liver can be reconjugated and reexcreted into the bile in the process of enterohepatic recycling. In addition to the physiological function of emulsifying lipids in the intestinal tract, bile acids are particularly important in respect of their ability to dissolve and transport cholesterol in the bile.

Structure and origin of bile acids: An overview

SummaryAn overview of the structure and the origin of naturally occurring bile acids is given. Most naturally occurring bile acids belong to the 5β-series, with hydroxyl groups in the A, B, and C ring of the steroid system. Hydroxyl groups are mostly found at the C3, C6, C7, C12 and C23 positions and are α- rather than β-oriented. In most bile acids, the A/B ring junction iscis (5β-series). However, the A ring can be usually present in the more stable (chair) or less stable (boat) conformation. Both B/C and C/D ring junction aretrans. With respect to the angular C9-methyl group, the hydrogen atoms at C5 and C8 are m-oriented whereas those at C9 and C14 aretrans-oriented. The archetypal bile acid is 5β-cholanic acid (3) from which all other CM bile acids can be derived. In addition to the bile acids with 24 carbons, some naturally occurring C27 bile acids have been identified including di-, tri- and tetra-hydroxy derivatives of coprostanic acid isolated from bile of several reptile species. The most dominant bile acids and their natural sources are given and a selection of naturally occurring bile acids with unusual structures which have been mostly isolated from the bile of reptiles and amphibians is described.

Chemical and metabolic transformations of selected bile acids.

SummaryThis article surveys chemical transformations of selected bile acids. Chemical transformations were initially carried out with the aim of determining the structure of bile acids. More recently they have been concerned with bile acid interconversions as well as with the synthesis of steroid hormones, vitamins and thérapeute agents. Studies of similarities and differences in the biosynthesis of bile acids from cholesterol have occupied many researches. However, this article reviews only papers dealing with the synthesis of potential intermediates in the biosynthesis of bile acids. Steroid hormones such as pregnenolone, progesterone and testosterone are synthesized from methyl thiodeoxycholate whereas cortisone is synthesized from methyl deoxycholiate. Numerous papers and patents devoted to the synthesis of ursodeoxycholic acid from cholic or chenodeoxycholic acid testify to its effectiveness in the treatment of cholelithiasis. Chenodeoxycholic acid appears to be an excellent precursor in the synthesis of steroid plant growth regulators, as well as in the synthesis of metabolites and vitamin D analogues. Chirality of bile acids has been exploited in the synthesis of cyclic and acyclic receptors and solvents. Cholic and deoxycholic acids have been used to create new macrocyclic structures which show different capacities to bind and transport other compounds. Another important trend in the chemistry of bile acids is their application in combinatorial chemistry.

Biochemical changes in sugar beet lines in dependence on soil moisture

Nine sugar beet lines were grown in a glasshouse on chernozem soil watered to 35, 50 and 65 % of maximal soil water capacity. After 12 d, plant water potential, net photosynthetic rate, contents of soluble proteins, DNA and RNA, proportion of ribulose-1,5-bisphosphate carboxylase/oxygenase (RUBISCO) protein, and carbonic anhydrase activity were measured. As soil moisture decreased, the leaf water potential and net photosynthetic rate decreased. DNA and RNA content and carbonic anhydrase activity decreased under moderate drought, and increased with severe drought. RUBISCO protein proportion did not change significantly under decreased soil moisture, while the total soluble protein content decreased.

Alleviation of cadmium toxicity by naphthenate treatment

The work is concerned with the effect of low concentrations (10−7 mol dm−3) of sodium naphthenate on total content of Cd and its particular forms in the intercellular space and inside cells, as well as on some physiological and biochemical parameters of young soybean plants grown in the presence of 1 mmol dm−3 solution of cadmium chloride. Presence of naphthenate reduced in average by 40 % content total and intracellular Cd in root, stem and leaves and alleviated the harmful effect of Cd on activity of nitrate reductase and content of photosynthetic pigments.