Identification and characterization of a new family of high-affinity receptors for Escherichia coli heat-stable enterotoxin in rat intestinal membranes.

Novel high-affinity, low-capacity binding sites in intestinal membranes for the heat-stable toxin produced by Escherichia coli have been defined. The appearance of these sites is observed in the presence of physiological concentrations of NaCl in binding reactions. Scatchard analyses of equilibrium binding in the absence of NaCl demonstrated a single class of binding sites with KD = 1.9 x 10(-9) M and Bmax = 0.75 pmol/mg of protein. In contrast, similar experiments in the presence of NaCl demonstrated, in addition to the previously described low-affinity site, a high-affinity site with a KD of 2.1 x 10(-11) M and a Bmax of 73 fmol/mg of protein. Confirmation of the presence of high- and low-affinity sites was obtained in studies of the kinetics of ST binding. These sites exhibited similar dissociation but markedly different association kinetics. Determination of the association and dissociation constants permitted calculation of the KD's for the high- and low-affinity sites, which were 1.15 x 10(-11) M and 1.89 x 10(-9) M, respectively. These data agree closely with those obtained in studies of equilibrium binding. Furthermore, similar values for the KD's of these sites were obtained in experiments of competitive displacement of labeled ST, confirming the presence of two receptors for this toxin. Binding of ST to high-affinity sites is completely reversible and does not appear to be coupled to activation of particulate guanylate cyclase. In contrast, binding of ST to low-affinity sites appears to be partially reversible and may be coupled to activation of guanylate cyclase.(ABSTRACT TRUNCATED AT 250 WORDS)     

Are there seasonal variations of trace element concentrations (Cd,Pb, Zn) in wood of Fagus trees in Germany?

Concentrations of Cd, Pb and Zn were determined in stem wood of beech trees (Fagus sylvatica L.) from 3 sites in northern Germany. Distinct radial distribution patterns of the elements were observed in the xylem. Concentrations of Cd and Pb increased from the youngest, outermost annual rings towards the center of the stem. With Zn intermediate concentrations were observed in the sapwood and higher levels at the center of the stem.Temporal and spatial stability of such distribution patterns in the trunks was investigated. Wood samples taken from the same individual tree in different months of the year were analysed. Marked seasonal variations of mineral concentrations were observed. Also the shape of the distribution patterns of the elements varied with the season. Such variations were larger than those observed with samples taken simultaneously from different sides of the trunk. Furthermore, Pb concentrations in the stem showed variations with height above ground.The results indicate, that radial distribution patterns of Cd, Pb and Zn in xylem rings of beech are not stable. Biomonitoring trace element pollution levels by analysis of beech wood is, thus, questionable. To obtain a reliable historical record of pollution from tree rings, the distribution patterns should be stable over a long period of time. This basic requirement of the dendroanalytical method does not hold for the examined beech. Still, with other tree species and under more favourable conditions the dendroanalytical biomonitoring method may prove valuable.Presented as symposium paper at the V International Congress of Ecology, Yokohama, August 23–30, 1990.     

Dehydrins associated with the development of frost resistance of Asian white birch

Seasonal changes in the content of dehydrins in Asian white birch (Betula platyphylla Sukacz.) growing under extreme cold conditions of Eastern Siberia (Central Yakutia) were studied for the first time by SDS-PAGE and immunoblotting. Several polypeptides, including putative storage proteins, which content was higher in winter than in other periods, were observed. Intraspecies polymorphism of dehydrins was detected during plant dormancy. The two groups of dehydrins were found: dehydrins with mol wts of 56-73 kD, which were present year-round, and dehydrins with mol wts of 15–21 kD, evidently related to the development of frost resistance because they were absent in summer but present in large amounts in winter. Under low winter temperatures, the highest level of dehydrins coincided with the lowest content of water in buds, which was accompanied by increased plant frost resistance to the highest values.     

Production of Ricinoleic Acid from Castor Oil by Immobilised Lipases

Abstract

Porcine pancreatic lipase (PPL), Candida rugosa lipase (CRL), and Castor bean lipase (CBL) were immobilized on celite by deposition from aqueous solution by the addition of hexane. Lipolytic performance of free and immobilized lipases were compared and optimizations of lipolytic enzymatic reactions conditions were performed by free and immobilized derivatives using olive oil as substrate. Afterwards, the influence on lipolysis of castor oil of free lipases and immobilized lipase derivatives have been studied in the case of production of ricinoleic acid. All of the lipases performances were compared and enzyme derivative was selected to be very effective on the production of ricinoleic acid by lipolysis reaction. Various reaction parameters affecting the production of ricinoleic acid were investigated with selected the enzyme derivative.

The maximum ricinoleic acid yield was observed at pH 7–8, 50°C, for 3 hours of reaction period with immobilized 1,3-specific PPL on celite. The kinetic constants K_m and V_max were calculated as 1.6 × 10^?4 mM and 22.2 mM from a Lineweaver–Burk plot with the same enzyme derivative. To investigate the operational stability of the lipase, the three step lipolysis process was repeated by transferring the immobilized lipase to a substrate mixture. As a result, the percentange of conversion after usage decreased markedly.     

Effect of 900-, 1800-, and 2100-MHz radiofrequency radiation on DNA and oxidative stress in brain

Ubiquitous and ever increasing use of mobile phones led to the growing concern about the effects of radiofrequency radiation (RFR) emitted by cell phones on biological systems. The aim of this study is to explore whether long-term RFR exposure at different frequencies affects DNA damage and oxidant-antioxidant parameters in the blood and brain tissue of rats. 28 male Sprague Dawley rats were randomly divided into four equal groups (n = 7). They were identified as Group 1: sham-control, Group 2: 900 MHz, Group 3: 1800 MHz, and Group 4: 2100 MHz. Experimental groups of rats were exposed to RFR 2 h/day for 6 months. The sham-control group of rats was subjected to the same experimental condition but generator was turned off. Specific absorption rates (SARs) at brain with 1 g average were calculated as 0.0845 W/kg, 0.04563 W/kg, and 0.03957, at 900 MHz, 1800 MHz, and 2100 MHz, respectively. Additionally, malondialdehyde (MDA), 8-hydroxydeoxyguanosine (8-OHdG), total antioxidant status (TAS), and total oxidant status (TOS) analyses were conducted in the brain tissue samples. Results of the study showed that DNA damage and oxidative stress indicators were found higher in the RFR exposure groups than in the sham-control group. In conclusion, 900-, 1800-, and 2100-MHz RFR emitted from mobile phones may cause oxidative damage, induce increase in lipid peroxidation, and increase oxidative DNA damage formation in the frontal lobe of the rat brain tissues. Furthermore, 2100-MHz RFR may cause formation of DNA single-strand breaks.     

The imprinting mechanism of the Prader-Willi/Angelman regional control center

The 2 Mb domain on chromosome 15q11-q13 that carries the imprinted genes involved in Prader-Willi (PWS) and Angelman (AS) syndromes is under the control of an imprinting center comprising two regulatory regions, the PWS-SRO located around the SNRPN promoter and the AS-SRO located 35 kb upstream. Here we describe the results of an analysis of the epigenetic features of these two sequences and their interaction. The AS-SRO is sensitive to DNase I, and packaged with acetylated histone H4 and methylated histone H3(K4) only on the maternal allele, and this imprinted epigenetic structure is maintained in dividing cells despite the absence of clearcut differential DNA methylation. Genetic analysis shows that the maternal AS-SRO is essential for setting up the DNA methylation state and closed chromatin structure of the neighboring PWS-SRO. In contrast, the PWS-SRO has no influence on the epigenetic features of the AS-SRO. These results suggest a stepwise, unidirectional program in which structural imprinting at the AS-SRO brings about allele-specific repression of the maternal PWS-SRO, thereby preventing regional activation of genes on this allele.     

Seasonal changes in the content of photosynthetic pigments in perennial grasses of cryolithic zone

Seasonal changes in the content of photosynthetic pigments and their ratio were studied in perennial gramineous (Psathyrostachys juncea Tzvel., Elymus sibiricus L., Bromopsis inermis Leys) and equisetaceous (Equisetum variegatum Schleich. ex Web., E. scirpoides Michx.) plants grown under conditions of cryolithic zone in Central and North-East Yakutia (the Pole of Cold). Simultaneously with growth in chlorophyll (Chl) content in all the studied species of perennial grasses of Central Yakutia, a considerable increase in the content of carotenoids (Car) of the xanthophyll cycle (XC) pigments was observed with the beginning of autumn temperature fall. Similar dependence was found in horsetails of the Pole of Cold region: the content of Car was increased by 1.2–3.0 times comparing to the summer values, whereas at the same time the concentration of oxygen-containing forms increased and the ratio of Chl to xanthophylls and Car decreased. In general, horsetails had typically a lower content of Chl and Car, which was determined not only by extremely harsh climatic conditions of the region but also by specific characteristics of Equisetaceae plants. Closer to winter time, the formation of secondary Car, rhodoxanthin, was also observed in the studied horsetail species. In this case, rhodoxanthin presumably plays more important antioxidant role than XC, which can not operate at constantly low temperatures. The obtained data suggest that one of the characteristics of cold hardening of perennial grasses under conditions of cryolithic zone of Yakutia is, presumably, the high level of primary, as well as secondary, Car with most profound antioxidant properties.     

The effects of dietary boric acid and borax supplementation on lipid peroxidation,antioxidant activity,and DNA damage in rats

The aims of this study were to clarify the effects of high dietary supplementation with boric acid and borax, called boron (B) compounds, on lipid peroxidation (LPO), antioxidant activity, some vitamin levels, and DNA damage in rats. Thirty Sprague Dawley male rats were divided into three equal groups: the animals in the first group (control) were fed with a standard rodent diet containing 6.4 mg B/kg, and the animals in the experimental group were fed with a standard rodent diet added with a supra-nutritional amount of boric acid and borax (100 mg B/kg) throughout the experimental period of 28 days. The B compounds decreased malondialdehyde (MDA), DNA damage, the protein carbonyl content (PCO) level in blood, and glutathione (GSH) concentration in the liver, Cu–Zn superoxide dismutase (SOD), and catalase (CAT) activity in the kidney. The B compounds increased GSH concentration in blood and the vitamin C level in plasma. Consequently, our results demonstrate that B supplementation (100 mg/kg) in diet decreases LPO, and enhances the antioxidant defense mechanism and vitamin status. There are no differences in oxidant/antioxidant balance and biochemical parameters except for serum vitamin A and liver GSH concentration, between the boron compounds used in this study.     

A microscopic study of kidney tissue in Familial Mediterranean Fever patients

Familial Mediterranean Fever (FMF) is an autosomal recessive hereditary disease leading mostly to renal failure and nephrotic syndrome. The ultrastructure of kidney has not been fully investigated in FMF associated renal disease. The aim of this study is to provide further evidence on the ultrastructure of kidney in patients with FMF who suffer from renal disease. Renal biopsies obtained from two patients who were diagnosed with FMF renal disease complications were examined. Examination of renal tissue by light and electron microscopy identified degenerations both in tubules and the filtration barrier. Foot processes were partly effaced. Amorphous material was found in thickened glomerular basement membranes. Fibrous material deposits in thick Bowman's capsule wall were also seen. Finally, degeneration in the form of folding of plasma membrane and vacuolization as well as fusion in mitochondria cristae, was observed. Accumulation of tissue remnants in the lumen was also found in tubules.