Screening of microbial secondary metabolites inhibiting cholesterol biosynthesis with the use of hepatoblastoma G2]

The culture of hepatoblastoma G2 (Hep G2) cells is proposed as an effective model for screening of microbial metabolites--inhibitors of sterol biosynthesis. This model can be applied at early stages of screening procedures and is quite effective for testing of crude extracts of producers' culture broth. The test is based on measurement inhibition of the radiolabelled precursors incorporation in cholesterol and separate fractions of lipids by microbial metabolites in Hep G2 cells. That allows not only to reveal inhibitors of cholesterol biosynthesis, but also to evaluate mechanism of action, including ability to inhibit the synthesis of cholesterol ethers. The cholesterol biosynthesis inhibition was tested at 150 microbial cultures (actinomycetes and imperfect fungi), isolated from soil. The ability to inhibit 14C-acetate incorporation into cholesterol was found in 15-20% of microbial cultures possessing antifungal activity of extracts (culture broth and mycelium).     

Interpretation of FISH Results in the Case of Nonuniform Internal Radiation Exposure of Human Body with the Use of Model Approach

Russian Journal of Genetics - The fluorescence in situ hybridization (FISH) technique allows for the estimation of the number of stable chromosomal aberrations (translocations) in human blood...     

Chronic hypoxia abolishes posthypoxia frequency decline in the anesthetized rat

In anesthetized rats, increases in phrenic nerve amplitude and frequency during brief periods of hypoxia are followed by a reduction in phrenic nerve burst frequency [posthypoxia frequency decline (PHFD)]. We investigated the effects of chronic exposure to hypoxia on PHFD and on peripheral and central O2-sensing mechanisms. In Inactin-anesthetized (100 mg/kg) Sprague-Dawley rats, phrenic nerve discharge and arterial pressure responses to 10 s N2 inhalation were recorded after exposure to hypoxia (10 +/- 0.5% O2) for 6-14 days. Compared with rats maintained at normoxia, PHFD was abolished in chronic hypoxic rats. Because of inhibition of PHFD, the increased phrenic burst frequency and amplitude after N2 inhalation persisted for 1.8-2.8 times longer in chronic hypoxic (70 s) compared with normoxic (25-40 s) rats (P < 0.05). After acute bilateral carotid body denervation, N2 inhalation produced a short depression of phrenic nerve discharge in both chronic hypoxic and normoxic rats. However, the degree and duration of depression of phrenic nerve discharge was smaller in chronic hypoxic compared with normoxic rats (P < 0.05). We conclude that after exposure to chronic hypoxia, a reduction in PHFD contributes to an increased duration of the acute hypoxic ventilatory response in anesthetized rats. Furthermore, after exposure to chronic hypoxia, the central network responsible for respiration is more resistant to the depressant effects of acute hypoxia in anesthetized rats.     

Analysis of DNA status in bone marrow cells of mice internally irradiated with 90Sr

The response of bone marrow cells of CBA mice injected with 22.2, 222 and 592 kBq/animal to additional gamma-irradiation (3 Gy) for testing purposes was evaluated using SCG (Comet assay). A decrease in induction of DNA damage right after additional gamma-irradiation was determined. It correlated with bone marrow cell quantity and the tail length before additional gamma-irradiation. The results support the suggestion about the activation of DNA repair in bone marrow cells under exposure to 90Sr in vivo.     

Genetic analysis of response regulator activation in bacterial chemotaxis suggests an intermolecular mechanism

Response regulator proteins of two-component systems are usually activated by phosphorylation. The phosphorylated response regulator protein CheY-P mediates the chemotaxis response in Escherichia coli. We performed random mutagenesis and selected CheY mutants that are constitutively active in the absence of phosphorylation. Although a single amino acid substitution can lead to constitutive activation, no single DNA base change can effect such a transition. Numerous different sets of mutations that activate in synergy were selected in several different combinations. These mutations were all located on the side of CheY defined by alpha4, beta5, alpha5, and alpha1. Our findings argue against the two-state hypothesis for response regulator activation. We propose an alternative intermolecular mechanism that involves a dynamic interplay between response regulators and their effector targets.     

Fetal dose assessment for the offspring of the Techa Riverside residents

Massive releases of fission products from the plutonium facility Mayak (Southern Urals, Russia) resulted in the contamination of the Techa river and its floodlands (1949–1956). The results of long-term studies of exposure populations have been used for different purposes of retrospective dosimetry. The riverside residents were exposed via various pathways and their progeny were exposed in utero both to external radiation and to internal radiation from radionuclides ingested by the mothers prior to conception and during pregnancy. Fetal doses due to ⁹⁰Sr from the maternal skeleton were estimated and compared with doses from other pathways. Individual red bone marrow (RBM) doses to the late fetuses were calculated on the basis of ⁹⁰Sr contents measured in the maternal skeleton and the ⁹⁰Sr transfer coefficients (TC) to the fetal skeleton were determined on the basis of the Techa river data. Two values of TC were assumed depending on the mothers’ maturation status in the period of maximum releases: TC=0.2 for group 1 (adulthood of mothers in 1950) and TC=0.02 for group 2 (mothers not yet in menarche age in 1950). Fetal doses in both groups that resulted from the different TC values varied by a factor of about 5–8. Furthermore, the fetal RBM doses due to ⁹⁰Sr from the maternal skeleton were found to depend on the distances from the site of release and the time after major ⁹⁰Sr intake. The average fetal RBM doses in the population of the upper, middle and lower Techa riverside regions were close to the ratio 3:2:1 and 20 years after the onset of contamination, the fetal doses have decreased by a factor of about 3–5. The distance from the site of release determined the relative contribution of different pathways to the fetal dose. For the settlements that are located closest to the site of release, the external exposure was of major concern up to the date of evacuation. For the non-evacuated settlements, the contribution of internal doses was higher and after 1956, external exposure is assumed to be negligible. From 1956 on, ⁹⁰Sr that has been transferred from the maternal skeleton was practically the single source of in utero exposure. Received: 8 December 2000 / Accepted: 1 July 2001     

Carboxyl methylation regulates phosphoprotein phosphatase 2A by controlling the association of regulatory B subunits

Phosphoprotein phosphatase 2A (PP2A) is a major phosphoserine/threonine protein phosphatase in all eukaryotes. It has been isolated as a heterotrimeric holoenzyme composed of a 65 kDa A subunit, which serves as a scaffold for the association of the 36 kDa catalytic C subunit, and a variety of B subunits that control phosphatase specificity. The C subunit is reversibly methyl esterified by specific methyltransferase and methylesterase enzymes at a completely conserved C-terminal leucine residue. Here we show that methylation plays an essential role in promoting PP2A holoenzyme assembly and that demethylation has an opposing effect. Changes in methylation indirectly regulate PP2A phosphatase activity by controlling the binding of regulatory B subunits to AC dimers.     

The characteristics of 90Sr accumulation and elimination in residents of the Urals region in the period of 1957-1988

A number of radiation accidents occurred at the "Mayak" plutonium production complex (Southern Urals, Russia) in 1950-60s that resulted in environmental contamination of large territories in the Southern and Middle Urals with long-lived radionuclides (90Sr, 137Cs). The results of long-term radiological monitoring were compiled into special databases at the Urals Research Center for Radiation Medicine, which allow the study of 90Sr behaviour in the environment and its accumulation in the human body. Retrospective analysis of the data on postmortem measurements of 90Sr in bones performed on residents of the Urals region over 1957-1988 were conducted for the first time. A time-dependence of 90Sr accumulation in the human body upon its density in soils was observed. Age and gender characteristics of 90Sr retention in bones were studied. It was shown that the levels of 90Sr in residents of large territories north and north-east of Chelyabinsk region, and of bordering territories of Sverdlovsk region were several times higher than the average values for the Russian Federation. Based on the measurements the direct dependence of 90Sr content in the skeleton on 90Sr density in soils was observed 30 years after the 1957 accident (the so called "Kyshtym accident"). However, for individual settlements this dependence may be weakened as a result of the influence of different biological and social factors. No influence of gender on 90Sr accumulation in the skeleton was found in the study. The analysis of age characteristics of 90Sr accumulation in bone, as influenced by skeleton growth and maturation, showed maximal levels of 90Sr from global fallout to be accumulated in bones of the persons born in 1949-1955.