Liver cytochrome P-450 system in rats with a varying susceptibility to the development of experimental alcoholism

The cytochrome P-450-dependent monooxygenase system of the liver was studied in laboratory noninbred male rats selected according to the intensity of their initial alcohol motivation and the dynamics of these parameters was followed up during 10-day alcoholisation. It was shown that in the animals inclined to the development of alcoholism the activity of the monooxygenase system (cytochrome P-450, B5; enzymes: aminopyrine N-demethylase, aniline p-hydroxylase, NADPH-cytochrome c-reductase) is higher than in the animals noninclined to the development of this disease. 10-day alcohol consumption in the free-choice situation between water and 15% ethanol solution did not change the parameters investigated. The only exception was NADPH-cytochrome c-reductase: its activity grew in both the groups of the animals by 40-75%.     

Immunohistochemical analysis of development of suspension and tissue neurotransplants

Engraftment and development of suspension and whole tissue allografts from the mouse brain embryonal tissue, in which cells of the green fluorescent protein (GFP) are synthesized, have been studied. The transgenic mouse cells with synthesized GFP can be used for neurotransplantation. Whole tissue and suspension transplants are able to survive more than 30 days without rejection in the brain of adult mice. It was found that the cells with synthesized GFP are able to differentiate in the neuronal and glial directions in both whole tissue and suspension transplants. The results of immunohistochemical analysis showed the reciprocal ingrowth of fibers between the cells of donor and recipient in both cases. A study of proliferation and differentiation of the cells showed the higher ability of tissue transplants for development.     

Radiation biology of structurally different Drosophila melanogaster genes. Report I. The vestigial gene: molecular characteristic of "point" mutations

The screening of PCR-detected DNA alterations in 9 spontaneous and 59 gamma-ray-, neutron - or neutron + gamma-ray-induced Drosophila vestigial (vg) gene/"point" mutations was carried out. The detected patterns of existence or absence of either of 16 overlapping fragments into which vg gene (15.1 kb, 8 exons, 7 introns) was divided enable us to subdivide all mutants into 4 classes: (i) PCR+ (40.7%) without the detected changes; (ii) "single-site" (33.9%) with the loss of a single fragment; (iii) partial detections (15.2%) as a loss of 2-9 adjacent fragments and (iv) "cluster" mutants (10.2%) having 2-3 independent changes of(ii) and/or (iii) classes. All spontaneous mutants except one were found to be classified as (ii) whereas radiation-induced mutants are represented by all 4 classes whose interrelation is determined by the dose and radiation quality. In particular, the efficacy of neutrons was found to be nine times as large as that of gamma-rays under the "cluster" mutant induction. Essentially, the distribution of DNA changes along the gene is uneven. CSGE-assay of PCR+-exon 3 revealed DNA heteroduplexes in 5 out of 17 PCR+-mutants studied, 2 of which had small deletions (5 and 11 b) and 3 others made transitions (A --> G) as shown by the sequencing. Therefore, gamma-rays and neutrons seem to be significant environmental agents increasing the SNP risk for the population through their action on the germ cells. The results obtained are also discussed within the framework of the track structure theory and the notion of quite different chromatin organization in somatic and germ cells.     

Cytokines and resistive breathing

The results of recent experimental studies indicating the involvement of the immune system in the control of breathing are analyzed and summarized. The hypothesis on the role of cytokines in the mechanisms of respiratory muscle fatigue and a decrease in the ventilatory sensitivity to hypercapnia during respiration with the added resistive load is justified. Possible ways of implementation of the respiratory cytokine effects are discussed.     

Heterogeneity of the Stearoyl-CoA desaturase-1 (SCD1) Gene and Metabolic Risk Factors in the EPIC-Potsdam Study

Background

Stearoyl-CoA desaturase-1 (SCD1) is an enzyme involved in lipid metabolism. In mice and humans its activity has been associated with traits of the metabolic syndrome, but also with the prevention of saturated fatty acids accumulation and subsequent inflammation, whereas for liver fat content inconsistent results have been reported. Thus, variants of the gene encoding SCD1 (SCD1) could potentially modify metabolic risk factors, but few human studies have addressed this question.

Methods

In a sample of 2157 middle-aged men and women randomly drawn from the Potsdam cohort of the European Prospective Investigation into Cancer and Nutrition, we investigated the impact of 7 SCD1 tagging-single nucleotide polymorphisms (rs1502593, rs522951, rs11190480, rs3071, rs3793767, rs10883463 and rs508384) and 5 inferred haplotypes with frequency >5% describing 90.9% of the genotype combinations in our population, on triglycerides, body mass index (BMI), waist circumference (WC), glycated haemoglobin (HbA1c), high-sensitivity C-reactive protein (hs-CRP), gamma-glutamyltransferase (GGT), alanine aminotransferase (ALT) and fetuin-A.

Results

No significant associations between any of the SNPs or haplotypes and BMI, WC, fetuin-A and hs-CRP were observed. Associations of rs10883463 with triglycerides, GGT and HbA1c as well as of rs11190480 with ALT activity, were weak and became non-significant after multiple-testing correction. Also associations of the haplotype harbouring the minor allele of rs1502593 with HbA1c levels, the haplotype harbouring the minor alleles of rs11190480 and rs508384 with activity of ALT, and the haplotype harbouring the minor alleles of rs522951, rs10883463 and rs508384 with triglyceride and HbA1C levels and GGT activities did not withstand multiple-testing correction.

Conclusion

These findings suggest that there are no associations between common variants of SCD1 or its inferred haplotypes and the investigated metabolic risk factors. However, given the results from animal models, heterogeneity of human SCD1 warrants further investigation, in particular with regard to rare variants.     

Neural stem and progenitor cells of human embryos and fetuses as a basis of biomedical new technologies

Isolation and cultivation of stem and progenitor cells of human embryos and fetuses at the age of 7-12 weeks of gestation have been described. The embryonic cells of human brain formed neurospheres with heterogenous composition. Cell differentiation took place not only in the presence of serum or as a result of attachment of neurosphere to a sublayer, but also in floating neurospheres in the presence of mitogens. In most neurospheres, the nestin-immunopositive cells were located near the surface while the cells stained for beta-tubulin III and glial fibrillar acid protein, as compact groups inside the neurospheres.     

Presence and activation of nuclear phosphoinositide 3-kinase C2beta during compensatory liver growth

Highly purified liver nuclei incorporated radiolabeled phosphate into phosphatidylinositol 4-phosphate (PtdIns(4)P), PtdIns(4,5)P(2), and PtdIns(3,4,5)P(3). When nuclei were depleted of their membrane, no radiolabeling of PtdIns(3,4,5)P(3) could be detected showing that within the intranuclear region there are no class I phosphoinositide 3-kinases (PI3K)s. In membrane-depleted nuclei harvested 20 h after partial hepatectomy, the incorporation of radiolabel into PtdIns(3)P was observed together with an increase in immunoprecipitable PI3K-C2beta activity, which is sensitive to wortmannin (10 nm) and shows strong preference for PtdIns over PtdIns(4)P as a substrate. On Western blots PI3K-C2beta revealed a single immunoreactive band of 180 kDa, whereas 20 h after partial hepatectomy gel shift of 18 kDa was noticed, suggesting that observed activation of enzyme is achieved by proteolysis. When intact membrane-depleted nuclei were subjected to short term (20 min) exposure to micro-calpain, similar gel shift together with an increase in PI3K-C2beta activity was observed, when compared with the nuclei harvested 20 h after partial hepatectomy. Moreover, the above-mentioned gel shift and increase in PI3K-C2beta activity could be prevented by the calpain inhibitor calpeptin. The data presented in this report show that, in the membrane-depleted nuclei during the compensatory liver growth, there is an increase in PtdIns(3)P formation as a result of PI3K-C2beta activation, which may be a calpain-mediated event.     

Development of a nutrient medium to culture microorganisms of the genus Campylobacter

The nutrient media developed in the USSR and CSR for the isolation of campylobacteria are multi-purpose as they enable the isolation of campylobacteria not only from material derived from patients with acute intestinal infection, but also from the internal organs of healthy and diseased animals and poultry as well as from a variety of objects in the environment.