The role of the cytosolic fraction and of initiation factor eIF-2 for changes of the rate of protein synthesis during liver regeneration

The protein synthesis-stimulating activity of the cytosolic fraction from regenerating rat liver was tested in a cell-free system using washed polysomes from normal rat liver. This activity undergoes significant changes during liver regeneration after partial hepatectomy (p.h.). An initial decrease until 16 h after p.h. is followed by a significant increase until 24 h after p.h. Beyond 32 h after p.h. the activity begins to decline again. Evidence is presented that these changes of the cytosolic activity may not be due to alterations in the distribution of protein synthesis-stimulating factors between the microsomal and the cytosolic fraction. The Met-tRNAf-binding activity of the cytosolic fraction changes during liver regeneration analogously to the protein synthesis-stimulating activity measured in the polysomal assay. This indicates that initiation factor eIF-2 is involved in the observed changes of the cytosolic activity. This conclusion could be confirmed by addition of purified eIF-2 to the polysomal assay system. Addition of eIF-2 to cytosolic fractions of low endogenous protein synthesis-stimulating activity (16 h after p.h.) enhances amino-acid incorporation to a significantly higher extent than addition to highly active cytosolic fractions (24 h and 32 h after p.h.). From these results it is concluded that changes in eIF-2 plays an essential role in the described alterations of the cytosolic activities during liver regeneration.     

Isolation of antiserum against rat transcortin and characteristics of specific antibodies

Repeated chromatography of rat plasma protein on DEAE-cellulose, hydroxylapatite and subsequent gel-filtration through Sephadex G-200 were used to obtain a pure rat transcortin homogeneous upon polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. The molecular weight of transcortin was about 66 kDa as determined by SDS-polyacrylamide gel electrophoresis. Immunization of a rabbit with the homogeneous preparation of rat plasma transcortin caused development of antibodies to transcortin. It was shown that the antibodies of rabbit antisera in the experiments made in vitro and in vivo neutralized 60 and 65% of 3H-corticosterone-transcortin complexes, respectively. Specific antibodies to the transcortin were isolated from the homogeneous fraction of IgG by affinity chromatography on transcortin-sepharose 4B. 125J-labelled antibodies were adsorbed by protein A-sepharose; IgG can be eluted by IM acetic acid as a sharp peak. The SDS-polyacrylamide gel electrophoresis demonstrated that affinity-eluted material contains 25 and 50 kDa polypeptides.     

Characteristics of the expression of globin genes in blood cells of patients with homozygotic beta-thalassemia from Tadzhikistan

The synthesis of globin protein in blood reticulocytes of patients from Tajikistan suffering from homozygous beta-thalassemia was studied. Beta-thalassemia has been revealed in all cases, with synthesis of beta-globin being retained though essentially reduced. It was shown that, unlike homozygous beta+-thalassemia of other populations, beta +thalassemia with sharp inhibition of the beta-globin protein synthesis is most representative for the region (alpha/beta greater than 10).     

Effect of exogenous DNAses on various stages of DNA replication in microorganisms

Effect of DNAase 1 on DNA synthesis and cell division was studied in microorganisms deficient in some stages of DNA replication initiation. The DNA synthesis induced by exogenous DNAase was found to be a replicative origin since it was registered from the "origin" of chromosomal replication under the conditions of initiation of proteins functioning. Stimulation of DNA synthesis in bacterial cells having mutations in DNA B and DNA G genes by DNAase 1 indicates that exogenous DNAases participate in replicative fork during the DNA synthesis.     

Influence of a prolonged tamoxifen administration on steroidogenesis by incubated rat testes

Tamoxifen was administered i.m. for 9 days to adult male rats in a daily dose of 100 micrograms or 1 mg. The treatment resulted in a significant reduction of the plasma levels of testosterone and LH, without modification of the plasma levels of FSH and of the testes weight. Upon incubation, the testes from the tamoxifen-treated rats produced less testosterone and 7 alpha-hydroxytestosterone, but metabolized [4-14C]testosterone in the same way as the control animals. Small doses of hCG (0.5 i.u. for 9 days) were unable to modify the tamoxifen effect on the testicular function, while tamoxifen significantly inhibited the increase of the plasma levels of testosterone induced by the administration of moderate doses of hCG (1.5 i.u. or 2.5 i.u. for 9 days) to hypophysectomized rats. Tamoxifen treatment, however, did not modify significantly the reactivity of the testes towards high doses of hCG (10 i.u.), administered either 2 h before sacrifice or for 9 days. It is concluded that a prolonged administration of tamoxifen in the rat has, besides an indirect effect resulting from a decrease of the LH levels, a direct inhibitory influence on the testicular testosterone formation, which can be reversed by high doses of hCG.     

Kinetics of Pseudomonas fluorescens growth under different conditions of culturing

The dynamics of Pseudomonas fluorescens VKM-1472 growth was studied under the conditions of batch and continuous cultivation, and the behaviour of the organism was investigated upon nutrition shifts and starvation. At D greater than 0.375 h-1, just as in the batch culture with a substrate excess, the strain realised excessive metabolism [15]: the yield in terms of the substrate fell down (38% for the batch culture and 40% for the continuous culture), the titre of viable cells decreased to a considerable extent, and maintenance expenditures increased (3 times for qgluc and 7.5 times for qO2). When the culture was incubated under the oligotrophous conditions, the biomass yield decreased fourfold within 8 days and the titre of viable cells dropped down twofold within the same period of time. However, the organism was still capable of oxidising a wide range of substrates (17 substrates were studied). As soon as a substrate was added, it was oxidised at a high rate and changes in the macromolecular composition were characteristic of an "up-jump" in the growth rate [11]. It is possible that the growth and behaviour of this organism are associated with its ecological niche occupied in natural systems.     

Effect of the cytoplasm of irradiated cells on chromatin degradation in mouse thymocytes and hepatocytes

The addition of a cytoplasmic fraction, isolated from cells 3h after irradiation of mice, to exposed or intact thymocyte nuclei causes a 2- or 3-fold acceleration of chromatin degradation in the nuclei incubated in conditions optimum for activity of Ca2+,Mg2+-dependent endonuclease to be manifest. In contrast to thymocytes, no chromatin degradation products are found in liver cells of irradiated mice. The cytoplasmic fraction isolated from hepatocytes of irradiated animals fails to activate chromatin degradation in thymocyte nuclei.     

Suppression of the initiation of DNA synthesis as an index of cellular radiosensitivity

A gamma-radiation dose (Di) suppressing DNA synthesis initiation by 35% in primary suspension cultures of mammalian cells, is nearly the same as D0 for survival of clonogenic cells of the same lines and tissues. The extent of DNA synthesis suppression is assessed by impulse 3H-thymidine incorporation in the acid-insoluble fraction of irradiated cells. The values of Di determined in this way for HeLa cells, Ehrlich ascites tumor cells, mouse bone marrow and thymus cells are 2.0, 1.5, 1.5, and 1.0 Gy, respectively; as determined by clonogenic capacity of these cells, Di = 1.9, 2.0, 1.3, and 1.0 Gy, respectively.     

Catecholamine content and acetylcholinesterase activity in the brain of rats exposed to lasers

The catecholamine content was quantitated and acetylcholinesterase activity determined in rat brain locally exposed to laser radiation. It was shown that the adrenaline/noradrenaline ratio changed in the tissues under study and the cholinergic system was involved in the abscopal effect of laser radiation.     

On the preservation of avian blood cells

The functional state of erythrocytes from hen during their conservation with a preserving solution for 24 days at 4 degrees C, has been estimated by studying some biochemical and hemorheological parameters. Results show an initial phase in the preservation period (4-5 days) in which red blood cells maintain their values at levels similar to those at the beginning of the experience, except for osmotic resistance. Furthermore a progressive erythrocyte deformability loss, linked to ATP depletion (with rise in inorganic phosphate levels) as well as a gradually higher rate of hemolysis, were detected.