Immunogenic properties of neomycin conjugates with macromolecular carriers]

To obtain diagnostic antibodies to neomycin, immunogenic properties of the neomycin conjugates with macromolecular carriers were studied. Bovine serum albumin and copolymers of N-vinylpyrrolidone with crotonic acid and N-hydroxyphthalimide ether of crotonic acid were used as carriers. It was shown that immunization of mice by the conjugates in combination with Freund's adjuvant resulted in production of neomycin antibodies, the titer being 1/80 to 1/130. When the antibiotic conjugates with the copolymers of N-vinylpyrrolidone were used and not the neomycin conjugates with the carrier of the protein nature, the neomycin antibodies were produced in the absence of Freund's full adjuvant. With the use of the isolated antibodies to neomycin a method for indirect solid phase enzyme immunoassay of neomycin was developed at the minimum detectable level of 25 ng/ml.     

Release of mtDNA from mitochondria and activation of its replication in tissues of irradiated mice

A nessessary condition for normal functioning of mitochondria is the maintenance of certain numbers of intact mtDNA molecules. In the present study, we investigasted changes in the number of mtDNA copies in brain and spleen cells of mice subjected to irradiation. For the first time, we observed the irradiation-induced output of mtDNA fragments into brain and spleen cell cytosol. In the cytosol of these cells, examined in mice 5 h after 5 Gy irradiation, 1841 h.p. mtDNA fragments were detected able to persist for at 3 weeks. In addition, larger fragments of mtDNA (10,090 b.p.) were detected in the cytosol of brain cells of irradiated mice. The occurrence of mtDNA fragments in the cytosol of brain cells is accompanied with an increase in the number of mtDNA copies in the mitochondrial matrix. The induction of mtDNA replication in brain cells of irradiated animals may be considered as a compensatory reaction in response to mtDNA damage. A sharp decrease in the amount of mtDNA copies in the mitochondrial matrix of spleen cells on the first day after irradiation may be considered as apoptosis development. However, the compensatory reaction in brain cells was also noticed but in later terms.     

Restriction Endonuclease Sst12I from a Strain of Streptomyces Recognizing the Nucleotide Sequence 5"-CTGCAG-3"

A new restriction endonuclease Sst12I belonging to type II and recognizing the sequence 5"-CTGCAG-3" was isolated from the bacterial strain Streptomycessp. St-12. The enzyme hydrolyzes DNA between adenine and guanine residues; thus, it is a true isoschizomer of restrictase PstI. In contrast to PstI, the restriction endonuclease Sst12I hydrolyses DNA both at 37 and 55°C and remains active after long-term storage.     

Changes in Resistance to Salinity and Temperature in Some Species of Polychaetes from the White Sea in Early Ontogenesis

We studied the effects of different salinities on plankton larvae of some polychaetes in the White Sea. It has been found that the salinity resistance of Alitta virens (Nereidae) increases during ontogenesis. Successful fertilization and further larval development in this species occur at the salinity of 22 to 34; embryos taken into the experiment at the stage of 32 blastomeres, trochophores, and early nektochaetes could survive and normally develop at the salinity of 16–32, 14–45, and 12–45 respectively. The rate of settling and metamorphosis in late nektochaetes of A. virens at normal or lowered (down to 14) salinity is dependent on temperature in the range of 5 to 23°C. It is found that the larvae of Harmothoe imbricata (Polynoidae) show the greatest salinity resistance at the stage of nektochaeta, whose lower limit of salinity is 14. Later larval stages of these species can survive in a wide range of salinity due to the development of a provisory nephridial system. The eurybionty of larvae of Spirorbis spirorbis ready for metamorphosis was higher than that in the larvae of Circeus spirillum (Spirorbidae). Under salinity reduced down to 10 the larvae of S. spirorbis die in 8–14 days, whereas more stenohaline larvae of C. spirillum die by the 3-rd day of the experiment. At water temperatures under 5°C the survival of S. spirorbis was the highest at three examined values of salinity, whereas C. spirillum showed the highest survival only under normal salinity.     

Calcium-dependent proteinases of some invertebrates and fishes

In comparative-evolutionary aspect, the experimental data are considered about activity, biochemical properties, and peculiarities of structural organization of proteins of the calpain family in some invertebrates and fishes. Peculiarities of calpain-like proteins of invertebrates--the predecessors of calpains of higher animals are revealed. By the example of the studied taxons, there is traced complication of the structural organization and mechanisms of control of the calpain activities, which reflects stages of molecular evolution of the protein family.     

Anticoagulant activity of fucoidans from brown algae

The anticoagulant activity of polysaccharide fucoidans from 11 species of brown algae was studied. The anticoagulant activity was measured by the activated partial thromboplastin time (APTT), prothrombin time, and thrombin time. Inhibitory action of these fucoidans significantly varied from one species to another. Fucoidans from Laminaria saccharina and Fucus distichus exhibited high anticoagulant activity, while fucoidans from Cladosiphon okamuranus and Analipus japonicus were almost inactive. Other fucoidans exhibited intermediate inhibitory activity. The inhibitory effect of fucoidans on thrombin and factor Xa was investigated in the presence or in the absence of natural thrombin inhibitor, antithrombin III (AT III). In contrast to the best-studied anticoagulant, heparin, most of these fucoidans inhibited thrombin in the absence of AT III. In the presence of AT III the inhibitory effect of fucoidans considerably increased. In contrast to heparin, fucoidans weakly influenced factor Xa activity in the presence of AT III and their inhibitory effect was not observed in the absence of AT III. There was no correlation between the anticoagulant activities of this series of fucoidans and their anti-inflammatory action, studied earlier. It is suggested that these two types of fucoidan activities depend on different structural features of fucoidans. Results of this study demonstrate a possibility of preparation of fucoidans with high anti-inflammatory activity but low anticoagulant activity. Anticoagulant activity of the fucoidans did not exhibit direct dependence on the content of fucose, the other neutral sugars and sulfates; no dependence was also found between the anticoagulant activity and the structure of the backbone of their molecules.     

Activities of proteinases in invertebrate animals--potential objects of fish nutrition. Effects of temperature, pH, and heavy metals

Differences in the degree of separate and combined effects of temperature, pH, and heavy metals (zinc, copper) on the trypsin- and chymotrypsin-like proteinase activities have been established in the whole body of some invertebrate animals - potential objects of fish nutrition: pond snail Lymnaeae stagnalis, orb snail Planorbis purpura, zebra mussel Dreissena polymorpha, oligochaetae Tubifex sp. and Lumbriculus sp. in total, chironomid larvae Chironimus sp. and Ch. riparus, as well as crustacean zooplankton. It has been shown that enzymes of the potential victim at a low temperature can compensate low activity of intestinal proteinases of fish bentho- and planktophages.     

The increase in mitochondrial DNA copy number in the tissues of gamma-irradiated mice

Changes in the number of mitochondrial DNA (mtDNA) copies in the brain and spleen tissues of gamma-irradiated (3 Gy) mice were studied by comparative analysis of the long-extension PCR products of mtDNA (15.9 kb) and a fragment of the cluster nuclear beta-globin gene (8.7 kb) amplified simultaneously in one and the same test-tube within total DNA. The analysis showed that, compared to the nuclear beta-globin gene, an increase in mtDNA copy number (polyploidization) took place in the brain and spleen cells of mice exposed to gamma-radiation. This data led to the suggestion that the major mechanism for maintenance of the mitochondrial genome, which is constantly damaged by endogenous ROS and easily affected by ionizing radiation or other exogenous factors, is the induction of synthesis of new mtDNA copies on intact or little affected mtDNA templates because the repair systems in the mitochondria function at a low level of efficiency.