Interaction of chondroitin sulfate with enzymes

Chondroitin sulphate forms noncovalent electrostatic biocatalyst-glycosaminoglycan complexes in the solutions of enzymes. Chondroitin sulphate also interacts with enzymes developing complexes after carbodiimide activation of its carboxylic groups. Relatively low-molecular weight of biocatalysts (chymotrypsin, superoxide dismutase) forms stable noncovalent conjugates with the additional interaction as compared with electrostatic complexes. High-molecular weight of enzymes (acid phosphatase) develops covalent conjugates. Chondroitin sulphate is proposed for covalent binding of large proteins and multi-enzymatic complexes to obtain their stabilized derivatives for medical application.     

Effect of heparin on the antiradiation activity of cystamine, decreasing with a reduction in radiation intensity

It was shown that a single injection of heparin (250 units/kg) 15 min and 24 h before irradiation potentiated a slight radioprotective effect of cystamine (dichlorohydrate, 170 mg/kg) which was registered after the administration thereof to mice 30 min before irradiation with an absolutely lethal dose at a dose rate of 0.0025 Gy/c.     

Characterisation of new substrate specificities of Escherichia coli and Saccharomyces cerevisiae AP endonucleases

Despite the progress in understanding the base excision repair (BER) pathway it is still unclear why known mutants deficient in DNA glycosylases that remove oxidised bases are not sensitive to oxidising agents. One of the back-up repair pathways for oxidative DNA damage is the nucleotide incision repair (NIR) pathway initiated by two homologous AP endonucleases: the Nfo protein from Escherichia coli and Apn1 protein from Saccharomyces cerevisiae. These endonucleases nick oxidatively damaged DNA in a DNA glycosylase-independent manner, providing the correct ends for DNA synthesis coupled to repair of the remaining 5′-dangling nucleotide. NIR provides an advantage compared to DNA glycosylase-mediated BER, because AP sites, very toxic DNA glycosylase products, do not form. Here, for the first time, we have characterised the substrate specificity of the Apn1 protein towards 5,6-dihydropyrimidine, 5-hydroxy-2′-deoxyuridine and 2,6-diamino-4-hydroxy-5-N-methylformamidopyrimidine deoxynucleotide. Detailed kinetic comparisons of Nfo, Apn1 and various DNA glycosylases using different DNA substrates were made. The apparent K_m and k_cat/K_m values of the reactions suggest that in vitro DNA glycosylase/AP lyase is somewhat more efficient than the AP endonuclease. However, in vivo, using cell-free extracts from paraquat-induced E.coli and from S.cerevisiae, we show that NIR is one of the major pathways for repair of oxidative DNA base damage.     

A Neurophysiological Picture of the Genesis of Fatigue, Chronic Fatigue, and Overfatigue in Human Operators

The present widespread conceptual model of the neurophysiological picture of human fatigue, chronic fatigue, and overfatigue is limited to the picture of hypoexcitability and development of inhibitory processes in the CNS. The results of this study show that, when fatigue deepens and is followed by chronic fatigue and overfatigue, the inhibitory processes that developed earlier weaken. The speed of nervous processes and their lability increase in this period, exceeding the initial normal baseline values; the differential inhibition strengthens. Both main nervous processes weaken; the excitatory process begins to relatively predominate. The equalization and paradoxical phases manifest themselves in the CNS functioning. As fatigue deepens and overfatigue develops, excitability of nervous structures continues decreasing. Interhemispheric asymmetry increases.     

Hijacking of the human alkyl-N-purine-DNA glycosylase by 3,N4-ethenocytosine, a lipid peroxidation-induced DNA adduct

Lipid peroxidation generates aldehydes, which react with DNA bases, forming genotoxic exocyclic etheno(epsilon)-adducts. E-bases have been implicated in vinyl chloride-induced carcinogenesis, and increased levels of these DNA lesions formed by endogenous processes are found in human degenerative disorders. E-adducts are repaired by the base excision repair pathway. Here, we report the efficient biological hijacking of the human alkyl-N-purine-DNA glycosylase (ANPG) by 3,N(4)-ethenocytosine (epsilonC) when present in DNA. Unlike the ethenopurines, ANPG does not excise, but binds to epsilonC when present in either double-stranded or single-stranded DNA. We developed a direct assay, based on the fluorescence quenching mechanism of molecular beacons, to measure a DNA glycosylase activity. Molecular beacons containing modified residues have been used to demonstrate that the epsilonC.ANPG interaction inhibits excision repair both in reconstituted systems and in cultured human cells. Furthermore, we show that the epsilonC.ANPG complex blocks primer extension by the Klenow fragment of DNA polymerase I. These results suggest that epsilonC could be more genotoxic than 1,N(6)-ethenoadenine (epsilonA) residues in vivo. The proposed model of ANPG-mediated genotoxicity of epsilonC provides a new insight in the molecular basis of lipid peroxidation-induced cell death and genome instability in cancer.     

Identification of the ORC Complex Subunits That Can Interact with the ENY2 Protein of <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

The interaction of the Drosophila ENY2 protein with the ORC complex subunits was investigated. It is found that ORC4 and ORC6 subunits directly interact with ENY2.     

Resource density regulates the foraging investment in higher termite species

1. Resource density can regulate the area that animals use. At low resource density, there is a conflict in terms of balance between costs of foraging and benefits acquired. The foraging of the higher termite Nasutitermes aff. coxipoensis consists of searching throughout trails and a building galleries phase. 2. In this study, a manipulative field experiment was used to test the hypothesis that colonies of N. aff. coxipoensis forage towards a more profitable balance between the establishment of trails and gallery construction at low resource density. 3. The experiment was conducted in north‐eastern Brazil. Seven experimental plots were established with a continuous increase in resource density (sugarcane baits). Entire colonies of N. aff. coxipoensis were transplanted from their original sites to the experimental plot, totalling 35 nests. The number, branches and total length of trails and galleries were quantified. 4. The results show that N. aff. coxipoensis optimises its foraging output, intensifying the establishment of trails at the cost of gallery construction when resource density is low. The number of trails, the number of trail branches and the total length of trails decreased with increasing resource density. Interestingly, at low resource density, the search effort was concentrated on forming longer and a greater number of trails, a small proportion of which were converted into galleries. The opposite relationship was observed at high resource density. 5. These results suggest an optimisation of search efforts during foraging depending on resource density, a mechanism that may help researchers to understand the use of space by higher termite species.