Hypoxia and recovery perturb free radical processes and antioxidant potential in common carp (Cyprinus carpio) tissues

The effects of hypoxia exposure and subsequent normoxic recovery on the levels of lipid peroxides (LOOH), thiobarbituric acid reactive substances (TBARS), carbonylproteins, total glutathione levels, and the activities of six antioxidant enzymes were measured in brain, liver, kidney and skeletal muscle of the common carp Cyprinus carpio. Hypoxia exposure (25% of normal oxygen level) for 5h generally decreased the levels of oxidative damage products, but in liver TBARS content were elevated. Hypoxia stimulated increases in the activities of catalase (by 1.7-fold) and glutathione peroxidase (GPx) (by 1.3-fold) in brain supporting the idea that anticipatory preparation takes place in order to deal with the oxidative stress that will occur during reoxygenation. In liver, only GPx activity was reduced under hypoxia and reoxygenation while other enzymes were unaffected. Kidney showed decreased activity of GPx under aerobic recovery but superoxide dismutase (SOD) and catalase responded with sharp increases in activities. Skeletal muscle showed minor changes with a reduction in GPx activity under hypoxia exposure and an increase in SOD activity under recovery. Responses by antioxidant defenses in carp organs appear to include preparatory increases during hypoxia by some antioxidant enzymes in brain but a more direct response to oxidative insult during recovery appears to trigger enzyme responses in kidney and skeletal muscle.     

The effect of various chemical compounds and storage conditions on the rate of non-enzymatic deamidation of protein preparations

The effect of different substances partly used as preservatives for the blood storage and at different stages of manufacturing of human blood preparations on the dynamics of nonenzymatic deamidation of commercial protein preparations and on their heat stability was being studied. Albumin and gamma-globulin preparations in the solutions of 60% glycerol, 60% ethylene glycol, 40% beta-alanine, aspartic and glutamic acids in physiological concentrations, 40% glucose and 40% sucrose after 2-hour thermal denaturation (100 degrees) were incubated under (or close to) physiological conditions (pH 7, 37 degrees) for 0.7; 14, 21, 28 and 90 days. The protein preparations in the saline were used as control. After precipitation of protein with TCA, the contents of free ammonia and TCA-soluble products were measured by the Lowry technique. The precipitate washed by organic solvents was used to determine the amide fractions. No reliable difference in increasing the TCA-soluble Folin-positive products was observed. All substances studied but glycerol sped up the deamidation of albumin and gamma-globulin preparations both during thermal denaturation and incubation. On the contrary, glycerol slowed down the deamidation of proteins. Possible reasons of the observed phenomena are discussed.     

Influence of exogenous proteins on the mutagenic

Mutagenic factors of biological origin on the example of carbohydrate-binding proteins are observed. An attempt to summarize the existing data concerning participation of exogenous macromolecules in the mutagenic process is made. Mechanisms of the influence of exogenous macromolecules on mutagenesis, proliferation, and surviving of mammalian cells are discussed.     

Effects of Some Growth Factors and Cytokines on the Expression of the Repair Enzyme MGMT and Protein MARP in Human Cells In Vitro

The inducible repair enzyme O⁶-methylguanine-DNA methyltransferase (MGMT) eliminates O⁶-methylguanine adducts in DNA and protects the cells from damaging effects of alkylating agents. We have found that anti-MGMT antibodies recognize both the MGMT protein with a mol. weight?~?24 kDa and a protein with a mol. weight?~?48 kDa, which was named MARP (anti-methyltransferase antibody recognizable protein). A number of growth factors and cytokines were shown to regulate the expression of MGMT and MARP proteins. The ranges of concentrations of several growth factors and cytokines that caused increasing or decreasing protein amounts in human cell cultures were determined. The results of special biological experiments have allowed us to assume a possible role of MARP in the repair of alkyl adducts in human cells.     

Miscanthus as a Productive Biofuel Crop for Phytoremediation

There are many locations where soil quality improvements would be beneficial because of contamination, erosion, flooding, or past human activities. Miscanthus, a C-4 grass related to sugarcane, grows well in mildly contaminated soil and on sites where soil quality is poor, particularly with respect to nitrogen. Because of its high biomass yield, it is of interest as an energy crop, and as a plant to use for simultaneous crop production and phytoremediation. Here we review recent literature on using miscanthus for combined biomass production and phytoremediation of contaminated and marginal lands. We analyze both advantages and disadvantages for production of this crop along with phytoremediation of sites contaminated with metals and petroleum hydrocarbon. Reports of laboratory and field investigations, which use Miscanthus spp. for stabilizing and removing metals are considered. The potential for growing miscanthus commercially at contaminated and marginal sites in the regions of Central and Eastern Europe as well as the United States appears to be good because large quantities of biomass can be produced and effective phyto-stabilization can be achieved with very slow metal removal over time. In addition, soil quality is improved in many cases.     

The Fibroblast Growth Factor 14·Voltage-gated Sodium Channel Complex Is a New Target of Glycogen Synthase Kinase 3 (GSK3)

The FGF14 protein controls biophysical properties and subcellular distribution of neuronal voltage-gated Na⁺ (Nav) channels through direct binding to the channel C terminus. To gain insights into the dynamic regulation of this protein/protein interaction complex, we employed the split luciferase complementation assay to screen a small molecule library of kinase inhibitors against the FGF14·Nav1.6 channel complex and identified inhibitors of GSK3 as hits. Through a combination of a luminescence-based counter-screening, co-immunoprecipitation, patch clamp electrophysiology, and quantitative confocal immunofluorescence, we demonstrate that inhibition of GSK3 reduces the assembly of the FGF14·Nav channel complex, modifies FGF14-dependent regulation of Na⁺ currents, and induces dissociation and subcellular redistribution of the native FGF14·Nav channel complex in hippocampal neurons. These results further emphasize the role of FGF14 as a critical component of the Nav channel macromolecular complex, providing evidence for a novel GSK3-dependent signaling pathway that might control excitability through specific protein/protein interactions.     

Microparticles in physiological and in pathological conditions

Chronic diseases pose a severe burden to modern National Health Systems. Individuals nowadays have a far more extended lifespan than in the past, but healthy living was only scantily extended. As much as longer life is desirable, it is saddened by chronic diseases and organ malfunctions. One contributor to these problems was recognized to be represented by microparticles (MPs). Our purpose is to better understand MPs, to contrast their ominous threat and possible clinical importance. For this intent we correlated MPs with thrombotic pathologies, hemophilia, malaria, diabetes, cardiovascular diseases, endothelial dysfunctions, pulmonary hypertension, ischemic stroke, pre-eclampsia, rheumatologic diseases-rheumatoid arthritis, polymyositis-dermatomyositis, angiogenesis and tumor progression-cancer; we listed the possibilities of using them to improve transfusion methods, as a marker for acute allograft rejection, in stem cell transplantation, as neuronal biomarkers, to understand gender-specific susceptibility for diseases and to improve vaccination methods and we presented some methods for the detection of MPs.     

Change in the MGMT gene expression under the influence of exogenous cytokines in human cells in vitro

The influence of cytokines LIF, SCF, IL-3, and EMAP II and the Laferobion (IFN-a2b) drug on the MGMT gene expression in human cell cultures has been studied. It was shown that exogenous cytokines can modulate the MGMT gene expression at the protein level. EMAP II is able to increase or decrease the MGMT level, depending on the experimental conditions. Cytokines LIF, SCF, IL-3 and Laferobion decreased the MGMT expression level in human cells in vitro. Some conditions leading to the destruction of the MGMT protein complex were identified.