MANIFESTATIONS OF OXIDATIVE STRESS AND MOLECULAR DAMAGES IN OVARIAN CANCER TISSUE.

Indices of oxidative stress are recognized molecular markers and prognostic criteria for malignant transformation of tissue, but their value depends on the type of tumor and the stage of its development. The goal of this study was to clarify the relationship between the characteristics of the oxidative stress system including metal-associated ones and the cytotoxicity manifestations in neoplastically transformed human ovarian tissue. The highest level of Mn-superoxide dismutase activity (by 630%) and metallothionein protein (MT, 100%) has been estimated for the first time in malignant ovarian tissue compared to normal ovarian tissue. The researchers have also found a much higher level of oxy-radical formation (by 332%), a lower activity of catalase (by 49%) and a lower level of reduced glutathione (by 46%) and its redox index (0.84 versus 0.89 in the control) in tumor tissue. Under the relatively stable content of zinc, copper and cadmium in MTs, the content of zinc and especially copper in a form non-binding with MTs was significantly lower in the malignant tissue compared to normal one while the content of cadmium was higher. A discriminant analysis of all definable parameters revealed that the higher content of the products of oxidative destruction of proteins, lipids, fragmented DNA and the activity of cathepsin D, especially in its free form (by 235%), are the main characteristic signs of malignant ovarian tissue.

[Trace elements storage peculiarities and metallothionein content in human thyroid gland under iodine deficiency euthyroid nodular goiter].

Accumulation of iodine and copper in the node, paranodular and contralateral (not affected tissue by node) tissues of thyroid gland in relation to the level of metal-binding proteins, potential antioxidants and oxidative changes in tissue was investigated. To assess the severity of the pathological process the molecular markers of cytotoxicity were used. The reduction of total iodine (by 19.5%), increase of inorganic iodine fraction (by 82.4%) and total copper content (twice) in paranodular and nodular tissues compared with contrlateral part have been established. Excess of copper in goitrous-changes tissue was partially accumulated in the metallothioneins. The level of metal-binding form of metallothioneins and reserve of free thiols of these proteins was higher two-three times and lower content of reduced glutathione in node-affected tissue compared to the contralateral part. Signs of cytotoxicity among them: higher cathepsine D free activity (up to 84.6% and 134.4% in paranodular tissue and node respectively) and higher level of DNA strand breaks in the node (up to 22.6%) were observed. In paranodular tissue the range of indices variability compared with parenchyma of contralateral part is shorter than in the node. Thus, under low level of iodine organification and high copper level in goitrous-modified tissue of thyroid gland metallothionein may provide a partial compensatory effect on prooxidative processes.

[Functions of metallothioneins and a system of antioxidant defense under the effect of Co- and Zn-containing nanocomposites on crucian carp (Carassius auratus gibelio)].

The effect of metal-nanocomposites (Me-NC) of cobalt and zinc (Co- and Zn-NC, correspondingly) synthecized on the basis of vinylpyrrolidone (PS) on the metal-accumulative proteins with antioxidant potential metallothioneins (MT) in crucian carp (Carassius auratus gibelio) was studied. Fish was subjected to the effect of Co-NC, Zn-NC, Co2+, Zn2+ or polymer carrier (PC) in the concentrations correspondent to 50 microg x Co/l or 100 microg x Zn/l during 14 days. It was shown that the MTs response is highly specific for the nature of metal, both in ion and Me-NC form: the effect of Co and Co-NC provoked the elevation of total MT concentration (MT-SH) and activation of antioxidant defence, whereas Zn and Zn-NC induced the decrease of the concentration of MT-SH and the inhibition of antioxidant defense. All the exposures provoked the decrease of the concentration of immunoreactive chelating MT form (MTi) and reduced glutathione, activation of anaerobiosys and Mn-superoxide dismutase, and also decrease of the concentration of proteins and lipids oxidative injury products. It was accompanied by the increase of the content of erythrocytes with nuclear abnormalities but did not cause the decrease of choline esterase activity. According to the rate of MT-SH and MTi concentrations, antioxidant potential of MTs is determined by its apoform. Our data indicate that partial biodegradation of Me-NC occurs in the organism of crucian carp.

[Population-related peculiarities of molecular stress-responsive systems of bivalve mollusk under the effect of tetrazine pesticide].

Tetrazine pesticides are widely used for the treatment of crops in most EU countries and USA. However, data about the effect of environmentally realistic concentrations of biocides on the molecular stress response system in non-target organisms are absent. The aim of our study was the comparison of adaptive capability of bivalve mollusk Anodonta cygnea from two populations under the effects of commercial pesticide Apollo in terms of biochemical parameters of the digestive gland. The differences between parameters of oxidative stress and glutathione transferase activity in specimens of control groups from clean (I group) and polluted (B group) areas have been shown. Under the effect of Apollo, the level of protein carbonyls and microsomal oxidation processes increased, and the level of metallothioneins and oxyradical formation decreased in the specimens from both populations. However, the treatment provoked the activation of antioxidant processes in the I group and their inhibition in B group. Potentially the injury of cellular thiols, glutathione and metallothioneins, seems to be key point of tetrazine pesticides toxicity.

[Metal-binding functions and antioxidant properties in human thyroid gland under iodine deficient nodular colloidal goiter].

Copper and zinc levels in the tissue of thyroid gland (TG) and their metal-binding proteins metallothioneins (MT) as well as state of the antioxidant system in persons that had no thyroid disease and patients with endemic iodine deficiency nodular colloidal goiter has been investigated. In the patients with thyroid disease, oxidative damage was indicated despite elevated levels of MT-SH and glutathione, and elevated copper and decreased zinc concentration in TG tissue. MTs partly bound the excess of copper but its concentration in the unbound to MT form was two-fold compared to the control value.

[Evaluation of biological effects of cobalt-nanocomposites with the use of biochemical markers of bivalve mollusk Anodonta cygnea].

Intensive implementation of nanomaterials requires development of novel methods for evaluation of their potential ecotoxicity. The aim of our study was to identify specific characteristics of the effect of cobalt-nanocomposite (Co-NC) on the molecular stress-responsive system in the digestive gland of bivalve mollusk Anodonta cygnea. Nanocomposite was synthesized by mixing alcohol solution of copolymer N-vinylpirrolidone, 5-(tret-butylperoxy)-5-methyl-1-hexene-3-yne and dimethylaminoethylmetacrylate and cobalt (II) chloride. After 14 days of the mollusk exposure in the presence of Co-NC, CoCl, or corresponding polymer substance it was shown that the Co-NC, in contrast to other agents, does not cause an oxidative stress due to the superoxide dismutase activity, metallotioneins (MTs) level, glutathione redox index and oxyradical production. Multivariate analysis confirmed specific features of the Co-NC's effect related to an enhanced expression of MTs, while CoCl2 activated lactate dehydrogenate and oxyradical production, and polymer substance enhanced glutathione transferase activity.

Reactivity of metallothioneins of frog Rana ridibunda treated by copper and zinc ions.

The metal-buffering and stress proteins metallothioneins (MTs) of frog are characterised by unusually high content of copper as for vertebrate animals and instability that was shown in our previous studies. They easily lost copper and especially zinc under unfavourable conditions. The aim of this study was to examine the reactivity of SH groups in the MTs from the liver of frog Rana ridibunda after the effect of Cu2+ (0.01 mg/l) and Zn2+ (0.1 mg/l) ions on the organism during 14 days. The alpha- and beta-domains of MTs with molecular weights of about 4 kDa were separated by the size-exclusion chromatography on Sephadex G-50. Unlike higher vertebrates, frogs demonstrated higher reactivity of alpha-domain than beta-domain with the Ellman's reagent (DTNB). The signs of partial oxidations in beta-domain included the creation of by-products with molecular weight about 12 kDa, low reactivity of SH-groups, and typical of -S-S-bonds peculiarities of UV-spectra. The effect of both metal ions on frog provoked the elevation of SH-groups reactivity in a-domain with the appearance of by-product with molecular weight of 16 kDa and its reduction in beta-domain. The incubation of MTs of control animals with 0.5 and 5.0 mM of H2O2 did not affect its chromatographic characteristics. In the frogs loaded by Cu2+ and Zn2+ the effect of 5.0 mM H2O2 on MTs provoked the release of 4 kDa product. So the alpha-domain is responsible for the increased release of metals from injured MTs in frogs, whereas extremely high oxidizability of beta-domain makes its participation in the exchange of metals elusive and provokes the aggregation of MTs.

[The effect of pesticide acetamiprid on biochemical markers in tissues of fresh water bivalve mussels Anodonta cygnea L. (Unionidae)].

The effects of ecological real concentration of pesticide "Mospilan" (active substance - acetamiprid) on the status of oxidative stress biomarkers, neurotoxicity and metallothioneins' (MTs) characteristics of freshwater bivalve mussels Anodonta cygnea were investigated. The prooxidative changes (decrease of superoxide dismutase activity, reduced glutathione content, increase of content of lipid and proteins peroxidation products) and decrease of the MTs content in gills and especially in digestive gland in the experimental group were reflected, whereas acetylcholinesterase activity was not changed. The effect of acetamiprid induces the appearance of additional chromatographic MTs form.

[Effect of zinc ions, manganese and lead ions on the thermostable proteins of carp liver]. / Vplyv ioniv tsynku, marhantsiu ta svyntsiu na termostabil'ni bilky pechinky koropa.

Effect of comparatively low concentration of the zinc, manganese, lead ions (0.1, 0.13 and 0.01 mg/l respectively) on the thermostable proteins (TP) of the carp liver after fish incubation for 14 days was studied. It is revealed that about 30 of zinc, 40% of manganese and 6% of lead were bound in the TP solution. The results of chromatography on Sephadex G-75 show, that the metallothionein-like proteins (MT, M-6 kDa) were the primary metal-binding proteins among the TP for zinc and manganese, and the heavy fraction of TP (M-60 kDa)--for lead. Under the action of each of the metallions its content in the TP is significantly increased. The distribution of metals among TP compounds is similar to that in the control. The metals action causes specific changes of the heavy fraction properties (UV spectrum, SH-groups content). The properties of MT do not change significantly under the control value.

[Regulatory mechanisms of oxygen transport function of hemoglobin in irradiated animals]. / Issledovanie mekhanizmov reguliatsii kislorodtransportnoi funktsii gemoglobina u obluchennykh zhivotnykh.

The oxygen affinity of hemoglobin of both common and 2,3-diphosphoglycerate concentration-purified erythrocyte hemolysates, hemoglobin stability, 2,3-diphosphoglycerate concentration in erythrocytes, morphological blood content were studied in rabbits irradiated with X-rays in a dose of 0.155 C/kg. An increase in the oxygen affinity of common hemolysate hemoglobin at a higher 2,3-diphosphoglycerate concentration on the 1st, 4th day after irradiation and a decrease in the oxygen affinity of both common and purified hemolysate hemoglobin on the 18th-30th day after irradiation were observed. A conclusion is made that disturbance of the hemoglobin function control by 2,3-diphosphoglycerate in irradiated animals is determined by alteration of protein physicochemical properties.