Disturbances of fertilizing potential of spermatozoa are closely associated with dysfunction of ion-transporting ATPases, in particular Ca2+, Mg2+-АТРase. Reduced activity of tapsigargin-resistant and tapsigargin-sensitive Ca2+, Mg2+-АТРase leads to disruption of Ca2+-homeostasis and is characteristic for abnormal spermatozoa (pathoospermia). In order to study the peculiarities of action of Ca2+, Mg2+-АТРase, we determined the initial reaction rate, the maximum (plateau) amount of the reaction product and the cha­­racteristic reaction time. To determine these kinetic parameters of Ca2+, Mg2+-dependent hydrolysis of ATP catalyzed by Ca2+, Mg2+-ATPase, the dynamics of product accumulation of the ATP-hydrolases reaction was studied. The obtained curves were linearized in the coordinates {P/t; P}. Analyzing the changes in the activity of Ca2+, Mg2+-ATPase, the kinetics of primary-active transport of calcium ions through the plasma membrane and membranes of intracellular Ca2+-stores in saponin-permeabilized spermatozoa of infertile men was studied. It was shown that in normozoospermic samples, the transport of Ca2+ ions through the plasma membrane is characterized by a higher capacity than through the membranes of intracellular Ca2+-stores, but it occurs with practically the same initial velocity and characteristic reaction time. It was found that in pathospermic samples, transport of Ca2+ ions with the participation of both components of Ca2+, Mg2+-ATPase occurs less intensively and is characterized by a lower capacity compared to spermatozoa of men with preserved fertility. Specific changes in the kinetic parameters of Ca2+, Mg2+-dependent hydrolysis of ATP lead to inhibition of tapsigargin-resistant and tapsigargin-sensitive Ca2+, Mg2+-ATPase activity and cause a decrease in fertilizing potential of spermatozoa. Keywords: Ca2+, Mg2+-ATPase, ATP hydrolysis, spermatozoa, male infertility, pathospermia

Glyphosate is the worldwide used herbicide of the first priority. However, its biochemical effects in the aquatic animals are studied scantly. The ex vivo approach has been recently proposed to provide the express evaluation of the adverse impact without the treating of the organisms. The aim of this study was to verify this approach for the assessment of the toxicity of glyphosate to the bivalve mollusk. The samples of the gills and digestive gland tissues of freshwater bivalve Unio tumidus mollusk were exposed to a range of the concentrations of glyphosate (commercial formulation Roundup MAX) at the concentrations 13.3, 26.7, 66.8 and 133.6 µg×L-1 during 2 h at 20 °C followed by 15 h at ~ 2–4o C. The markers of oxidative injury (total antioxidant activity, end-products of lipid peroxidation (TBARS) and protein carbonyls (PC)), cellular low weight thiols GSH/GSSG and metallothionein (MT), and cholinesterase activity as the index of neurotoxicity were analyzed. We also assayed the index of cell vitality as the lysosomal membrane stability from the Neutral Red Retention (NRR) test. The results have shown that the lowest concentrations of glyphosate caused the most prominent changes of the indices: the decrease of MT concentration (by ~ two times) and cholinesterase activity. The total antioxidant activity was decreased substantially in all exposures correspon­dingly to a decrease of the MT and/or GSH concentrations. However, the levels of TBARS and PC were not changed comparing to control detecting the early stage of the injury. Surprisingly, NRR increased in the exposures to higher concentrations of glyphosate, probably due to strong chelating ability of glyphosate or other compounds of formulation. This study allows us to detect the earlier biological effects of glyphosate in the low environmentally realistic concentrations. Further validation of this approach needs the comparison of the results in the ex vivo and in vivo experiments.Keywords: Roundup; bivalve mollusk; cytotoxicity; ex vivo

The review analyzes and summarizes information on the history of development and the current state of methodological approaches to the qualitative and quantitative determination of phytohormones in plant tissues. Plant hormones play an indispensable role in many physiological processes during a plant life cycle, from seed’s germination to senescence. The determination of the endogenous hormones concentration is essential for elucidating the role of a particular hormone in any physiological process. A sensitive and quick analytical method of a simultaneous quantitative estimation of the main classes of phytohormones is essential for the investigation of signaling control networks in specific developmental pathways and physiological responses. In plant tissues, phytohormones are present in very low concentrations (from 10-9 M to 10-6 M); hence, the availability of highly effective, comprehensive and reliable analytical techniques for their identification is extremely important. The article gives a brief description of the main classes of plant hormones and outlines their functional activity. The importance of methods for hormones identification in plant tissues and their use in plant physiology and agricultural practice is discussed. The study presents a valid and tested sequence of procedures for the extraction of plant hormones, a methodology for purification of the obtained extracts from interfering substances and an up-to-date method of plant hormones quantification (indole-3-acetic, abscisic, gibberellic, salicylic acids and five forms of cytokinins). The quantification method combines a high-performance liquid chromatography with a mass spectrometry. Four chromatographic methods for the sepa­ration and detection of substances in aliquots as well as ionization conditions for hormones in a mass spectrometer are described. The presented analytical technique is adapted for scientific laboratories in Ukraine.Keywords: HPLC/MS-analysis, plant hormones, sample preparation

The influence of ferrum (III) citrate added to the cultivation medium, on the reduction of sulfate, nitrate, and nitrite ions by sulfate-reducing bacteria Desulfovibrio desulfuricans IMV K-6, Desulfovibrio sp. Yav-6, and Desulfovibrio sp. Yav-8 isolated from Yavorivske Lake was studied. It was established that ferrum (III) citrate inhibits the biomass accumulation, SO42- reduction, and H2S production by the bacteria after addition of 1.74–3.47 mM Na2SO4×10 H2O and 1.74–10.41 mM FeC6H5O7 to the medium, in comparison with the growth and level of the reduction of sulfate ions by bacteria in the medium supplemented with only Na2SO4×10 H2O. At conditions of the bacteria cultivation in the presence of an equimolar amount (3.47 mM) of Na2SO4×10 H2O and FeC6H5O7, they reduced 2.5−2.7 times more Fe(ІІІ) than SO42- with Fe2+ production at a concentration 2.4−2.7 times greater than H2S. FeC6H5O7 inhibited growth, NO3- or NO2- reduction and NH4+ production by the bacteria in the presence of 1.74−3.47 mM NaNO3 or NaNO2 and 1.74−10.41 mM FeC6H5O7 in the medium, compared to the growth and level of nitrate or nitrite ions reduction in the medium with only NaNO3 or NaNO2. In the medium with the same initial content of 3.47 mM NaNO3 and 3.47 mM FeC6H5O7, the bacteria reduced 1.4 times more NO3- than Fe(ІІІ), with NH4+ production at concentration 1.1 times higher than that of Fe2+. In the medium with 3.47 mM NaNO2 and 3.47 mM FeC6H5O7, the cells reduced 1.4−1.6 times more Fe(ІІІ) than NO2-, with Fe2+ production at concentra­tion 1.5−1.6 times higher than NH4+. Ferrum (III) citrate had more inhibitory effect on the dissimilatory reduction of sulfate by the bacteria than of nitrate and nitrite ions, since the SO42- reduction by the bacteria at its presence in the medium decreased 2.0−4.7 times. The reduction of NO3- and NO2- decreased only 1.3−1.9 and 1.7−3.1 times, respectively, as compared with their reduction in the media with only Na2SO4×10 H2O, NaNO3 or NaNO2. Despite the fact that the reduction by cells of 1.74−10.41 mM Fe(III) in the media with Na2SO4×10 H2O, NaNO3 or NaNO2 decreased 1.1−2.1, 1.6−2.7 and 1.1−2.5 times, respectively, compared with its reduction in the medium with only FeC6H5O7. The investigated strains of bacteria were resistant to high concentrations of ferrum (III) citrate and, therefore, can be applied in the technologies of complex environment purification from pollution with ferrum, sulfur, and nitrogen compounds.Keywords: Desulfovibrio, ferrum, sulfates, nitrates, nitrites