Visualization and Cytotoxicity of Fluorescence-Labeled Dimeric Magnetite-Gold Nanoparticles Conjugated with Prostate-Specific Membrane Antigen in Mouse Macrophages.

We demonstrated the possibility of penetration of magnetite-gold nanoparticles conjugated with prostate-specific membrane antigen into mouse macrophages. It was found that after 3-h incubation with nanoparticles in a concentration of 15 mg/liter at 37oC, they were seen in only 13% macrophages. In about 90% cells, the nanoparticles were detected within the cytoplasm. Under these conditions, membrane damage was revealed in 25% cells. These results should be taken into account in further development and application of nanomaterials for diagnostic and therapeutic purposes in oncology.

Low Concentrations of Hydrogen Peroxide Activate the Antioxidant Defense System in Human Sperm Cells.

The effect of low concentrations of hydrogen peroxide (10-100 µM) on sperm motility and on the activity of the sperm enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDS) was investigated. Incubation of semen samples with 10 and 100 µM hydrogen peroxide increased the content of spermatozoa with progressive motility by 20 and 18%, respectively, and enhanced the activity of GAPDS in the sperm cells by 27 and 20% compared to a semen sample incubated without additions. It was also found that incubation with 10 µM hydrogen peroxide increased the content of reduced glutathione (GSH) in sperm cells by 50% on average compared to that in the control samples. It is supposed that low concentrations of hydrogen peroxide activate the pentose phosphate pathway, resulting in NADPH synthesis and the reduction of the oxidized glutathione by glutathione reductase yielding GSH. The formed GSH reduces the oxidized cysteine residues of the GAPDS active site, increasing the activity of the enzyme, which in turn enhances the content of sperm cells with progressive motility. Thus, the increase in motile spermatozoa in the presence of low concentrations of hydrogen peroxide can serve as an indicator of normal functioning of the antioxidant defense system in sperm cells.

[Effect of the incubation medium pH on damage to macrophages plasma membranes].

In this paper we showed the pH-dependent change in the sensitivity of the membranes of murine peritoneal macrophages to UV-radiation. This relationship is discussed in terms of lipid bilayer membrane stability modification to the action of ROS and lipid peroxidation process (LPP) at different pH. Iron-ascorbate reinforced LPP also led to pH-dependent membranes damage. The increase of the cells incubation medium temperature up to 37 degrees C, which also stimulated LPP, did not change the picture of the pH-dependent damage. Decrease of the incubation medium pH did not reduce H2O2-induced cell damage. Increase of the pH intensified the cells damage.

[Effect of hydroxylated fullerene C60(OH)25 on macrophage plasma membrane integrity].

Our study has shown that the damaging effect of hydroxylated fullerene C60(OH)25 on mouse peritoneal macrophage plasma membranes increased when we enlarged the concentration of fullerene in the incubation media (from 0.005 to 0.5 mg/ml), the incubation temperature (from 22 degrees C to 37 degrees C) and the time of incubation (from 30 to 90 min). In conditions of the H2O2-induced membrane damage, fullerene was observed to intensify the H2O2-induced damaging effect at a concentration of 0.05 mg/ml and reduce it at a concentration of 0.5 mg/ml. In conditions of the UV-induced membrane damage, it was discovered that the damaging effect of UV increased when C60(OH)25 nanoparticles were added to the incubation media before irradiation and decreased when they were added after irradiation. Eventual participation of ROS in damaging effects of C60(OH)25 was discussed.

[Damaging effect of "Poviargol" on mice peritoneal macrophage plasma membrane].

The damaging effect of "Poviargol", a substance containing silver nanoparts, was studied. It was shown that the damaging effect of "Poviargol" took place from the concentration of 2 mkg/ml and got its maximum at 10-12 microg/ml. Decrease of the incubation temperature from 30 to 4 degreesC led to amplification of the membrane-acting effect of "Poviargol"; however, inverse relation was observed in the range from 37 to 30 degreesC. The damaging effect of "Poviargol" increased when pH of the incubating medium was raised to 8.4 and also when the concentration of calcium ions in the incubation medium was raised to 8 mmol/l. The damaging effect decreased when pH of the incubation medium was reduced to 6.3, as well as in the presence of radioprotector serotonin. Our study allows us to suppose that reactive oxygen species and lipid peroxidation make a substantial contribution to the damaging effect of "Poviargol" on the macrophage plasma membrane.

[Alteration of UV-sensitivity of mice peritoneal macrophage plasma membranes by calcium ions].

Effect of Ca2+ ions on UV-induced mice peritoneal macrophage plasma membrane damage has been studied. Drop of the extracellular Ca2+ concentration has been found to result in a reduced expression of this damage. On the contrary, a raised intra- and extra-cellular Ca2+ level is associated with a higher number of cells with damaged plasma membranes. These findings make it possible to suggest that this change in the plasma membrane photosensitivity might be a result of alterations in the membrane lipid matrix electrical stability owing to UV-induced lipid photo-peroxidation. This study has also shown that free radical peroxidation of membrane lipids plays a significant part in UV-induced cell damage.

[The influence of hydrogene ion concentrations on peritoneal macrophage plasma membrane's photosensitivity].

It was demonstrated on macrophages treated with UV irradiation dose 9 J/sm2 (lamda max =306 nm) that intra- or extracellular pH reducing lead to decrease the number of cells with damaged membranes in macrophage population. An intra- or extracellular pH elevation leads to increase of UV-irradiation membrane-acting effect. It was also shown that pH-dependence of UV-irradiation damage effect has been lost after preliminary osmotic swelling of cells. The cells survived after UV-irradiation in doses 8 and 10 J/sm2 (lamda max =297 nm) have an intracellular pH lower than non-irradiated cells.

[Damaging effect of digitonin on macrophage plasma membranes exposed to UV-irradiation].

It was shown that macrophage irradiation in 4.6 J/cm2 (lambda(max) = 306 nm) dose leads to small quantity of damaged cells in cell population, which doesn't change substantially during 60 min of incubation in darkness. So as detergent digitonin treatment (without irradiation) in 3 mkg/ml concentration doesn't lead to substantial cell damage. Also the result of combined influence of UV-irradiation and digitonin added after irradiation, 15 min before the damaged cells counting, has been got. It was shown that macrophage incubation for 15 minutes leads to cell damaging twice as much sum of UV (4.6 J/cm2) and digitonin (3 mkg/ml) damaging. However the level of cell damaging obtained 30 minutes later after finishing of irradiation doesn't exceed the sum of separate effects of this factors. Further increase of postradiation time leads to synergic effect again.

[Effect of mineral water on metabolic processes].

The paper comparatively analyzes the effect of the mineral water "Penta" on the biochemical parameters of blood and urine, which characterize the functional activity of renal metabolic processes. A group comprising 10 examinees without renal disease took water by the routine mineral water scheme for 4 days. Comparison of the biochemical parameters before and after water taking revealed a significant reduction in azotemia and uric acid levels with its simultaneously enhanced excretion, as well as an increasing tendency for the excretion of oxalates, i.e. the most important parameters determining the formation of urate and oxalate calculi. These findings allow use "Penta", to a certain degree, in the treatment of renal disease, urolithiasis in particular, and in the prevention of stone formation.

[Influence of Ca2+ ions on UV-induced damage to mice peritoneal macrophage plasma membranes]. / Vliianie ionov na UF-indutsirovannoe povrezhdenie plazmaticheskikh membran peritoneal'nykh makrofagov myshei.

It was shown that UV-irradiation caused damage to mice peritoneal macrophage plasma membranes. A decrease in extracellular Ca2+ leads to a decrease of the damaging effect. An increase in extracellular Ca2+ or adding of calcium ionophore A23187 to the medium is accompanied by an increase in a number of damaged cells. These data allow us to suppose that modification of the damaging effect of UV-irradiation by Ca2+ ions can be bound with changing of electric stability of membrane lipid matrix.

[Influence of intracellular pH on character of mice peritoneal macrophage response to red light irradiation]. / Vliianie vnutrikletochnogo pH na kharakter otveta peritoneal'nykh makrofagov pri deistvii krasnogo sveta.

It was detected that exposure of macrophages to red light (600-740 nm) led to the changes in their intracellular pH and hydrolytic activity. The character of these changes depends on the initial level of pH in the cells. The maximum effect of irradiation is detected if the initial pH level is low. It is possible that Na/H-exchanger takes part in normalizing effect of red light on intracellular pH level.

[Alteration of intracellular pH of macrophages after UV-irradiation]. / Izmenenie vnutrikletochnogo pH makrofagov posle UF-oblucheniia.

It was shown that in vitro exposuse of mice peritoneal middle-wave ultraviolet radiation (lambdamax = 306 nm) in doses which don't damage to cause plasma membrane caused dose-dependent decreasing of their intracellular pH. After exposure of cells to 0.5 J/cm2 it was detected an acidification of intracellular contents followed by an increase of intracellular pH up to control level (after 40 min of incubation) and then above it (on 45 min of incubation). An increase of irradiation dose was accompanied by more evident reduction of intracellular pH and lack of its restoration on 45 min of postradiational incubation under irradiation with a dose of 3 J/cm2.

[Photodynamic damage of yeast subcellular fraction induced by elevated levels of endogenous protoporphyrin IX]. / Fotodinamicheskoe povrezhdenie subkletochnykh struktur drozhzhei s indutsirovannym nakopleniem éndogennogo protoporfirina IX.

The 2,2'-dipyridyl-induced accumulation of protoporphyrin IX in Saccharomyces cerevisiae cells was shown to be accompanied by the photoinhibition of cell respiration and the enhancement of the photoinduced permeability of plasma membranes to the fluorescent dye primuline. The visible-light illumination (at 400-600 nm) of the mitochondria and plasma membranes isolated from yeast cells with a high level of endogenous protoporphyrin IX intensified lipid peroxidation in these subcellular organelles. Comparative studies showed that the rad 52 mutant cells, which are deficient in the postreplicative recombinational DNA repair system, are considerably more sensitive to the inactivating action of visible light than are the wild-type cells and the rad 3 mutant cells, which are deficient in the excision DNA repair system. The contribution of photodynamic damage to the yeast subcellular organelles to the lethal photodynamic effect is discussed.

[Effect of short-term hypoxia and re-oxygenation on mice peritoneal macrophages in vitro]. / Deistvie kratkovremennoi gipoksii i reoksigenatsii na peritoneal'nye makrofagi myshei in vitro.

Microfluorimetry of single cells could help to analyze their morphology and function state during changes of gas environment. It is very important to have a possibility of the cell visual control during hypoxia and collection of dynamic fluorimetric data in digital form. The effects of short-term pO2 decrease were studied. For estimating the effects of hypoxia and reoxygenation we used the mice peritoneal macrophages, which are very sensitive to physical, chemical and regulatory stimuli. A special small chamber for fluorimetric measurements during pO2 changes, was developed. The level of active oxygen forms, intracellular pH, and cell membrane instability were investigated during replacement of air by nitrogen or argon (of the basal level decreased to 20% of basic level) and in subsequent reoxygenation. The increase of active oxygen forms was shown during 30 min of hypoxia and their level continued to rise immediately after reoxygenation. A short-term decrease and subsequent increase of pO2 in the medium led to an increase of intracellular pH level. The shifts of measured cell indices were stabilized after 30-40 min of pO2 changes thus suggesting a fast comprehension of countermeasure cell mechanisms. No macrophages with membrane disorders were found despite the rise of the active oxygen forms level during hypoxia and reoxygenation in vitro. There were no significant differences between nitrogen and argon used for replacement of air in the medium. The data obtained suggest a high resistance of macrophages against pO2 changes and an involvement of the antioxidative mechanisms for cell protection especially during reoxygenation period.

[Damage to macrophage membranes by exposure to middle-wave ultraviolet radiation]. / Povrezhdenie membran makrofagov pri deistvii srednevolnovogo ul'trafioletovogo izlucheniia..

An influence of middle-wave ultraviolet radiation (lambda max = 306 nm) on plasma membranes of mice peritoneal macrophages was studied by microfluorimetry analysis. It was found that a percentage of cells with damaged plasma membranes in the irradiated macrophage population reliably increased with the UVB dose starting with 6 J/cm2. Irradiation of cells with 4.2 J/cm2 UVR dose which does not cause evident damage to plasma membranes led to the latent damage which was detected by treatment with detergent digitonin (4.5 micrograms/ml).

[Modification of damaging effect of ultraviolet radiation on peritoneal macrophage membranes in mice]. / Modifikatsiia povrezhdaiushchego deistviia ul'trafioletovogo izlucheniia na membrany peritoneal'nykh makrofagov myshei.

Different factors modificating damaging effect of middle-wave ultraviolet radiation (lambda max = 306 nm) on mice peritoneal macrophage plasma membranes were studied. It was shown that damaging effect of ultraviolet declined when the cells were simultaneously treated by red light (lambda max = 713 nm). This protective effect increased when a red component of light became greater and achieved almost 100% when it consisted 28.6%. It was also found that the decrease in intra- and extracellular pH led to the decrease in damaging effect of UVR. The increase of pH bring to an elevation of UVR destructive effect.

[Effect of argon and nitrogen on the peritoneal macrophages in mice and their resistance to the UV damaging effect in vitro]. / Vliianie argona i azota na peritoneal'nye makrofagi myshei i ikh ustoichivost' k povrezhdaiushchemu vosdeistviiu UF-oblucheniia in vitro.

Explored were effects of argon and nitrogen on intracellular pH in peritoneal macrophages in mice and resistance of cellular membranes to the UV damaging effect in vitro. Blasting argon or nitrogen along the surface of cell cultures in airtight chamber for 20 minutes was shown to decrease 5-folds the oxygen content of solution as compared with initial level with culture pH unchanged. Ten-minute blasting argon or nitrogen through the incubation chamber slightly elevates intracellular pH in macrophages. The standard cell incubation conditions recovered following approximately 60 minutes in hypoxic atmosphere, the ability of macrophages to build up fluorescein was degraded and they increased intracellular pH no matter the indifferent gas yet more marked in case of nitrogen in use. It was demonstrated that the normobaric gas environment with oxygen partly replaced by nitrogen or argon protects plasmatic membranes of cells from UV-induced damage.

[Effect of a low-frequencey magnetic field on esterase activity and change in pH in wheat germ during swelling of wehat seeds]. / Vliianie nizkochastotnogo magnitnogo polia na aktivnost' ésteraz i izmenenie pH u zarodysha v khode nabukhaniia semian pshenitsy.

The role of nonsteady phenomena determined by a low velocity of ion movements in a weak external field is considered in relation to their possible nonlinear effects on processes occurring in boundary layers near the membrane, particularly, on the release of membrane-bound proteins and pH value. It is shown that a short-term treatment of wheat seeds with low-frequency magnetic field at the stage of esterase activation during seed swelling enhances the activation of esterases; the effect observed at final stages of activation depends on the time after the treatment with electromagnetic field. Treatment of seeds with electromagnetic field at this stage changed qualitatively the time course of the release of reaction products into the medium: the reaction rate increased initially and then decreased below the control level. At earlier stages of swelling in treated seeds and at all stages in control seeds, the time course of the product release was linear. The retardation of the release of the reaction products at terminal stages of esterase activation is presumably related to the release of proteins and their complexes under the action of electromagnetic field and the resulting restoration of the barrier properties of membranes. Treatment with electromagnetic field also caused a noticeable acceleration of proton flow form the medium, which was judged from pH changes in the bulk medium and in the vicinity of germ surface. The difference between the treated and control samples after 23-24 h of imbibition became statistically significant and was as high as 0.4 pH units. By taking into account the nonsteady phenomena occurring upon action of low-frequency electromagnetic field, it is possible to explain unusual dependences of biological effects on the amplitude of the electromagnetic field, including the atypical enhancement of these effects by the action of weak low-frequency fields.

[The study of role of membrane-bound Ca2+ in the regulation of relationship between neuron and glia during rhythmic excitation]. / Issledovanie roli membranosviazannogo Ca2+ v reguliatsii vzaimootnoshenii neirona i glii pri ritmicheskom vozbuzhdenii.

The role of membrane-bound Ca2+ in the regulation of Ca2+ transport through voltage-gated Ca2+ channel, and NMDA-glutamate and n-acetylcholine receptors upon interaction of a neuron with glia during rhythmic excitation was studied. It was found that the redistribution and transport of Ca2+ play a crucial role in the conductance of rhythmic excitation in both a "neuron-neuron" system and the processes providing the maintenance of a stationary level of rhythmic excitation in the system "neuron-glia".

[The use of microfluorimetric analysis for the studies of the influence of changed gravity on isolated lymphocytes in the spleen of the mouse]. / Ispol'zovanie mikrofluorimetricheskogo analiza dlia izucheniia vliianiia izmenennoi sily tiazhesti na izolirovannye limfotsity selezenki myshei.

Effects of simulated changed gravity on isolated splenetic lymphocytes of mouse were evaluated with the microfluorimetic technique enabling observation of probe fluorescence in individual cells. It was stated that 5 and 60 min of clinostatting stimulated fluorochrome accumulation in cells whereas centrifugation, particularly for 60-minutes, decreased the ability of cells to accumulate fluorescein. Also, alteration of the gravity force had the opposite effect on the ability of cells to retain fluorescein. The most significant drain of the dye immediately after short hypogravity may be indicative of the functional lability of cells during clinostatting. However, as judged by slight changes in intracellular pH, metabolic and regulatory functions were not affected. Hypergravity for 15 min decreased intracellular pH. The increased period of centrifugation stabilized the parameter. Absence of changes in the proliferative activity in all the test exposures also backs up the conclusion that short-term changes in gravity do not produce any substantial shifts in the morphofunctional state of cells in vitro.