Opposite effects of low intensity light of different wavelengths on the planarian regeneration rate.

Planarian freshwater flatworms have the unique ability to regenerate due to stem cell activity. The process of regeneration is extremely sensitive to various factors, including light radiation. Here, the effect of low-intensity LED light of different wavelengths on regeneration, stem cell proliferation and gene expression associated with these processes was studied. LED matrices with different wavelengths (red (λmax = 635 nm), green (λmax = 520 nm) and blue (λmax = 463 nm), as well as LED laser diodes (red (λmax = 638.5 nm), green (λmax = 533 nm) and blue (λmax = 420 nm), were used in the experiments. Computer-assisted morphometry, whole-mount immunocytochemical study and RT-PCR were used to analyze the biological effects of LED light exposure on the planarian regeneration in vivo. It was found that a one-time exposure of regenerating planarians with low-intensity red light diodes stimulated head blastema growth in a dose-dependent manner (up to 40%). The green light exposure of planarians resulted in the opposite effect, showing a reduced head blastema growth rate by up to 21%. The blue light exposure did not lead to any changes in the rate of head blastema growth. The maximum effects of light exposure were observed at a dose of 175.2 mJ/cm2. No significant differences were revealed in the dynamics of neoblasts' (planarian stem cells) proliferation under red and green light exposure. However, the RT-PCR gene expression analysis of 46 wound-induced genes revealed their up-regulation upon red LED light exposure, and down-regulation upon green light exposure. Thus, we have demonstrated that the planarian regeneration process is rather sensitive to the effects of low-intensity light radiation of certain wavelengths, the biological activity of red and green light being dictated by the different expression of the genes regulating transcriptional activity.

Internal gravity waves in the energy and flux budget turbulence-closure theory for shear-free stably stratified flows.

We have advanced the energy and flux budget turbulence closure theory that takes into account a two-way coupling between internal gravity waves (IGWs) and the shear-free stably stratified turbulence. This theory is based on the budget equation for the total (kinetic plus potential) energy of IGWs, the budget equations for the kinetic and potential energies of fluid turbulence, and turbulent fluxes of potential temperature for waves and fluid flow. The waves emitted at a certain level propagate upward, and the losses of wave energy cause the production of turbulence energy. We demonstrate that due to the nonlinear effects more intensive waves produce more strong turbulence, and this, in turn, results in strong damping of IGWs. As a result, the penetration length of more intensive waves is shorter than that of less intensive IGWs. The anisotropy of the turbulence produced by less intensive IGWs is stronger than that caused by more intensive waves. The low-amplitude IGWs produce turbulence consisting up to 90% of turbulent potential energy. This resembles the properties of the observed high-altitude tropospheric strongly anisotropic (nearly two-dimensional) turbulence.

Accuracy of the First Stage of Hepatotoxicity Testing with the Use of Drosophila melanogaster Test System.

The genotoxic effects of heterocyclic compounds were evaluated on the basis of genetic and toxicological characteristics of a biological model of Drosophila melanogaster. Analysis of the viability parameters (fertility, progeny mortality) showed that of 6 tested substance, substance No. 3 exhibited minimum toxicity. After application of substances No. 1 and No. 5 in the studied concentrations, the number of survived flies was insufficient for further analysis, which attested to high toxicity of these substances. The intensity of apoptosis was studied in response to substances Nos. 2, 4, and 6. Substance No. 4 proved to be optimal by the parameter toxicity/apoptosis (low toxicity/high apoptosis), while substance No. 3 exhibited low toxicity, which manifested in low apoptosis intensity.

Bag-breakup fragmentation as the dominant mechanism of sea-spray production in high winds.

Showing the record strengths and growth-rates, recent hurricanes have highlighted needs for improving forecasts of tropical cyclone intensities most sensitive to models of the air-sea interaction. Especially challenging is the nature of sea-spray supposed to strongly affecting the momentum- and energy- air-sea fluxes at strong winds. Even the spray-generation mechanisms in extreme winds remained undetermined. Basing on high-speed video here we identify it as the bag-breakup mode of fragmentation of liquid in gaseous flows known in a different context. This regime is characterized by inflating and consequent bursting of the short-lived objects, bags, comprising sail-like water films surrounded by massive liquid rims then fragmented to giant droplets with sizes exceeding 500 micrometers. From first principles of statistical physics we develop statistical description of these phenomena and show that at extreme winds the bag-breakup is the dominant spray-production mechanism. These findings provide a new basis for understanding and modeling of the air-sea exchange processes at extreme winds.

[Cytogenetic indices for somatic mutagenesis in mammals exposed to chronic low-dose irradiation].

We used cytogenetic analysis in the studies of the biological effects of a radiation factor of natural and artificial origin (under conditions ofthe 30-km exclusion zone ofthe Chernobyl experimental landfills in Ukraine, Belarus and Russia). The studies have been performed on various types of mammals: domestic animals--cows, pigs, horses and rodents--root voles, the Af mouse line, and yellow necked field mouse, bank voles. We found significant changes in the level of MN and chromosomal aberrations in the animals that were exposed to the conditions of chronic low-dose radiation for a long time (bothin the habitat and upon exposure in the Chernobyl zone) regardless of the type of animal and nature of contamination.

[Morphometric and cytogenetic studies of the follicular epithelium thyroid of small mammals under chronic exposure to small doses].

We have detected the genotoxic effect (increase of micronuclei) after chronic exposure to low-level ionizing radiation of the thyroid follicular epithelium of the voles living for many generations in the radioactive territory. At the same time, we also discovered the stimulating effect of chronic low-level ionizing radiation on the activity of morphogenetic processes in the thyroid parenchyma of the animals from natural populations (increased micro-follicles). These data were confirmed by the experiments; chronic irradiation of the thyroid gland of rats at the doses of 5 and 50 cGy revealed an increased induction of micronuclei and the increase in the number of small follicles. An increased proliferative potential of thyroid may be regarded as a non-specific adaptive response to chronicbody exposure to ionizing radiation.

Stat5 gene dosage in T cells modulates CD8+ T-cell homeostasis and attenuates contact hypersensitivity response in mice.

BACKGROUND: Contact hypersensitivity assay (CHS) faithfully models human allergies. The Stat5 transcription factors are essential for both lymphocyte development and acute immune responses. Although consequences of Stat5 ablation and transgenic overexpression for the lymphocyte development and functions have been extensively studied, the role of Stat5 gene dosage in contact allergies has not been addressed. OBJECTIVE: We investigated the effect of Stat5 gene dosage modulation in contact allergies using CHS in mice. METHODS: Transgenic animals heterozygous for the germline Stat5 null allele were subjected to CHS. To dissect cell type sensitive to Stat5 gene dosage, animals with Stat5 haplo-insufficiency in T cells, where one Stat5 allele was removed by Lck-Cre-mediated deletion (Stat5(ΔT/+)), were tested by CHS. Frequency of T cells, B cells, and monocytes were analyzed in Stat5(ΔT/+) and wild-type animals by flow cytometry. Proliferation of Stat5(ΔT/+) CD8(+) T cells was studied in vitro by stimulation with IL-4 and IL-2 cytokines, and changes in the expression of Stat5 target genes were assayed by quantitative real-time PCR assay. RESULT: Haplo-insufficiency of Stat5 in T cells leads to the reduction in CD8(+) T cells in all lymphoid organs and attenuates CHS response. Stat5(ΔT/+) CD8(+) T cells failed to fully activate Stat5-dependent expression of cell cycle/survival target genes, such as Bcl2 and Pim1, and to proliferate efficiently in response to IL-2 and IL-4 cytokine. CONCLUSION: Our data identify Stat5 as a dose-dependent regulator of CD8(+) T-cell functions in contact allergies and suggest that modulation of Stat5 dosage could be used to target contact allergies in humans.

[Study of possible involvement of MEK mitogen-activated protein kinase and TGF-ß receptor in planarian regeneration processes using pharmacological inhibition analysis].

Possible involvement of MEK mitogen-activated protein kinase and TGF-ß receptor in the processes of regeneration and morphogenesis in freshwater planarian flatworms Schmidtea mediterranea was studied using a pharmacological inhibitor analysis. It was found that pharmacological inhibitors of these kinases significantly inhibit the regeneration of the head end of the animals and that this effect is realized due to inhibition of proliferative activity of neoblasts, planarian stem cells. It is shown that that the inhibition of the studied protein kinases in regenerating planarians markedly disturbs stem cell differentiation and morphogenesis.

[A role of some intracellular signaling cascades in planarian regeneration activated under irradiation with low-temperature argon plasma].

Using inhibitory analysis the role of some intracellular signaling pathways in activation of planarian regeneration under the influence of low-temperature argon plasma (LTAP) has been investigated. Inactivation of specific inhibitors of intracellular signaling enzymes such as the receptor tyrosine kinase (EGFR), TGF ß receptor, calmodulin, adenylate cyclase, phospholipase A2, phospholipase C, cyclin-dependent protein kinase, JAK2-protein kinase, JNK-protein kinase MEK-protein kinase led to inhibition of the head growth during its regeneration in planarians. Pretreatment with LTAP irradiation provided no inhibitory action of some cascades regulating proliferation. However, the inhibitors of the key regulators of regeneration: TGF ß receptor, calmodulin and MEK-protein kinase completely suppressed the activating effect of plasma. Thus, by the example of regenerating planarians it is shown, that biological activity of low-temperature argon plasma LTAP is caused by modulation of a plurality of cellular signaling systems.

[Morphometric analysis of follicular structure of the thyroid gland after chronic low-dose gamma-irradiation].

A quantitative study of follicle average cross-sectional diameter distribution was conducted in the thyroid gland (TG) of mouse like rodents (25 tundra voles, 24 CBA mice, 16 Wistar rats) after chronic exposure to low-level external y-radiation both in the environment and under the experimental condition (absorbed dose range 0.05-0.5 Gy). Spectrum analysis of TG follicle cross-sectional diameter distribution in the irradiated animals has demonstrated a universal regularity: in comparison with the unirradiated animals there was a significant (1.3-1.7-fold) increase in content of small follicles (with a cross-sectional diameter lower than 36-41 microm in the studied animal species). A similar phenomenon was reproduced in the model experiments (TG regeneration in rats after hemithyroidectomy). The observed activation of the folliculogenesis processes after chronic low-dose irradiation in small doses may be interpreted as a nonspecific adaptive reaction of TG to radiation induced damage of its parenchyma.

[Function of the thyroid gland of tundra voles living on sites with the increased natural radioactivity].

We studied a functional condition of the thyroid gland of tundra voles living on sites with the increased level of a natural radioactivity (in Republic Komi areas in Russia). We have revealed that the functional condition of the thyroid gland depends on the phase of the population cycle of animals. The level of thyroid hormones in the irradiated animals differed from the norm; a high concentration of hormones in the irradiated animals was observed in the phase of the maximum number of animals, while males and females showed multidirectional differences from the control in the phase of the decreased number of animals.

[Comparative morphological analysis of peripheral endocrine glands in small mammals inhabiting areas with high levels of radioactive pollutants and exposed to chronic irradiation in laboratory experiments].

The comparative study of cellular-tissue reactions in endocrine organs (thyroid and suprarenal glands, ovary) of rodents exposed to radiation in natural conditions (Radium station in Komi Republic and 30-km zone of Chernobyl APP) and experimental conditions modeling the chronic exposure has been conducted. There is evidence that chronic irradiation in low doses causes morphological disorders in different levels of structural organization (cellular-tissue, organism and population levels). The experimental results showed that observed variations in thyroid, suprarenal glands and ovary by morphometric parameters reflect the natural changes in their functional activity (within the physiological limits). These changes are directed at the homeostasis maintenance in changed conditions and have a compensatory and adaptation character. The effects of low dose radiation influence with combination of other agents may be amplified at the cellular-tissue reactions level. In comparison with experimental results, the natural conditions (high level of radioactivity with alpha- and beta-emitters, high natural radionuclides, toxic elements and extreme climatic factors) induce more expressed changes as a significant increasing of chromosomal and genes mutations in cells, destructive processes in organs of endocrine system and disorders of reproductive functions.

[Biological effects of phytoecdysteroids and chronic low-dose irradiation].

The influence of serpistene in dose of 5 and 50 mg/kg on chronic low-dose gamma-irradiation (22.6 cGy) effects on cytogenetic (abnormal sperm cell, marrow bone micronucleus) and function and morphology (thyroid and adrenal glands) parameters of mice was estimated. The serpistene modifies effects of gamma-irradiation depends on the administration regime and a dose of the substance. The most expressive radioprotective effect on endocrine organs after serpistene prophylactic administration was found. The prophylactic dose was 5 mg/kg for adrenal gland and both doses--for thyroid gland. The most expressive radioprotective effect on marrow bone cells after serpistene therapeutic administration in a dose of 5 mg/kg was found. The most expressive antimutagenic effect on somatic and germinal cells of prophylactic and therapeutic administration in a dose of 50 mg/kg was found.

[Clinico-morphological features of chronic hepatopathy with concomitant viral infection and chronic narcotic intoxication].

The aim of this work was to evaluate liver function in subjects suffering from viral hepatitis and concomitant narcotic dependence. Clinico-biochemical, virologic, and morphological studies involving 92 patients revealed a number of specific features of chronic viral hepatitis complicated by drug addiction. However, the majority of them could be reliably identified only by a quantitative microscopic method. Worthy of special note are high frequency of lymphoid follicle formation in the portal tracts and signs of developing micronodular liver cirrhosis.

[Assessment of human health risk upon acute inhalational exposure to chemical substances].

To provide the safety of the population and environment with the occurrence of man-caused catastrophes due their increase is now an urgent problem. Toxic hazard coefficients were calculated for hydrogen sulfide in the settlements located in a possible chemical pollution area in the Orenburg gas-chemical complex. Assessment of the hazard of toxic effects to the population upon exposure to hydrogen sulfide has indicated that there may be toxic effects in the inhabitants of the study nearby inhabited localities. The findings do not characterize the possible severity of intoxication so it is necessary to develop a system of criteria for the differentiation of the severity of negative effects in order to evaluate the hazard of acute chemical exposures.

[Retinoic acid as a regulator of planarian morphogenesis].

The effect of retinoic acid on regeneration of two species of asexual planarian races, Girardia tigrina and Schmidtea mediterranea, was studied. It was established that retinoic acids at physiological concentrations (10(-7)-10(-10) M) inhibit the regeneration of the head part of planarians but have no effect on tail blastema growth. It is shown that regeneration of the head part is inhibited as a result of arrest of the cell cycle of neoblasts, proliferating stem cells, during the transition from the G1/G0 to the S phase. Thus, the morphogenetic role of retinoic acids in planarians, primitive bilaterally symmetrical animals, has been demonstrated.

[Melatonin effect on the regeneration of the flatworm Girardia tigrina].

The melatonin effect on the anterior and posterior ends of a free-living flatworm Girardia tigrina was studied, as well as the variability of the mitotic activity of the stem cells (neoblasts) in the anterior and posterior postblasteme. This hormone may inhibit the regeneration of the anterior end of the animal in the physiologic-friendly concentrations of 10(-10)-10(-15) M by suppressing the mitotic activity of the neoblasts. This hormone does not affect the posterior end's regeneration; thus, its regeneration effect is significantly elective.

[Cytogenetic effects in follicular epithelium of thyroid gland under prolonged exposure to gamma-radiation at low-doses].

The forming processes of micronucleated follicular thyrocytes in thyroid gland of mature Wistar rats under exposure to the prolonged low-intensity gamma-radiation with 5 and 50 cGy (dose rates: 25, 400 microGy/h; duration: 55, 80 days) were investigated. The chronic exposure to low-intensity gamma-radiation in both doses invokes the frequency of micronuclated thyrocytes three times higher in comparison with control animals. As a result of the small-sized micronuclei formation prevalence in irradiated group, the average size of micronuclei was 1.4-2 times lower than control values. This phenomenon can be reproduced in model experiments with hemithyroidectomized animals exposured to acute gamma-radiation with 2-4 Gy. The obtained results show high sensitivity of micronucleus test to the early determination of radiation-induced genetic damages in follicular epithelium of thyroid gland.

Immunostimulating complexes incorporating Eimeria tenella antigens and plant saponins as effective delivery system for coccidia vaccine immunization.

Immunostimulating complexes (ISCOMs) are unique, multimolecular structures formed by encapsulating antigens, lipids, and triterpene saponins of plant origin, and are an effective delivery system for various kinds of antigens. The uses of ISCOMs formulated with saponins from plants collected in Kazakhstan, with antigens from the poultry coccidian parasite Eimeria tenella, were evaluated for their potential use in developing a vaccine for control of avian coccidiosis. Saponins isolated from the plants Aesculus hippocastanum and Glycyrrhiza glabra were partially purified by HPLC. The saponin fractions obtained from HPLC were evaluated for toxicity in chickens and chicken embryos. The HPLC saponin fractions with the least toxicity, compared to a commercial saponin Quil A, were used to assemble ISCOMs. When chicks were immunized with ISCOMs prepared with saponins from Kazakhstan plants and E. tenella antigens, and then challenged with E. tenella oocysts, significant protection was conveyed compared to immunization with antigen alone. The results of this study indicate that ISCOMs formulated with saponins isolated from plants indigenous to Kazakhstan are an effective antigen delivery system which may be successfully used, with low toxicity, for preparation of highly immunogenic coccidia vaccine.

[Hepatopathies in concomitance of chronic drug and alcoholic intoxications].

Liver biopsy specimens were morphologically investigated in 87 patients, including 30 with drug (marijuana and poppy straw) addiction, 23 with chronic alcoholic intoxication, and 25 abused narcotics and alcohol concomitantly. All the patients were not found to be infected with viruses of hepatitis B, C, G, TTV, or CMN; the clinical manifestations and biochemical blood parameters were studied over time. In patients who simultaneously used drugs and alcohol, the specific features of hepathopathy were shown to be more pronounced and more rapidly progressive changes as fatty hepatosis and diffuse liver tissue fibrosis than in those used either drugs alone or alcohol alone. This is favored by chronic inflammation and cholestasis that is accompanied by bile duct proliferation. Concomitance of these processes leads to permanent liver tissue atrophy and death and to the relatively early development of hepatic monolobular cirrhosis. The mechanisms responsible for the patho- and morphogenesis of combined toxic hepatopathy are discussed.