Age-related adaptive responses of mitochondria of the retinal pigment epithelium to the everyday blue LED lighting.

The effect of everyday blue light (λ = 440-460 nm) on mitochondria of the retinal pigment epithelium of different age groups of Japanese quail was studied using electron microscopy, morphometric methods, and biochemical analysis. We have found a significant increase in the number of mitochondria, including those modified, mainly in young birds. In addition, cell metabolic activity increased in response to blue lighting. These changes are assumed to reflect an adaptive response of mitochondria aimed at neutralizing the phototoxic effect of blue light caused by accumulation of lipofuscin granules.

Photoinduced changes in subcellular structures of the retinal pigment epithelium from the Japanese quail Coturnix japonica.

Fifteen-week-old sexually mature female Japanese quails (Coturnix japonica) grown under various lighting conditions were used in the study. It was found that the number of mitochondria and phagosomes was increased by 1.5-fold in the retinal pigment epithelium from birds reared for 95 days under blue light (440-470 nm) vs. reduced blue light component conditions. Also, it was found that egg production was increased by 15% in birds reared under blue light compared to other lightning conditions. Thus, we concluded that blue light conditions resulted in elevating metabolic activity and accelerating pace of life in Japanese quails. It is assumed that the blue light-induced effects are probably due to inhibition of melatonin synthesis.

[The estimation of age-related sensitivity of the retinal pigment epithelium of Japanese quail (Coturnix japonica) to the light damage].

The effect of blue light damage (445-455 nm, 4 J/cm2) to retinal pigment epithelium (RPE) subcellular structures was investigated in 4 age risk groups (9, 25, 40 and 52 weeks) of Japanese quail Coturnix japonica by light and electron microscopy. The indicator of biological aging of RPE was age-related accumulation of lipofuscin granules: 5-6-fold increase in their quantity increasing by 5-6 times in quails at 52 weeks. The main photo-induced changes observed after 24 h of the photo radiation were located in the blood-retinal barrier, such as loss of homogeneity of Bruch's membrane, disorganization of basal processes, deformations of the nuclei and mitochondria shapes. Those effects ofphotobleaching were more expressed in young birds. But for the older 52-week age birds it was not so noticeable, because their retinal pigment epithelium structures had disorders which were similar to those in younger birds after photodamage.

[Dynamic of myocarditis development in rats after injection of cardiac myosine combined with IFA].

Myocarditis development was investigated after immunization rats with single subcutaneous injection of cardiac myosin (800 microg/kg) with incomplete Freund's adjuvant (IFA) (M + IFA group). Control group received equal volume of IFA alone or nothing (intact group). On days 4, 14, and 21 after injection, light and electron microscopy of heart sections, morphometric analysis, estimation of proinflammatory cytokines (IL-1p, IL-6, VEGF, TNFa and iNOS) expression were used to evaluate inflammatory response in myocardium. In addition, we estimated cardiac myosin antibody levels in blood serum and nitrite and nitrate levels in blood serum. Our data showed that immunization with cardiac myosin combined with IFA led to inflammatory response in the rat myocardium. Acute inflammation (i.e. lymphocyte infiltration of myocardium and increase of proinflammatory cytokines level) in M + IFA group occurred on 21 days after immunization.

[Dynamics of autoimmune myocarditis development in rats].

The development of autoimmune myocarditis in rats after a single hypodermic injection of rat myosin mixed with a complete Freund's adjuvant (CFA) (400 microg/kg in 200 microl) was studied. The rats from the control group were injected with only CFA. The titer of antibodies to myosin, infiltration of lymphocytes into the myocardium, ultrastructural damage of myofibrils, mitochondria, and nuclei of cardiomyocytes were maximally pronounced on days 14-21 after the immunization with myosin, which indicates a peak of the inflammatory reaction. The content of nitrites and nitrates in the blood serum and myocardium of immunized rats were also studied. A certain contribution to the development of the inflammation is made by CFA: in rats injected with only CFA, morphological signs of myocarditis were found, but to a much lesser degree than in the group immunized with myosin.

[Electron microscopy description of cardiomyocytes from the left ventricle of rat heart after apoptosis induction by isoproterenol].

Changes in cardiomyocytes from the left ventricle of rat heart were studied by light and electron microscopic and morphometric methods in the myocardial regions neighboring necrotic foci formed after the injection of 80 mg/kg beta adrenomimetic isoproterenol. TUNEL assay was used to detect apoptotic cardiomyocytes. Three types of cardiomyocytes (A, B, and C) differing by the ultrastructure of the nucleus and the degree of mitochondrial changes were identified at all studied stages of necrotic focus development (4-48 h). B and C type cardiomyocytes could represent cells at different stages of apoptosis. The apoptotic changes in cardiomyocytes proved to prevail in early lesion foci (4-18 h), while cardiomyocytes at later stages were prone to necrosis; cardiomyocytes can exhibit signs of apoptosis and necrosis at the same time.

[Adaptive responses of the secretory cardiomyocytes of the right atrium during simulation of the long-term and repeated gravity by antiorthostatic suspension of rats]. / Adaptivnye reaktsii sekretornykh kardiomiotsitov pravogo predserdiia pri modelirovanii dlitel'nogo i povtornogo deistviia ponizhennoi sily tiazhesti metodom antiortostaticheskogo vyveshivaniia krys.

Reactive changes in right atrium cardiomyocytes during antiorthostatic tail suspension of rats commonly used to simulate low gravity have been studied by electron microscopy and morphometry. A 14 day suspension proved to increase contractile and secreting activities of cardiomyocytes. At the same time, signs of depleted activity are observed in some cells. Elongation of the experiment to 30 days leads to development of adaptive compensatory responses and increases their secreting capacity. A 30 day return to normal orthostatic position does not completely restores the structure and functioning of cardiomyocytes and leads to accumulation of internal secretion. A repeated 14 day suspension to a certain extent facilitates cardiomyocyte adaptation to altered conditions as compared to a single exposure; apparently, secretion release decreases while its production is activated.

[Changes in mitochondria and inter-mitochondrial junctions of the rat cardiomyocytes upon adrenergic stimulation by isoproterenol]. / Izmenenie mitokhondrioma i mezhmitokhondrial'nykh kontaktov v kardiomiotsitakh serdtsa krys pri adrenergicheskoi stimuliatsii izoproterenolom.

Changes in rat cardiomyocytes and their mitochondria and intermitochondrial junctions (IMJs) upon beta-adrenoreceptor stimulation with isoproterenol were studied by the methods of light and electron microscopy and computer-aided morphometry. It was found that isoproterenol injections (0.3 mg/kg for eight days) resulted in myocardial hypertrophy, which was more pronounced in the right than in the left ventricle. In the hypertrophied cardiomyocytes of both ventricles, an adaptive response of mitochondria was observed: their ultrastructure, size, and number changed, and the number and average length of IMJs increased. A positive correlation between the degree of cell hypertrophy and the number of IMJs was revealed. The reactive properties of mitochondria, including IMJ formation, differed depending on their location in the cell (i.e., in the paranuclear, intermyofibrillar, or subsarcolemmal regions). These results suggest that the rates and intensities of adaptive compensatory processes developing in the mitochondria of cardiomyocytes exposed to beta-adrenoreceptor stimulation differ in the left and right ventricles.

Influence of 2G-hypergravity on the right atrium secretory cardiomyocytes of rats.

For elucidation of secretory cardiomyocytes adaptation to prolonged and repeated influence of changed gravity the ultrastructural and morphometrical analysis of right atrium cells was made in rats after they were exposed to primary and repeated 2G-centrifugation in ground-based experiment. It was found prolonged 2G-hypergravity resulted in hypertrophy of right atrium cardiomyocytes and stimulated of their mitochondrial associative activity and granular secretion; repeated influence induced changes differed from those after single influence.

[Anticancer properties of flagellate protozoan Trypanosoma cruzi Chagas, 1909]. / Protivorakovye potentsii zhgutikovogo prosteishego Trypanosoma cruzi Chagas, 1909.

The anticancer activity of Trypanosoma cruzi has been confirmed by the example of seven strains. Five virulent strains induced the infection, which inhibited sarcoma-180 growth 1.5-22.0 times. The parasites featured tumortropism; i.e., the successfully developed in cancer cells and even preferred them to normal cells. This taxis-based phenomenon was particularly pronounced at cocultivation of the normal and cancer cells. Cultures of the seven (avirulent and virulent) strains can produce an anticancer agent that selectively damages human cancer cells in vitro. The long-term anticancer effect of T. cruzi or preparations from it, as well as possible its cancer preventing effect, has been demonstrated. Three problems are discussed on the basis of the obtained and recently published data: (1) the mechanism of T. cruzi anticancer effect; (2) the nature of the anticancer agent; and (3) the distribution of the considered phenomenon among trypanosomatides. The anticancer activity of T. cruzi may be due to a combination of surface cellular antigens and an inhibiting or lysing factor.

[Compensatory function of submandibular salivary glands in diabetes and prospects for stimulating them with isoproterenol]. / Kompensatornaia funktsiia sliunnykh podcheliustnykh zhelez pri dibete i vozmozhnost' ee stimuliatsii izoproterenolom.

A brief review is presented of experimental studies conducted over several years (1970s-1990s) on the incretory potential of salivary glands, specifically on their role in the maintenance of carbohydrate homeostasis; a review is also presented of data on the stimulation of this potential by isoproterenol, a beta-adrenergic agonist.

[Oscillatory changes in cells of the submandibular glands in mice during the secretory cycle]. / Kolebatel'nye izmeneniia v kletkakh podnizhnecheliustnykh zhelez myshei v techenie sekretornogo tsikla.

By means of 3H-leucine radioautography, ultrastructural and morphometrical analysis, it has been demonstrated that in the cells of the acinar (Ac) and granular (Gr) parts of the submandibular gland (SMG) in mice with a synchronized for 3 h nutrition cycle there are endogenic fluctuations (for about 1 h) of the secretory process. They are demonstrated as alterations in intensity of protein synthesis in the cells, their areas and ultrastructure. In the Ac cells at the moment of feeding and in 50, 110, 150 min after feeding maximal leucine incorporation is observed. The changes in the cell area during the first hour occurs with 10 minutes' overtake of the incorporation intensity, and during the following 2 h an equilibrium is reached between these two parameters, when the maximal contents of the label corresponds to the minimal area of the cells. In the Gr cells one maximum of 3H-leucine incorporation occurs in 60-80 min after feeding. The minimal content of the label takes place in 10 and 120 min after feeding. During the time mentioned inverse ratio between the intensity incorporation of the labelled isotope and the change of the cell volume is observed. This is connected with autoregulatory processes in the cells. It is possible that in the Ac cells together with removal of the secretory granules by means of exocytosis (in 10-20 min after feeding) the secretory material is removed before it is formed as granules by parvivesicular extrusion. In the Gr cells apocrinic and endocrinic secretion takes place. Presence of certain specific formations (secretory lysosomes) explains peculiarities of the secretory process in the SMG.

[Rhythmic changes in protein synthesis in mouse pancreatocytes after food stimulation, in alloxan diabetes and in isoproterenol exposure]. / Ritmicheskie izmeneniia sinteza belka v pankreatsitakh myshi posle pishchevoi stimuliatsii, pri alloksanovom diabete i pri vozdeistvii izoproterenolom.

Radioautography revealed data on protein synthesis dynamics in pancreacytes of intact mice, subjected to alloxan diabetes and isoproterenol (ISP) injection after synchronized secretion. Rhythmic fluctuation of protein synthesis with maximum uptake of 3H-leucine 40 and 80 min after treatment was observed in intact animals. The above maximums correlated with the increase in the cell size. In alloxan diabetes, injections of ISP and their combination the rhythm of protein synthesis is retained, the total intensity of isotope uptake increases significantly, but at the same time secretion periods, particularly in the latter case decrease. The results obtained revealed the existence of endogenic rhythms in protein synthesis with certain secretion plasticity within pancreacytes.

[The distribution of insulin-containing cells in the developing tissues of the common frog (Rana temporaria)]. / Raspredelenie insulinsoderzhashchikh kletok v razvivaiushchikhsia tkaniakh travianoi liagushki (Rana temporaria).

Monoclonal antibodies to pig insulin were used to follow the changes in localization of insulin-containing cells during the ontogenesis of the brown frog, Rana temporaria L., from stage 20 (Dabagian, Sleptsova, 1975) until completion of metamorphosis and beginning of active nutrition. Insulin-containing were localized in brain, surface epithelium, intestine, olfactory epithelium, taste teats, kidney tubules, ciliated epithelium of the oral cavity, pancreas, Jacobson and interjaw glands was found out. Localization of insulin-containing cells and type of their specific fluorescence varied at different stages of development. Formation of the insulin-containing system of amphibians on the whole is typical for the development of diffuse endocrine system in vertebrates.

[Immunofluorescent and immunoenzymatic analysis of an insulin-like protein of the mouse submandibular glands using monoclonal antibodies to insulin]. / Immunofliuorestsentnyi i immunofermentnyi analiz insulinopodobnogo belka podcheliustnykh zhelez myshei s pomoshch'iu monoklonal'nykh antitel k insulinu.

As has been reported earlier, submandibular glands of animals and man contain insulin-like protein (ILP), similar in some of its properties to pancreatic insulin. An immunocytochemical method with polyclonal anti-insulin serum has shown that ILP is contained in cells of the submandibular gland granular ducts. This paper is concerned with the determination of localization of ILP and estimation of its structural identity with insulin by using monoclonal antibodies to pig insulin. ILP and insulin were extracted from mouse submandibular glands and pancreas and partially purified. Solid-phase immunoenzymatic analysis was used to determine the capacity of mouse ILP and pancreatic insulin to bind specifically with monoclonal antibodies to pig insulin indicating the structural identity of ILP and insulin. The absence of cross-reaction of antibodies with the nerve growth factor and kallikrein (biologically active substances contained in the submandibular gland granular ducts) was established. An immunofluorescent indirect method with the use of monoclonal antibodies to insulin has shown that ILP is localized in cells of the submandibular gland granular ducts. It confirms the previously reported results.

[Concentration and distribution of insulin and insulin-like protein in the organs of normal and diabetic mice]. / Soderzhanie i raspredelenie insulina i podobnogo emu belka v organakh myshi v norme i pri diabete.

Presence of insulin or insulin-like protein has been studied in mouse liver, kidneys, lungs, duodenum, jejunum, submandibular and parotid salivary glands, in femoral, diaphragmal and abdominal wall muscles by means of the immunofluorescent method. In order to understand the role of the extrapancreatic insulin for compensation of the insular insufficiency, corresponding organs have been examined in mice with alloxan diabetes. The immunoreactive insulin is proved to be present only in cells of the granular parts of the salivary tubules of the submandibular and striated ducts of the parotid glands. As demonstrates microfluorometry, a relative amount of insulin in the submandibular gland cells is 1.5 times and in the parotid gland cells--2 times as small as in beta-cells of the pancreatic glands. Under alloxan diabetes insulin content in the salivary gland cells decreases by 1.3-1.9 times (in the beta-cells--by 2.7 times). This may designate that the extrapancreatic insulin (or insulin-like protein) participates in compensation of hypoinsulinemia. In mice with alloxan diabetes, immunoreactivity of insulin is also revealed in hepatocytes.

[Changes in mouse submaxillary glands after injection of isoproterenol in intact and depancreatized animals]. / Izmeneniia podcheliustnykh zhelez myshei posle in"ektsii izoproterenola intaktnym i depankreatizirovannym zhivotnym.

The changes in the relative weight, cell area and ultrastructure of the submaxillary glands (SMG) of CBA/C57BL mice and those in the content of immunoreactive insulin-like protein (IRILP) occurring in the organ were studied. The changes were examined on day 6 after isoproterenol (ISP) injection (23 mg/100 g bw) to healthy animals (group I), on day 13 after pancreatectomy and on day 6 after injection of the indicated dose of ISP (group II), and on day 13 after pancreatectomy without ISP injection (group III). Intact animals served as control. In all the three experimental groups, the ratio between the SMG weight and the total weight of animals showed a tendency towards increase. The highest increase was recorded in group II, where the weight of the SMG was 46.1% higher than the control value. In groups II and III, the area of acinar cells increased by 49.1 and 12.5%, respectively. The area of salivary tube granular ducts decreased by 12.6% in group II and slightly increased (by 4.7% much greater than in group III. Electron microscopy demonstrated that secretory activity of the granular duct cells was enhanced in all the three groups and that secretory extrusion occurred via the apocrine (in group II animals via the holocrine as well) cells. The radioimmunoassay data suggest that IRILP content in the SMG of controls and animals entering groups I-III was 685 +/- 50, 1125 +/- 125, 914 +/- 120 and 302 +/- 66 mu units/g weight, respectively. It is concluded that the synthesis and accumulation of IRILP in the SMG after ISP injection are activated and that the formation and extrusion of IRILP after ISP injection to diabetic animals are overtly activated. Presumably ISP injection to diabetic mice facilitates the stimulation of the compensatory function of the SMG as an IRILP-producing organ.

[Ratio of "light" and "dark" cells in heterogineous cell populations of experimental tumors and their metastases]. / Sootnoshenie "svetlykh" i "temnykh" kletok v geterogennykh kletochnykh populiatsiiakh nekotorykh éksperimental'nykh opukholei i ikh metastazov.

The number of undifferentiated "light" cells and of differentiated "dark" cells was calculated in heterogeneous tumour cell populations of sarcoma 45 and Walker carcinosarcoma of rats at various stages of tumour growth and metastases.

[Metabolic study of sarcoma 45 and Walker carcinosarcoma cells during the process of growth and exposure to anticoagulants (an enzyme cytochemical analysis)]. / Issledovanie metabolizma kletok sarkomy 45 i kartsinosarkomy Uoker v protsesse ikh rosta i pri antikoaguliantnom vozdeistvii (énzimotsitokhimicheskii analiz).

The enzyme activities of lactate dehydrogenase (LDH), succinate dehydrogenase (SDH) and acid phosphatase (AP) in the rat sarcoma 45 (S. 45) cells and Walker carcinosarcoma (WCS) cells were estimated from the histochemical study. No significant changes were found in the histochemical reaction intensity during the tumour growth but at the late stage of WCS growth the LDH activity increased. WCS metastases were distinguished by the elevated enzyme activity of LDH as compared with the tumour. During the anticoagulant treatment the LDH and SDH activity in S. 45 and WCS cells falls, but the AP activity increases.