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Biomedical subjects

G Csaba

Publications and source records attributed to G Csaba.

At least 19 recordsLinked to original sources

In vitro effect of biogenic amines on the hormone content of immune cells of the peritoneal fluid and thymus. Is there a hormonal network inside the immune system?

Immune cells contain different hormones and hormone-like molecules, such as insulin, endorphin, triiodothyronine (T3) histamine, serotonin. In earlier in vitro experiments insulin down-regulated histamine, serotonin and T3 content of thymus cells. Now we studied the effect of biogenic amines on the endorphin, T3, serotonin and histamine content of rat peritoneal and thymic cells. Cells were obtained from male rats of 100g body weight. 100 ng/ml serotonin or 300 ng/ml histamine was added for 30 min. After that the cells were prepared for flow cytometric analysis with antibodies to endorphin, T3, histamine and serotonin as primary antibodies and anti-rabbit IgG as secondary antibody. Finishing the measurements the cells were also studied by confocal microscopy. T3 concentration (binding of anti-T3 antibody) increased in peritoneal mast cells after serotonin treatment and in the monocyte-macrophage-granulocyte group after histamine treatment. Thymocytes' T3 content radically decreased after both treatments. Serotonin and histamine treatment also radically reduced the amine content of each other. Endorphin level was resistant to hormonal treatments. The results call attention to a possible hormonal network inside the immune system in which hormones produced by the immune cells themselves can influence each other.

Animals↗

How does the unicellular Tetrahymena utilise the hormones that it produces? Paying a visit to the realm of atto-and zeptomolar concentrations.

Changes in the hormone content of Tetrahymena pyriformis GL were investigated during histamine, serotonin or insulin treatment at concentrations of 10(-6)M to 10(-21)M for 30 min. The immunologically demonstrable hormone content was studied by using specific antibodies, flow cytometry and confocal microscopy. Histamine at the higher ranges elevated the serotonin content of Tetrahymena, whereas serotonin at the lower ranges (down to 10(-21)M) decreased its histamine levels. Insulin did not affect its serotonin content, whereas serotonin increased its insulin content at each concentration studied (down to 10(-21)M). Insulin between 10(-6)M and 10(-21)M increased the histamine levels of Tetrahymena, although histamine influenced its insulin level only at 10(-6)M. Our results call attention to the presence of hormonal interactions even at "low" levels of phylogeny and to the extreme sensitivity of the hormone receptors of Tetrahymena. These data might explain (1) the requirement of Tetrahymena for (vertebrate) hormone production and hormone receptors and (2) the way that it uses these hormones under natural conditions.

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Perinuclear localization of biogenic amines (serotonin and histamine) in rat immune cells.

EDAC fixation, which polymerizes biogenic amines, and clorgyline, which blocks monoamine oxydase, were used to demonstrate the perinuclear and nuclear localization of histamine and serotonin (5-HT) of immune cells in rat peritoneal fluid and thymus. For detection, an immunocytochemical method combined with confocal microscopy was used. In peritoneal cells (lymphocytes) 5-HT formed spots around the nucleus, while histamine was localized diffusely. In thymocytes, 5-HT was distributed in patches, while histamine formed a garland around the nuclear envelope. Clorgyline enhanced the fluorescence and increased the number of fluorescent positive cells only for histamine, and then histamine-positive nuclei were also present. The results show that: (1) EDAC fixation demonstrates the presence of biogenic amines in immune cells, (2) using this fixation, the perinuclear localization of biogenic amines can be demonstrated, (3) the localization of the two amines is different and (4) the nuclear localization of histamine in thymocytes can be surmised and in mast cells (described previously) is proven again. The potential importance of the nuclear and near-nuclear localization of biogenic amines is discussed.

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Effects of extremely low concentrations of hormones on the insulin binding of Tetrahymena.

FITC-insulin binding to previously hormone-treated Tetrahymena was studied by flow cytometry and confocal microscopy. Hormones produced by Tetrahymena were chosen for study and the hormone concentrations were administered between 10(-6) and 10(-21)M for 30 min. Endorphin, serotonin and insulin significantly reduced the hormone binding however histamine did not influence it at all. Endorphin, serotonin and insulin were significantly effective down to 10(-18)M and the effect of insulin and endorphin suggest a similar mechanism. The results call attention to the efficacy of very low hormone concentrations, which can influence the hormone content (earlier experiments) and receptor binding capacity (present study) of a unicellular organism. This seems to be very important, as in wild (natural) conditions the dilution of signaling materials secreted by a water-living protozoan is very high. In addition, the results point to the selectivity of response, as not all of the hormones that deeply influence other physiological indices (e.g. histamine) have an effect on insulin content or insulin receptors.

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Influence of paraformaldehyde and EDAC fixation on the demonstrability of hormones (histamine, endorphin, triiodothyronine) in rat immune cells: an immunocytochemical comparative analysis.

The amount and localization of three hormones (histamine, endorphin and triiodothyronine [T(3)]) was measured in male and female rat peritoneal cells (lymphocytes, mast cells, monocyte-macrophage-granulocyte group [mo-gran]) using flow cytometry as well as confocal microscopy after paraformaldehyde (PFA) or EDAC fixation. In the EDAC fixed lymphocytes and mo-gran of female animals two-magnitude higher levels of histamine were measured after EDAC fixation and one magitude higher in mast cells. The amount of T(3) was almost four-fold in lymphocytes and 2.5-4-fold in mast cells and mo-gran. Endorphin content was not altered by the type of fixation. In each cell type in males one magnitude higher levels of histamine and T(3) were measured after EDAC fixation and a small, but significant, elevation of endorphin. Confocal microscopy supports the quantitative data. The results show that (1) the fixation with the crosslinking molecule, EDAC, is more suitable for immunocytochemical studies of amino-acid type hormones in immune cells, (2) more histamine and T(3) are present in the immune cells than it was supposed previously when studying PFA-fixed preparations, (3) the estimation of the amount of peptide hormones seems to be accurate after PFA fixation, (4) there is a quantitative difference comparing the results of PFA and EDAC fixation between males and females.

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EDAC fixation increases the demonstrability of biogenic amines in the unicellular Tetrahymena: a flow cytometric and confocal microscopic comparative analysis.

Earlier experiments demonstrated the presence of hormones of the higher ranked animals in Tetrahymena. In the present experiments two fixatives, paraformaldehyde, which is commonly used and a carbodiimide, EDAC that was recommended by Panula et al. for the immunocytochemistry of histamine, are compared in Tetrahymena for the demonstration of histamine and serotonin by using flow cytometry and confocal microscopy. Both hormone levels were significantly higher after EDAC fixation; serotonin almost doubled and histamine was more than fivefold. The confocal microscopic pictures were clearer and the hormones' localization was easier. The results support earlier observations on the presence of these hormones in Tetrahymena and points to the advantage of EDAC fixation for demonstrating these hormones immunocytochemically.

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Influence of in vitro and in vivo insulin treatment on the hormone (histamine, serotonin, endorphin and triiodothyronine) content of thymus and spleen cells.

Thymic and spleen cells were treated in vitro or in vivo with insulin. The in vitro treatments were done with 10(-6), 10(-9), 10(-12) and 10(-15) M concentrations for 30 min and after that histamine, serotonin, endorphin and triiodothyronine (T3) content of the cells were detected by using antibodies to the hormones and flow cytometry as well as confocal microscopy. For in vivo treatment 1 IU/kg insulin was given for adult rats and 1 h after that the target hormone contents were determined by the same manner. Histamine and T3 content radically decreased in the thymus after in vitro treatment independent on the insulin concentrations administered. In vivo treatment halved histamine and T3 content. Serotonin content also decreased after in vitro treatment with the two higher concentrations, however the in vivo treatment did not cause a change. Histamine content was elevated after in vitro treatment in the spleen, independent on the insulin concentration. Endorphin level was not influenced at all. The experiments demonstrate that insulin is a factor which regulates the content (production, storage, secretion?) of some immunologically important molecules of the immune cells. Since each hormone molecule studied has important immunomodulatory role, the experiment points to the indirect immunomodulatory role of insulin.

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Majority logic gate for magnetic quantum-dot cellular automata.

We describe the operation of, and demonstrate logic functionality in, networks of physically coupled, nanometer-scale magnets designed for digital computation in magnetic quantum-dot cellular automata (MQCA) systems. MQCA offer low power dissipation and high integration density of functional elements and operate at room temperature. The basic MQCA logic gate, that is, the three-input majority logic gate, is demonstrated.

Journal Article↗

Prolonged effect of stress at weaning on the brain serotonin metabolism and sexuality of female rats.

Weanling female rats were stressed (by water and food deprivation for two days) and three months later the following indexes were studied: 5-HT and 5-HIAA levels in five brain regions, blood plasma and cerebrospinal fluid (CSF), sexual activity and nocistatin level of the plasma and CSF. The 5-HIAA content of hypothalamus and brainstem was significantly decreased (in the brainstem with one third) and in the striatum significantly increased. Plasma nocistatin level was significantly increased. Meyerson index and lordosis quotient were similar to control, but the estrus frequency almost doubled in the stressed animals. Much more defense reactions were observed in the stressed females during trials of mating. The results demonstrate that, 1) the perinatal period is not only sensitive to the remote-effects of stress but later could also be stress-sensitive critical periods, and 2) the continuously differentiating (e.g. bone marrow) cells are sensitive to late imprinting by stress, as well as to the brain and the sexual system.

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Comparison of the binding of anti-tubulin antibody and the fluorescent taxol derivative Flutax-1 to the microtubular system of Tetrahymena.

Using confocal microscopic analysis, FITC-labelled anti-alpha-tubulin antibody and the fluorescent taxol derivative Flutax-1 in fixed and living Tetrahymena pyriformis GL, longitudinal microtubules, oral and somatic cilia, deep fibers, and contractile vacuole pores were equally labeled. While the antibody stained transversal microtubules, these were not labeled by Flutax-1. At the same time, oral cilia were more intensely stained by Flutax-1, than by the antibody. There were no differences in the staining of fixed preparations and living cells. The observations suggest (i) the difference between the MAPs of longitudinal and transversal microtubules which allow or inhibit the binding of the indicator molecules, and (ii) the different functions of these two types of microtubules.

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Effect of the inhibition of triiodothyronine (T3) production by thiamazole on the T3 and serotonin content of immune cells.

Triiodothyronine (T3) and serotonin are present in the cells of immune system (blood, peritoneal and thymic lymphocytes, monocytes and granulocytes of the blood and peritoneal fluid, mast cells). In the present experiments the effect of thiamazole, an antithyroid drug was studied on the content of these two hormones in immune cells after one and two weeks of continuous treatment (by drinking water, containing 30 mg/100 ml thiamazole, ad libitum) in adult male rats, using flow cytometric and confocal microscopic analysis. In thymic lymphocytes both hormone contents significantly increased in both time points. A significant decrease of T3 was observed in peritoneal monocytes and granulocytes also in both time points, in peritoneal mast cells after one week and in blood lymphocytes after two weeks. Serotonin content in addition to the elevated thymic values (in both measurements) was significantly reduced in blood lymphocytes after two weeks. Confocal microscopy demonstrated heterogeneous cell populations with disparate hormone content and mostly diffuse localization The experiments call attention to the presence of T3 in the immune cells and to its variable concentration under the effect of a thyrostatic drug as well, as to the T3-serotonin relationship in the cells of the immune system.

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Alcohol consumption during gestation reduces the histamine and triiodothyronine content of rat immune cells.

OBJECTIVE: Different factors acting during pregnancy can cause non-morphological alterations of cells which are manifested later, in adulthood. We studied the effect of maternal alcohol consumption for one day in early pregnancy on the hormone content of immune cells in the adult rat. METHODS: Lactating dams were given 15% ethanol in the drinking water for 24 h on the 3rd day post partum, exposing their pups to ethanol in the breast milk. Some of the same dams had been successfully mated on the day of delivery, so that they were also 3 days pregnant on the treatment day, exposing embryos to alcohol on the third day of pregnancy. In 4 month old pups histamine and triiodothyronine (T(3)) content of citrate elicited peritoneal immune cells (lymphocytes, monocyte-macrophage-granulocyte group, mast cells) as well as thymic cells were determined by flow cytometry and confocal microscopy using specific antibodies. RESULTS: Alcohol treatment during pregnancy decreased highly significantly the content of both hormones in peritoneal cells of the 4 month old adult animals while it was ineffective by breast feeding after birth. Thymic cells did not show any changes. CONCLUSION: Since the immune system had not developed at the time of treatment (3rd day of pregnancy), stem cells were presumably imprinted. Our results indicate the deleterious effects of early maternal alcohol consumption on the hormone content of the immune system.

Alcohol Drinking↗

Effect of a single early-gestational alcohol consumption on the insulin binding by immune cells of adult rats.

Rat dams were exposed to 15% ethanol containing water ad libitum for 24 h at the 3rd day of gestation. Adult progenies' immune cells (peritoneal, blood and thymic lymphocytes, monocyte-macrophage-granulocyte group and mast cells) were studied by flow cytometry for binding of FITC-labelled insulin (compared to the appropriate cells of untreated animals). All of the cell types studied bound significantly less insulin, except mast cells. The results call attention to the life-long effect of occasional alcohol consumption in early pregnancy manifested in the immune system.

Alcohol Drinking↗

Hormonal interactions in Tetrahymena: effect of hormones on levels of epidermal growth factor (EGF).

Tetrahymena pyriformiswas treated with insulin, histamine or serotonin for 30 min and epidermal growth factor (EGF) level was studied inside the cells using specific antibodies and flow cytometry as well as confocal microscopy. The EGF concentration was highly significantly elevated after hormone treatment, regardless of the hormone used. EGF was localized mainly in the cortical region (mucocysts) and in vesicles and this localization did not differ in untreated and treated cells. The results call attention to the possibility of interactions between hormones at unicellular level and points to the presence of a hormonal system in Tetrahymena that includes receptors, hormones and signal transduction pathways as well as hormonal interactions. This could be the basis of further evolution to the hormonal system of multicellulars.

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Presence of hormones (triiodothyronine, serotonin and histamine) in the immune cells of newborn rats.

Five hormones in lymphatic cells from the spleen and thymus of newborn rats were measured using flow cytometry and confocal microscopic immunofluorescence. Immunoreactive histamine, serotonin and triiodothyronine (T(3)) were present in the cells. However, growth hormone and insulin were not observed, except for low levels of insulin in a few spleen cells. Serotonin content was modest compared to the histamine and T(3) levels. The localization of each hormone in the cells was different. The importance of these observations in the light of functional data is discussed.

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Three-generation investigation on serotonin content in rat immune cells long after beta-endorphin exposure in late pregnancy.

Female rats were treated with beta-endorphin on the 19th day of pregnancy. Serotonin content of immune cells (peritoneal lymphocytes, monocyte-macrophage-granulocyte group (mo-gran), mast cells, blood lymphocytes, granulocytes and monocytes, thymus lymphocytes) were studied in the mothers (P-generation four weeks after delivery), in the male offspring (F1) generation (at seven weeks), in the female offspring (four weeks after their own delivery) and in their offspring (F2 generation, at seven weeks). P-mother cells' serotonin content was not influenced by endorphin treatment, while F1 generation's mo-gran and blood lymphocyte serotonin content was reduced (in contrast, histamine content of mo-gran increased). Four weeks after delivery, an increase in serotonin content was observed in the F1 generation in the peritoneal lymphocytes and mast cells as well as in blood lymphocytes. In contrast, serotonin content was reduced in blood granulocytes and monocytes. In the F2 (grandson) generation, a reduction in mast cell serotonin content and sensitization of blood and thymic lymphocytes to repeated endorphin treatment was provoked. The significant changes were more expressed in the F2 generation compared to F1, also appearing earlier. The results unequivocally suggest that the increase in endorphin levels during late pregnancy can cause permanent changes in the F1 and F2 generations, which means that the imprinting effect can be transgenerationally transmitted.

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Prolonged effect of stress (water and food deprivation) at weaning or in adult age on the triiodothyronine and histamine content of immune cells.

We used two days of total water and food deprivation as stress for female rats at weaning (three weeks old) and at adult age (two and a half months old). Triiodothyronine (T3) and histamine content of immune cells (lymphocytes, mast cells and monocyte-macrophage-granulocyte group in peritoneal fluid; lymphocytes, granulocytes and monocytes in blood; and lymphocytes in thymus) were studied three weeks after stress application using specific antibodies for flow cytometry and confocal microscopy. The stress at weaning increased T3 content of thymus lymphocytes. In case of adult T3, there was a cell type independent significant effect of stress, decreasing values in peritoneal fluid and slightly increasing effect in the blood. Histamine content of granulocytes was also significantly elevated. The experiments demonstrate that not only fetal or neonatal stress has long-lasting consequences, but also stress events in later periods of life in cells (organs) that are continuously differentiating. We will go on to discuss the importance of T3 and histamine in connection with stress and immunity.

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