The psoralen-pyrimidine base complexes resulting from interstrand cross-linking through $C_4$-cycloaddition is studied by ab initio and DFT methods. The results indicate that in the case of the molecular complex formation between psoralens and pyrimidine base, the most probable photocycloadditions are 8-MOP< >Thy, Ps< >Cyt and Ps< >Thy. The geometries of photoadducts were optimized at the HF levels and ${\Delta}{G^{\circ}}$ were calculated. The photocycloadduct was inferred to be a C4-cycloaddition product with the stereochemistry of trans-syn 8-MOP< >Thy, trans-anti Ps(3, 4)< >Cyt, trans-anti Ps(12, 13)< >Cyt, trans-syn Ps(3, 4)< >Thy, trans-syn Ps(12, 13)< >Thy, trans-anti Ps(12, 13)< >Ps(12, 13), cis syn, cis anti Thy< >(3, 4)Ps(12, 13)< >Thy.
This experiment was carried out to elucidate the effects of the thyroid function on lactation in female rats. One hundred and five female rats, whose body weight was approximately 250g with normal parturition, were divided into 3.5 THY, 35 PTU, and 35 CON. The $30{\mu}g$ L-thyroxine per rat was administered subcutaneously for the THY group with 3-days intervals arid 0.03% propylthiouracil solution was drunk for the PTU group. After the treatments body weight, thyroid weights and prolactin levels in serum, and histological changes in thyroid and mammary gland were investigated for 3 weeks with 3-days interval. The results obtained were as follows 1. The body weights of PTU group were lower than those of CON arid THY groups. The changes in body weights were significant between 3 and 6 days and between 15 and 18 days. 2. Differences in thyroid weight among the groups were significant after 9 days, The thyroid weights of PTU group were much higher than those of CON and THY group. 3. The follicular epithelia of PTU group after 6 days showed cuboidal phenomena which were accompanied by hypertrophy and hyperplasia, and this phenomena continued until post weaning period. Those of THY group after 9 days showed a squamous degeneration together with pyknosis. 4. The prolactin concentrations of THY group were higher than the other groups after 12 days. and those of CON group were higher than the others after 18 days. However, those of PTU group were lower all through the period. 5. The secretary epithelial cells of THY and CON groups became cuboidal after 12 days, but after 15 days the differentiation of mammary tissue was progressing in THY group faster than CON group. The degeneration of mammary tissue were observed in PTU group as time lapses, so after 15 days the exfoliation of secretory epithelium and atrophy of alveolus were recognized. 6. The body weights of offspring for all experimental groups were increasing as time lapses, but the values for PTU group were markedly lower than the others.
Vakili, A.R.;Khorrami, Behzad;Mesgaran, M. Danesh;Parand, E.
Asian-Australasian Journal of Animal Sciences
/
v.26
no.7
/
pp.935-944
/
2013
Essential oils have been shown to favorably effect in vitro ruminal fermentation, but there are few in vivo studies that have examined animal responses. The objective of this study was to evaluate the effects of thyme (THY) and cinnamon (CIN) essential oils on feed intake, growth performance, ruminal fermentation and blood metabolites in feedlot calves fed high-concentrate diets. Twelve growing Holstein calves ($213{\pm}17kg$ initial BW) were used in a completely randomized design and received their respective dietary treatments for 45 d. Treatments were: 1-control (no additive), 2-THY (5 g/d/calf) and 3-CIN (5 g/d/calf). Calves were fed ad libitum diets consisting of 15% forage and 85% concentrate, and adapted to the finishing diet by gradually increasing the concentrate ratio with feeding a series of transition diets 5 wk before the experiment started. Supplementation of THY or CIN did not affect DMI and ADG, and feed efficiency was similar between treatment groups. There were no effects of additives on ruminal pH and rumen concentrations of ammonia nitrogen and total VFA; whereas molar proportion of acetate and ratio of acetate to propionate decreased, and the molar proportion of propionate increased with THY and CIN supplementation. Rumen molar concentration of butyrate was significantly increased by adding CIN compared to control; but no change was observed with THY compared with control group. No effects of THY, or CIN were observed on valerate, isobutyrate or isovalerate proportions. Plasma concentrations of glucose, cholesterol, triglyceride, urea-N, ${\beta}$-hydroxybutyrate, alanine aminotransferase and aspartate aminotransferase were not changed by feeding THY or CIN. Results from this study suggest that supplementing a feedlot finishing diet with THY or CIN essential oil might be useful as ruminal fermentation modifiers in beef production systems, but has minor impacts on blood metabolites.
The mammary gland contains a subpopulation of epithelial cells with large proliferative potentials which are the likely targets for carcinogens. These clonogenic cells can proliferate and differentiate into functional glandular structures. Rat mammary epithelial cells (RMEC) were isolated and characterized in vitro. By flow cytometry of RMEC stained with fluorescein isothiocyanate-peanut agglutinin(PNA) and phycoerythrin anti-Thy-1.1 monoclonal antibody, it was possible to four cell subpopulations from 7-8 week old F344 female rat mammary glands: cells negative to both reagents (B-), PNA-positive cells (PNA+), Thy-1.1-positive cells (Thy-1.1+), and cells positive to both reagents (B+). When single PNA+ cells were isolated and cultured in Matrigel with irradiated (∼50 Gray) 3T3 fibroblast feeder layer, they gave rise to multicellular clonal structures of three types: alveolar, foamy alveolar, and squamous colonies. The developed structures were similar to the mammary glands in vivo. These results suggest that some of PNA+ cells possesses many of the characteristics of multipotent clonogenic stem-like cells.
Background : The immune responses mediated by CD8+T cells are known to be significant in controlling M. tuberculosis infections. In order to determine the role of cytotoxic CD8+T cells in the protective immune mechanism in latently infected subjects, this study examined whether or not the cytotoxic immune responses of CD8+T cells specific to the M. tuberculosis somatic antigens are induced in BCG vaccinated healthy subjects. Methods : Cytotoxicity and $IFN-{\gamma}$ elispot assays were used to investigate the activities of CD8+T cells specific for the $thyA_{30-38}$ peptide epitope in circulating peripheral blood mononuclear cells (PBMC) from BCG-vaccinated HLA-A*0201 and A*0206 subjects. Results : The results indicate the cytotoxic and $IFN-{\gamma}$ immune responses of CD8+T cells specific for $thyA_{30-38}$ were induced in BCG vaccinated healthy subjects. Conclusion : The cytotoxic and $IFN-{\gamma}$ responses by CD8+T cells specific for the M. tuberculosis somatic antigens are induced in BCG-vaccinated subjects, and appear to be involved in the protective immune mechanism in latently infected people against a M. tuberculosis infection.
The conceptus which are resulted by mating between two genetically non-identical partners can be considered to be an allograft to the mother science which is not rejected by the mother's immunological attack. The present studies have been, therefore, attempted in order to elucidate the mechanism by which protection of the fete-placental allograft, between the C3H/HeJ female mouse and DBA/2 male mouse occurred. For this purpose, firstly systemic immunity was investigated by measuring T and B lymphocytes subsets. Natural killer cell activity in maternal splenic tissue and by observing the effects of pregnancy serums, progesterone and hCG on immune systems. Secondly, local immunity also investigated by measuring T lymphocytes subsets, natural killer cell activity in lymph nodes draining the uterus. The subsets of Thy-1.2$^+$ cells and L 3T4$^+$ cells decreased slightly while the subsets of Ly2$^+$ cell increased significantly compared with those of the control group beyond the mid-gestational stage. The subsets of B cell gradually in-creased from the mid-gestational stage untill delivery. The natural killer cell activity in the maternal splenic tissue significantly increased during the period of 5th to 8th day of gestation. The natural killer cell activity was significantly suppressed by the pregnancy serums and non-pregnant serums compared with those of serum-free group. The treatment of hCG significantly suppressed natural killer cell activity in the dose dependent manner (1 unit/ml-1000 unit/ml) while pro-gesterone increased the natural killer cell activity at phamarcological dose only. In the lymph nodes draining the uterus, the subsets of Thy-1.2$^+$ cells significantly increased during the period of implantation and L3T4$^+$ cell subsets slightly increased during the mid-gestational stage. The subsets of Ly2$^+$ cell increased significantly during the mid-gestational stage, but decreasing slightly be-fore delivery. The natural killer cell activity was significantly elevated after the implantation period in the lymph nodes draining the uterus. The natural killer cell activity of the lymph nodes draining the uterus was higher than those of splenic tissue during the same periods of gestation. It is therefore, concluded that during the pregnancy, the phenomena which the fete-placental allograft has not been rejected and rather protected from the maternal immunological attack might be due to local immune suppression in fete-maternal interface tissues rather than systemic immune suppression. And the subsets of Thy-1.2$^+$ cells and L3T4$^+$ cells mainly contribute to accepting allograft in early stage of pregnancy, while the subsets of Ly2$^+$ cell and the subsets of B cell increased significantly compared with those of the control group beyond the mid-gestational stage, so their role in systemic immunity and local immunity gradually increased from the mid-gestational stage until delivery.
Proceedings of the Korea Contents Association Conference
/
2018.05a
/
pp.551-552
/
2018
K/BxN serum can induce arthritis in normal mice because of abundant autoantibodies that trigger an innate inflammatory response in joints. To determine whether IL-17 is involved in the pathogenesis of serum-induced arthritis, we injected wild-type and $IL-17^{-/-}$ mice with K/BxN serum and evaluated them for signs of arthritis. Unlike wild-type mice, $IL-17^{-/-}$ mice did not show any signs of arthritis. IL-17 was produced predominantly by $CD3^-CD4^-gdTCR^-NK1.1^-Sca1^{int}Thy1^{hi}$ cells residing in the inflamed synovial tissue. When synovial cells extracted from normal joints were stimulated with IL-23 or autoantibody-containing immune complexes, a substantial fraction of $Sca1^{int}Thy1^{hi}$ cells produced IL-17. Thus, we have identified a novel population of IL-17-producing innate synovial cells that play a crucial role in the development of K/BxN serum-induced arthritis.
Journal of the Korean Society of Food Science and Nutrition
/
v.24
no.3
/
pp.470-486
/
1995
Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.
Effects of Paragonims westermai infection were observed in mice on the change of serum IgE level, the number of peripheral eosinophils and the distribution of Thy $1.2^{+}{\;}(CD3),{\;}L3T4^{+}{\;}(CD4),{\;}and{\;}Lyt-2^{+}$ (CD8) splenic T Iymphocytes without mitogen serum IgE increased at 3 weeks after the infection and reached a peak on week 4 and maintained high levels of IgE until the 23r6 week. Peripheral eosinophil numbers Increased at the second week and attained peak level on week 9. The frequency of $L3T4^{+}$(CD4) and $Lyt-2^{+}$ (CD8) T Iymphocytes decreased slightly until 4 weeks after the infection, but not significantly. Absolute number of $L3T4^{+}{\;}and{\;}Lyt-2^{+}$ T Iymphocytes, and the ratio of L3T4/Lyt-2 were not markedly changed over the period of observation. The frequency of Thy $1.2^{+}$ (CD3) T lymphocytes in the infected group slightly decreased until 4 weeks after the infection and showed significant reductions at the 2nd and 4th week of the infection (p < 0.05).
Ki, Min-Hyo;Paik, Kee-Joo;Lee, Ji-Hyeon;Chung, Hae-Young;Lee, Kyung-Hee;Kim, Kyu-Won;Kim, Nam-Deuk
Archives of Pharmacal Research
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v.21
no.3
/
pp.298-304
/
1998
Retinoids are applied to not only cancer prevention but also cancer chemotherapy by stimulating differentiation of cells. We studied differentiation inducing effect of all-trans retinoic acid (ATRA) by studying proportion of high dense fractions of stem-like cells and the size of S phase fraction in cell cycle. From mammary organoids obtained from 7- to 8-week old F344 female rat mammary gland, we cultured rat mammary epithelial cells (RMEC) and treated physiological doses of $10^{-6}$, $10^{-7}$, and $10^{-8}$ M ATRA from the first day and then cultured for 4, 7, and 14 days. After that, immunostaining was performed using peanut agglutinin (PNA) and anti-Thy-1.1 monoclonal antibody (Thy-1.1) that can be used as markers of differentiation. We separated four different cell subpopulations by flow cytometry: cells negative to both reagents (B-), PNA-positive cells (PNA+), Thy-1.1-positive cells (Thy-1.1+), and cells positive to both reagents (B+). We observed continuous decreases of high dense fractions of stem-like cells (PNA+ subpopulations) for 14 days and as much decreases as high doses of ATRA, which were thought to be proportional to doses of ATRA. We labeled RMEC with bromodeoxyuridine and investigated cell cycle fractions that went through S phase. We observed a tendency of decrease of S phase fraction with time in culture, which, is thought to be related to continuous decreases of PNA+ subpopulations and inhibitory role of ATRA on cell cycle. These results suggest that physiological doses of ATRA could stimulate differentiation of RMEC and convert stem-like RMEC to differentiated cells in SFM for a relatively long period of 14 days.
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