• Title/Summary/Keyword: $\alpha$-CD

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Expression of the Promoter for the Maltogenic Amylase Gene in Bacillus subtilis 168

  • Kim Do-Yeon;Cha Choon-Hwan;Oh Wan-Seok;Yoon Young-Jun;Kim Jung-Wan
    • Journal of Microbiology
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    • v.42 no.4
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    • pp.319-327
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    • 2004
  • An additional amylase, besides the typical $\alpha-amylase,$ was detected for the first time in the cytoplasm of B. subtilis SUH4-2, an isolate from Korean soil. The corresponding gene (bbmA) encoded a malto­genic amylase (MAase) and its sequence was almost identical to the yvdF gene of B. subtilis 168, whose function was unknown. Southern blot analysis using bbmA as the probe indicated that this gene was ubiquitous among various B. subtilis strains. In an effort to understand the physiological function of the bbmA gene in B. subtilis, the expression pattern of the gene was monitored by measuring the $\beta-galactosidase$ activity produced from the bbmA promoter fused to the amino terminus of the lacZ struc­tural gene, which was then integrated into the amyE locus on the B. subtilis 168 chromosome. The pro­moter was induced during the mid-log phase and fully expressed at the early stationary phase in defined media containing $\beta--cyclodextrin\;(\beta-CD),$ maltose, or starch. On the other hand, it was kept repressed in the presence of glucose, fructose, sucrose, or glycerol, suggesting that catabolite repression might be involved in the expression of the gene. Production of the $\beta-CD$ hydrolyzing activity was impaired by the spo0A mutation in B. subtilis 168, indicating the involvement of an additional regu­latory system exerting control on the promoter. Inactivation of yvdF resulted in a significant decrease of the $\beta-CD$ hydrolyzing activity, if not all. This result implied the presence of an additional enzyme(s) that is capable of hydrolyzing $\beta-CD$ in B. subtilis 168. Based on the results, MAase encoded by bbmA is likely to be involved in maltose and $\beta-CD$ utilization when other sugars, which are readily usable as an energy source, are not available during the stationary phase.

소자약침(蘇子藥鍼)의 천식억제(喘息抑制) 및 면역조절효과(免疫調節效果)에 대(對) 실험적(實驗的) 연구(硏究)

  • Ko, Seung-Kyoung;Hong, Kwon-Eui
    • Journal of Pharmacopuncture
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    • v.9 no.3 s.21
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    • pp.47-59
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    • 2006
  • Objectives : The purpose of this study was to confirm the suppressive effect against asthma and immune regulatory effect of Perillae Fractusher Herbal-acupuncture at Chok-samni(ST36) on ovalbumin-induced asthma in mice. Methods : C57BL/6 mice were sensitized and challenged with OVA(ovalbumin) for 12 weeks. The mice in the PF-HA group were treated with PF-HA at ST36 for the later 8 weeks(3 times a weeks). The mice in the OVA-Needle-Prink(NP) group were treated with single prick with an injection needle at ST36 for the later 8 weeks(3 times a weeks). Results : 1. The lung weight of the mice treated with PF-HA at St36 were decreased significantly compared with that of the control group. 2. The total leukocytes and eosinophils in BALF of the group treated with PF-HA were decreased significantly compared with those of the control group. 3. Eosinophils in BALF and the collagen accumulation in lung of the mice treated with PF-HA were decreased significantly compared with those of the control group. 4. The Concentrations of IgE, IL-4 and IL-5 in BALF, and IL-4, IL-5 and IL-13 in serum of the PF-HA group were decreased significantly compared with those of the control group. 5. The numbers of CD3-/CCR3+, Gr-1+/CD11b+,CD4+, and CD3e+/CD69+cells in lung of the mice group treated with PF-HA at St36 were decreased significantly compared with those of the control group. 6. The mRNA expressions of TNF-${\alpha}$, IL-4, IL-5, IL-13 in lung of the mice group treated with PF-HA at St36 were decreased significantly compared with those of the control group.

Effects of Beryllium on Human Serum Immunoglobulin and Lymphocyte Subpopulation

  • Kim, Ki-Woong;Kim, DaeSeong;Won, Yong Lim;Kang, Seong-Kyu
    • Toxicological Research
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    • v.29 no.2
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    • pp.115-120
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    • 2013
  • To investigate the effects of short-term exposure of beryllium on the human immune system, the proportion of T-lymphocytes such as CD3+, CD4+, CD8+, CD95, and NK cells, and the proportion of B cells and $TNF{\alpha}$ level in peripheral blood and immunoglobulins in the serum of 43 exposed workers and 34 healthy control subjects were studied. External exposure to beryllium was measured by atomic absorption spectrometer as recommended by the NIOSH analytical method 7300. T lymphocyte subpopulation analysis was carried out with flow cytometer. The working duration of exposed workers was less than 3 months and the mean ambient beryllium level was $3.4{\mu}g/m^3$, $112.3{\mu}g/m^3$, and $2.3{\mu}g/m^3$ in molding (furnace), deforming (grinding), and sorting processes, respectively (cited from Kim et al., 2008). However, ambient beryllium level after process change was non-detectable (< $0.1{\mu}g/m^3$). The number of T lymphocytes and the amount of immunoglobulins in the beryllium-exposed workers and control subjects were not significantly different, except for the total number of lymphocytes and CD95 (APO1/FAS). The total number of lymphocytes was higher in the beryllium-exposed individuals than in the healthy control subjects. Multiple logistic regression analysis showed lymphocytes to be affected by beryllium exposure (odd ratio = 7.293; p<0.001). These results show that short-term exposure to beryllium does not induce immune dysfunction but is probably associated with lymphocytes proliferation.

The effects of HYT on various immunological factors related to pathogenesis of allergic dermatitis in NC/Nga mice induced by Biostir AD (아토피피부염 동물 병태모델에서의 형개연교탕(荊芥連翹湯)의 면역조절작용)

  • Kang, Ran-Yi;Park, Bo-Kyoung;Gim, Seon-Bin;Choi, Hak-Joo;Kim, Dong-Hee
    • Journal of Haehwa Medicine
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    • v.18 no.2
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    • pp.47-62
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    • 2009
  • Atopic dermatitis induced NC/Nga mice were used to investigate the efficacy of HYT on the recovery of dermatitic symptoms by how HYT influenced the immune related factors. The results are as below: 1. Compared to the control, HYT treated group showed recovery of atopic dermatitis by the naked eye observation, and significant reduction of dermatits index was observed after 14 weeks. 2. HYT treated group showed significant decrease of the ratio of CCR3+, B220+/IgE+, and Gr-1+/CD11b+ immune cells in dorsal skin by 40.5%, 34.2%, and 48.1%, respectively. 3. HYT treated group showed increase in the ratio of CD19+ immune cells within PBMC by 10.8%, as well as decrease in CD3+, CD3+/CD69+, NKT+ ratios by 5.3%, 35.2%, and 44.9%, respectively. 4. HYT treated group showed increase in the expression of IFN-$\gamma$ in serum by 589.3%, whereas the expression of IL-4, IL-5, IL-6, IL-13, TNF-$\alpha$, MCP-1 and RANTES were decreased by 31.4%, 82.1%, 97.1%, 39.5%, 83.7%, 26.1%, 48.6%, respectively. A 47.2% decrease in IgE expression was also observed. The results above strongly supported the improvement of atopic dermatitis by HYT treatment through immune modulation. Further studies on the synergistic effect of each ingredients of HYT and therapeutic effects according to the dosage of each ingredient should be followed for clinical applications. This work was (partly) supported by the RIC program of MKE(Ministry of Knowledge Economy) in Daejeon University.

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The effects of HYGB on various immunological factors related to pathogenesis of allergic dermatitis in NC/Nga mice (형개연교탕가미방의 아토피 피부염 병태모델에서의 면역조절능에 관한 연구)

  • Park, Sung-Hee;Park, Bo-Kyoung;Gim, Seon-Bin;Choi, Hak-Joo;Kim, Dong-Hee
    • Journal of Haehwa Medicine
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    • v.18 no.2
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    • pp.63-79
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    • 2009
  • Atopic dermatitis induced NC/Nga mice were used to investigate the efficacy of Hyunggaeyunkyotanggamibang(HYGB) on the recovery of dermatitic symptoms through its influence on the immune related factors. The results are as below: 1. HYGB treated group showed improvement of atopic dermatitis with naked eye observation, and significant decrease of dermatitis index was observed after 14 weeks. 2. HYGB treated group showed significant decrease of the ratio of CCR3+, B220+/IgE+, and CD11b+/Gr-1+ immune cells in dorsal skin by 41.7%, 21.5%, and 23.8%, respectively. 3. HYGB treated group showed an increase of CD19+ immune cells by 10.3% in PBMC, whereas CD3+, CD3+/CD69+, NKT+ immune cells were decreased by 4.3%, 42.9%, and 21.7%, respectively. 4. HYGB treated group showed an increase in the expression of IFN-$\gamma$ in the serum by 514.3%. However, the expressions of IL-4, IL-5, IL-6, IL-13, TNF-$\alpha$, MCP-1 and RANTES were decreased by 21.2%, 69.8%, 90.5%, 28.7%, 72.2%, 26.1%, and 19.9%, respectively. Also, the expression of IgE was decreased by 44.3%. 5. HYGB treated group showed a decrease of the expression of IL-4 and IL-5 by 43% and 44.3%. The results above indicated that HYGB clinically used for atopic dermatitis treatment has objective validity, and therefore can be provided as the basic data for EBM(Evidence based medicine) construction for anti-allergic and anti-inflammatory studies.

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Effect of Dietary Taurine or Glycine Supplementation on Plasma and Liver Free Amino Acid Concentrations in Rats (식이내의 타우린 또는 글라이신 보강이 흰쥐의 혈장과 간의 유리아미노산 농도 및 패턴에 미치는 영향)

  • 박정은
    • Journal of Nutrition and Health
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    • v.31 no.2
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    • pp.126-134
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    • 1998
  • Our previous study demonstrated that dietary taurine or glycine supplementation significantly lowered plasma and hepatic cholesterol and triglyceride concentrations in rats fed a cholesterol-free diet. In the present study, the effect of long term dietary taurine or glycine supplementation, for the purpose of preventing and/or treating of hyperlipidemia and other known biological functions, on plasma and hepatic free amino acid concentrations and profiles were evaluated in rats. Three groups of male rats(110-130g) were fed a control diet(CD), taurine-supplemented diets(TSD ; CD+ 1.5% taurine) or glycine-supplemented diet(GSD ; CD+1.5% glycine) for 5 weeks. Plasma and hepatic free amino acid concentrations were determined by an automated amino acid analyzer based on ion-exchange chormatography. The feeding of TSD for 5 weeks yielded a 444% higher plasma taurine concentration , and the feeding GSD for the same period resulted in a 143% higher plasma glycine level in rats compared to those fed DB. Hepatic taurine concentration was significantly higher in rats fed TSD(145% increase) compared to the control rats. However, hepatic glycine concentration was not influenced by dietary glycine supplementation , which implies that the massive dose of glycine entering the body was more rapidly metabolized or excreted than taurein. Dietary taurine or glycine supplementation resulted in similar changes in plasma free amino acid concentrations, except in levels of taurine and glycine. Plasma levels of histidine, lysine, phenylalanine , alanine, proline, hydroxypoline, $\alpha$-aminogutyric acid, cystathionine and ethanolamine were significantly higher in rats fed TSD or GSD than those fed GD. Glycine supplementation did not change hepatic free amino acid concentrations as compared to CD. Concentrations of most hepatic free amino acids were not influenced by dietary taurine supplementation with the exception of significantly higher levels of asparate and tyrosine(56-63% increase) and lower levels of histidine and glutamate(33-34% decrease) compared to the control rats. These results suggest long-term dietary taurine or glycine supplementation resulted in increases in most plasma free amino acid levels, but did not cause a characteristic change in plasma aminogram pattern compared to rats fed CD.

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Effect of Herbal-acupuncture with Erythrinae Cortex Solution at the Joksamni($ST_{36}$) of Mice with Collagen-induced Arthritis (해동피약침(海桐皮藥鍼)이 Collagen-induced Arthritis 억제 및 면역조절에 미치는 영향)

  • Kim, Young-Wah;Lee, Hyun
    • Journal of Acupuncture Research
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    • v.24 no.5
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    • pp.171-183
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    • 2007
  • Objectives : The purpose of this study was to analyze the effects of Erythrinae Cortex herbal-acupuncture solution(EC-HAS) at the Joksamni($ST_{36}$) of mice with collagen II-induced arthritis(CIA). Methods : The author performed several experimental procedures to observe the effects of the EC-HAS at the arthritis. The severity of arthritis, changes of cytokine level and antibody level, histological changes of the CIA mouse joint were analyzed. Results : 1. The incidence of arthritis and arthritis index were significantly decreased in the cases which were treated with the EC-HA. 2. Cartilage destruction and synovial cell proliferation were reduced, and the expression of the collagen fibers was similar with that of the normal group. 3. The levels of IL-6, $IFN-{\gamma}$, $TNF-{\alpha}$, $IL-1{\beta}$ in serum of the CIA mice which were treated with the EC-HA were significantly decreased compared with those of the normal group. 4. The levels of IgG, IgM and anti-collagen II in serum of CIA mice when they treated with the EC-HA were significantly decreased compared with those of the normal group. 5. The expression ratio of $CD4^+$ to $CD8^+$ cells in the EC-HA treated mice were maintained as much as the normal group of the lymph nodes in the CIA mice. 6. The $CD3e^+CD69^+$ and $CD11b^+Gr-1^+$ cell populations in the knee joint were significantly decreased in the EC-HA treated group. Conclusions : These results suggest that the EC-HA at the ST36 may be responsible roles to control on the synovial cell proliferation and to prevent the cartilage destruction in rheumatoid arthritis. These results will be important supporting evidence for the practical use of the EC-HA at rheumatoid arthritis clinic in the future.

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Cross Talk Experiment with Two-element CdTe Detector and Collimator for BNCT-SPECT

  • Manabe, Masanobu;Ohya, Ryosuke;Saraue, Nobuhide;Sato, Fuminobu;Murata, Isao
    • Journal of Radiation Protection and Research
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    • v.41 no.4
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    • pp.328-332
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    • 2016
  • Background: Boron Neutron Capture Therapy (BNCT) is a new radiation therapy. In BNCT, there exists some very critical problems that should be solved. One of the severest problems is that the treatment effect cannot be known during BNCT in real time. We are now developing a SPECT (single photon emission computed tomography) system (BNCT-SPECT), with a cadmium telluride (CdTe) semiconductor detector. BNCT-SPECT can obtain the BNCT treatment effect by measuring 478 keV gamma-rays emitted from the excited state of $^7Li$ nucleus created by $^{10}B(n,{\alpha})$ $^7Li$ reaction. In the previous studies, we investigated the feasibility of the BNCT-SPECT system. As a result, the S/N ratio did not meet the criterion of S/N > 1 because deterioration of the S/N ratio occurred caused by the influence of Compton scattering especially due to capture gamma-rays of hydrogen. Materials and Methods: We thus produced an arrayed detector with two CdTe crystals to test cross talk phenomenon and to examine an anti-coincidence detection possibility. For more precise analysis for the anti-coincidence detection, we designed and made a collimator having a similar performance to the real BNCT-SPECT. Results and Discussion: We carried out experiments with the collimator to examine the effect of cross talk of scattering gamma-rays between CdTe elements more practically. As a result of measurement the coincidence events were successfully extracted. Conclusion: We are now planning to carry out evaluation of coincidence rate from the measurement and comparison of it with the numerical calculations.

Tumor Cell Clone Expressing the Membrane-bound Form of IL-12p35 Subunit Stimulates Antitumor Immune Responses Dominated by $CD8^+$ T Cells

  • Lim, Hoyong;Do, Seon Ah;Park, Sang Min;Kim, Young Sang
    • IMMUNE NETWORK
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    • v.13 no.2
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    • pp.63-69
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    • 2013
  • IL-12 is a secretory heterodimeric cytokine composed of p35 and p40 subunits. IL-12 p35 and p40 subunits are sometimes produced as monomers or homodimers. IL-12 is also produced as a membrane-bound form in some cases. In this study, we hypothesized that the membrane-bound form of IL-12 subunits may function as a costimulatory signal for selective activation of TAA-specific CTL through direct priming without involving antigen presenting cells and helper T cells. MethA fibrosarcoma cells were transfected with expression vectors of membrane-bound form of IL-12p35 (mbIL-12p35) or IL-12p40 subunit (mbIL-12p40) and were selected under G418-containing medium. The tumor cell clones were analyzed for the expression of mbIL-12p35 or p40 subunit and for their stimulatory effects on macrophages. The responsible T-cell subpopulation for antitumor activity of mbIL-12p35 expressing tumor clone was also analyzed in T cell subset-depleted mice. Expression of transfected membranebound form of IL-12 subunits was stable during more than 3 months of in vitro culture, and the chimeric molecules were not released into culture supernatants. Neither the mbIL-12p35-expressing tumor clones nor mbIL-12p40-expressing tumor clones activated macrophages to secrete TNF-${\alpha}$. Growth of mbIL-12p35-expressing tumor clones was more accelerated in the $CD8^+$ T cell-depleted mice than in $CD4^+$ T cell-depleted or normal mice. These results suggest that $CD8^+$ T cells could be responsible for the rejection of mbIL-12p35-expressing tumor clone, which may bypass activation of antigen presenting cells and $CD4^+$ helper T cells.

Inhibition of Langerhans cell function by UVB radiation

  • Okamoto, Hiroyuki;Mizuno, Kana;Horio, Takeshi
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.190-193
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    • 2002
  • The functional disruption of Langerhans cells (LC) by UVB radiation is involved in antigen-specific immunosuppression of contact hypersensitivity. We tested whether UVB radiation inhibits the endocytotic activity of LC, which leads to impaired subsequent migration and maturation. Human monocyte-derived LC that took up lucifer yellow (L Y) or FITC-dextran (Fd) exclusively migrated in response to 6Ckine and matured. Exposing LC to 10-40 mJ/cm$^2$ of UVB radiation reduced their endocytotic activity in fluid phase pinocytosis (measured by uptake of LY) and in receptor-mediated endocytosis (measured by uptake of Fd). Membrane ruffling and CD32 expression were also suppressed by UVB radiation. UVB-irradiated, endocytosing LC had less movement towards 6Ckine, expressed less CD54 and CD86, and had less effective stimulatory activity in allo-MLR than nonirradiated, endocytosing LC. Endocytosis up-regulated TNF-$\alpha$ production by LC, but prior UVB radiation inhibited this enhancement. The finding that impaired endocytosis of LC by UVB radiation inhibits subsequent migration and maturation was also confirmed in murine epidermal cells obtained from unirradiated and 2OmJ/cm$^2$ of UVB-irradiated skin.

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