• Title/Summary/Keyword: P19 cells

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Improved Purification Process for Cholera Toxin and its Application to the Quantification of Residual Toxin in Cholera Vaccines

  • Jang, Hyun;Kim, Hyo-Seung;Kim, Jeong-Ah;Seo, Jin-Ho;Carbis, Rodney
    • Journal of Microbiology and Biotechnology
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    • v.19 no.1
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    • pp.108-112
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    • 2009
  • A simplified method for the purification of cholera toxin was developed. The 569B strain of Vibrio cholerae, a recognized hyper-producer of cholera toxin, was propagated in a bioreactor under conditions that promote the production of the toxin. The toxin was separated from the bacterial cells using 0.2-${\mu}m$ crossflow microfiltration, the clarified toxin was passed through the membrane into the permeate, and the bacterial cells were retained in the retentate. The 0.2-${\mu}m$ permeate was then concentrated 3-fold and diafiltered against 10 mM phosphate buffer, pH 7.6, using 30-kDa crossflow ultrafiltration. The concentrated toxin was loaded onto a cation exchange column, the toxin was bound to the column, and most of the impurities were passed unimpeded through the column. The toxin was eluted with a salt gradient of phosphate buffer, pH 7.0, containing 1.0 M NaCl. The peak containing the toxin was assayed for cholera toxin and protein and the purity was determined to be 92%. The toxin peak had a low endotoxin level of $3.1\;EU/{\mu}g$ of toxin. The purified toxin was used to prepare antiserum against whole toxin, which was used in a $G_{M1}$ ganglioside-binding ELISA to determine residual levels of toxin in an oral inactivated whole-cell cholera vaccine. The $G_{M1}$ ganglioside-binding ELISA was shown to be very sensitive and capable of detecting as little as 1 ng/ml of cholera toxin.

Effects of Gamgung-tang on Proliferation and cAMP Accumulation of Thyroid Cells (감궁탕이 갑상선세포의 증식과 cAMP 축적에 미치는 영향)

  • Kim, Mee-Kyung;Shon, Yun-Hee;Nam, Kyung-Soo;Son, Ok-Lye;Kim, Cheorl-Ho;Jeon, Byung-Hun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.3
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    • pp.623-627
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    • 2005
  • Abnormal thyroid cell proliferation has a very important role in hypothyroidism. Thyroid stimulating hormone (TSH) stimulates proliferation and maintains differentiated function in thyroid follicular cells. A functioning rat thyroid cell line (FRTL) was used to study the effect of Gamgung-tang (GGT, Glycyrrhizae Radix, black beans, Angelicae Radix and Cnidii Rhizoma) on proliferation and cAMP accumulation of thyrocytes. Proliferation of cell was assessed by DNA synthesis and incorporation of $[^3H]thymidine$ into DNA. The concentration of cAMP was measured simultaneously with growth assessment. Extract of GGT ($0.15{\sim}0.9\;mg/ml$ increased DNA synthesis in a dose-dependent manner. GGT at 0.6 (p<0.05) and 0.9 mg/ml (p<0.01) significantly increased $[^3H]thymidine$ incorporation. A comparable effect was observed with TSH. GGT also enhanced cAMP accumulation. These results indicate that GGT increases the proliferation of thyrocytes and may be considered a promising agent for the treatment of autoimmune thyroid disease.

Effect of Licochalcone A on the Production of Cytokines in LPS-Activated RAW264.7 Macrophage Cells (Licochalcone A가 대식세포주의 사이토카인 생성에 미치는 영향)

  • Lee, Gi-Sae;Lee, Sung-Ho;Cho, Young-Chang;Yoon, Goo;Cheon, Seung-Hoon;Kang, Bok-Yun
    • YAKHAK HOEJI
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    • v.53 no.6
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    • pp.321-327
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    • 2009
  • Licochalcone A is a chalcone isolated from the roots of Glycyrrhiza inflate. In this study, we examined the effects of licochalonce A on the production of cytokines in LPS-activated macrophages. Licochalcone A inhibited the secretion of proinflammatory cytokines such as IL-1$\beta$, IL-6, and TNF-$\alpha$. The reduced secretion of proinflammatory cytokines is related to the differences in the mRNA expression of IL-1$\beta$, IL-6, and TNF-$\alpha$. Moreover, licochalcone A inhibited the mRNA expression of IL-12p40, IL-18, and IL-23p19. To investigate its mechanism, we performed gel shift assay. Licochalcone A reduced nuclear NF-${\kappa}B$ binding activity in LPS-activated RAW264.7 cells. Taken together, these results suggest that licochalcone A has anti-inflammatory effects in LPS-activated macrophages and its mechanism could be through the down-regulation of binding to the ${\kappa}B$ site.

Design of a 4-bit Digital Phase Shifter in Quasimillimeter Wave Band for Satellite Communication (준밀리미터파대 위성통신용 4-bit 디지털 위상변위기의 설계)

  • 신동환;임인성;김우재;민경일;오승엽
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.10 no.3
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    • pp.461-470
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    • 1999
  • This paper presents the description of a 4-bit digital p-i-n diode phase shifter that was designed for quasimillimeter wave band satellite receiver to use in phased-array systems. 180$^{\circ}$ and 90$^{\circ}$ cells are designed in reflection type that consists of a 3-dB rat-race hybrid coupler, 45$^{\circ}$ and 22.5$^{\circ}$ cells are designed in loaded-line type to reduce the size of circuit and the number of diode to be used. The 4-bit phase shifter uses eight p-i-n diodes mounted in the microstrip circuit. The average insertion loss for the 16 phase states is 6.92dB over the 19.8~20.3 GHz band and maximum phase error is 6.2$^{\circ}$ at 20 GHz.

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Isolation of Lipase Producing Yeast and Optimization of Cultivation Condition (Lipase 생산 효모균주의 분리 및 배양조건 최적화)

  • 박명훈;류현진;오경근
    • KSBB Journal
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    • v.19 no.2
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    • pp.148-153
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    • 2004
  • Lipase catalyzes the hydrolysis of glycerides into fatty acids and glycerol. The study of microbial lipases has been stimulated in resent years. It is due to the potential uses of lipases in esterification of oils to glycerol, alcohols and carbohydrates. Development of lipase producing yeast has been focused concerning to the utilization of yeast culture for animal feed. In this study, yeast like cells was isolated from a waste oil and sludge. A strain having higher lipase activity was selected by random mutagenesis using UV-radiation. The optimal cultivation conditions in submerged culture were examined in terms of lipase production. 2.0% of high fructose syrup, 1,0% of CSL, and 1.0% of olive oil were selected as the nutritional media for the production of lipase. The maximum lipase activity of 1.12 U/ml and viable cell number of 8.8${\times}$10$\^$7/ cells/mL were obtained at 27$^{\circ}C$ with an initial pH of 5.0.

Sleep deprivation induces structural changes in the adult rat testis: The protective effects of olive oil

  • Fatemeh Karimi;Ali Noorafshan;Saied Karbalay-Doust;Maryam Naseh
    • Clinical and Experimental Reproductive Medicine
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    • v.50 no.1
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    • pp.19-25
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    • 2023
  • Objective: Sleep deprivation (SD) is a common problem in today's stressful lifestyle and have physiological consequences, including reproductive dysfunction and infertility. As an antioxidant, olive oil may be effective in reducing testicular and spermatological damage by decreasing the production of free radicals. Methods: This study investigated the effects of olive oil on sperm quality and testicular structure using stereological methods to assess rats with SD. Results: When comparing SD group to grid floor+distilled water (GR) group, we found that the sperm count and motility, as well as the percentage of slow progressive sperm was significantly lower in SD group (p<0.05), but the percentage of immotile sperm was higher (p<0.01). However, no improvement was observed in sperm count or motility after concomitant treatment of SD group with olive oil. Stereological examinations revealed no significant change in the total volumes of the seminiferous tubules, interstitial tissue, and germinal epithelium in the study groups. Conversely, the total number of testicular cell types was significantly lower in SD group than in GR group. Although the total number of Sertoli and Leydig cells was significantly higher in the S +olive oil group than in the untreated SD group, no significant difference in the total number of other testicular cell types was observed between the two groups. Conclusion: SD potentially induced structural changes in testis that affected sperm count and motility. However, olive oil only improved the total number of Sertoli and Leydig cells in the animals with SD and did not improve sperm count and motility.

Effect of the mixed culture of heterofermentative lactic acid bacteria and acid-tolerant yeast on the shelf-life of sourdough (이상발효유산균과 내산성 효모와의 혼합배양이 사워도우의 저장성에 미치는 영향)

  • Lim, Eun-Seo
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.471-481
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    • 2016
  • The primary objective of this study was to investigate the effects of the bacteriocin-producing heterofermentative lactic acid bacteria (LAB) and acid-resistant yeast isolated from Mukeunji, a Korean ripened kimchi on shelf-life extension and quality improvement of sourdough. According to gene sequence analysis the heterofermentative LAB that showed the antimicrobial activity against bread-spoilage Bacillus strains were identified as Leuconostoc mesenteroides LAS112, Lactobacillus brevis LAS129, and L. mesenteroides subsp. dextranicum LAB137. In addition, the yeasts that were able to grow at acidic pH were identified as Pichia membranifaciens YS05, Pichia fermentans YS19, and Pichia anomala YS26. During sourdough fermentation the levels of acetic acid and bacteriocin produced by L. brevis LAS129 strain were higher than those of L. mesenteroides LAS112 and L. mesenteroides subsp. dextranicum LAS 137 strains, whereas LAS112 strain produced the highest levels of lactic acid. The maximum bacteriocin activity (640 AU/g) against Bacillus subtilis ATCC 35421 was obtained in sourdough fermented by mixed culture of L. brevis LAS129 and P. membranifaciens YS05 or P. anomala YS26. After 24 h of fermentation at $30^{\circ}C$, the viable cell counts of LAS129 ($10^9CFU/g$) in sourdough were higher than those of the YS05 or YS26 ($10^7CFU/g$). Meanwhile, the viable cells of bread-spoilage strain in sourdough fermented with these strains were significantly (P < 0.05) lower than the control group.

Effects of the Oriental Medicinal Prescriptions, Jahyulyangeuntang and Yanghyuljangeunkeonbohwan, on Calcium Absorption in the Human Colon Carcinoma Cell Line (Caco-2 Cells) (자혈양근탕 및 양혈장근건보환이 인체 소장 상피세포주 (Caco-2) 모델에서 칼슘 흡수에 미치는 영향)

  • 박태선;임현정;황귀서
    • Journal of Nutrition and Health
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    • v.35 no.4
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    • pp.446-453
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    • 2002
  • Effects of the two oriental medicinal prescriptions, Jahyulyangeuntang (JH) and Yanghyuljangeunkeonbohwan (YH), on intestinal calcium absorption were examined in the human colon carcinoma tell line, Caco-2 cells. Intestinal calcium absorption was evaluated at the level of Ca uptake into the cells across the brush border membranes, as well as at the level of net Ca transport (implying the amount of intestinal Ca transported into the blood stream). When the Caco-2 cells were incubated for 4, 8, 16 and 24 days post seeding, the cells were differentiated continuously, and showed progressively increased activities of Ca uptake (1.13 $\pm$ 0.04, 1.19 $\pm$ 0.02, 1.94 $\pm$ 0.03, and 2.40 $\pm$ 0.12 nmole.mg protein$^{-1}$ .30 min$^{-1}$ , respectively). Pretreatment of confluent Caco-2 cells with 50 $\mu\textrm{g}$/ml of YH for 24 hours resulted in a 30% increase in Ca uptake (p < 0.07), while pretreatment of the cells with the same concentration of JH for 6 hours resulted in a 24% increase (p < 0.05) in Ca uptake, compared to the value for the control cells (2.34 $\pm$ 0.10 nmole.mg protein$^{-1}$ .30 min$^{-1}$ ). When the cells were pretreated with varied concentrations (5-100 $\mu\textrm{g}$/ml) of the test samples for 6 hours, maximal increases in Ca uptake were observed in the cells pretreated with 100 $\mu\textrm{g}$/ml of YH (a 23% increase), and 50 $\mu\textrm{g}$/ml of JH (a 28% increase), respectively : however, no influence was seen on the net Ca transport activity. These results show that pretreatment with JH or YH, the two oriental medicinal prescriptions commonly used for improvement of bone metabolism, could possibly increase Ca accumulation inside the cells. but not the intestinal Ca net transport in vitro.

Properties of stretch-activated $K^+$ channels in an G292 osteoblast-like cell (G292 세포에서 세포막 신장으로 활성화되는 $K^+$통로의 특성)

  • Lee, Sang-Gook;Jung, Dong-Keun;Suh, Duk-Joon;Park, Soo-Byung
    • The korean journal of orthodontics
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    • v.30 no.2 s.79
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    • pp.197-204
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    • 2000
  • [$K^+$]-selective ion channels were studied in excised inside-out membrane patches from human osteoblast-like cells (G292). Three classes of $K^+$channels were present and could be distinguished on the basis of conductance. Conductances were $270\pm27\;pS,\;113\pm12\;pS,\;48\pm8\;pS$ according to their approximate conductances in symmetrical 140 mM KCl saline at holding potential of -80 mV It was found that the small conductance (48 pS) $K^+$channel activation was dependent on membrane voltage. In current-voltage relationship, small conductance $K^+$channel showed outward rectification, and it was activated by the positive potential inside the membrane. In recordings, single channel currents were activayed by a negative pressure outside the membrane. The membrane pressure increased $P_{open}$ of the $K^+$ channel in a pressure-dependent manner. In the excised-patch clamp recordings, G292 osteoblast-like cells have been shown to contain three types of $K^+$ channels. Only the small conductance (48 pS) $K^+$channel is sensitive to the membrane stretch. These findings suggest that a hyperpolarizing current, mediated in part by this channel, may be associated with early events during the mechanical loading of the osteoblast. In G292 osteoblast-like cells, $K^+$channel is sensitive to membrane tension, and may represent a unique adaptation of the bone cell membrane to mechanical stress.

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Activity of Cytokines and Expression of CD62L in Patients with Bronchial Asthma (기관지 천식환자에서 CD62L의 발현 및 싸이토카인의 변화)

  • Song, Kwang-Seon;Lee, Won-Yeon;Hong, Ae-Ra;Kim, Hee-Sun;Yong, Suk-Joong;Shin, Kye-Chul
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.1
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    • pp.90-98
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    • 1998
  • Background : The CD4+ T-helper cells comprise functionally distinct subsets of Th1 and Th2 cells that are distinguished on the basis of differential cytokines production Th1 cells secrete interferon-$\gamma$, lymphotoxin, interleukin-2. Th2 cells produce interleukin-4, interleukin-5, interleukin-10. A previous study shown that Th2 cells and their cytokines increased in patients with atopic asthma. We compared cytokines(IL-4, IFN-$\gamma$) activity and subpopulation of T-lymphocytes in peripheral blood from atopic asthmatics versus non-asthmatics. Method: Fifteen patients with atopic asthma(nine men, six women), twelve patients with chronic bronchitis(six men, six women), five healthy persons(three men, two women) were studied. Activity of IL-4, IFN-$\gamma$ and T-cell subpopulation in peripheral blood were estimated. Results: Patients had a median age of 55yr. The mean activity of IL-4 of asthmatics was significantly increased(control $0.75{\pm}1.1pmol/L$, atopic asthmatics $3.50{\pm}0.75pmol/L$, chronic bronchitis $2.01{\pm}1.2pmol/L$), but IFN-$\gamma$ was not significantly increased. In the T lymphocyte sunsets the percent of CD62L+ T-lymphoeytes of asthmatics was not significantly increased (control $16.7{\pm}16.4%$, atopic asthmatics $24.8{\pm}23.6%$, chronic bronchitis $17.0{\pm}16.9%$). Conclusion: In this study elevated production of IL-4 was observed in atopic asthmatics. CD62L+T-lymphoeytes was not increased in atopic asthma.

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