• 제목/요약/키워드: secretory production

검색결과 144건 처리시간 0.024초

Enhanced macrophage function of red ginseng acidic polysaccharide(RGAP) in combination with IFN-$\gamma$

  • Choi, Hye-Sook;Kwak, Yi-Seong;Park, Jong-Dae;Pyo, Suhk-Neung
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 2006년도 춘계학술대회
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    • pp.83-88
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    • 2006
  • In this study we examined the potential for the synergistic augmentation of the activity of inflammatory mouse peritoneal macrophages by stimulation with RGAP combined with IFN-$\gamma$. The moderate augmentative effect induced by preincubation with RGAP was observed in the production of IL-1, IL-6 and NO but not TNF-$\alpha$. In addition, IFN-$\gamma$ had a low activating effect. Following preincubation with both RGAP A and IFN-$\gamma$, a marked enhancement of secretory activity and tumoricidal activity was noted in mouse peritoneal macrophages. Treatment of peritoneal macrophage with combination increased the generation of IL-1, IL-6, TNF-$\alpha$ and NO, whereas the production of reactive oxygen species were not altered. These results demonstrate the synergistic effects on macrophage function of RGAP in combination with IFN-$\gamma$ and suggest that the ability of IFN-$\gamma$ to prime macrophages to produce secretory molecules in response to RGAP may have implications for immunotherapy with this combination.

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Secretory Expression of Human $\alpha_{s1}$-Casein in Saccharomyces cerevisiae

  • Kim, Yoo-Kyeong;Yu, Dae-Yeul;Kang, Hyun-Ah;Yoon, Sun;Chung, Bong-Hyun
    • Journal of Microbiology and Biotechnology
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    • 제9권2호
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    • pp.196-200
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    • 1999
  • A recombinant human $\alpha_{s1}$-casein was expressed as a secretory product in the yeast Saccharomyces cerevisiae. Three different leader sequences derived from the mating factor $\alpha$l (MF$\alpha$l), inulinase, and human $\alpha_{s1}$-casein were used to direct the secretion of human $\alpha_{s1}$-casein into the extracellular medium. Among the three leader sequences tested, the native leader sequence of human $\alpha_{s1}$-casein was found to be the most efficient in the secretory expression of human $\alpha_{s1}$-casein, which implies that the native leader sequence of human $\alpha_{s1}$-casein might be used very efficiently for the secretory production of other heterologous proteins in yeast. The recombinant human $\alpha_{s1}$-casein was proteolytically cleaved as the culture proceeded. Therefore, an attempt was made to produce human $\alpha_{s1}$-casein using a S. cerevisiae mutant in which the YAP3 gene encoding yeast aspartic protease 3 (YAP3) was disrupted. After 72 h of culture, most of the human $\alpha_{s1}$-casein secreted by the wild type was cleaved, whereas more than 70% of the human $\alpha_{s1}$-casein secreted by yap3-disruptant remained intact. The results suggest that YAP3 might be involved in the internal cleavage of human $\alpha_{s1}$-casein expressed in yeast

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인삼의 생약학적 연구 (Pharmacognostical Study on the Panax ginseng C. A. Meyer)

  • 도원임;이유진;박종희
    • 생약학회지
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    • 제35권1호통권136호
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    • pp.45-51
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    • 2004
  • Panax ginseng C. A. Meyer (Araliaceae) is listed in Shen Nung Pentsao Ching(神農本草經) as the remedy for replenishing the primodial qi(氣), restoring pulse, treating collapse, tonifing the spleen and lag, promoting the production of body fluid to quench thirst, tranquilizing the mind, and improving the function of brain. The prescriptions of In Sam Tang(人參揚) are also recorded in many other Chinese medical books. The identification of the age of Panax ginseng is very important in commercial market as well as in research field. However, any reports about it have not been clearly established yet. To clarify the criterion, the morphological and anatomical characteristics of the roots of various age Panax ginseng cultivated in Korea were studied. The characteristics of cork layer, secretory canal, and vessel were shown to bε useful keys to confirm the age of Panax ginseng.

Superoxide Anion Production by Human Neutrophils Activated by Trichomonas vaginalis

  • Song, Hyun-Ouk;Ryu, Jae-Sook
    • Parasites, Hosts and Diseases
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    • 제51권4호
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    • pp.479-484
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    • 2013
  • Neutrophils are the predominant inflammatory cells found in vaginal discharges of patients infected with Trichomonas vaginalis. In this study, we examined superoxide anion ($O^{\cdot}_{2^-}$) production by neutrophils activated by T. vaginalis. Human neutrophils produced superoxide anions when stimulated with either a lysate of T. vaginalis, its membrane component (MC), or excretory-secretory product (ESP). To assess the role of trichomonad protease in production of superoxide anions by neutrophils, T. vaginalis lysate, ESP, and MC were each pretreated with a protease inhibitor cocktail before incubation with neutrophils. Superoxide anion production was significantly decreased by this treatment. Trichomonad growth was inhibited by preincubation with supernatants of neutrophils incubated for 3 hr with T. vaginalis lysate. Furthermore, myeloperoxidase (MPO) production by neutrophils was stimulated by live trichomonads. These results indicate that the production of superoxide anions and MPO by neutrophils stimulated with T. vaginalis may be a part of defense mechanisms of neutrophils in trichomoniasis.

Enhanced Immune Cell Functions and Cytokine Production after in vitro Stimulation with Arabinoxylans Fraction from Rice Bran

  • Choi, Eun-Mi;Kim, Ah-Jin;Hwang, Jae-Kwan
    • Food Science and Biotechnology
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    • 제14권4호
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    • pp.479-486
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    • 2005
  • Arabinoxylan, a complex polysaccharide in cereal cell walls, has recently received research attention as a biological response modifier. The immunomodulating effect of arabinoxylans from rice bran (AXrb) was studied using a combined process of extrusion and commercial hemicellulase treatment in order to elucidate the augmentation mechanism of cell-mediated immunity in vitro. The cytotoxicity of mouse spleen lymphocytes against YAC-1 tumor cells was significantly enhanced by treatment with AXrb at $10-100\;{\mu}g/mL$. In an attempt to investigate the mechanism by which AXrb enhance NK cytotoxicity, we examined the effect of AXrb on cytokine production by spleen lymphocytes. Culture supernatants of the cells incubated with AXrb were collected and analyzed for IL-2 and IFN-${\gamma}$ synthesis by ELISA. IL-2 and IFN-${\gamma}$ production were increased significantly. These results suggest that AXrb may induce Th1 immune responses. Macrophages play an important role in host defenses against tumors by killing them and producing secretory products, which protect against bacterial, viral infection and malignant cell growth. AXrb were examined for their ability to induce secretory and cellular responses in murine peritoneal macrophages. When macrophages were treated with various concentrations ($10-100\;{\mu}g/mL$) of AXrb, AXrb induced tumoricidal activity, as well as increasing phagocytosis and the production of NO, $H_2O_2$, TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. These results indicate that reactive oxygen species, reactive nitrogen species, and inflammatory cytokines are likely to be the major mediators of tumoricidal activity in AXrb-treated macrophages. Therefore, AXrb may be useful in cancer immunotherapy and it is anticipated that AXrb obtained using extrusion and subsequent enzyme treatment can be used as an ingredient in nutraceuticals and cereal-based functional food.

토끼 아래눈물샘의 미세구조에 관한 전자현미경적 연구 (Electron Microscopic Study on the Rabbit Inferior Lacrimal Glands)

  • 박영희;안의태;고정식;박대균;김명수;박경호
    • Applied Microscopy
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    • 제37권1호
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    • pp.23-33
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    • 2007
  • 눈물샘은 주위환경으로부터 눈의 표면을 보호해주는 눈물막(tear film)을 구성하는 데 중요한 작용을 한다. 포유동물의 종에 따라 눈물샘의 구조와 구성요소는 차이가 있다. 포유류의 눈물샘에는 크게 두 가지 종류 즉, 위눈물샘 (superior lacrimal gland)과 아래눈물샘 (inferior lacrimal gland)으로 나눌 수 있다. 위눈물샘은 사람, 토끼, 개, 말, 낙타, 소등에 있는데, 안구의 위쪽에 위치하며, 아래눈물샘은 설치류, 토끼, 식충류 등에 존재하며, 안구의 아래쪽에 위치하고 있다. 토끼는 두 종류의 눈물샘 즉, 위눈물샘과 아래눈물샘을 모두 갖고 있는 데, 본 실험에서는 아래 눈물샘의 전자현미경적 구조를 관찰하였다. 토끼의 아래 눈물샘을 광학현미경으로 관찰한 일반구조는, 눈물샘은 전체적으로 피막에 싸여있었고, 샘실질은 결합조직에 의해 소엽으로 나뉘어지고, 소엽에는 분비부, 소엽속관 및 소엽사이관이 있다. 샘포세포는 아주 좁은 속공간을 가지고 있으며, 사이관(intercalated duct)은 샘포와 소엽속관을 연결한다. 소엽속관은 단층의 입방상피로 구성되어 있으며, 배출관인 소엽사이관이 결합조직 안에 잘 발달되어 있다. 눈물샘의 미세구조는 눈물샘 샘포세포는 피라밋형으로, 세포사이는 잘 발달된 연접복합체로 연접하였으며, 좁은 꼭대기 부분은 약간의 미세융모들이 샘의 내강쪽으로 향해 있다. 세포들의 가쪽면 사이에는 세포사이세관이 형성되었고, 그 속에도 상당수의 미세융모들이 관찰된다. 눈물샘분비세포는 세 종류로서 장액세포(serous cell), 장점액세포(seromucous cell)와 점액세포(mucous cell)로 이루어졌다. 장액세포는 샘포의 바닥 쪽에 많고, 세포질에는 전형적인 골지복합체와 전자밀도가 높은 분비과립이 있으며, 둥글둥글한 소포상의 과립형질내세망이 특징적인 구조이다. 한편 장점액세포는 전형적인 납작한 과립형질내세망이 조밀하고, 리보소체도 많아서 매우 어둡게 보이며, 분비과립에는 전자밀도가 높은 장액분비과립, 미세먼지같은 중등도의 전자밀도를 지닌 과립, 전자밀도가 낮으며 가끔은 융합되는 점액 분비과립을 갖고 있다. 점액세포의 과립세포질세망은 세포질전체에 퍼져 있으며, 관모양의 사립체들이 드물게 관찰된다. 점액세포에서는 세 가지 종류의 분비과립 즉, 전자밀도가 낮은 분비과립, 전자밀도가 비교적 높으며 빈도가 낮은 과립과 중간정도의 전자밀도를 가진 과립이 있으며, 전자밀도가 낮은 과립은 서로 융합된 모습을 보이기도 한다. 과립세포질세망은 주로 세포의 바닥부분에 위치하며, 골지복합체는 세포의 중간부분인 핵상부에 주로 위치한다. 점액세포의 가쪽벽에는 세포사이공간이 있어 가쪽주름(lateral fold)과 내강에 미세융모를 지닌 세포사이세관(intercellular canaliculi)이 관찰된다. 분비관을 구성하고 있는 세포들에는 세포질이 어두운 세포와 밝은 세포가 있었으며, 세포질내에는 전자밀도가 높은 분비과립이 관찰되었다. 전체적인 특징은 눈물샘분비세포 중 장액세포의 것과 비슷하였으나, 과립의 크기는 작았다. 분비관을 구성하는 세포들 사이에도 연접복합체가 매우 잘 발달되어 있었다. 샘포에서 사이관으로 이행되는 곳에서도 샘포세포와 사이관세포 사이에서도 연접복합체가 관찰되었다. 분비관세포의 분비과립 가운데는 중심부분에 전자밀도가 더 높은 중심을 가진 다른 모양의 과립이 관찰되기도 하였다.

Inhibitory Effects of Ginsenoside-Rb2 on Nicotinic Stimulation-Evoked Catecholamine Secretion

  • Lim, Hyo-Jeong;Lee, Hyun-Young;Lim, Dong-Yoon
    • The Korean Journal of Physiology and Pharmacology
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    • 제18권5호
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    • pp.431-439
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    • 2014
  • The aim of the present study was to investigate whether ginsenoside-Rb2 (Rb2) can affect the secretion of catecholamines (CA) in the perfused model of the rat adrenal medulla. Rb2 ($3{\sim}30{\mu}M$), perfused into an adrenal vein for 90 min, inhibited ACh (5.32 mM)-evoked CA secretory response in a dose- and time-dependent fashion. Rb2 ($10{\mu}M$) also time-dependently inhibited the CA secretion evoked by DMPP ($100{\mu}M$, a selective neuronal nicotinic receptor agonist) and high $K^+$ (56 mM, a direct membrane depolarizer). Rb2 itself did not affect basal CA secretion (data not shown). Also, in the presence of Rb2 ($50{\mu}g/mL$), the secretory responses of CA evoked by veratridine (a selective $Na^+$ channel activator ($50{\mu}M$), Bay-K-8644 (an L-type dihydropyridine $Ca^{2+}$ channel activator, $10{\mu}M$), and cyclopiazonic acid (a cytoplasmic $Ca^{2+}$-ATPase inhibitor, $10{\mu}M$) were significantly reduced, respectively. Interestingly, in the simultaneous presence of Rb2 ($10{\mu}M$) and L-NAME (an inhibitor of NO synthase, $30{\mu}M$), the inhibitory responses of Rb2 on ACh-evoked CA secretory response was considerably recovered to the extent of the corresponding control secretion compared with the inhibitory effect of Rb2-treatment alone. Practically, the level of NO released from adrenal medulla after the treatment of Rb2 ($10{\mu}M$) was greatly elevated compared to the corresponding basal released level. Collectively, these results demonstrate that Rb2 inhibits the CA secretory responses evoked by nicotinic stimulation as well as by direct membrane-depolarization from the isolated perfused rat adrenal medulla. It seems that this inhibitory effect of Rb2 is mediated by inhibiting both the influx of $Ca^{2+}$ and $Na^+$ into the adrenomedullary chromaffin cells and also by suppressing the release of $Ca^{2+}$ from the cytoplasmic calcium store, at least partly through the increased NO production due to the activation of nitric oxide synthase, which is relevant to neuronal nicotinic receptor blockade.

거미(Nephila clavata L. Koch) 견사선 분비관에서의 큐티클 전구체 생성에 관한 연구 (Study on Production of Cuticle Precursor within Silk Gland Duct of the Spider, Nephila clavata L. Koch)

  • 문명진
    • Applied Microscopy
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    • 제25권3호
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    • pp.20-32
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    • 1995
  • Ultrastructural aspects on the production of the duct cuticle and formation of cuticular precursors within silk glands of the orb web spider, Nephila clavata L. Koch(Araneae: Araneidae), were studied using transmission electron microscope. Four kinds of silk glands(ampullate glands, tubuliform glands, flageliform glands, and aggregate glands), which connected with large spinning tubes(spigots) of the spinnerets, were examined and discussed in terms of cuticle precursor production. Inner cuticular intima which composed of three layers of cuticles-subcuticle, endocuticle and exocuticle- were commonly originated from duct epithelial cells surrounding the cuticle. The morphology and internal textures of each cuticle precursors were very diverse according to the types of silk glands. However several common features were observed. These cuticle precursors were first produced from the rough endoplasmic reticulum and next concentration was accomplished through the Golgi complex. After this step, cuticle precursors were released to the cuticle layer as a form of secretory granule by the mechanism of merocrine secretion commonly.

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High-Level Expression and Secretion of Bacillus pumilus Lipase B26 in Bacillus subtilis Chungkookjang

  • Lee, Mi-Hwa;Song, Jae-Jun;Choi, Yoon-Ho;Hong, Seung-Pyo;Rha, Eu-Gene;Kim, Hyung-Kwoun;Lee, Seung-Goo;Poo, Har-Young;Lee, Sang-Chul;Seu, Young-Bae;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • 제13권6호
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    • pp.892-896
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    • 2003
  • High-level expression of the lipase B26 gene from Bacillus pumilus was achieved using Bacillus subtilis Chungkookjang isolated from the Korean traditional fermented bean paste, Chungkookjang. For the secretory production of recombinant lipase B26 in a Bacillus host system, pLipB26 was constructed by ligating the lipase B26 gene into the recently designed Escherichia coli-Bacillus shuttle vector, pLipSM, and that was then transformed into B. subtilis Chungkookjang. Among the various vector, medium, and host combinations, B. subtilis Chungkookjang harboring the pLipB26 exhibited the highest lipase activity in PY medium, and B. subtilis Chungkookjang secreted two times more enzymes than B. subtilis DB 104 under the same condition. When B. subtilis Chungkookjang harboring the pLipB26 was cultured in a 5-1 jar-fermentor containing 21 of a PY medium, the maximum lipase activity (140 U/ml) and production yield (0.68 g/l) were obtained during the late exponential phase from a cell-free culture broth. Although B. subtilis Chungkookjang also secreted extracellular proteases at the late exponential phase, these results suggested the potential of B. subtilis Chungkookjang as a host for the secretory production of foreign proteins.

Production of Cellulosic Ethanol in Saccharomyces cerevisiae Heterologous Expressing Clostridium thermocellum Endoglucanase and Saccharomycopsis fibuligera β-glucosidase Genes

  • Jeon, Eugene;Hyeon, Jeong-eun;Suh, Dong Jin;Suh, Young-Woong;Kim, Seoung Wook;Song, Kwang Ho;Han, Sung Ok
    • Molecules and Cells
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    • 제28권4호
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    • pp.369-373
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    • 2009
  • Heterologous secretory expression of endoglucanase E (Clostridium thermocellum) and ${\beta}$-glucosidase 1 (Saccharomycopsis fibuligera) was achieved in Saccharomyces cerevisiae fermentation cultures as an ${\alpha}$-mating factor signal peptide fusion, based on the native enzyme coding sequence. Ethanol production depends on simultaneous saccharification of cellulose to glucose and fermentation of glucose to ethanol by a recombinant yeast strain as a microbial biocatalyst. Recombinant yeast strain expressing endoglucanase and ${\beta}$-glucosidase was able to produce ethanol from ${\beta}$-glucan, CMC and acid swollen cellulose. This indicates that the resultant yeast strain of this study acts efficiently as a whole cell biocatalyst.