• Title/Summary/Keyword: SGT

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Medium Optimization and Application of Affinity Column Chromatography for Trypsin Production from Recombinant Streptomyces griseus

  • Chi, Won-Jae;Song, Ju-Hyun;Oh, Eun-A.;Park, Seong-Whan;Chang, Yong-Keun;Kim, Eung-Soo;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1191-1196
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    • 2009
  • The production of Streptomyces griseus trypsin (SGT) by S. griseus IFO13350 transformed with the expression vector pWHM3-TR1R2, containing sprT encoding SGT and the two positive regulatory genes sgtR1 and sgtR2, was investigated in various media. Cultivation in Ferm-0 gave 1.4 times more trypsin activity than in C5/L medium. In addition, replacement of 2% glucose and 1% skim milk in Ferm-0 with 2% dextrin and 1% tryptone (designated Ferm-II) enhanced trypsin activity 4.1-fold. To simplify the purification process, the supernatant from the S. griseus transformant cultured in Ferm-II medium was fractionated with ammonium sulfate (25-55%), then subjected to Hitrap Benzamidine FF affinity column chromatography. The specific activity of SGT purified by one-step chromatography was 69,550 unit/mg protein and the overall purification yield was above 8%, indicating that this method is more effective than those previously reported. Purified SGT was most active at pH 8.0 and $50^{\circ}C$, and it maintained activity between pH 7.0 and 9.0 and at temperatures up to $70^{\circ}C$. These enzymatic properties are very similar to those of authentic eukaryotic trypsin purified from bovine pancreas.

Decreased Serum Monocyte Chemoattractant Protein-1 in Salivary Gland Tumor Patients

  • Mardani, Maryam;Andisheh-Tadbir, Azadeh;Khademi, Bijan;Melekzadeh, Mahyar;Vaziri, Lida
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3601-3604
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    • 2016
  • Background: The monocyte chemoattractant protein-1 (MCP-1/CCL2) is a potent chemoattractant for natural killer cells, monocytes, and memory T lymphocytes. However, any role in the genesis of salivary gland tumors (SGT) is unknown. To assess the diagnostic relevance of chemokines in SGT, MCP-1 levels in the serum of patients were investigated in association with tumor progression and clinical aggressiveness. Materials and Methods: Using an ELISA kit, we assessed and compared the circulating levels of MCP-1 in blood serum of 70 SGT patients with 44 healthy control samples. Results: The results of this study showed that the concentration of MCP-1 was significantly lower in patients with benign ($463.8{\pm}158.5pg/ml$, P=0.033) and malignant ($454.8{\pm}190.4pg/ml$, P=0.007) SGT than in healthy subjects ($645.7{\pm}338.9$). No significant difference in mean serum levels of MCP-1 was observed between the benign and malignant group (p=0.9). While MCP-1 levels were lower in patients with an advanced clinical stage, advanced tumor size, higher tumor grade, or lymph node involvement, but the mean MCP-1 level between groups showed no statistically significant difference (p>0.05). Conclusions: MCP-1 levels in the serum of patients with SGT were decreased, indicating that this might a good marker for discriminating patients with SGT from healthy people. However, no clear-cut relationship was detected between MCP-1 levels and clinicopathologic factors, and MCP-1 is not a good marker for evaluating tumor dissemination.

Fine Needle Aspiration Cytology of Subacute Granulomatous Thyroiditis -A Clinico-Cytological Review of 10 Cases with Immunocytochemical Analysis- (아급성 육아종성 갑상샘염의 세침흡인 세포소견 -아급성 육아종성 갑상샘염 10예의 임상-세포소견 및 면역세포염색 소견 분석-)

  • Kim, Do-Kyung;Pyo, Ju-Yeon;Park, Jong-Pil;Jin, Lian-Hua;Jeong, Woo-Hee;Son, Eun-Ju;Hong, Soon-Won
    • The Korean Journal of Cytopathology
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    • v.19 no.1
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    • pp.27-33
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    • 2008
  • Although subacute granulomatous thyroiditis(SGT) is usually diagnosed clinically, papillary carcinoma or other thyroid conditions must be considered in the differential diagnosis. We retrospectively reviewed the clinical and fine-needle aspiration(FNA) cytologic findings seen in 10 SGT cases to decide what are the most reliable cytologic findings and the most helpful molecular tools for reaching a confident cytologic diagnosis. The most representative smear slides were retrieved to perform immunocytochemistry for cytokeratin 19(CK19) and Ret protein. Five papillary carcinomas(PTCs) were included as controls. The constant and typical cytologic findings of SGT were multinucleated giant cells(MGCs) (100%), epithelioid granulomas(90%), an inflammatory dirty background(90%) and plump transformed follicular cells(80%) without fire-flare cells, oncocytic cells or transformed lymphocytes. The immunoreactivities for CK19(37.5%) and Ret(10%) of the follicular cells of SGT were less than those(CK19 and Ret:100%) of PTC. CK19 immunoreactivity of the MGCs was seen in only one case of PTC. There was no significant difference between CK19 and Ret immunocytochemical staining for the MGCs of both SGT and PTC. The results of this study demonstrate that the cytological diagnosis of SGT can be improved by employing a combination of the typical and constant diagnostic cytological features and immunocytochemical results.

Gene Expression Profiling of Human Salivary Gland Carcinogenesis with cDNA Microarray (인간 타액선 암발생에서 cDNA Microarray를 이용한 유전자발현 Profile연구)

  • Kim, Eun-Cheol;Shin, Min;Lee, Dong-Geun;Lee, Ju-Seok;Park, Myung-Hee
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.23 no.4
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    • pp.306-323
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    • 2001
  • 종양발생에서 유전자 발현을 확인하고 profile 변화를 monitor하는 것은 병리학적 변화의 원인뿐 아니라 질병탐지와 진료의 새로운 목표를 확인하기 위한 새로운 기회를 제공해준다. cDNA microarray는 수천개의 유전자 발현을 동시에 연구할 수 있는 최신의 방법으로 피부, 유방, 간을 비롯한 다른 인체장기에서는 일부 이루어졌으나 array를 이용해 타액선 종양 연구에서는 전혀 이루어지지 않았다. 인간의 타액선 세포의 악성형질전환을 조절하는 분자적 상태를 연구하기 위해 본 연구는 약 2,000개의 유전자가 print된 cDNA microarray를 이용하여 인간 타액선 도관상피세포주(HSG)와 악하선에서 기원한 미분화 선암종(SGT)간에 비교연구를 하였다. Cy3와 Cy5 dye로 각각의 세포주에서 얻은 RNA와 reciprocal hybridize시키고 GenePix 4000 scanner로 스캔하고 GenePix Pro로 분석한 후 log2로 평균발현비율을 전환시켜 최소 2배이상의 발현을 보이는 유전자를 분석대상으로 하였다. 90%이상의 유전자가 비슷한 발현을 보였으며 2배이상의 발현을 보이는 경우 HSG가 SGT에 비해 72개 유전자가, SGT가 HSG에 비해 111개의 유전자 발현이 up-regulation되어 총 10%미만의 발현차이를 보였고 반복된 hybridization 으로부터 얻은 선택된 spot의 Pearson 상관계수는 -0.85이였다. HSG에서는 6번 p 염색체에서 과발현되는 유전자가 가장 많았고, SGT에서는 11번 q 염색체에서 가장 많았는데 HSG에서는 SGT에 비해 9, 13, 17, 18, 20, 21, 22염색체에서 과발현 되는 유전자 수가 많았고, SGT에서는 HSG에 비해 2, 7, 10, 15 염색체에서 유전자 발현 증가가 관찰되었다. HSG와 SGT간의 유전 발현을 기능별로 분석한 결과 몇 가지 주요 경로가 세포악성에 관련됨을 발견하였고, 타액선 도관상피세포에서 선암종을 구별하는데 기여하는 관련된 몇종의 과다 발현된 유전자를 찾았는데 전사인자, 성장인자 및 수용기, 세포골격 및 세포외기질 단백, 세포내 신호전달조절자 및 인자, 세포표면 항원등의 그룹으로 분류할 수 있었다. 따라서 이러한 microarray를 이용한 분자학적 표지자 연구가 악성 타액선 종양 발생과정에서 큰 도움을 줄 수 있을 뿐 아니라 유전자 조절에 의한 진단, 예후, 치료에서의 정확성을 개선시킬 수 있으리라 여겨진다.

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Study in the Hepatoprotective Effect of Sipyimiguanjung-tang and Osuyubujaijung-tang (십이미관중탕(十二味寬中湯)과 오수유부자리중탕(吳茱萸附子理中湯)의 간손상(肝損傷) 보호작용(保護作用)에 대한 연구)

  • Kim, Hyoung-Soon;Bae, Young-Chun;Lee, Sang-Min;Kim, Kyung-Yo;Won, Kyung-Sook;Lee, Kyung-Seong
    • Journal of Sasang Constitutional Medicine
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    • v.15 no.1
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    • pp.90-108
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    • 2003
  • Osuyubujaijung-tang(OBT) and Sipyimiguanjung-tang(SGT) has been developed as prescriptions for the Soyeumin constitution. The hepatoprotective effect of the water extract of Osuyubujaijung-tang(OBT) and Sipyimiguanjung-tang(SGT) was investigated against carbon tetrachloride (CCl4)-induced hepatic damage. A single intra-peritoneal injection of CCl4 produced liver damage in rats as manifested by the significant rise of aspartate aminotransferase(AST), alanine aminotransferase(ALT), and alkaline phosphatase(ALP) in serum as compared to those of untreated normal group. Pretreatments of rats with Osuyubujaijung-tang(OBT) and Sipyimiguanjung-tang(SGT) 500 mg/kg for 7 days) were significantly reduced AST, ALT, and ALP levels compared with CCl4-treated control group. Treatment of rats with CCl4 led to significantly increase in lipid peroxidation and significantly decrease in cytochrome P450 and P450 reductase. The oral administration of Osuyubujaijung-tang(OBT) and Sipyimiguanjung-tang(SGT) water extract significantly inhibited the accumulation of microsomal thiobarbituric acid reactive substance (TBARS) and increased the cytochrome P450 and P450 reductase activity. All these biochemical alterations resulting from CCl4 administration were inhibited by the pretreatment with Osuyubujaijung-tang(OBT) and Sipyimiguanjung-tang(SG1) extract. These results suggest that Osuyubujaijung-tang(OBT) and Sipyimiguanjung-tang(SGT) water extract can be useful as a hepatoprotective agent. And the effect of NO modulation by NO synthesis or precursors, and Osuyubujaijung-tang(OBT) and Sipyimiguanjung-tang (SGT) water extract was researched on chronic liver damage induced by CCl4 administration. It was observed that endogenous NO protected the liver from lipid peroxidation, fibrosis, and damage. Osuyubujaijung-tang(OBT) and Sipyimiguanjung-tang(SGT) water extract showed the hepatoprotective effect on the chronic liver cirrhosis model and relationship with NO modulation.

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Several Growth Properties of Pes-gallinaceua (Corydalis, Fumariaceae) Group (현호색속 Pes-gallinaceua절 집단의 생장특성)

  • 민병미
    • The Korean Journal of Ecology
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    • v.26 no.4
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    • pp.181-188
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    • 2003
  • Frequencies of each size class, annual change of tuber state and above ground/belowground rate in sect. Pes-gallinaceua group of Corydalis (Fumariaceae) were studied to clarify spring ephemeral´s growth characteristics from 1999 to 2000 in Namhansansung area. Shoot grew from the tuber corresponding with soil thawing in the early growth season, and the deeper tuber was located in soil, the later shoot germinated. Shoot appeared in middle March and died early in May, and the period of total growth season was 45 days. Frequencies were 40% (in total dry weight) or 53% (in tuber´ dry weight) in the smallest size class, and 93% (in total dry weight) or 96% (in tuber´ dry weight) below medium size, therefore, almost all of plants were small. Specific gravity of tuber (SGT) gradually decreased from late growth season to the time that leaf unfolded in next year. At this time, the value of SGT was 0.14. Thereafter, SGT increased to 0.42 at withering time of aboveground part. During growth season, the SGT of flowering plants was lower than that of non-flowering plants. And SGT difference between flowering plants and non-flowering plants was significant at 1% level. In the late growth season, SGT decreased along the increase of tuber volume. The tuber volume roughly increased along the leaf area but there was no a conspicuous trend. Shoot/tuber rate had nothing to do with tuber volume. SGT decreased and leaf area increased along the tuber dry weight, and tendencies of increase or decrease were conspicuous. Therefore, the smaller the plants were, the larger organic matter accumulated in tuber during a growth season. Shoot/tuber rates were various along each plant.

Molecular Cloning and Analysis of the Genes in the Vicinity of Streptomyces griseus Trypsin (SGT) Gene from Streptomyces griseus ATCC10137 (Streptomyces griseus ATCC10137에서 Trypsin 유전자 sprT의 주변 유전자군 분석)

  • Chi Won-Jae;Kim Mi-Soon;Kim Jong-Hee;Kang Dae-Kyung;Hong Soon-Kwang
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.255-261
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    • 2005
  • A 6.7kb DNA fragment containing the sprT gene encoding Streptomyces griseus trypsin (SGT) was cloned from Streptomyces griseus ATCC 10137, and the complete nucleotide sequence was determined. Nucleotide sequence and deduced amino acid or the EcoRI-HindIII fragment revealed the presence or the six complete ORFs containing the sprT gene and one incomplete ORF, which were named ORF1, SGT, ORF2, ORF3, ORF4, ORF5, and ORF6, respectively. ORF1 has homology with the oxidoreductases from several organisms. ORF2 and ORF3 show similarity with unknown proteins and transcription regulator that belongs to the ArsR family, respectively. ORF4 and ORF5 show homology with the peptidoglycan bound protein with LPXTG motif from Listeria monocytogenes and the membrane protein with transmembrane helix from several organisms, respectively. The last ORF, ORF6, shows homology with the lipoprotein from Streptomyces avermitilis.

Effects of Growth Conditions on Structural and Optical Properties of ZnS Nanoclusters (용액성장법의 성장조건이 ZnS 나노클러스터의 구조적, 광학적 특성에 미치는 영향)

  • 이상욱;이종원;조성룡;김선태;박인용;최용대
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2001.07a
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    • pp.558-561
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    • 2001
  • In this study, the ZnS nanosized thin films were grown by the solution growth technique (SGT), and their structural and optical properties were examined. X-ray diffraction patterns showed that the ZnS thin film obtained in this study had the cubic structure ($\beta$-ZnS). With decreasing growth temperature and decreasing concentration of precursor solution, the surface morphology of film was found to be improved. In particular, this is the first time that the surface morphology dependence of ZnS film grown by SGT on the ammonia concentration is reported. The energy band gaps of samples were shown to vary from 3.69 eV to 3.91 eV, demonstrating that the quantum size effect of SGT grown ZnS is remarkable. Photoluminescence (PL) peaks were observed at the positions corresponding to the lower energy than that to energy band gap, illustrating that the surface states were induced by the ultra-fineness of grains in ZnS films.

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Simultaneous Analysis of Liquiritin and Glycyrrhizin in Sagunja-tang by HPLC-PDA (HPLC-PDA에 의한 사군자탕 중 Liquiritin과 Glycyrrhizin의 동시분석)

  • Seo, Chang-Seob;Kim, Jung-Hoon;Shin, Hyeun-Kyoo
    • Korean Journal of Pharmacognosy
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    • v.42 no.3
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    • pp.240-245
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    • 2011
  • A high-performance liquid chromatography (HPLC) method was developed for quantitative analysis of liquiritin and glycyrrhizin in Sagunja-tang (SGT, Sijunzi-tang in Chinese), a traditional Korean medicine. HPLC analysis was performed using a Gemini C18 column operating at $40^{\circ}C$, and photodiode array (PDA) detection at 254 nm and 280 nm for quantification of the two components in SGT. The mobile phase using a gradient flow consisted of two solvent systems. Solvent A was 1.0% (v/v) aqueous acetic acid and solvent B was acetonitrile with 1.0% (v/v) acetic acid. Calibration curves were acquired with $r^2$ values > 0.9998, and the relative standard deviations (RSDs, %) for intra- and inter-day precision were not exceed 4.0%. The recovery of each component was in the range of 91.85 - 108.62%, with a RSD less than 4.0%. The contents of the two components in SGT were 7.94 - 13.83 mg/g.

Sagantang-induced Apoptotic Cell Death is Associated with the Activation of Caspases in AGS Human Gastric Carcinoma Cells (사간탕 처리에 의한 AGS 인체 위암세포의 caspase 활성 의존적 apoptosis 유발)

  • Park, Cheol;Hong, Su Hyun;Choi, Sung Hyun;Lee, Se-Ra;Leem, Sun-Hee;Choi, Yung Hyun
    • Journal of Life Science
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    • v.25 no.12
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    • pp.1384-1392
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    • 2015
  • Sagantang (SGT), a Korean multiherb formula comprising six medicinal herbs, Paeonia lactiflora Pall., Belamcanda chinensis (L.) DC, Gardenia jasminoides Ellis, Poria cocos Wolf, Cimicifuga heracleifolia Komarov, and Artractylodes japonica Koidzumi, was recorded in “Dongeuibogam.” The present study investigated the anticancer potential of SGT in AGS human gastric carcinoma cells. The results indicated that SGT treatment significantly inhibited the growth and viability of AGS cells in a dose-dependent manner, which was associated with the induction of apoptotic cell death, as evidenced by the formation of apoptotic bodies, in addition to chromatin condensation and DNA fragmentation, and the accumulation of annexin-V positive cells. The induction of apoptotic cell death by the SGT treatment was associated with up-regulation of Fas protein expression, truncation of Bid, and down-regulation of the anti-apoptotic Bcl-2 protein. The SGT treatment also effectively induced the loss of mitochondrial membrane potential, which was associated with the activation of caspases (caspase-3, -8, and -9) and degradation of poly (ADP-ribose) polymerase. However, a pan-caspase inhibitor significantly blocked the SGT-induced apoptosis and growth suppression in AGS cells. This study suggests that SGT induces caspase-dependent apoptosis through an extrinsic pathway by upregulating Fas, as well as through an intrinsic pathway by modulating Bcl-2 family members in AGS cells. The results suggest that SGT may be a potential chemotherapeutic agent for the control of human gastric cancer cells. However, further studies will be needed to confirm the potential of SGT in cancer prevention and therapy in an in vivo model and to identify biological active compounds of SGT.