• 한국생물공학회:학술대회논문집
• /
• 2005.04a
• /
• pp.327-333
• /
• 2005

Dihydroxy-acid dehydratase (DHAD, 2,3-dihydroxy-acid hydrolyase, EC 4.2.1.9) is one of the key enzymes involved in the biosynthetic pathway of the branched chain amino acid isoleucine and valine. Although the enzyme have been purified and characterized in various mesophiles including bacteria and eukarya, the biochemical properties of DHAD has bee not yet reported from hyperthermophilic archaea. In this study, we cloned, expressed, and purified a DHAD homologue from the thermoacidophilic archaeon Sulfolobus solfataricus P2, which grows optimally at $80\;^{\circ}C$ and pH 3, in E. coli. Characterization of the recombinant S. solfataricus DHAD (rSso_DHAD) revealed that it is the dimeric protein with a subunit molecular weight of 64,000 Da in native structure. rDHAD showed the highest activity toward 2,3-dihydroxyisovaleric acid among 17 aldonic acid substrates Interestingly, this enzyme also displayed 50 % activities toward some pentonic acids and hexonic acids when compared with the activity of this enzyme to the natural substrate. Moreover, rSso_DHAD indicated relatively higher activity toward D-gluconate than any other hexonic acids tested in substrates. $K_m$ and $V_{max}$ values of rSso_DHAD were calculated as $0.54\;{\pm}\;0.04\;mM$ toward 2,3dihydroxyisovalerate and $2.42\;{\pm}\;0.19\;mM$ toward D-gluconate, and as $21.6\;{\pm}\;0.4\;U/mg$ toward 2,3-dihydroxyisovalerate and $13.8\;{\pm}\;0.4\;U/mg$ toward D-gluconate, respectively. In the study for biochemical properties, the enzyme shows maximal activity between $70^{\circ}C$ and $80^{\circ}C$, and the pH range of pH 7.5 to 8.5. The half life time at $80^{\circ}C$ was 30 min. A divalent metal ion, $Mn^{2+}$, was only powerful activators, whereas other metal ions made the enzyme activity reduced. $Hg^{2+}$, organic mercury, and EDTA also strongly inhibited enzyme activities. Particularly, the rSso_DHAD activity was very stable under aerobic condition although the counterparts reported from mesophiles had been deactivated by oxygen.

• Culinary science and hospitality research
• /
• v.9 no.4
• /
• pp.113-122
• /
• 2003

This study was conducted to examine the antagonism of useful microbes against soybean sprout rotting pathogens and their effect on the growth of soybean sprouts. The antagonism against soybean sprout rotting pathogens and the effect on the growth of soybean sprouts were examined by using P. areofacience 14H-3, P. fluorescens R1-12 and B. cereus Yell, bacteria were shown to inhibit mycellial growth of Rhizotonia solani strongly. The results of this study are summarized as follows. P. areofacience 14H-3 and B. cereus Yell were highly antagonistic against Rizoctonia solani, while they were especially highly antagonistic against bacterial diseases. The effect of inhibiting the proliferation of soybean sprout rotting pathogens was also examined by adding the culture solution for antagonistic bacteria to the PDA. Both P. areofacience 14H-3 and P. fluorescens Rl-12 showed the inhibition rate of 78.8%, while B. cereus Yell did 52.9%. The fresh weight and length of soybean sprouts were measured after raising them with added antagonistic microbes and culture medium. Soybean sprouts treated with B. cereus Yell showed increased higher, compared with those not treated with it. Soybean sprouts were also raised in the culture solution with antagonistic bacteria to examine the growth of soybean sprouts. Soybean sprouts treated with the culture solution of 200 times showed better growth than those not treated with it. Analyze proximate composition in soybean sprout showed that moisture, ash, total sugar did not appear difference, but in case of crude protein B. cereus Yell(8.9%) increased about 2 times than control(3.6%), but occasion of crude fat and crude fiber were P. areofacience 14H-3, P. fluorescens Rl-12 increased about each 2 times than control. In occasion of vitamin, bacterial antagonist(9.4∼10.8mg%) was more higher than control(9.9mg%).

• Development and Reproduction
• /
• v.7 no.1
• /
• pp.15-22
• /
• 2003

The Fas antigen (Fas) as a cell-surface receptor protein which mediates apoptosis-inducing signals plays an important role in the immune system. Expression of Fas mRNA is detected not only in lymphoid organs but also in the nonlymphoid organs. In the ovary, most of the follicles is known to undergo atreisa through apoptosis. However, the exact mechanism of atresia was not elucidated yet. Therefore, the purposes of the present study were to investigate the expression of Fas and Fas ligand in mouse ovary and to clarify the relationship between expression of Fas and Fas ligand and atresia of follicle. The result of RT-PCR demonstrated that Fas and Fas ligand mRNA was expressed in ovary, especially granulosa cells and oocytes. The immunohistochemistry showed that the granulosa cells and oocytes in growing follicles were stained for Fas and Fas ligand, but primordial follicles were not. Furthermore, Fas and Fas ligand were intensively stained in the atretic follicles As results of TUNEL staining to detect apoptotic cells in the ovaries, the number of TUNEL-positive (apoptotic) granulosa cells and oocytes increased in the atretic follicles compared to the healthy normal follicles. These results demonstrate that there is the positive relationship between expression of Fas and Fas ligand in granulosa cells and oocyies and apoptosis of them leading to atresia of follicles. It suggests that expression of Fas and Fas ligand could be associated with atresia of follicles in mouse ovary.

• Journal of Life Science
• /
• v.18 no.2
• /
• pp.180-186
• /
• 2008

In this study, we examined the effects of the fermented soybeans by Bacillus subtilis (FSB) and the novel three-step fermented soybeans (TFS) on the expression and activity of COX-2 in human leukemic U937 cell model. Treatment of phorbol 12-myristate 13-acetate (PMA) significantly induced pro-inflammatory mediators such as COX-2 expression and prostaglandin $E_2\;(PGE_2)$ production, whereas the levels of COX-1 remained unchanged. However, pre-treatment with FSB and TFS significantly attenuated the PMA-induced COX-2 protein as well as mRNA, which was associated with inhibition of $PGE_2$ production. Moreover, TFS exerts a much better inhibitory activity than FSB against PMA-induced activation of COX-2 and production of $PGE_2$ in U937 cells. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-inflammatory activity of FSB and TFS.

• Food Science and Preservation
• /
• v.11 no.3
• /
• pp.342-346
• /
• 2004

Chemical components in the peel and flesh of full riped trifoliate oranges(Poncirus trifoliata) were investigated. Contents of crude protein, crude fat and ash in peel and flesh were 5.15 and 3.31$\%$, 3.82 and 6.65$\%$, 2.62 and 2.09$\%$, respectively. Vitamin C contents were 4.70 mg$\%$ in peel and 70.93 mg$\%$ in flesh. Free sugars were fructose, glucose and sucrose, the level of each or total free sugars was twice higher in peel than in flesh. Organic acids were citric acid, malic acid and oxalic acid, the total contents in peel and flesh were 0.35 and 1.25$\%$, respectively. Free amino acids were aspartic acid, histidine, tyrosine, arginine, valine, lysine, ammonia, cysteine, alanine, glutamic acid, isoleucine, leucine in peel, and lysine, valine, ammonia, arginine, tyrosine, isoleucine, methionine, leucine, histidine, phenylalanine in flesh.

• Applied Biological Chemistry
• /
• v.36 no.5
• /
• pp.321-325
• /
• 1993

The anchovy-based powder for instant soup packed in twenty tea bag were repacked with (Product B) or without oxygen absorber (Product A) in laminated film bag $(PVDC/OPP,\;thickness:\;100.5\;{\mu}m,\;size:22{\times}18cm)$, and then stored at ambient temperature $(25{\pm}3^{\circ}C)$. Moisture, crude protein and crude lipid contents in anchovy-based powder for instant soup were 12.1%, 54.7% and 2.9%, respectively. Moisture content showed little changes during storage in both product (A) and (B). pH and saturated fatty acid such as 20 : 5 and 22 : 6 decreased, while volatile basic nitrogen content, carbonyl value, thiobarbituric acid value, monoenoic fatty acid such as 16 : 0, brown pigment formation and Hunter values increased during storage of product (A). But, these values showed a little change during storage of product (B). It is concluded that anchovy-based powder for instant soup can be handily and safely used during storage of 150 days.

• IMMUNE NETWORK
• /
• v.13 no.4
• /
• pp.116-122
• /
• 2013

This study was conducted to determine whether CD4 T cell responses to citrullinated fibrinogen occur in patients with rheumatoid arthritis (RA), especially in HLA-DR4-positive subjects. Whole peripheral blood mononuclear cells (PBMCs) of RA patients and control subjects were stimulated with citrullinated fibrinogen peptides, and T-cell production of proliferation and proinflammatory cytokines, such as interferon-${\gamma}$(IFN-${\gamma}$) and interleukin-17A (IL-17A), were measured. In addition, CD4 T cells from RA patients were stimulated with the citrullinated fibrinogen peptide, $Fib-{\alpha}$ R84Cit, identified as a DRB1*0401-restricted T cell epitope in HLA-DR4 transgenic mice, and the degree of T cell activation was examined similarly. No proliferative responses to the citrullinated fibrinogen peptides were observed in whole PBMCs or CD4 T cells from RA patients. Furthermore, no increased production of IFN-${\gamma}$ or IL-17A was found in whole PBMCs or CD4 T cells stimulated with the citrullinated fibrinogen peptides, although these cells responded to recall antigen, a mixture of tetanus toxoid, purified protein derivative (PPD) from Mycobacterium tuberculosis, and Candida albicans. The results of this study indicate that anti-citrulline immunity in RA patients may be mediated by fibrinogen because there is no evidence of CD4 T cell-mediated immune responses to citrullinated fibrinogen peptides.

• Maxillofacial Plastic and Reconstructive Surgery
• /
• v.30 no.1
• /
• pp.41-51
• /
• 2008

Mucoepidermoid carcinoma (MEC) is the most common malignant salivary gland tumor which compromises about 6$\sim$8% of all tumors followed by the adenoid cystic carcinoma (ACC) and adenocarcinoma. Most deaths from salivary carcinomas are caused by recurrent or metastatic lesions that are resistant to conventional therapy. Therefore, knowledge of cellular properties and tumor-host interactions that influence the vascular metastasis is important for the design of more effective therapy of salivary carcinomas. Neoangiogenesis is essential for tumor growth, which is postulated to be fundamentally dependent on the induction of stromal neovascularization. However, how neovascularization takes place in live tissue has not been fully established, especially in recruitment and differentiation of endothelial cells in the salivary gland tumors. Vascular endothelial growth factor (VEGF) is a heparin-binding, dimeric polypeptide growth factor known to exert its mitogenic activity specifically on endothelial cells. VEGF has been shown th be directly involved in angiogenesis, which in essential for the pathogenesis of many solid tumors. von Willebrand factor (vWF) is a large multimeric protein synthesized by megakaryocytes and endothelial cells that enable platelets to adhere to exposed subendothelium and, as well, to respond to changes in the blood flow. Recent studies suggest that increased levels of vWF correlate with progression of disease, metastasis, or survival time and thus may have a prognostic significance. vWF is explained as an acute phase proteins which is increased in cancer or as a result of increased endothelial cell synthesis associated with tumor-induced angiogenesis. Due to adhesive properties of vWF, its increased concentrations may also contribute metastasis of tumor. In this study, we determined the mRNA expression of VEGF and vWF in salivary ACC, MEC and pleomorphic adenoma by in situ hybridization. As a result, stronger expression of VEGF and vWF was seen in salivary ACC and MEC which has more invasive nature than the salivary benign tumor.

• Journal of Periodontal and Implant Science
• /
• v.28 no.4
• /
• pp.769-786
• /
• 1998

The purpose of the present study is to examine the effect of cell proliferation and alkaline phosphatase activity in osteoblastic cells and to compare the bone healing ability of rat calvarial defects between the control group and the safflower seed extract treated group. Osteoblastic cells were obtained from calvariae of a fetal rat. Cells were cultured containing DMEM and safflower seed extract ($10^{-6}g/ml$, $10^{-3}g/ml$) at $37^{\circ}$ with 5% $CO_2$ in 100% humidity for 3 days. MTT was performed to examine the viability of the cells, and alkaline phosphatase activity was analyzed to examine the mineralization in vitro. Rat calvarial defects($5{\times}5mm$) in 250g Sprague-Dawly were made using round bur. Rats were administrated with safflower seed extract(0.35g/kg/day) for experimental periods. Calvarial defects were studied histopathologically and immunohistochemically at 1,4, and 8 weeks. High concentration group($10^{-3}g/ml$) of safflower seed extract significantly increased in the cell proliferation and alkaline phos phatase synthesis in osteoblastic cells. The infiltration of inflammatory cells and osteoclastic activities were decreased in the safflower seed extract treated group as compared with control group. Bone maturation was accelerated in the safflower seed extract treated group as compared to control group. No difference in osteoinductive process was observed between the control and the safflower seed extract treated group. Immunohistochemical observation revealed that protein expression of TGF-$\beta$and osteonectin during early healing phase in the safflower seed extract treated group was slightly increased as compared to control group. These results indicate that safflower seed extract promotes the healing process in bony defect of rat calvariae, and retains a potential applicability as an adjuvant therapeutic modality for regeneration of periodontal bony defect.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.34 no.3
• /
• pp.468-472
• /
• 2007

The Wiskott-Aldrich Syndrome (WAS) is an inherited immunodeficiency caused by a variety of mutations in the gene encoding the WAS protein (WASp). First described in 1937 by Wiskott, the incidence of WAS has so far been estimated at 4 in 106 live births. The Wiskott-Aldrich Syndrome is an X-linked condition characterized by 1) an increased tendency to bleed caused by a reduced number of platelets, 2) recurrent bacterial, viral and fungal infections, and 3) eczema of the skin. The purpose of this report is to present cases highlighting the clinical features of the syndrome and the required considerations in the treatment of patients. The report consists of two particular cases: a 2-year-11-month-old boy seen for a routine oral examination prior to his bone marrow transplantation and a 2-year-6-month-old boy with herpes gingivostomatitis and teeth discoloration.