• 한국식품과학회지
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• 제30권1호
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• pp.22-34
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• 1998

고정화효소와 산소전극 시스템을 이용한 효소센서를 제작하여 식품 중의 당, 유기산, 알코올 성분을 동시 측정 하였다. 효소가 기질과 반응하여 소비한 산소의 변화량이 전압차이로 나타나므로 시간당 전압 감소량이 최대인 값으로부터 각 성분의 농도를 측정하였으며, 이때 1분내에 최대기울기를 구할 수 있어 신속한 측정이 가능하였다. 효소의 고정화 지지체로는 nylon cloth를 사용하였고, asymmetrical coupling 방법에 의하여 기질 작용 순으로 위치하도록 효소를 고정화하였다. 한 개의 양극과 6개의 음극으로 제작된 multiple cathode system으로 포도당, 젖산, 에탄올 성분을 동시 측정할 수 있는 효소 센서를 제작하였다. 위의 센서 제작을 위하여 mutarotase과 glucose oxidase/lactate oxidase/alcohol oxidase와 catalase가 각기 사용되었다. 이들 효소센서의 최적조건은 $pH\;7.0,\;40^{\circ}C$의 0.1 M 인산완충용액이었으며 각 효소 센서의 방해물질을 알아 보기 위하여 여러 가지 당과 각종 유기산, 알콜류에 대한 효소 감응도를 살펴 본 결과 포도당 센서에서 유기산의 영향을 제외하고는 10% 내외였다. 따라서 포도당과 유기산을 동시 측정하기 위하여 포도당/젖산의 영향을 고려한 적절한 보정관계식을 도입하여 순수한 유리당과 유기산의 값을 측정할 수 있었다. 제작된 효소센서의 검증을 위하여 분광광도법. HPLC, GC를 이용한 결과, 분석방법간에 높은 상관관계를 보여 주었다. 아울러 각 효소센서의 안정성을 살펴본 결과 알코올 센서를 제외하고는 30일 이후에도 80%이상 효소감응도가 유지되었다.

• 셀메드
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• 제4권1호
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• pp.6.1-6.12
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• 2014

Anthrax is the deadly disease for human being caused by Bacillus anthracis. Instantaneous research work on the mode of infection of the organism revealed that different proteases are involved in different steps of pathogenesis. Present study reports the in silico characterization and the detection of pathogenic proteases involved in anthrax infection through protein-protein interaction. A total of 13 acid, 9 neutral, and 1 alkaline protease of Bacillus anthracis were selected for analysing the physicochemical parameter, the protein superfamily and family search, multiple sequence alignment, phylogenetic tree construction, protein-protein interactions and motif finding. Among the 13 acid proteases, 10 were found as extracellular enzymes that interact with immune inhibitor A (InhA) and help the organism to cross the blood brain barrier during the process of infection. Multiple sequence alignment of above acid proteases revealed the position 368, 489, and 498-contained 100% conserved amino acids which could be used to deactivate the protease. Among the groups analyzed, only acid protease were found to interact with InhA, which indicated that metalloproteases of acid protease group have the capability to develop pathogenesis during B. anthracis infection. Deactivation of conserved amino acid position of germination protease can stop the sporulation and germination of B anthracis cell. The detailed interaction study of neutral and alkaline proteases could also be helpful to design the interaction network for the better understanding of anthrax disease.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.45-45
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• 2003

Human papillomavirus type 16(HPV16) has been known to the major factor for the development of uterine cervical carcinomas. We have extended these studies to investigate the in vivo activities of HPV-16 E6/E7 when expressed in squamous epithelia of transgenic mice. Grossly, hK14HPV16E6/E7 transgenic mice had multiple phenotypes, including wrinkled skin that was apparent prior to the appearance of hair on neonates, thickened ears, and loss of hair in adults. In the transgenic mice, the wrinkled skin phenotype on the body and legs died at the age of 3∼4 weeks. Histological analysis of demonstrated that E6/E7 causes epidermal hyperplasia in multiple transgenic lineages with high penetrance. This epithelial hyperplasia was characterized by an expansion of the proliferating compartment and an expansion of the keratinocyte and was associated with hyperkeratosis. These transgenic mice expressed E6/E7 transgene mainly in skin, heart, pancreas and kidney. Hyperplasia was found at the skin. The enzyme activities of GR, GPx and CuZnSOD were measured from the transgene cause keratinocyte at the skin. The specific enzyme activities were significantly higher in transgenic mice skin compared to the normal mice skin. Thus these transgenic mice may be useful for the develpment of antioxidant enzymes or other therapies for HPV-associated hyperkeratosis.

• Asian Pacific Journal of Cancer Prevention
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• 제15권22호
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• pp.9791-9795
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• 2014

To study the gene expression change and possible signal pathway during androgen-dependent prostate cancer (ADPC) becoming androgen-independent prostate cancer (AIPC), an LNCaP cell model of AIPC was established using flutamide in combination with androgen-free environment inducement, and differential expression genes were screened by microarray. Then the biological process, molecular function and KEGG pathway of differential expression genes are analyzed by Molecule Annotation System (MAS). By comparison of 12,207 expression genes, 347 expression genes were acquired, of which 156 were up-ragulated and 191 down-regulated. After analyzing the biological process and molecule function of differential expression genes, these genes are found to play crucial roles in cell proliferation, differntiation, cell cycle control, protein metabolism and modification and other biological process, serve as signal molecules, enzymes, peptide hormones, cytokines, cytoskeletal proteins and adhesion molecules. The analysis of KEGG show that the relevant genes of AIPC transformation participate in glutathione metabolism, cell cycle, P53 signal pathway, cytochrome P450 metabolism, Hedgehog signal pathway, MAPK signal pathway, adipocytokines signal pathway, PPAR signal pathway, TGF-${\beta}$ signal pathway and JAK-STAT signal pathway. In conclusion, during the process of ADPC becoming AIPC, it is not only one specific gene or pathway, but multiple genes and pathways that change. The findings above lay the foundation for study of AIPC mechanism and development of AIPC targeting drugs.

• Journal of Microbiology and Biotechnology
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• 제17권11호
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• pp.1826-1832
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• 2007

We describe the fabrication of poly(ethylene glycol) diacrylate (PEG-DA) hydrogel microstructures with a high aspect ratio and the use of hydrogel microstructures containing the enzyme ${\beta}$-galactosidase (${\beta}$-Gal) or glucose oxidase (GOx)/horseradish peroxidase (HRP) as biosensing components for the simultaneous detection of multiple analytes. The diameters of the hydrogel microstructures were almost the same at the top and at the bottom, indicating that no differential curing occurred through the thickness of the hydrogel microstructure. Using the hydrogel microstructures as microreactors, ${\beta}$-Gal or GOx/HRP was trapped in the hydrogel array, and the time-dependent fluorescence intensities of the hydrogel array were investigated to determine the dynamic uptake of substrates into the PEG-DA hydrogel. The time required to reach steady-state fluorescence by glucose diffusing into the hydrogel and its enzymatic reactions with GOx and HRP was half the time required for resorufin ${\beta}$-D-galactopyranoside (RGB) when used as the substrate for ${\beta}$-Gal. Spatially addressed hydrogel microarrays containing different enzymes were micropatterned for the simultaneous detection of multiple analytes, and glucose and RGB solutions were incubated as substrates. These results indicate that there was no cross-talk between the ${\beta}$-Gal-immobilizing hydrogel micropatches and the GOx/HRP-immobilizing micropatches.

• Journal of Pharmaceutical Investigation
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• 제6권2호
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• pp.69-82
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• 1976

Tablet product design problem was structured as constrained optimization problem and subsequently solved by multiple regression analysis and Lagrangian method of optimization. We used Lagrangian method for the purpose of finding the reason of the previous results. Biodiastase and cellulase were the enzymes, chosen, $Avicel{\circledR}$ and corn starch or calcium carboxy methyl cellulose were the binder and disintegrant, respectively. The effect of the dry binder and disintegrant concentration on tablet hardness, friability, volume, disintegration time was recorded. Optimization of this parameter was studied by using the constrained optimization method. In addition to finding a optimal condition of the enzyme tablets, the application of sensitivity analysis studies to such problems was also illustrated. In order to get a stable preparations of the enzyme tablets, accelerated test of coating tablets was carried out in this study. the results are as follows. 1) The minimum disintegration time, such that the average tablet volume did not exceed 0.0154 cubic inch and the average friability value did not exceed 0.62%, was 6.6 minutes and then $Avicel{\circledR}$ and corn starch were 15.4% and 17.2%, respectively. 2) The multiple-correlation coefficients for the regression models of tablet hardness, friability, disintegration time and volume were with in the 95% confidence range. 3) According to the test results, calcium carboxymethyl cellulose can be used as a disintegrant instead of corn starch.

• 동의생리병리학회지
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• 제22권3호
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• pp.580-584
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• 2008

In the present study, Radix clematidis extract (RCE) was evaluated to determine if it could protect pancreatic ${\beta}$ cells against multiple low dose streptozotocin (MLDS)-induced diabetes. Injection of mice with MLDS resulted in hyperglycemia and hypoinsulinemia, which was confirmed by immunohistochemical staining. However, the induction of diabetes by MLDS was completely prevented when mice were pre-administrated with RCE. Generation of oxidative stress is implicated in MLDS, a ${\beta}$ cell specific toxin-induced islet cell death. In this context, to elucidate the mechanisms of protective effects in RCE pre-administrated diabetic mice, we investigated the expression of heme oxygenase-1 (HO-1), which is one of the anti-oxidant enzymes. MLDS-induced HO-1 expressions were significantly reduced in MLDS-treated mice. However, the decrease of HO-1 by MLDS were protected by pretreatment of RCE. The molecular mechanism by which RCE inhibits diabetic conditions by MLDS appears to involve inhibition of HO-1 expression. Taken together, these results reveal the possible therapeutic value of RCE for the prevention of type 1 diabetes progression.

• 대한약리학회지
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• 제32권2호
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• pp.233-241
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• 1996

Previous studies have shown that glucocorticoid suppresses microsomal epoxide hydrolase(EH) gene expression and that EH expression is altered during pregnancy. The effects of progesterone on the expression of rat EH and certain glutathione S-transferase(GST) genes were examined in this study. Northern RNA blot analysis revealed that progesterone was effective in increasing hepatic EH mRNA levels at 12 h to 48 h after treatment with a maximal 9-fold increase being noted at 12 h time point. Nonetheless, multiple daily treatment with progesterone rather caused minimal relative increases in EH mRNA levels. GST Ya and Yb1/2 mRNA levels were also transiently elevated at 12 h after progesterone treatment, followed by gradual decreases from the maximal Increases at day 1, 2 and 5 post-treatment. These changes in EH and GST mRNA levels were noted only at a relatively high dose of progesterone. Furthermore, immunoblot analyses showed that rats treated with progesterone for 5 days failed to show EH or GST induction, indicating that progesterone-induced alterations in EH and GST mRNA levels do not reflect bona fide induction of the detoxifying enzymes. Concomitant progesterone treatment of rats with the known EH inducers including ketoconazole and clotrimazole failed to additively nor antagonistically alter EH mRNA levels. In contrast, dexamethasone substantially reduced ketoconazole- or clotrimazole-inducible EH expression. These results showed that progesterone stimulates the EH, GST Ya and Yb1/2 gene expression at early times followed by marked reduction in the RNA levels from the maximum after multiple treatment and that the changes in mRNA do not necessarily reflect induction of the proteins.

• 한국균학회지
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• 제18권4호
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• pp.209-217
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• 1990

Aspergillus niger에 있어서 여러가지 섬유질 기질이 세포내외의 섬유질 분해효소계의 생합성 및 동질효소의 양상에 미치는 영향을 조사하였다. Cellulase 및 xylanase 효소계의 생합성은 사용된 기질에 따라 큰 차이가 있었으며, 특히 cellulase 효소계의 CMCase와 ${\beta}-glucosidase$는 혼합기질의 사용시 효소활성이 증진되는 기질의 공조효과를 보였다. 또한 기질에 따라 섬유질분해효소계 동질효소의 생성 양상이 다르게 나타났으나 세포내외간 동질효소 양상의 차이는 발견되지 않았으며 배양시간에 따른 동질효소의 변화도 없었다. 이러한 결과들은 A. niger의 cellulase 및 xylanase효소계의 생합성은 기질에 의한 효소의 유도 수준에서 상호 연관적으로 조절되어짐을 보여주며, 세포외 효소에서 나타나는 multiple-isozyme의 형성이 합성되어진 효소의 post-secretional modification에 의한 결과라기 보다는 각각의 유전자 발현 결과에 의한 것임을 시사한다.

• Journal of Microbiology and Biotechnology
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• 제17권8호
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• pp.1344-1352
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• 2007

An opportunistic human pathogen, Pseudomonas aeruginosa, contains the major catalase KatA, which is required to cope with oxidative and osmotic stresses. As an attempt to uncover the $H_2O_2$-dependent regulatory mechanism delineating katA gene expression, four prototrophic $H_2O_2$-sensitive mutants were isolated from about 1,500 TnphoA mutant clones of P. aeruginosa strain PA14. Arbitrary PCR and direct cloning of the transposon insertion sites revealed that one insertion is located within the katA coding region and two are within the coding region of oxyR, which is responsible for transcriptional activation of several antioxidant enzyme genes in response to oxidative challenges. The fourth insertion was within PA3815 (IscR), which encodes a homolog of the Escherichia coli iron-sulfur assembly regulator, IscR. The levels of catalase and SOD activities were significantly reduced in the iscR mutant, but not in the oxyR mutant, during the normal planktonic culture conditions. These results suggest that both IscR and OxyR are required for the optimal resistance to $H_2O_2$, which involves the expression of multiple antioxidant enzymes including KatA.