• Journal of Life Science
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• v.30 no.5
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• pp.420-427
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• 2020

A repebody, an artificial non-immunoglobulin protein scaffold, is expected to be a solution in the search for faster, cheaper, and customizable antibodies. However, the production of medical repebodies remains difficult due to their low yield and the complex purification processes required. The Pseudomonas fluorescens ABC transporter system has been suggested as an efficient and cost-effective method for repebody production, but the total yield is low because of the secreted protein's positive charge; thus, a repebody with a high isoelectric point needs to be changed into a more negatively charged protein for better secretion. To achieve this, we first attached oligo-aspartic acids to the N- and C-terminals of the repebody, but secretion efficiency was not enhanced significantly. Subsequently, we devised an alternative method for improved secretion efficiency by engineering fifteen positively charged amino acids to aspartic acid in the non-antigen binding sites of the repebody to give a high net negative charge. As a result, secretion efficiency was greatly enhanced from 21.2% (wildtype) to 58.5% (negatively supercharged). The negatively supercharged repebody was succussfully produced extracellularly by ABC transporter secretion system in P. fluorescens.

• Journal of Life Science
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• v.30 no.8
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• pp.731-741
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• 2020

Coronavirus disease 19 (COVID-19) is caused by SARS-CoV-2 (Severe Acute Respiratory SyndromeCoronavirus 2). To date, seven coronaviruses that can infect humans were reported. Among them, infections with four coronavirus strains (HCoV-229E, HCoV-OC43, HCoV-NL63, and HCoV-HKU1) resulted in mild symptoms such as common cold, whereas SARS-CoV and MERS-CoV caused severe symptoms and epidemics in 2002 and 2012, respectively. In the most recent, SARS-CoV-2 was first reported in Wuhan, China in December 2019 and became a notorious cause of the ongoing global pandemics. To diagnose, treat, and prevent COVID-19, the development of rapid and accurate diagnostic tools, specific therapeutic drugs, and safe vaccines essentially are required. In order to develop these powerful tools, it is prerequisite to understand a phenotype, a genotype, and life cycle of SARS-CoV-2. Diagnostic techniques have been developing rapidly around world and many countries take the fast track system to accelerate approval. Approved diagnostic devices are rapidly growing facing to urgent demand to identify carriers. Currently developed commercial diagnostic devices are divided into mainly two categories: molecular assay and serological & immunological assay. Molecular assays begins the reverse transcription step following polymerase chain reaction or isothermal amplification. Immunological assay targets SARS-CoV-2 antigen or anti-SARS-CoV-2 antibody of samples. In this review, we summarize the phenotype, genome structure and gene expression of SARS-CoV-2 and provide the knowledge on various diagnostic techniques for SARS-CoV-2.

• Pediatric Infection and Vaccine
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• v.17 no.2
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• pp.137-147
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• 2010

Purpose : The purpose of this study is to evaluate the usefulness and limitation of QuantiFERON-TB Gold (QTB) for diagnosis of latent tuberculosis in children. Methods : The medical records of 112 children who were tested by tuberculin skin test (TST) and QTB for detection of latent tuberculosis (TB) in Eulji General Hospital during the period from January 2007 to June 2009 were retrospectively reviewed. Results : Of the 112 participants, the clinical TB group included 15 (13.4%), the contact group included 43 (38.4%), and the non-contact group included 54 (48.2%). Positive rates of TST and QTB were 100% and 80% in the clinical TB group, 64% and 14% in the close contact group, 27% and 0% in the casual contact group, and 52% and 2% in the non-contact group, respectively. Sensitivity of QTB was 80.0% and specificity was 92.6%. Agreement between QTB and TST was poor ($\kappa$=0.209). We also confirmed that the positive rate of QTB increased as the age of the patient increased (P=0.011). A QTB indeterminate result was observed in 11 (9.8%) subjects. QTB was retested in 15 subjects. In 5 of the 6 subjects who had positive results initially, positive results persisted for a median 2.2 months after termination of treatment. Conclusion : Although QTB was associated with several problems, including low sensitivity and a high rate of indeterminate results, it had clinical importance due to its high specificity. We found good correlation with regard to exposure and QTB positivity, including that of young children under 5 years of age. However, clinical application of interferon-$\gamma$ releasing assay for young children for diagnosis of active and latent tuberculosis will require additional prospective studies.

• Journal of Life Science
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• v.11 no.2
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• pp.181-189
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• 2001

Cytokines are hormone-like proteins which mediate and regulast inflammatory and immune responses. Transforming growth factor -$\beta$1(TGF-$\beta$) plays an important role in the control of the immune response and wound healing, and in the development o various tissues and organs, Nitric oxide(NO) is major messenger molecule regulating immune function and blood vessel dilation and serving as a neurotransmitter in the brain and peripheral nervous system. Also, NO is to be a potent mutagen that cause mutation in the p53 tumor suppressor gene in early phases of human gastric carcinogenesis. The purpose of this study was to investigate the effect of Helicobacter phlori lystes, lipopolysaccharide (LPS), and Staphylococcus enterotoxin B(SEB) on production of TGF-$\beta$1 and NO by human fibroblasts. Primary cultured human fibroblasts were incubated with H. pylori lysates(Hp), LPs, SEB, Hp+LPS, Hp+SEB, Hp+LPS+SEB. Cultured supernatants that were collected at 24, 48 and 72 hr were assessed for TGF-$\beta$1 by enzyme-linked immunosorbent assay and NO production by quantification of nitrite ion. TGF-$\beta$1 production in fibroblasts exposed with Hp, LPS or SEB for 48 hrs was enhanced, but for 72 hrs inhibited. Its production by doble exposure such as Hp+LPS, Hp+SEB, Hp+LPS+SEB was lowered in comparison with single exposure of Hp in cases of 24 and 48 hrs incubation, but for 72 hrs decreased in Hp vaculoating toxin(+), increased in Hp vacuolating toxin(-). No production in fibroblasts increaed at all doses of LPS. But its production by exposure of SEB increased or decreased according to dose and incubation time. Also, NO production by Hp vacuolating toxin(+) increased at all doses, but its production by Hp vacuolating toxin(-) decreased. Its production by doble exposure such as Hp+LPS, Hp+SEB, Hp+LPS+SEB decreased in comparison with single exposure Hp Therefore, quantities pf TGB-$\beta$1 and NO released by human fibroblasts shows differences according to kinds of stimulants. Also, in care stimulated with same kinds of stimulants, its productions exhibit quantitative differences according to exposure times. These results suggest that the decreased of TGF-$\beta$1 in fibroblasts by mixed exposure with Hp producing vacuolating toxin and bacterial toxins such as LPS and SEB may effect negatively in healing of host tissue and increased of NO by infection oh H. pylori may related to the increased susceptibility for human gastric carcinogenesis.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.2
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• pp.89-100
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• 2009

SLE 등의 자가면역질환 병인론에 있어서 $CD4^+$ T 조력 림프구 (T helper cell)의 기능과 항상성 조절의 이상이 중요하다는 것은 알려진 사실이지만 $CD4^+$ T 조력 림프구 (T helper cell)의 발달과 기능에 대한 성호르몬의 정확한 역할에 대하여는 아직 확립되어 있지 않은 실정이다. 에스트로겐의 $CD4^+$ T 림프구에 대한 여러 방면의 작용에 대한 결과는 에스트로겐 매개 신호가 전달되는 정황에 매우 의존적이라고 할 수 있으며, 또한 에스트로겐이 B 림프구와 항원발현세포들 같이 T 림프구 기능을 변화시킬 수 있는 다른 분획 세포들의 기능에 대해서도 영향을 미칠 수 있다는 사실을 고려한다면, 에스트로겐이 $CD4^+$ T 림프구매개반응에 미치는 작용은 매우 복잡하다고 할 수 있다. 현재로선 이런 과정들과 관련된 에스트로겐의 정확한 기전에 대한 연구 성과는 현재로선 아직걸음마 단계에 지나지 않는다고 볼 수 있다. 일부 분자학적 표적들의 발견에도 불구하고 ER-$\alpha$와 ER-$\beta$, 또는 이런 표적들의 조절에 대한 유전자적 또는 비유전자적 경로의 상대적 역할에 대해서는 현재로선 알려진 바가 많지 않은 것 또한 사실이며, T 림프구 자체의 미세 환경과 ER 매개 전사반응에 영향을 미칠 수 있는 방법 역시 아직까진 알려진 바가 없다. 따라서 에스트로겐의 $CD4^+$ T 림프구에 대한 다각적 영향을 규명하는 데 있어 이런 경로들에 대한 보다 체계적이고 상세한 연구가 반드시 필요하다. 최근 골다공증의 병인에 있어 에스트로겐에 의한 TNF 및 RANK-RANKL계 억제에 대해 연구가 심도 있게 진행됨으로써 결과적으로 다른 시토카인과 면역세포들에 대한 영향이 간접적으로 규명되고 있는 점은 매우 고무적인 현상이라고 여겨진다. 이러한 연구들은 추후 에스트로겐의 면역 및 염증체계에 대한 특징적 작용을 규명하는데 있어서 밑거름이 될 수 있을 것이다. 나아가 호르몬 자체, 또는 SERM 같은 준호르몬제제들이 면역계에 미치는 영향을 분자학적으로 규명함으로써 향후 이러한 지식들이 또한 류마티스성 자가면역질환이나 만성염증성질환, 또는 전신성 감염질환에 있어 면역글로불린이나 기타 다른 기존 치료 약제들을 대체 보완할 수 있는 호르몬 치료로 이어질 수 있는 결정적인 단서를 제공할 수 있을 것이라 기대할 수 있을 것이다.

• Journal of Yeungnam Medical Science
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• v.21 no.2
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• pp.191-197
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• 2004

Background: This study was performed to reconsider the efficacy of transrectal ultrasonography (TRUS) in diagnosing prostate cancer by analyzing the results of a digital rectal examination (DRE), serum prostate-specific antigen (PSA) and a transrectal ultrasonography in patients with prostate specific antigen levels of 10 ng/ml or less. Materials and Methods: One-hundred and eighty one men with PSA levels of 10 ng/ml or less, who had a TRUS-guided tissue biopsy performed, were included in this study. The detection rate of prostate cancer was compared according to the TRUS result and the presence or absence of nodularity and the consistency of the prostate on DRE. Results: In a total 181 patients, there were 73 patients with PSA levels of 4 ng/ml or less and 4 of them had prostate cancer. Thre were 108 patients with PSA levels of 4-10 ng/ml and 18 of them were prostate cancer. TRUS was performed in 152 patients and 16 out of 58 patients diagnosed with prostate cancer, 3 out of 39 diagnosed with suspicious prostate cancer, and 2 out of 55 patients diagnosed as having no prostate cancer were found to have prostate cancer. In 40 patients, a nodule was palpated on DRE and 8 of them were found to have prostate cancer. Five out of 19 patients with a stony hard consistency, 3 of 12 with a firm to hard consisency, 12 of 129 with a firm consistency, 0 of 13 with a soft to firm consistency, and 2 of 8 with a soft consistency were prostate cancer. In the prostate cancer patients, there were 4 patients with PSA levels of 4 ng/ml or less and all these patients were diagnosed with prostate cancer or suspicious prostate cancer on TRUS but the nodule was not palpated in all patients. Two were soft and 2 were firm consistency on DRE. Conclusion: In patients with serum PSA levels of 10 ng/ml or less, TRUS is a more useful supporting method than DRE and a more active application of TRUS may lead to an early diagnosis and pertinent treatment of prostate cancer.

• Korean Journal of Plant Resources
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• v.27 no.5
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• pp.415-420
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• 2014

Toxoplasmosis is an important cause of foodborne, inflammatory illnesses, as well as congenital abnormalities. Currently available therapies are ineffective for persistent chronic disease and congenital toxoplasmosis or have severe side effects which may result in life-threatening complications. There is an urgent need for safe and effective therapies to eliminate or treat this cosmopolitan infectious disease. The aim of this study was to investigate the in vitro anti-Toxoplasma activities of Suaeda maritime, one of the halophytes, using tachyzoit of T. gondii RH strain infected HeLa cells. As the results, the selectivity of Suaeda maritime extract was 6.63, which was higher than Sulfadiazine selectivity (2.06). Also, we perfomed the cell proliferation inhibition test and the morphological study to evaluate the anti-T. gondii activity of Suaeda maritime extract with HeLa cells. As the results, the inhibition rate of the Suaeda maritime extract was high inhibition rate. This indicates that the Suaeda maritime extract may be used for new anti-T. gondii agent.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.1
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• pp.23-28
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• 2004

The aim of this study is to assess the anti-irritation activities of mung bean (Phaseolus aureus) extract against various irritants used in cosmetics. For its antidotal activity, mung bean has been used as a medicinal or cosmetic material since ancient times. However, there have been few reports describing the biological activities of these beans and no comprehensive surveys of the constituents. We obtained an ethanolic extract of mung bean and isolated the major constituents, such as vitexin and isovitexin. And we previously reported that the mung bean extract containing vitexin and isovitexin had excellent antioxidant and anti-inflammatory activities. To investigate the mechanisms of anti-inflammatory activity of mung bean extract, we examined the inhibitory effects on histamine release from rat peritoneal mast cells and lipoxygenase activity. Mung bean extract inhibited histamine release in a concentration dependent manner but showed no inhibitory activity in the 5-lipoxygenase assay. And, clinical studies were conducted to evaluate the anti-irritation effects of mung bean extract against various irritants used in cosmetics such as lactic acid, retinol, and preservatives. When 2.0％ of mung bean extract was applied to cosmetic formulae containing each of irritants, it revealed considerable anti-irritation efficacy. Our results of the human patch test with 20 volunteers showed that this extract reduced skin irritations caused by 5.0％ lactic acid, 4000 IU retinol, and 1.0％ preservative mixture by about 60％, 30％, and 50％ respectively. The stinging potential test for assessing subjective irritation also showed that the extract reduced the unpleasant sensations by about 50∼30％. Finally, we performed a double-blind usage test with 30 subjects to compare formulae containing mung bean extract with placebo. From the results of questionnaires for 4 weeks of use, we confirmed the excellent anti-irritation effect of mung bean extract. Conclusively, we could discover new material that had anti-irritation effects and apply this mung bean extract to the final cosmetic products successfully.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.54 no.3
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• pp.265-269
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• 2009

Technology development for sufficient dry matter production of Chinese milk vetch (CMV) is most important in CMV-rice cultivation system in order to provide sufficient nutrients to rice plants. However, when the CMV plants are dominated by the weed, especially Alopecurus aequalis, the CMV growth could be reduced due to light and nutrient competition. In addition, A. aequalis is potential host of the rice dwarf virus disease. Therefore, control of A. aequalis is necessary to enhance the biomass production of CMV plants when CMV stands are insufficient. The use of chemical like sethoxydim (20%, ai) showed the highest control rate of 84% at early stage and was reduced as application was delayed. A. aequalis control did not change the CMV seedling stand before and after herbicide treatment and the reseeding stand in fall was rather increased 2.2 to 2.6 times. On the other hand, in untreated control, the CMV stand at May 15 and reseeding stand in fall was significantly reduced as compared with the before herbicide treatment. Control of A. aequalis increased the CMV dry matter production by 164% for 50% CMV coverage rate and 63% for 25% CMV coverage rate. This is equivalent to $12.3{\sim}16.4\;kgN$/10a which is greater than the recommended nitrogen rate of 9kg/10a. The result indicates that the control of A. aequalis is an efficient way to enhance dry matter production in CMV-rice cultivation system especially when CMV stand is poor.

• Development and Reproduction
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• v.5 no.2
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• pp.167-173
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• 2001

To characterize the difference in glycoconjugates of mouse epididymis, lectin labeling of the tissue section was conducted using Ulex europaeus agglutinin I(UEA I), succinylated wheat germ agglutinin(sWGA), and Griffonia simplicifolia lectin-I(GSL-I). UEA I which binds to outer $\alpha$-L-fucose residue that is a terminal sugar of the side chain branched from oligosaccharide chain gave the labeling in the proximal caput epithelia exclusively. Lumen was commonly labeled in all of the organ. It suggested that the glycoconjugates bearing outer $\alpha$ -L-fucose residue were largely expressed in the initial segments ot epididymis and subjected to secretion. GSL-I which binds to terminal $\alpha$ -D-galactosyl residue of glycoconjugates gave the labeling in the cytoplasm of clear cells and basal cells, and cilia in corpus and cauda regions but not in the caput region. There was no vast difference in labeling pattern by sWGA which binds to N-acetyl-glucosamine residue among the epididymal regions. Clear cells in corpus and cauda epithelia showed more intense labeling by sWGA compared to principal cells, suggesting the functional specialization of this type of cells. The labeling intensities of luminal content by UEA I and sWGA decreased in cauda region compared to corpus region suggesting the presence of enzymatic activities responsible for processing the $\alpha$-L-fucose and N-acetyl-glucosamine residues from secreted glycoconjugates. In summary, the difference in glycoconjugates bearing the $\alpha$-L-fucose, $\alpha$-D-galactose, and N-acetyl-glucosamine residues according to the type of epithelial cells and epididymal segments suggests functional specialization and different roles of each segment in the processing of sperm surface antigens during the epididymal transit.