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Construction of fluorescent red silk using fibroin H-chain expression system (누에 형질전환에 의한 견사선에서의 적색형광단백질 발현)

  • Kim, Sung Wan;Yun, Eun Young;Choi, Kwang-Ho;Kim, Seong Ryul;Park, Seung Won;Kang, Seok Woo;Kwon, O-Yu;Goo, Tae Won
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.87-92
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    • 2012
  • We constructed the fibroin H-chain expression system to produce Discosoma sp. red fluorescent protein variant2 (DsRed2) in transgenic silkworm cocoon. Fluorescent cocoon could be made by fusing DsRed2 cDNA to the heavy chain gene and injecting it into a silkworm. The DsRed2 fusion protein, each with N- and C-terminal sequences of the fibroin H-chain, was designed to be secreted into the lumen of the posterior silk glands. The expression of the DsRed2/H-chain fusion gene was regulated by the fibroin H-chain promoter. The use of the 3xP3-driven EGFP cDNA as a marker allowed us to rapidly distinguish transgenic silkworms. The EGFP fluorescence became visible in the ocelli and in the central and peripheral nervous system on the seventh day of embryonic development. A mixture of the donor and helper vector was micro-injected into 1,020 Kumokjam, bivoltin silkworm eggs. We obtained 6 broods. The cocoon was displayed strong red fluorescence, proving that the fusion protein was present in the cocoon. Accordingly, we suggest that the DsRed2 fluorescence silk will enable the production of novel biomaterial based on the transgenic silk.

Relative Association of Overhydration and Muscle Wasting with Mortality in Hemodialysis Patients: Assessment by Bioelectrical Impedance Analysis (혈액투석 환자에서 Bioelectrical Impedance Analysis를 활용하여 측정한 과수분량과 근육량 감소와 사망률의 상관관계)

  • Kim, Eunju;Seo, Sang Oh;Choi, Yu Bum;Lee, Mi Jung;Lee, Jeong Eun;Kim, Hyung Jong
    • The Korean Journal of Medicine
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    • v.93 no.6
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    • pp.548-555
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    • 2018
  • Background/Aims: Assessment of fluid status in hemodialysis patents is very important. Overhydration in hemodialysis is associated with generalized edema, cardiovascular complications, and hypertension. The aim of this study was to determine the factors correlated with mortality of hemodialysis patients, assessing body muscle mass and fluid status using bioelectrical impedance analysis (BIA). Methods: This study enrolled 93 patients who underwent hemodialysis between January 2010 and May 2015 at CHA Bundang Medical Center. Medical records of enrollees up to June 2017 were reviewed retrospectively. These included laboratory results (serum albumin, C-reactive protein [CRP], lipid profile, etc.) and BIA data (extracellular water, intracellular water, total body water, soft lean mass, fat free mass, skeletal muscle mass, etc.). Results: Eleven of 93 patients had expired by May 2017. Among the surviving subjects, mean age was younger, CRP levels were lower, albumin levels were higher, and extracellular water/total body water (ECW/TBW) ratios were lower than in the expired patient group. Kaplan-Meier survival analysis revealed that overhydration (ECW/TBW > 0.4) was associated with higher mortality. Conclusions: In hemodialysis patients, overhydration is an important factor in mortality, and BIA could be a reliable modality in its assessment. We suggest that, for hemodialysis patients, overhydration is more of a risk factor for mortality than is muscle wasting.

Biological Roles of the Glycan in the Investigation of the Novel Disease Diagnosis and Treatment Methods (신개념 질병 진단 및 치료 연구에 있어서의 당사슬의 생물학적 역할)

  • Kim, Dong-Chan
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1379-1385
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    • 2018
  • Glycans are attached to proteins as in glycoproteins and proteoglycans. They are found on the exterior surface of cells. O- and N-linked glycans are very common in eukaryotic cells but may also be found in prokaryotes. The interaction of cell surface glycans with complementary glycan binding proteins located on neighboring cells, other cell types, pathogens like virus, or bacteria is crucial in biologically and biomedically important processes like pathogen recognition, cell migration, cell-cell adhesion, development, and infection. Their implication in pathological condition, suggests an important role for glycans as disease markers. In addition, a great amount of research has been shown that appropriate glycosylation of a recombinant therapeutic protein is critical for product solubility, stability, pharmacokinetics and pharmacodynamics, bioactivity, and safety. Besides, cancer-associated glycosylation changes often involve sialic acid in glycan branch which play important roles in cell-cell interaction, recognition and immunological response. This review aims at giving a comprehensive overview of the glycan's biological function and describing the relevance among the glycosylation, disease diagnosis and treatment methods. Furthermore, the high-throughput analytic methods available to measure the profile changing patterns of glycan in the blood serum as well as possible underlying biochemical mechanisms.

Development of an apple/pear pomace fermented with Lentinus edodes Mycelia (사과/배 부산물 및 표고버섯균사체를 이용한 발효물 개발)

  • Kim, Jin-Kyeong;Jo, Seong-Wha;Kim, Eun-Ji;Jeong, Do-Youn
    • Korean Journal of Food Science and Technology
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    • v.51 no.3
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    • pp.286-294
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    • 2019
  • The purpose of this study was to investigate the possibility of enhancing the functional compounds in apple and pear pomace (APP) by fermentation with mycelia from the mushroom Lentinula edodes. A 30% (w/v) APP with added rice bran and Biji was fermented with L. edodes at $24^{\circ}C$ and 80% humidity. The cellulase and pectinase activities in the fermented APP (FAPP) were higher than those in the non-fermented control. In addition, the physiological activities of the FAPP, including DPPH, ABTS radical scavenging, and SOD-like activity, as well as the total polyphenol and ${\beta}-glucan$ contents were higher than those in the control. FAPP treatment significantly reduced LPS-induced nitric oxide (NO) levels in Raw 264.7 cell. Therefore, FAPP treatment was considered to more effectively suppress cell injury caused by inflammatory cytokines through inhibition of LPS-induced NO production. These results suggest that the levels of functional components in APP can be increased by fermentation with this mushroom mycelium. However, further studies are needed before it can be used as a functional material.

The Effect of Low Molecule Collagen Peptide on Skin Anti-glycation and Collagen Synthesis as a Skin Aging Therapy (피부 노화 치료로서 저분자콜라겐펩타이드의 피부 항당화와 콜라겐 합성 효과)

  • Kim, Hong Seok;Hong, Won Kyu;Lee, Mun Hoe;Kim, Hyeong Min;Chung, Hee Chul;Lee, Jin Hee
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.47 no.2
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    • pp.147-153
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    • 2021
  • Collagen hydrolysate (CH) is known to prevent skin aging by stimulating skin dermal fibroblasts to promote synthesis of extracellular matrix such as collagen and elastin. Recently, among the various factors that cause skin aging, advanced glycation end products (AGEs) have received particular attention. However, the effect of CH on AGE accumulation has not been studied. Since CH could affect AGE accumulation by promoting production of skin structural proteins, clinical trial was performed using low molecule collagen peptide (LMCP), which were CH containing 25% tripeptide and 4% Gly-Pro-Hyp. Skin autofluorescence (SAF) values were measured using an AGE reader to evaluate accumulation of AGE in skin. As a result of applying 0.5% and 1.0% LMCP solutions to the subject's forearm for 8 weeks, the SAF value at the test site significantly decreased compared to the control site. Additionally, in vitro test was performed using CCD-986sk to evaluate the promotion of collagen synthesis in skin fibroblasts by LMCP. As a result, 800 ㎍/mL of LMCP significantly increased synthesis of human pro-collagen Iα1 (COL1A1) in CCD-986sk. Through this study, we have confirmed that tropical LMCP applications can promote collagen synthesis to help anti-glycation effects, suggesting that LMCP has potential as an anti-aging cosmetic material.

Positive effect of Saengmaeg-san intake on blood lipid and arteriosclerosis index during high-intensity training (고강도 트레이닝 시 생맥산 섭취가 혈중 지질 및 동맥경화 지수에 미치는 긍정적인 영향)

  • Kwak, Jae-Jun;Lee, Jae-Hoon;Yook, Jang Soo;Lee, Sang-Ho;Ha, Min-Seong
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.6
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    • pp.1512-1523
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    • 2021
  • The purpose of this study was to investigate the effect of Saengmaeg-san intake on blood lipids and arteriosclerosis index in male tennis players. Seventeen male tennis players enrolled in university were assigned to a Saengmaeg-san intake group (n=9) and a placebo control group (n=8) according to whether they took Saengmaeg-san during 4-week high-intensity tennis training. Tennis training was conducted 5 times a week for 4 weeks, and the exercise intensity was performed at 70-90% of the spare heart rate. The intake of Saengmaeg-san was 110ml at a time before breakfast, before, during, during, and after exercise, and after dinner, a total of 7 times, 770ml per day. For all data, mean and standard deviation were used. Two-way repeated measures ANOVA was used to examine the effect of time and treatment, and Pearson's Correlation was used to examine the relevance of differences in blood lipid responses after treatment. As a result of this study, the Saengmaeg-san intake group significantly improved blood lipids (triglyceride, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol) and arteriosclerosis index, and there was a significant correlation appeared between ∆TG, ∆LDL-C, and ∆TC. As a result, the intake of Saengmaeg-san during high-intensity training of male college student tennis players can have a positive effect on blood lipids and arteriosclerosis index, suggesting that it can be an effective sports drink as an exercise supplement.

The Role of NF-${\kappa}B$ in the TNF-$\alpha$-induced Apoptosis of Lung Cancer Cell Line (폐암세포주의 TNF-$\alpha$ 유발 apoptosis에서 NF-${\kappa}B$의 역할)

  • Kim, J.Y.;Lee, S.H.;HwangBo, B.;Lee, C.T.;Kim, O.H.;Han, S.K.;Shim, O.S.;Yoo, C.G.
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.2
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    • pp.166-179
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    • 2000
  • Background: The main reason for the failure of anti-cancer chemotherapy is the build up of resistance by cancer cells to apoptosis. The activation of NF-${\kappa}B$ in many cancer cell lines is reported to be underlying mechanism behind the build up of resistance of cancer cells to apoptosis. However, this relationship varied depending on the cells used in the experiments. In this study, the role of NF-${\kappa}B$ activation in the TNF-$\alpha$-induced apoptosis in lung cancer cell line was evaluated. Methods: NCI-H157 cells were used in all experiments. Cells were exposed to a high dose of TNF-$\alpha$(20 ng/ml) for 24 or 48 hours with or without blocking NF-${\kappa}B$ activation. TNF-$\alpha$-induced activation of NF-${\kappa}B$ was inhibited either by overexpression of $I{\kappa}B{\alpha}$-super repressor($I{\kappa}B{\alpha}$-SR) or by pre-treatment with proteasome inhibitor. Cell viability and apoptosis were evaluated with MTT assay and Western blot analysis for PARP fragment, respectively. Results: Cell viability of NCI-H157 cells was not affected by TNF-$\alpha$ treatment alone; however, combined treatment with TNF-$\alpha$ and cycloheximide reduced cell viability significantly, indicating that resistance to TNF-$\alpha$ is mediated by the new proteins synthesized after TNF-$\alpha$ stimulation. To evaluate the role of NF-${\kappa}B$ in the transcription of anti-apoptotic proteins. delete NF-${\kappa}B$ activation was inhibited before TNF-$\alpha$ stimulation. as described above. $AD5I{\kappa}B{\alpha}$-SR-transduction inhibited TNF-$\alpha$-induced nuclear translocation of p65. TNF-$\alpha$-induced cell death and apoptosis increased after inhibition of TNF-$\alpha$-induced activation of NF-${\kappa}$ by methods. Conclusion: These results suggest that TNF-$\alpha$-induced activation of NF-${\kappa}B$ may be closely related to the acquisition of the resistance to TNF-$\alpha$-induced apoptosis in lung cancer cells. Therefore. blocking of NF-${\kappa}B$ pathway can be a useful therapeutic modality in the treatment of lung cancer.

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Antioxidant activity and sensory characteristics of rice cookies containing dandelion complex powder (민들레복합분말 첨가 쌀 쿠키의 항산화적, 관능적 품질특성)

  • Byeon, Yang Soo;Ra, Ha Na;Kim, Hae Young
    • Korean Journal of Food Science and Technology
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    • v.49 no.2
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    • pp.173-180
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    • 2017
  • This study evaluated the characteristics of rice cookies with varying amounts (0, 250, 500, and 750 mg) of dandelion complex powder. Dandelion powder is considered a functional food with skin-moisturizing and atopic skin improvement effects by KFDA. Quality characteristics of AF343 and rice cookies were measured by determining antimicrobial activities, physical characteristics, sensory evaluation values, and antioxidant activities. An antibiotic susceptibility test of the powder showed positive activities in Escherichia coli (O157:H7), Staphylococcus aureus, Bacillus cereus, and Listeria monocytogenes. Rice cookie containing 250-500 mg powder showed significantly increased antioxidant activities compared to controls (p<0.05). However, sensory intensities between the two sample groups were not significantly different. Thus, rice cookies containing 250-500 mg dandelion complex powder were successfully developed to improve antioxidant and antimicrobial qualities. These products may attract the attention of health-conscious consumers in the highly competitive cookie industry.

The effect of Nutrient Intake and Past Dairy Products Consumption on Bone Mineral Density of Postmenopausal of Korean Women (과거의 우유 및 유제품의 섭취가 한국노인의 골밀도에 미치는 영향)

  • 강은주
    • The Korean Journal of Food And Nutrition
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    • v.11 no.1
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    • pp.87-98
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    • 1998
  • This study was conducted to investigate Osteoporosis on the effect of dietary factors and past young age period's dairy products consumption on BMD in 170 postmenopausal Korean women without diagnosed disease. Bone mineral density(BMD) of lumbar spine was investigated by dual energy X-ray absorptiometry. Dietary assesment of the subjects were measured by Cognitive Food Frequency Recall method. The measurements of group average were 57.9kg of body weight, 36.73 of BMI(Body Mass Index), 2103.3㎉ of energy, 638.7mg of dietary calcium, 70.3g of protein, and 10.58mg of iron. Nutrient intake levels were similar to or more than the level of Korean Recommended Dietary Allowances. Bone Mineral Density(BMD) of Lumbar spine(L2-L4) of group average was 0.912g/$\textrm{cm}^2$, and under 50 yr's 1,02g/$\textrm{cm}^2$, 50~54 yr's 0.92g/$\textrm{cm}^2$, 55~59 yr's 0.85g/$\textrm{cm}^2$, over 60 yr's 0.85g/$\textrm{cm}^2$, had been getting low degree on aging. BMD of the Lumbar spine was positively correlated with calorie, body weight, dietary calcium, protein, phosphorus and serum albumin. Past dairy products consumption experiment was highly significant on BMD in teenage period(R square = 26, p-value 0.0031). Particularly in over 60 yr group, the correlations between BMD and past dairy products consumption in teenage period had shown highly significance(r=0.48, p<0.02). 55~59 yr age group had also positive correlation(R squae = 0.29, p<0.05). This results confirm that the most effective way of preventing osteoporosis and the fractures is to maximize peak bone mass in early life and to minimize bone loss through the balanced intake of Ca and other nutrients and regular physical activity.

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Characterization of Antibacterial Compounds from Bacillus polyfermenticus CJ6 and Its Growth Inhibition Effect on Food-Borne Pathogens (Bacillus polyfermenticus CJ6가 생산하는 항세균 물질의 특성 및 병원성 식중독 미생물의 성장 억제 효과)

  • Jung, Ji-Hye;Chang, Hae-Choon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.6
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    • pp.903-911
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    • 2011
  • In this study, Bacillus polyfermenticus CJ6 harboring antibacterial activity was isolated from meju. The antibacterial activity of Bacillus polyfermenticus CJ6 was stable in the pH range of 3.0~9.0, but it disappeared after culture at $70^{\circ}C$ for 24 hr. Antibacterial activity was inactivated by proteinase K, protease, and ${\alpha}$-chymotrypsin, indicating its proteinaceous nature. The growth inhibitory effects of B. polyfermenticus CJ6 culture on food-borne pathogens such as Staphylococcus aureus, Salmonella Typhi, Listeria monocytogenes, and Escherichia coli O157:H7 were examined in this study. Approximately 6~6.2 log CFU/mL of each pathogen was co-cultured with B. polyfermenticus CJ6 in a 50 mL culture volume for 24 hr. Growth of S. aureus and L. monocytogenes was completely inhibited after 3 hr of incubation. Growth of S. Typhi and E. coli O157:H7 was also completely inhibited after 6 hr of incubation. The antibacterial compounds from B. polyfermenticus CJ6 were purified by solid phase extraction (C18 Sep-pak cartridge), recycling preparative HPLC, and analytical HPLC. Ultra-high performance liquid chromatography and electrospray ionization tandem mass spectrometry analysis were used to identify the purified antibacterial compounds, which were confirmed to be five peptides (757.4153 Da, 750.3444 Da, 1024.5282 Da, 1123.6083 Da, and 1617.8170 Da).