• 한국미생물·생명공학회지
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• 제26권5호
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• pp.406-412
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• 1998

Trichoderma sp. C-4 유래의 endoglucanase 유전자(egl6)를 ADH1 promoter 하류에 연결하여 구성적 발현 plasmid인 pVT-C4를 제작하였다. 이를 3종의 S. cerevisiae숙주세포(YNN27, 2805, SEY2102)에 형질전환시켜 각 숙주세포당 80개의 형질전환체를 시험배양하여, 균체증식과 endoglucanase 발현량이 뛰어난 형질전환 효모 균주를 선별하였다. 3종의 재조합 효모를 YPD 배지에서 회분 배양했을 때, YNN27 균주가 가장 낮은 균체농도(OD$_{600}$=19)를, 2805 균주가 가장 높은 균체농도(OD$_{600}$=33)를 보였으며, 총 발현된 endoglucanase 활성은 균체농도에 의존적인 양상으로 나타나 YNN27의 경우 586 unit/l, SEY2102의 경우 1023 unit/l, 2805의 경우 1142 unit/l 순서로 증가하였다 비활성(균체농도당 endoglucanase활성 )은 세 균주 모두 31~34 unit/l/OD$_{600}$ 값을 보여 단위 균체당 발현능 차이는 거의 없었다. Plasmid 안정성은 배양 72시간에서 세 균주 모두 80% 이상의 높은 값을 보였다. 발현된 endoglucanase의 분비 국재성은 세포밖 배지에 약 80%, periplasmic space 잔존 활성이 11~13%, 세포질 분획에서의 활성이 8~10% 정도로 세 균주에서 모두 비슷한 분포를 보였다. 즉, egl6의 분비신호는 S. cerevisiae에서도 매우 효율적으로 분비 기능을 발휘하였다. 재조합 endoglucanase는 nondenaturing-PAGE 상에서 전형적인 당단백질의 disperse band를 두개 보였다. 세가지 숙주세포에 따른 재조합 endoglucanase활성 band들의 이동상의 차이는 거의 나타나지 않고, 당쇄부가 양상은 야생형보다 훨씬 더 복잡하고 불균일하게 발생했음을 알 수 있었다.수 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권2호
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• pp.224-233
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• 2012

The limiting sequence and relative ratio of lysine (Lys), methionine (Met), and threonine (Thr) for calves about 2 mo of age fed milk replacers (MR) containing soy protein are not clearly defined. The objective of the study was to investigate the effect of supplementing MR containing 22% CP, half from soy protein concentrate (SPC, 40.56% CP, flour) and half from whey proteins, with Lys, Met, and Thr to estimate amino acid (AA) sequence and their relative ratio for calves about 2 mo of age. A method of partial deduction of AA was adopted. Twenty-four newborn calves (half males and half females, $40.7{\pm}0.9$ kg of BW) were fed 1 of 4 MR diets for 56 d (n = 6/diet). The diets were supplemented with all (positive control) or with 2 of the 3 AAs: Lys, Met and Thr, (i.e., PC (22% CP, 2.34% Lys, 0.72% Met and 1.80% Thr), PC-Lys (22% CP, 1.64% Lys, 0.72% Met and 1.80% Thr), PC-Met (22% CP, 2.34% Lys, 0.50% Met and 1.80% Thr), and PC-Thr (22% CP, 2.34% Lys, 0.72% Met and 1.26% Thr)). Calves were fed thrice daily; starter (20% CP, 1.03% Lys, 0.30% Met and 0.69% Thr), hay (3.23% CP, 0.29% Lys, 0.12% Met and 0.23% Thr) and water were offered free choice. Starter and hay were only offered beginning on d 36 (after 5 wk) and d 43 (after 6 wk), respectively. BW, body size and blood samples measures were taken every two weeks. Three-day total collection of feed refusals, feces, and urine were recorded starting at d 33 and d 54 of age, respectively. From the results, the limiting sequence and relative ratio between the 3 AAs in calves with different diet structures were calculated. The limiting sequence of the 3 AAs were ranked as Lys, Met and Thr; the proper ratio was 100:29:70 for MR-only diet and 100:30:60 for diets consisted of MR, starter and hay. Nitrogen digestion and utilization and nutrient digestibility were negatively affected by AA deletion treatments. From the evidence of this experiment, it did not appear that the AA limiting sequence was selectively altered by differences in diet structures such as would be encountered in practice. The relative ratio between the 3 AAs varied with the offer of starter and hay to calves, and the average ratio was 100:29.5:65 for calves during 2 to 10 wk of age.

• Asian Pacific Journal of Cancer Prevention
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• 제15권23호
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• pp.10255-10262
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• 2015

High risk human papillomavirus (HR-HPV) E2 proteins play roles in transcriptional regulation and are commonly functionally disrupted when the HPV genome integrates into host chromosomes. Some 15-40% of cancer cases, however, contain an intact E2 gene or episomal HPV. In these cases, polymorphism of the E2 gene might be involved. This study aimed to determine polymorphisms of the E2 gene in episomal HPV16 detected in high grade squamous intraepithelial lesions and squamous cell carcinomas and altered functions compared to the E2 prototype. The E2 gene was amplified and sequenced. Two expression vectors containing E2 gene polymorphisms were constructed and transfected in SiHa and C33A cells, then E6 gene as well as Il-10 and TNF-${\alpha}$ expression was determined by quantitative RT-PCR. Expression vectors and reporter vectors containing the HPV16 long control region (LCR) were co-transfected and transcriptional activity was determined. The results showed that a total of 32 nucleotides and 23 amino acids were changed in all 20 cases of study, found in the transactivation (TA) domain, hinge (H) region and DNA binding (DB) domain with 14, 5 and 13 nucleotide positions. They mostly caused amino acid change. The expressing vectors containing different E2 gene polymorphisms showed E6 mRNA suppression, TNF-${\alpha}$ mRNA suppression and IL-10 induction but no statistically significant differences when compared to the E2 prototype. Moreover, promoter activity in HPV16 LCR was not affected by E2 protein with different gene polymorphisms, in contrast to nucleotide variations in LCR that showed an effect on transcription activity. These results demonstrated that E2 gene polymorphisms of episomal HPV16 did not affect transcriptional regulation and suggested that nucleotide variation as well as epigenetic modification of the LCR might play a role in inducing malignant transformation of cells containing episomal HPV16.

• Asian Pacific Journal of Cancer Prevention
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• 제15권6호
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• pp.2635-2640
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• 2014

Sera of cancer patients may contain antibodies that react with a unique group of autologous cellular antigens called tumor-associated antigens (TAAs). The present study aimed to determine whether a mini-array of multiple TAAs would enhance antibody detection and be a useful approach in esophageal cancer detection and diagnosis. Our mini-array of multiple TAAs consisted of eleven antigens, p53, pl6, Impl, CyclinB1, C-myc, RalA, p62, Survivin, Koc, CyclinD1 and CyclinE full-length recombinant proteins. Enzyme-linked immunosorbent assays (ELISA) were used to detect autoantibodies against eleven selected TAAs in 174 sera from patients with esophageal cancer, as well as 242 sera from normal individuals. In addition, positive results of ELISA were confirmed by Western blotting. In a parallel screening trial, with the successive addition of antigen to a final total of eleven TAAs, there was a stepwise increase in positive antibody reactions. The eleven TAAs were the best parallel combination, and the sensitivity and specificity in diagnosing esophageal cancer was 75.3% and 81.0%, respectively. The positive and negative predictive values were 74.0% and 82.0%, respectively, indicating that the parallel assay of eleven TAAs raised the diagnostic precision significantly. In addition, the levels of antibodies to seven antigens, comprising p53, Impl, C-myc, RalA, p62, Survivin, and CyclinD1, were significantly different in various stages of esophageal cancer, which showed that autoantibodies may be involved in the pathogenesis and progression of esophageal cancer. All in all, this study further supports our previous hypothesis that a combination of antibodies might acquire higher sensitivity for the diagnosis of certain types of cancer. A customized mini-array of multiple carefully-selected TAAs is able to enhance autoantibody detection in the immunodiagnosis of esophageal cancer and autoantibodies to TAAs might be reference indicators of clinical stage.

• 한국미생물·생명공학회지
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• 제25권4호
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• pp.386-390
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• 1997

The characteristics of alcohol dehydrogenase (ADH, EC 1.1.1.1, alcohol:NAD oxidoreductase) of thermotolerant alcohol-producing yeasts, Saccharomyces cerevisiae RA-74-2 and Kluyveromyces marxianus RA-912, were compared with that of mesophilic S. cerevisiae D, an industrial strain. Under anaerobic culture condition, both S. cerevisiae RA-74-2 and D had similar level of ADH activity at 30$\circ$C, and the activity of S. cerevisiae RA-74-2 at 37$\circ$C was the same level at 30$\circ$C. However, the level of ADH activity of S. cerevisiae D at 37$\circ$C decreased about 70% of that at 30$\circ$C. The level of enzyme activity of K. marxianus RA-912, which showed lower alcohol productivity than S. cerevisiae RA-74-2 and D, was about 43% of those strains at 30$\circ$C, and decreased somewhat at 37$\circ$C. The results showed a good correlation between the alcohol productivities and the level of ADH activities of these strains grown at 30$\circ$C and 37$\circ$C. And the higher heat stability of ADH of S. cerevisiae RA-74-2 than that of S. cerevisiae D seemed to reflect the ability of high temperature fermentation. Despite of its fermentation ability even at 45$\circ$C, however, the ADH of K. marxianus RA-912 showed lower heat stability than that of S. cerevisiae D. Both S. cerevisiae RA-74-2 and D showed similar patterns of two bands of ADH isozyme, and the low band of S. cerevisiae RA-74-2 moved slightly faster than that of S. cerevisiae D. The staining intensity of the bands of S. cerevisiae D at 37$\circ$C was weaker than those at 30$\circ$C. However, S. cerevisiae RA-74-2 showed no differences in total intensity of the bands of 30$\circ$C and 37$\circ$C. As the patterns of cellular proteins and ADH isozyme of K. marxianus RA-912 were different from S. cerevisiae RA-74-2 and D, K. marxianus might have its own characteristic ADH system.

• 한국수산과학회지
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• 제27권2호
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• pp.149-154
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• 1994

다시마는 식품으로서 우수한 기능과 영양을 가지고 있지만 조체가 강인하여 제대로 식생활에 이용되지 못하고 있다. 본 연구는 동해산 다시마의 원료학적 특성과 다시마 조체를 연화할 수 있는 방법을 연구하여 다양한 다시마가공품의 개발로 소비를 촉진시키고자 하였다. 동해산 다시마는 회분이 건조중량으로 $34.74\%$ 함유되어 있는 훌륭한 무기질원이며 탄수화물이 $50.5\%$로 매우 높고 거의 대부분이 비소화성 성분으로 훌륭한 다이어트 식품이 될 수 있다. 유리 아미노산으로 Glutamic acid가 $2340mg\%$으로 매우 높게 함유되어 있는 우수한 조미재료이다. 다시마 조체의 연화에 중조는 크게 효과가 없었으나 초산과 인산염이 효과가 컸다. 초산은 $0.3\%$ 농도에서 1시간 자숙에 0.160kg/mm(${\pm}0.0494$), 2시간 자숙에 0.099kg/mm(${\pm}0.0207$) 값을 보여 충분한 연화에는 1시간 이상이 요구되었다. 인산염은 $0.2\%$에서 30분 처리만으로도 0.165kg/mm(${\pm}0.0540$)값을 보여 탁월한 연화효과를 나타내었다. 이상의 결과를 종합하면 다시마는 장시간 가열에 의해서도 상당한 수준까지 연화시킬 수 있으나 장시간 자숙처리는 다시마 조체 중에 함유되어 있는 $K^+,\;Ca^{2+},\;Mg^{2+},\;Na^+$ 등의 미네랄의 손실을 초래하게된다(좌등 등, 1981). 따라서 식초, 인산염 등을 적절히 사용하여 단시간에 연화를 완료시키는 가공방법이 바람직할 것이다.

• Nutrition Research and Practice
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• 제14권3호
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• pp.175-187
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• 2020

BACKGROUND/OBJECTIVES: In this study, we investigated the beneficial effects of skate cartilage extracts containing chondroitin sulfate (SCS) on hyperlipidemia-induced inflammation and oxidative stress in high cholesterol diet (HCD)-fed mice in comparison with the effects of shark cartilage-derived chondroitin sulfate (CS). MATERIALS/METHODS: Low-density lipoprotein receptor knockout (LDLR-KO) mice were fed HCD with an oral administration of CS (50 and 100 mg/kg BW/day), SCS (100 and 200 mg/kg BW/day), or water, respectively, for ten weeks. RESULTS: The administration of CS or SCS reduced the levels of serum triglyceride (TG), total cholesterol (TC), and LDL cholesterol and elevated the levels of high-density lipoprotein cholesterol, compared with those of the control group (P < 0.05). Furthermore, CS or SCS significantly attenuated inflammation by reducing the serum levels of interleukin (IL)-1β and hepatic protein expression levels of nuclear factor kappa B, inducible nitric oxide synthase, cyclooxygenase-2, and IL-1beta (P < 0.05). In particular, the serum level of tumor necrosis factor-alpha was reduced only in the 100 mg/kg BW/day of SCS-fed group, whereas the IL-6 level was reduced in the 100 and 200 mg/kg BW/day of SCS-fed groups (P < 0.05). In addition, lipid peroxidation and nitric oxide production were attenuated in the livers of the CS and SCS groups mediated by the upregulation of hepatic proteins of antioxidant enzymes, such as superoxide dismutase, catalase, and glutathione peroxidase (P < 0.05). CONCLUSIONS: These results suggest that the biological effects of SCS, similar to those of CS, are attributed to improved lipid profiles as well as suppressed inflammation and oxidative stress induced by the intake of HCD.

• 한국미생물·생명공학회지
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• 제45권4호
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• pp.291-298
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• 2017

식품중의 영양성분을 잘 보존하고 섭취가 용이하며 소화가 잘 될 수 있는 가공방법의 최적화를 위하여 식품원료의 동결분쇄물과 일반분쇄물을 비교하였다. 단백질의 공급원으로 알려진 서리태는 동결분쇄한 것이 일반분쇄한 것에 비해 미세하나 에너지원으로서 영양성분 보존율(이하보존율)이 4.57% 높고, 미네랄은 큰 차이가 없었다. 특히, ${\beta}$-carotene의 경우, 동결분쇄가공분말에서 11.54% 높게 포함되어 있어 비타민 A의 전구물질인 공급에 유리함을 보였다. 아로니아의 총 안토시아닌은 동결분쇄가 453.49 mg으로 일반분쇄 158.98mg 보다 보존율이 높았다. 자몽에서 열에 약한 베타카로틴은 동결분쇄가 129.47%, 니아신은 439.39%로 동결분쇄가 보존율이 높았다. 발아현미는 동결분쇄에서 단백질, 탄수화물, 니아신이 107.74%, 103.87%, 156.52%로 각각 높았다. Protease에 의한 단백질 소화율은 일반분쇄에 비하여 아로니아가 120.5%, 서리태가 101.14%로 동결분쇄방법이 단백질 소화율이 높았다. 그러나, 자몽과 발아현미는 동결분쇄에서 94.06%와 93.14%로 소화율이 떨어졌다.

• 대한화장품학회지
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• 제30권4호
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• pp.471-477
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• 2004

피부 세포는 외부의 유해 요소 즉, 스트레스 환경에 노출되었을 때 세포 자신을 보호하기 위하여 다양한 방어 및 복구 체계를 가지고 있는데 그 중 하나가 보호단백질인 열충격단백질 70 kDa의 발현이다 Glucuronic acid를 피부세포에 다양한 농도로 전처리한 다음 유해자극(열, 활성산소)을 주었을 경우, western blottting을 통해 $0.12\%$ 농도에서 세포 내 열충격단백질이 발현됨을 알 수 있었다 그리고 confocal microscopy 및 세포생존율 실험을 이용하여 열 및 활성 산소에 대한 우수한 세포보호 효과를 확인하였다. 또한 마우스 피부를 이용하여 glucuronic acid 및 수중유(O/W)형 에멀젼에 적용하였을 때 경피 흡수 양상을 비교한 결과, glucuronic acid는 빠른 경피흡수거동을 보였다(투과속도 $0.83114 mg/cm^2/h,$ 지연시간 1.2 h, 분배계수 0.114). 에멀젼에서는 투과속도는 $0.04153{\;}{\mu}g/cm^{2}/h,$ 누적투과량은 $1456.25{\;}{\mu}g/cm^{2}$로 감소하였지만 지연시간은 2.48 h으로 증가하였고 지속적인 경피흡수(서방성)를 보였다.

• Journal of Chest Surgery
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• 제2권1호
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• pp.115-132
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• 1969

The normal physiological values of Korean mongrel dogs were studied, comparing them with foreign references, on the basis of control physiological data measured on 110 cases out of the dogs submitted to the experiments in our department. The hemodynamic measurements varied widely between the both limits of reported normal control data, with the mean value of heart rate $140.4{\pm}26.6$/min., and both the systolic and diastolic arterial blood pressures $137.2{\pm}31.7$mmHg and $104.7{\pm}14.4$ mmHg, as well as the venous pressure of $9.11{\pm}2.18$ cm.$H_2O$. Hematologically, the number of red blood cells $4,571,000{\pm}767,000$per cu.mm., the amount of hemoglobin $11.57{\pm}3.74$ gm/dl and the hematocrit $37.3{\pm}7.2$ per cent, were equally the values a little lower than the reported normal means. However, the white blood cells were within the reported normal limits both in number, $10,384{\pm}4,877$ per cu. mm and their differential counts with slightly broader ranges of variation. The platelet count was $149,800{\pm}47,000 $per cu. mm and was also far below the normal, while the coagulation time $9.03{\pm}2.69$min. and the prothrombin time $13.17{\pm}6.52$sec were within normal limits, though a little prolonged. The serum electrolytes, Na $146.6{\pm}10.44$mEq/L.,K $4.46{\pm}0.84$mEq/L., CI $118.3{\pm}7.88$mEq/L. and Ca $11.45{\pm}5.62$mg./dl, and the blood glucose level of $94.9{\pm}31.79$mg./dl were essentially not different from the reported normal values. The serum proteins, total protein $7.15{\pm}1.41$gm/dl., albumin $4.09{\pm}0.77$gm./dl. and globulin $3.18{\pm}0.88$gm/dl. were included near the higher limits of the reported normal levels. The thymol turbidity 0.1-3.04 units were normal in 10 cases, and the cephaline flocculation was within normal range except 2 cases out of 26 dogs, showing two positive in 24 hours. And the nitrogen series, NPN $34.61{\pm}10.29$mg/dl. and BUN $12.77{\pm}6.37$mg./dl. were normal. It may be concluded that from the point of view of hereby measured physiological data compared with the foreign references, the Korean mongrel dogs have a compatible laboratory data with only the special regards to their tendency toward anemia in red blood cell series.