• 대한화장품학회지
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• 제45권4호
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• pp.389-397
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• 2019

리소좀(lysosome)은 진핵세포에서 에너지 대사 및 세포 내 소화 작용에 관여하는 세포 소기관으로 protease, nuclease, glycosidase, lipase, phosphatase 들이 다수 존재한다. 우리는 선행 연구결과들을 통해 난백 리소좀의 멜라닌 색소 탈색능을 보고하였다[8]. 그러나 B16F10 melanocyte 세포주에서 난백 리소좀에 의한 멜라닌 함량 변화 및 피부장벽 조절 연구는 거의 보고되지 않았다. 따라서 우리는 계란 난백(egg white)으로부터 추출한 lysosome-related organelle extract (LOE)에 의한 세포 내 멜라닌 함량 변화 및 피부장벽 강화 효과를 규명하고자 하였다. 먼저 LOE의 미백 효능을 확인하기 위해 B16F10 세포주를 이용하여 세포독성 평가를 진행하였다. B16F10 세포주에서 LOE에 의한 세포독성은 0에서 20 mg/mL 농도에서 관찰되지 않았으나, 40 mg/mL 부터 세포독성이 관찰되어 이후 모든 실험에서 최대 농도값을 20 mg/mL로 설정하였다. 먼저 LOE를 이용한 melanin contents assay 결과, 음성 대조군인 α-MSH 처리군 대비 LOE 처리군 5, 10, 20 mg/mL 농도에서 61.5 ± 4.0%, 61.4 ± 7.3%, 58.3 ± 8.3%로 세포 내 멜라닌 함량이 감소되는 것을 확인하였고, 20 mg/mL 농도 조건에서 MITF 발현 억제도 관찰하였다. LOE의 피부 장벽에 미치는 영향을 관찰하기 위해 각질형성세포주(HaCaT)를 이용하여 TEER (trans-epithelial electrical resistance) assay를 수행한 결과, LOE에 의해 농도 의존적으로 TEER 저항값이 증가하여 LOE가 피부장벽 강화에도 효과가 있음을 알 수 있었다. 또한 피부 염증 유발을 위한 TNF-α 처리조건에서도 LOE는 TEER 저항값을 증가시켜 염증 유발 조건에서도 LOE에 의해 피부장벽이 정상적으로 회복되었음을 알 수 있었다. 마지막으로 cell migration assay를 통해 LOE에 의한 세포이동 촉진 효과를 관찰한 결과, LOE는 세포분열 및 세포이동을 촉진시켰다. 위 결과들을 통해 LOE는 미백 기능 뿐 아니라 피부재생 및 피부장벽 강화에도 효과를 나타내는 소재이며, 효소안정화 및 제형화 기술이 접목된다면 향후 새로운 미백 기능성 화장품 소재로도 개발될 수 있을 것이다.

• 대한치과교정학회지
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• 제33권3호
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• pp.169-183
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• 2003

교정력에 의한 치아 이동은 기계적인 힘에 의하여 압박측에는 다양한 구조를 가진 치주 조직에 혈류의 변화가 생기며 국소적으로 산소 장력에도 변화가 생겨 저산소 상태 가 유발됨은 이미 확인한 바 있다. 본 연구는 치아 주위 골격을 형성하는 조골세포를 대상으로 교정적 치아 이동과 유사한 시험관내 조건을 설정하여 저산소 상태 시 유발되는 조골세포 고사조절 기전을 규명하고자 시행하였다. 생리적인 저산소증의 실험조건으로 $2\%$ 산소상태를 설정하여 저산소 하에서 세포가 고사(apoptosis) 됨을 확인하였고, stress유발 시 많은 관련을 가진 것으로 알려진 p-38 MAPK의 활성을 관찰하였다. 또한 p-38 MAPK의 억제제인 SB203580의 전처치로 인하여 세포의 죽음이 억제됨을 확인하였고, 저산소 상태 시 활성형태로 분절되는 caspase-3, -6및 9등의 세포고사관련 효소들의 활성 형태로의 분절이 억제됨을 확인하였으며 이러한 caspase의 기질인 Lamin-A등의 분절 또한 억제됨을 밝혔다. 또한 마이토콘드리아 내의 cytochrome c의 세포질내로의 이동 또한 조절됨을 확인함으로써 p-38 MAPK의 조절단계를 시사하여 주고 있다. 본 연구로 치아 이동 시 유발되는 저산소 상태 하에서 발생하는 조골세포의 고사 조절에 p-38 MAPK가 관여함을 확인하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제31권6호
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• pp.888-898
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• 2018

Objective: The study was conducted to investigate the effects of replacing soybean meal (SBM) with different levels of detoxified Jatropha curcas kernel meal (DJM) in growing pig diets on growth performance, nutrients digestibility and meat edibility. Methods: A total of 144 pigs with initial body weight of $20.47{\pm}1.44kg$, were randomly allocated to 6 dietary treatments with 6 replications per treatment and 4 pigs per replication for a period of 79 days. Six diets (DJM0, DJM15, DJM30, DJM45, DJM60, and DJM75) were formulated using DJM to replace 0%, 15%, 30%, 45%, 60%, and 75% of SBM. From d 37 to 42, feces and urine were total collected from six barrows in each treatment. At day 79, thirty-six pigs were slaughtered for sampling. The feed intake and weight gain were recorded, while the intestinal morphology, digestive enzyme activities, nutrient digestibility and the content of residual phorbol esters in muscles were determined. Results: The results showed that increasing the replacement of SBM with DJM decreased the parameters including body weight, average daily gain, average daily feed intake, gain-to-feed ratio, weight and villus heights of duodenum, villus height and villus height/crypt depth of jejunum, digestive enzymes (protease, amylase, lipase, and trypsin) activities, and nutrients digestibility (nitrogen deposition, digestibility of nitrogen, energy digestibility, and total nitrogen utilization) (linear, p<0.05; quadratic, p<0.05) and there was no significant difference among DJM0, DJM15, and DJM30 in all measured indices. The highest diarrhea morbidity was observed in DJM75 (p<0.05). Phorbol esters were not detected in pig muscle tissues. Conclusion: The DJM was a good protein source for pigs, and could be used to replace SBM up to 30% (diet phorbol esters concentration at 5.5 mg/kg) in growing pig diets with no detrimental impacts on growth performance, nutrient utilization, and meat edibility.

• 농업과학연구
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• 제44권4호
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• pp.513-518
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• 2017

Soybean, one of most widely used swine feed component in the world, contains non-starch polysaccharides (NSP). The digestive system of weaned pigs is not yet fully developed, and thus weaned pigs cannot easily digest diets based on corn and soybean meal. Dietary exogenous enzymes supplementation has been intensively investigated to assist digestion of anti-nutritional factors, such as NSP. This experiment was conducted to investigate the effects of dietary enzyme cocktail on growth performance, intestinal morphology, and nutrient digestibility of weaned pigs. A total 36 weaned pigs ($5.92{\pm}0.48kg\;BW$; 28 d old) were randomly allotted to 2 dietary treatments (3 pigs/pen, 6 replicates/treatment) in a randomized complete block design. The dietary treatments were a typical diet based on corn and soybean meal (CON) and CON with 0.05% enzyme cocktail (Cocktail; mixture of xylanase, ${\alpha}-amylase$, protease, ${\beta}-glucanase$, and pectinase). Pigs were fed their respective diets for 6 wk. Growth performance, morphology of ileum, apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of dry matter, crude protein, and energy of weaned pigs were measured. No significant differences (p > 0.05) were observed for growth performance for the duration of the experimental period, and morphology of ileum, and nutrient digestibility between CON and Cocktail treatment groups. Therefore, the results from the current study indicated that enzyme cocktail supplementation in diets had no influence on growth performance, intestinal morphology, and nutrient digestibility of weaned pigs.

• 한국작물학회지
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• 제41권spc1호
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• pp.25-45
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• 1996

Soyfoods have potential roles in the prevention and treatment of chronic diseases, most notably cancer, osteoporosis, and heart disease. There is evidence that carcinogenesis are supressed by isolated soybean derived products in vivo such as a protease inhibitor, phytic acid, saponins and isoflavones. It is believed that supplementation of human diets with soybean products markedly reduces human cancer mortality rates. Especially, recent papers recognize the potential benefit of soybean isoflavone components for reducing the risk of various cancers. Isoflavones exhibit a multitude of medicinal effects that influence cell growth and regulation, which may have potential value in the prevention and treatment of cancer. In addition to potential biological effects, soybean isoflavones have the important physiological functions such as the induction of Bradyrizobium japonicum nod genes and the responses of soybean tissues to infection by Phytophthora megasperma as well as biochemical activities such as antifungal and antibacterial actions. Genistin, daidzin, glycitin and their aglycone (genistein, daidzein, glycitein) are the principal isoflavones found in soybean. Malonyl and acetyl forms have also been detected but they are thermally unstable and are usually transformed during the processing in glucoside form. Most soy products, with the exception of soy sauce, alcohol-extracted soy protein concentrate, and soy protein isolate, have total isoflavone concentrations similar to those in the whole soybean. Soybean-containing diets inhibit mammary tumorigenesis in animal models of breast cancer, therefore, it is possible that dietary isoflavones are an important factor accounting for the lower incidence and mortality from breast cancer. Of the total soybean seed isoflavones, $80\~90\%$ were located in cotyledons, with the remainder in the hypocotyls. The hypocotyls had a higher concentrations of isoflavones on a weight basis compared with cotyledons. Isoflavone contents were influenced by genetics, crop years, and growth locations. The effect of crop year had a greater impact on the isoflavone contents than that of location. The climate condition might be the attribution factor to variation in isoflavone contents. Also, while the isoflavone content of cotyledons exhibited large variations in response to high temperature during seed development, hypocotyls showed high concentration in isoflavone content. So, it is concluded that one of the factors affecting isoflavone content in soybean seeds is temperature during seed development. High temperature, especially in maturity stage, causes lower isoflavone content in soybean seed. It is also suggested that there may exist a different mechanism to maintain isoflavone contents between cotyledon and seed hypocotyls. In a conclusion, soy foods may be able to have a significant beneficial impact on public health.

• Journal of Acupuncture Research
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• 제21권2호
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• pp.41-55
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• 2004

Objective : To investigate the possible molecular mechanism (s) of melittin as a candidate of anti-cancer drug, we examined the effects of the compound on the growth of human lung carcinoma cell line A549. Methods : Growth inhibitory study, flow cytometry analysis, SDS-polyacrylamide gel electrophoresis and Western blot analysis, RT-PCR and in vitro caspases activity assay were performed. Results : Melittin treatment declined the cell viability of A549 cells in a concentration-dependent manner, which was associated with induction of apoptotic cell death. Melittin treatment down-regulated the levels of Bcl-XS/L mRNA and protein expression of A549 cells, an anti-apoptotic gene, however, the those of Bax, a pro-apoptotic gene, were up-regulated. Melittin induced the proteolytic cleavage and activation of caspase-3 and caspase-9 protease in a dose-dependent manner without alteration of inhibitor of apoptosis proteins family and Akt expression. Western blot analysis and RT-PCR data revealed that the levels of tumor suppressor p53 and cyclin-dependent kinase inhibitor p21 were also remained unchanged. Conclusions : Taken together, these findings suggest that melittin-induced inhibition of human lung cancer cell growth is associated with the induction of apoptotic cell death via regulation of several major growth regulatory gene products, and melittin may have therapeutic potential in human lung cancer.

• 미생물학회지
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• 제20권3호
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• pp.125-133
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• 1982

Mutational experiments were performed to imporve the cellulase productivity of Aspergillus phoenicis KU175, isolated from the southern part of Korea, as a high cellulase producer. By treatment ultra-violet light nad 4-NQO(4-Nitroquinoline-N-Oxide), mutation waas induced, and treatment ultra-violet light and 4-NQO (4-Nitroquinoline-N-Oxide), mutation was induced, and A.phoenicis KU175-115 was finally selected for its highest avicelase production. Avicelase production of the mutant was increased about 2 times compared with those of the wild strain. However, activities of other hydrolytic enzymes, such as amylase, protease and nuclease, of the mutant strain didn't show a marked difference compared with those of the nuclease, of the mutant strain didn't show a marked difference compared with the wild strain, except slight increase in ribonuclease activity and slight decrease in glucoamylase activity. Avicelases from the mutant strain selected were purified from wheat bran culture by successive salting out, followed by dialysis and column chromatography, and their charcteristics were compared with thosw of the wild strain. Avicelase was separated into three peaks in the mutant strain as well as in the case of wild strain. Avicelase II activity of the mutant strain was prominently higher than that of the wild strain, while avicelase I and III activities of those were equivalent. The optimal pH ranges and stability of avicelase II from the mutant strain were pH4-5 and pH3.5-6.0, respectively, as well as in the case of the wild strain. The optimal temperature and thermal stability of avicelase II from the mutant strain were $40{\sim}50^{\circ}C\;and\;20{\sim}55^{\circ}C$, respectively. These results were same as those of the wild strain. By the using of Eadie-Hofastee plot, $K_m\;and\;V_{max}$ of avicelase II from the mutant and the wild strain were calculated to be 2.29mg/ml and $4.84{\mu}g$ reducing sugar as glucose per min equally, from the line fitted to the data by the least square method. Activity of avicelase II from the mutant strain was slightly activated by $Mg^{++}\;but\;inhibited\;by\;Cu^{++}, \;Mn^{++}\;and\;Zn^{++}$, as well as in the case of the wild strain. Therefore, it was concluded that the mutant didn't induce the formation of another avicelase isozyme, or the changes in the properties of avicelase, but induce the changes in the productively of the same avicelase II by the action of regulatory gane.

• 동아시아식생활학회지
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• 제23권5호
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• pp.620-628
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• 2013

In this study, the condition for preventing abnormal fermentation was set by heating the nuruk extract, such that glycosyl enzymes maintain its activity and unnecessary microbes are removed. The total colony of microbes in the heated nuruk extract was highest in number at $25^{\circ}C$ and began to reduce at 50, $60^{\circ}C$ and sharply reduced over $70^{\circ}C$. Saccharogenic power (SP), glucoamylase and acidic protease activities were highest at $50^{\circ}C$, 10, 20, 30min and ${\alpha}$-amylase activity was lower at $50^{\circ}C$ than at $25^{\circ}C$. In the pHs of the nuruk extract, as the heat temperature became higher and treatment time was longer, the pHs were reduced significantly. The total acidities of heat treatments at 50, $60^{\circ}C$ were lower by 0.2% than at $25^{\circ}C$, where as the 70, $80^{\circ}C$ treatments showed a sharp rise from the early stage of fermentation. Soluble solids showed the same aspects with the glycosyl enzymes cases. In reducing sugar, 25, $50^{\circ}C$ treatments were sharply increased from the first day of fermentation while $60^{\circ}C$ treatments began to rise from second day 70, $80^{\circ}C$ were slightly increased after the fourth day. The normally alcohol fermented treatments were 25, 50 and $60^{\circ}C$ 30min. The $70^{\circ}C$ treatments almost did not alcohol fermentation. In the preference tests, taste and total acceptability were high at 25, $50^{\circ}C$ treatments. These results suggest that makgeolli using heat treated nuruk extract also has good taste as well as did not.

• 생명과학회지
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• 제17권8호통권88호
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• pp.1115-1120
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• 2007

본 연구에서는 다섯 종류의 phytochemical (resveratrol, genistein, epicatechin gallate, diaIly disulfide, caffeic acid phenetyl ester)과 sulindac sulfide가 암 억제 단백질 p53을 유도할 수 있는지에 대해 연구하였다. 처리한 모든 phytochemical에 의해 p53 단백질의 발현이 강하게 유도된 반면, sulindac sulfide에 의해서는 p53 단백질이 유도되지 않았다. 처리한 phytochemical 중 포도껍질이나 와인에 많이 들어있는 resveratrol에 의해 p53 단백질이 농도의존적 혹은 처리시간 의존적으로 증가 발현되는 것을 확인하였다. 암 억제 단백질인 p53 하위 단계의 유전자들만 집적되어 있는 membrane microarray를 이용하여 실험을 수행한 결과, 25개의 유전자가 up-regulation 된 반면, 2개의 유전자가 down-regulation 되는 것을 확인하였다. Up-regulation 되는 유전자중 4개를 선택하여, RT-PCR을 수행한 결과 모두 membrane microarray 실험의 결과와 일치하였다. 게다가 p53 null인 HCT116 세포주를 이용한 RT-PCR을 통하여 TSP-1 유전자의 발현은 p53 의존적이지 않은 반면, MASPIN 유전자는 p53 의존적임을 확인하였다. 이러한 연구 결과는 resveratrol에 의한 화학적 암 예방법의 분자생물학적 기전을 이해하는데 도움을 줄 것으로 기대된다.

• 동의생리병리학회지
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• 제17권2호
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• pp.394-402
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• 2003

This study was designed to investigate the protective effect of Bojungmyunyuk-dan(BJMY-Dan) on the cisplatin-induced cytotoxicity of primary rat astrocytes. BJMY-Dan is an oriental herbal prescription for its ability to recover protective effects against anti-cancer chemotherapies. After astrocytes were treated cisplatin, MTT assay was performed for cell viability test. To explore the mechanism of cytotoxicity, I used the several measures of apoptosis to determine whether this processes was involved in cisplatin-induced cell damage in astrocytes. Also, astrocytes were treated with BJMY-Dan and then, followed by the addition of cisplatin. Cisplatin decreased the viability of astrocytes in a dose and time-dependent manner. BJMY-Dan increased the viability of astrocytes treated cisplatin. Astrocytes treated cisplatin were revealed as apoptosis characterized by nuclear staining and flow cytometry. BJMY-Dan protected astrocytes from cisplatin-induced nuclear fragmentation and chromatin condensation. Also, caspase-3 and caspase-9 proteases were activated in astrocytes by cisplatin. BJMY-Dan inhibited the activation of caspase proteases in cisplatin-treated astrocytes. Cleavage of [poly(ADP-ribose) polymerase](PARP) was occurred at 12hr after treatment of cisplatin in astrocytes. BJMY-Dan recovered the cleavage of PARP in cisplatin-treated astrocytes. Also, BJMY-Dan inhibited the activation of pro-apoptotic factor, Bak by cisplatin. Lastly, astrocytes stained with JC-1 and Rhodamine 123 were photographed by fluorescence microscope to visualize changes of mitochondrial membrane permeability transition(MPT) during treatment with cisplatin for 24hr. BJMY-Dan recovered the change of MPT by cisplatin in astrocytes. According to above results, BJMY-Dan may protect astrocytes from cytotoxicity induced by chemotherapeutic agents, including cisplatin.