β-phenylethyl isothiocyanate(PEITC) is a component derived from cruciferous vegetables and has been demonstrated to fight many types of cancers through various molecular pathways. In the present study, we focused on its effect on the induction of apoptotic cell death to inhibit cell growth and its molecular mechanism in HSC-4 human oral cancer cells. A colorimetric MTS assay was used to examine cell viability. The apoptotic effect and was investigated using DAPI staining and the molecular target and mechanism of PEITC-mediated apoptosis were determined by Western blotting. The result showed that PEITC inhibited oral cancer cell growth and induced apoptosis via extrinsic signaling pathway evidenced by the activation of caspase 8, truncation of bid protein and induction of death receptor(DR) 5. DR5 protein level was increased through the activation of p38 and c-Jun N-terminal kinase(JNK). These results from this study strongly suggest that DR5 is a potential molecular target for PEITC-induced apoptosis in oral cancer via p38 and JNK.

Despite of the presence of estradiol-17β (E2) in ovarian follicles, its role(s) in in vitro maturation (IVM) is still largely unknown, especially in pigs. Thus, the current study was conducted to investigate the effect of E2 on in vitro maturation (IVM) of porcine oocytes and subsequent preimplantation development using in vitro fertilization (IVF)- or somatic cell nuclear transfer (SCNT)-derived embryos. To define the effects of E2 on IVM and early embryogenesis, porcine oocytes were matured in the presence or absence of E2, fertilized in vitro and cultivated to blastocyst stage. Compared to control group, the production of MII oocytes was significantly increased by treatment with E2, accompanying with the increase in MPF content and ERK phosphorylation, and monospermic fertilization and blastocyst development rates were also greatly elevated in the E2-treated oocytes. In addition, the advantageous role of E2 was also found in blastomere survival, which was further evidenced by both elevation of anti-apoptotic transcript Bcl-XL and decrease of pro-apoptotic transcript Bax. Furthremore, these positive effects of E2 were highly reproducible in early development of SCNT embryos. Collectively, the current study strongly suggests that E2 can be used as a efficient IVM supplement leading to successful nuclear/cytoplasmic maturatioin in pigs.

Nicotine, a major teratogen of cigarettes smoke induces embryonic abnormalities during the early stages of organogenesis. In this study, the protective effect of β-carotene against nicotine–induced embryos was evaluated by morphologic scoring, nile blue staining, lipid peroxidation, SOD activity assay and real-time PCR. The embryos exposed to nicotine (1 μM) revealed remarkable morphological anomalies compared to normal control group (p<0.05), but when β-carotene (1×10‒4 μM or 5×10‒4 μM) was added concurrently to the embryos exposed to nicotine, morphological parameters were significantly improved (p<0.05). Nicotine induced oxidative stress by increased lipid peroxidation, expression of proinflammatory cytokines (TNF-α and IL-1β), caspases-3 and decreased SOD activity. However, administration of β-carotene (1×10‒4 μM or 5×10‒4 μM) restored the SOD level and decreased oxidative damage in the embryos. These results indicate that β-carotene effectively counteracts the deleterious effects of nicotine on embryos and attenuates oxidative damage possibly through its antioxidant effects.

The extracellular GH11 β-1,4-xylanase (XylY) gene (633-bp) of Paenibacillus sp. strain KYJ-16 was molecularly cloned by repeated DNA walking and nested PCR method. The xylY gene was predicted to encode an extracellular protein consisting of 611 amino acids with a nesuced molecular mass of 23 kDa and a calculated pI of 9.55. Protein blast search revealed that the enzyme consisted of a putative catalytic domain, which is homologous to a catalytic GH11 domain. The highest sequence identity (92%) was obtained as the catalytic GH11 domain of XylY was compared to that of Paenibacillus sp. strain HGF5 (GenBank accession number: EGG35584) that has not yet been characterized. Enzymatic properties of the recombinant His-tagged enzyme (rXylY) overexpressed in E. coli BL21 harboring pET-28a(+)/xylY will be also presented.

The gene (2,304-bp) encoding a novel xylanolytic enzyme (XylD) with a catalytic domain, which is 70% identical to that of Cellulomonas flavigena DSM 20109 GH6 β-1,4-cellobiohydrolase, was identified from an earthworm (Eisenia fetida)-symbiotic bacterium, Cellulosimicrobium sp. strain HY-13. The enzyme consisted of an N-terminal catalytic GH6-like domain, a fibronectin type 3 (Fn3) domain, and a C-terminal carbohydrate-binding module 2 (CBM 2). XylDΔFn3-CBM 2 displayed high transferase activity (788.3 IU mg-1) toward p-nitrophenyl (PNP) cellobioside, but did not degrade xylobiose, glucose-based materials, or other PNP-sugar derivatives. Birchwood xylan was degraded by XylDΔFn3-CBM 2 to xylobiose (59.2%) and xylotriose (40.8%). The transglycosylation activity of the enzyme, which enabled the formation of xylobiose (33.6%) and xylotriose (66.4%) from the hydrolysis of xylotriose, indicates that it is not an inverting enzyme but a retaining enzyme. The endo-β-1,4-xylanase activity of XylDΔFn3-CBM 2 increased significantly by approximately 2.0-fold in the presence of 50 mM xylobiose.

Acyl-CoA synthetase 4(ACSL4) is an arachidonate-preferring enzyme abundant in steroidogenic tissues and postulated to modulate eicosanoid production. The human and mouse ACSL4 gene are mapped on chromosome X. The female mice heterozygous for ACSL4 deficiency became pregnant less frequently and produced small litters, with 40% of embryos surviving gestation. In this study, we examined the regulation of ACS4 by estradiol-17β and progesterone (P4) in the human endometrial cancer cell line HTB-1B. ACSL4 mRNA was increased in a dose-dependent manner. Also, expression of ACSL4 gene was up-regulated in a time-dependent manner in HTB-1B cells. However, combined treatment with progesterone and estradiol-17β modestly decreased the levels of ACS4L mRNA as compared with the estradiol-17β and progesterone respectively. Overall, these results suggest that the ACSL4 gene is regulated by progesterone and estradiol-17β in the HTB-1B cells.

The effects of β-lapachone on gastric secretion were investigated. The pylorus of male Sprague-Dawley rats was ligated and intraduodenally injected with β-lapachone, and the volume, pH, free HCl, and total acidity of gastric fluid were measured 6 hours after the operation. Treatment with β-lapachone resulted in dose-dependent inhibition of gastric secretion Gastric fluid was reduced to 42.9% of control level by 100 mg/kg of β-lapachone, leading to an increase of pH to 6.70 from 1.85 in the control group. In parallel with the increase of pH, at this dosage, free HCl and total acidity decreased to 16.7% and 12.0%, respectively, of control levels. β-Lapachone exhibited ED50 values of 72, 46, and 47 mg/kg for inhibition of gastric volume, free HCl, and total acidity, respectively, implying a superior efficacy on gastric acid to volume. In comparison, pantoprazole (30 mg/kg) reduced the volume, free HCl and total acidity of gastric fluid to 53.0%, 26.0%, and 25.0%, respectively, of control levels, resulting in an increase in pH to 6.36. In the current study, it was confirmed that β-lapachone at an appropriate dose (100 mg/kg) exerted a higher inhibitory effect on gastric secretion than pantoprazole (30 mg/kg), a well-known proton-pump inhibitor. Therefore, it is suggested that β-lapachone could be a candidate compound for prevention or treatment of gastric ulcers induced by diverse psychological and physical stimuli.

중고압 하에서 β-glucosidase 효소반응을 물리화학적 관점에서 연구하였다. 모델 기질 (p-nitrophenyl-β-D-glucopyranoside)에 대한 β-glucosidase 효소의 작용에 대한 압력 효과를 실험하였다. 즉, 압력 조건(25MPa, 50 MPa, 75 MPa, 100 MPa)과 시간 (10분, 60분, 1시간, 6시간, 24시간, 40시간)의 처리 조건에서 효소 활성도를 분광학적인 표준방법에 따라 측정하였다. 효소-기질 반응의 단계를 크게 kinetic 구간과 평형 구간으로 구분하여 물리화학적 모델을 적용하여, 정역반응속도 상수, 평형상수, 압력에 의한 부피 감소 등을 산출하였다. 대기압에서 100MPa까지 압력이 증가할수록 효소-기질 반응의 생성물이 더 많이 형성되었으며 전형적인 kinetic 구간과 평형 구간이 나타났다. 압력, 시간, 생성물농도 등의 데이터로부터 kinetic 구간과 평형에서의 생성물 예측 모델을 완성하였다. 결론적으로 중고압 처리에 의하여 효소-기질 반응이 촉진됨을 알 수 있었고, 임의의 압력 및 시간 조건에 따른 생성물의 농도를 예측할 수 있게 되었다.

The purpose of this study was to investigate the body composition, biochemical parameters, and consumption of convenience foods according to β-3 adrenergic receptor polymorphism in university students. A survey was conducted on a total of 486 students - 189 males and 297 females. Based on a self-reporting method, questionnaires were administered for over 20 minutes, and β-3 adrenergic receptor and blood samples were also analyzed. The genotype frequencies of β-3 adrenergic receptor polymorphism were Trp/Trp homozygote (73.0%) and Trp/Arg heterozygote (27.0%) in male students. For the female students, the distribution of genotypes was Trp/Trp (71.0%) and Trp/Arg (29.0%). There were no differences according to biochemical parameters (ALT, cholesterol, triglyceride, HDL-cholesterol, LDL-cholesterol, and hemoglobin) or body composition. Males with TT genotype frequently ate Ramyon (2.40±0.52), Cup Ramyon (2.37±0.39), Kimchi (2.23±0.61), and frozen meat (2.00±0.44), whereas males with TA genotype ate Fries (frozen food) (1.90±0.79), Smoked meat (1.67±0.81), and Canned fruit (1.64±0.81). Females with TT genotype frequently ate Frozen fries (2.21±0.35), Kimbab (2.12±0.44), and Ramyon (1.85±0.40), whereas females with TA genotype frequently ate Kimchi (1.73±0.98), Fries (frozen food) (1.46±0.26), and Cup Ramyon (1.30±0.34). When questioned about satisfaction about body shape, 22.8 and 60.8% of those with TT genotype answered that they were 'satisfied' or needed to 'lose weight', respectively, whereas 18.0 and 63.9% of those with TA genotype answered that they were 'satisfied' or needed to 'lose weight', respectively. In conclusion, this study found no significant effects in terms of β-3 adrenergic receptor polymorphism, which suggests that health-promoting education needs to be developed so that university students appropriately recognize their bodies and control their weight in desirable ways. Therefore, it is necessary to educate individuals with TT genotype how to buy reasonable foods by understanding the interrelationship between convenience foods and health care and by checking the nutrition index labels on convenience foods. Thus, it is recommended that a health-promoting program be developed for the promotion of healthy lifestyles.

생활수준이 향상되면서 건강식품을 통해 건강한 삶을 누리려는 경향이 증대되고 있다. 더불어 식물유래 천연물 성분에 대한 관심도 높아지면서 각종 영양소를 함유하고 있는 버섯에 대한 관심도 증가하고 있는 추세이다. 그 중 버섯에 함유 되어있는 β-glucan이 주목되고 있는데, β-glucan 은 면역 활성체의 기능, 항산화능, 생체조직 재생과 치유기능, 항생제, 항균, 항바이러스 및 대식세포를 자극하여 돌연변이 세포를 인식하고 공격하는 항종양 효과 등이 있다고 보고된 바 있다. 본 실험에서는 꽃송이 버섯(Sparassis crispa)과 표고 버섯(Lentinus edodes)을 대상으로 하여 다양한 배양방법에 따른 β-glucan 함량과 항산화 활성의 변화를 조사 하였다. 꽃송이 버섯, 표고 버섯의 β-glucan 함량은 메가자임사의 β-glucan assay kit(Megazyme International Ireland Limited, Wicklow, Ireland)를 사용하여 측정하였고, 항산화 활성의 검정은 DPPH(1,1-diphenyl-2-picrylhydrazyl) 활성감소와 nitric oxide radical 소거능 감소를 통해 확인하였다. β-glucan의 함량은 버섯 수확량과 반비례 관계를 나타내었으며 항산화 활성과는 상관관계가 있는 것으로 판단된다. 본 연구를 통하여 높은 β-glucan 함량을 지니며 항산화 활성능이 우수한 꽃송이 버섯과 표고버섯의 기능성 증대를 위한 배양방법을 제안할 수 있었다.

Edible mushrooms have been used and researched as medicinal ingredients. They improve immunity and contain excellent anticancer components with large amounts of minerals such as iron, calcium, and potassium. Due to this reason, it has been hailed as a raw material for functional foods. Especially, β-glucan, known to be contained in Ganoderma incidum Karst, Coriolus versicolor, and Phellinus linteus, was reported to inhibit proliferation of cancer cells by activating immune function (macrophages, natural killer cells and T-cells). In addition, mushroom polysaccharides dissolve in water but are undigested ingredients, resulting in stimulating the immune system as well as staying of parasympathetic nervous system in the stomach for a long time. As a result, they induce increase of T-cells and NK cell that attacks cancer cell and has effect on the discharge of body's waste products, blood purification, and constipation improvement. In this study, we report the culture characteristics of Sparassis crispa as to the medium growth compositions, yeast, and elicitor treatments to investigate the optimal condition for the highest β-glucan production in mushrooms.

xylanolytic gut bacterium isolated from Eisenia fetida, Cellulosimicrobium sp. strain HY-13, produced an extracellular glycoside hydrolase capable of efficiently degrading mannose-based substrates such as locust bean gum (LBG), guar gum, mannotetraose, and mannopentaose. The purified mannan-degrading enzyme (ManS, 34,926 Da) from strain HY-13 was found to have an N-terminal amino acid sequence of DEATTDGLHVVDD, which has not yet been identified. Under the optimized reaction conditions of 50℃ and pH 7.0, ManK exhibited extraordinary high specific activities of 7,109 IU/mg and 5,158 IU/mg toward LBG and guar gum, respectively, while the enzyme showed no effect on sugars substituted with p-nitrophenol and various non-mannose carbohydrates. ManK strongly attached to Avicel, lignin, β-cyclodextrin, and poly(3-hydroxybutyrate) granules, but not bound to chitin, chitosan, curdlan, or insoluble oat spelt xylan. The aforementioned characteristics of ManS suggest that it is a unique endo-β -1,4-mannanase with out additional carbohydrolase activities, which differentiates it from other well-known carbohydrolases.

To investigate the toxicological effects of β-glucan, we performed basic studying on β-glucan in Sprague-Dawley (SD) rats. Standard endpoints in this study included mortality, clinical observations, changes of body weights, analysis on food consumption, ophthalmoscopic examination, hematologic examination, serum biochemistry,analysis of organ weights, gross anatomic pathology and histopathology. No clinical signs and mortality were observed in animals treated with beta-glucan throughout the experimental period. The average body weight of each treatment groups showed similar levels at end of experiment. There were no treatment-related changes in mortality,body weights changes, food consumption, ophthalmoscopic examination. Although there were statistically significant differences between the control and treated groups in some relative and absolute organ weights, and hematological and biochemical analysis, the data were in biologically normal ranges and did not show a dose-dependent manner. In the morphological change, hepatic tissue were not showed ballooning degeneration and irregular arrangement of hepatic cell in β-glucan treatment groups with control group. Also, organs weights (liver, heart, kidney and stomach) and hematological indices (WBC, RBC, Hb, Hct and Platelet) did not show statistically significant differences among the experimental groups. In summary of these results, there were no changes in mortality, mean body weight, clinical signs, food consumption. There were no changes in ophthalmological examination, hematology, blood chemistry,necropsy and histopathology. In conclusion, although further investigation of glucan should be performed in the functions registered in many ancient literatures, β-glucan is physiologically safe and may have potential as candidate food for human health.

As a growth-template of ZnO nanorods (NR), a hexagonal β-Ni(OH)2 nanosheet (NS) was synthesized with the low temperature hydrothermal process and its microstructure was investigated using a high resolution scanning electron microscope and transmission electron microscope. Zinc nitrate hexahydrate was hydrolyzed by hexamethylenetetramine with the same mole ratio and various temperatures, growth times and total concentrations. The optimum hydrothermal processing condition for the best crystallinity of hexagonal β-Ni(OH)2 NS was determined to be with 3.5 mM at 95˚C for 2 h. The prepared Ni(OH)2 NSs were two dimensionally arrayed on a substrate using an air-water interface tapping method, and the quality of the array was evaluated using an X-ray diffractometer. Because of the similarity of the lattice parameter of the (0001) plane between ZnO (wurzite a = 0.325 nm, c = 0.521 nm) and hexagonal β-Ni(OH)2 (brucite a = 0.313 nm, c = 0.461 nm) on the synthesized hexagonal β-Ni(OH)2 NS, ZnO NRs were successfully grown without seeds. At 35 mM of divalent Zn ion, the entire hexagonal β-Ni(OH)2 NSs were covered with ZnO NRs, and this result implies the possibility that ZnO NR can be grown epitaxially on hexagonal β-Ni(OH)2 NS by a soluble process. After the thermal annealing process, β-Ni(OH)2 changed into NiO, which has the property of a p-type semiconductor, and then ZnO and NiO formed a p-n junction for a large area light emitting diode.

The phase Ti-Nb-Sn-HA bio materials were successfully fabricated by high energy mechanical milling and pulse current activated sintering (PCAS). Ti-6Al-4V ELI alloy has been widely used as biomaterial. But the Al has been inducing Alzheimer disease and V is classified as toxic element. In this study, ultra fine sized Ti-Nb-Sn-HA powder was produced by high energy mechanical milling machine. The phase Ti-Nb-Sn-HA powders were obtained after 12hr milling from phase. And ultra fine grain sized Ti-Nb-Sn-HA composites could be fabricated using PCAS without grain growth. After sintering, the microstructures and phase-transformation of Ti-Nb-Sn-HA biomaterials were analyzed by scanning electron microscope (SEM) and X-ray diffraction (XRD). The relative density was obtained by Archimedes principle and the hardness was measured by Vickers hardness tester. The -Ti phase was obtained after 12h milling. As result of hardness and relative density, 12h milled Ti-Nb-Sn-HA composite has the highest values.

E ffe ct of aluminum (al) on the plasma vite llogenin (VTG) , alkaline-labile phosphorus (ALP P ) , Calcium (Ca) , glutamate pyruvate transaminase (GP T) and hepatosomatic index (H SI ) we re examined in the e stradiol-17β (E 2)-administe red immature rockfish, Sebaste s schlege li. F ish we re intrape r itone ally inje cted with E 2 (5 ㎎/㎏ body we ight (B .W.) ) and/ or Al (0, 0.1, 1, 5 and 10 ㎎/㎏ B .W.) and then plasma we re extracted at 7 days. Thick VTG band was de te cted at a mole cularwe ight position of about 170 kD a in SD S-P AGE using the plasma of E 2-administe red rockfish. This band was reduced by the administration of Al to rockfish. Al reduced the concentrations of ALP P and Ca in a concentration-dependent way and a significant reduction occur red at Al concentrations gre ate r than 5 ㎎/㎏ and 10 ㎎/㎏, re spe ctive ly. The concentration of GP T was incre ased in concentration-dependent way at all the group of Al-administe red rockfish. H SI was reduced in a concentration-dependent way. The se re sults sugge st that Al inte r fe re with E 2-induced vite llogene sis by be ing toxic to hepatocyte .

A phytoremediation study has been conducted to see if some known aquatic plants can remove the pesticides, endosulfan-α, β and fenitrothion which are frequently used in the crop protection and golf course management, and are likely to exist as residual pollutants in the aquatic ecosystems. Among the five aquatic plants tested in the microcosms, water lily Nymphaea tetragona Georgi showed the highest degradation efficacies (85~95%) for the three pesticides as opposed to the control(13~26%). The efficacies for the other plants were in the range of 46~80% in the order of Pistia stratiotes, Cyperus helferi, Eichhornia crassipes, and Iris pseudoacorus. Fenitrothion, an organo-phosphorus pesticide, was much more vulnerable to the phytoremediation than the organo-chlorine pesticides, endosulfan-α and endosulfan-β. The kinetic rate constants (min-1) for removal of the three pesticides were more than 10 times higher than the control (non-planting) in case of Nymphaea tetragona Georgi. This aquatic plant showed kinetic rate constants about 2 times as much as the lower kinetic rate constants shown by Iris pseudoacorus. The reason for the highest degradation efficacy of water lily would be that the plant can live in the sediment and possess roots and broad leaves which could absorb or accumulate and degrade more pollutants in association with microbes. These results indicate that some of the selected aquatic plants planted near the agricultural lands and wetlands could contribute to remediation of pesticides present in these places, and could be applicable to protection of the aquatic ecosystems.

β-Glucosidase 효소활성이 높은 균주를 선발하기 위하여 다양한 김치에서 분리된 젖산균의 β-glucosidase 활성을 탐색하였다. 김치에서 분리된 156개의 젖산균 중 134개의 균주만이 cellobiose를 탄소원으로 대사하였으며, 세포내 β-glucosidase 활성이 세포외 활성보다 현저히 높았다. 배추김치에서 분리된 W. cibaria KFRI88010 균주가 3.7±0.5 unit/mg protein으로서 가장 높은 세포내 β-glucosidase 효소활성을 나타내었으며, 효소활성은 pH 5, 37oC 반응조건에서 가장 높게 나타났다. Mn2+를 비롯한 금속이온은 효소활성을 크게 저해하였다. W. cibaria KFRI88010 균주를 배양할 때 사용한 탄소원 중, fructose는 cellobiose나 glucose와 비교하여 약 2.5배 이상의 높은 세포내 β-glucosidase 효소활성을 나타내었다.