• Asian Pacific Journal of Cancer Prevention
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• 제16권3호
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• pp.915-921
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• 2015

Context: In the last half century, discovering, developing and introducing of clinical agents from marine sources have seen great successes, with examples including the anti-cancer compound trabectedin. However, with increasing need for new anticancer drugs, further exploration for novel compounds from marine organism sources is strongly justified. Objective: The major aim of this study was to evaluate the antitumor and antioxidant potential of Sargassum tenerrimum J.Agardh (Sargassaceae) on Ehrlich ascites carcinoma (EAC) in Swiss albino mice. Materials and Methods: An ethanol extract of S. tenerrimum (EEST) from whole algae was used to evaluate cytotoxicity followed by in vivo assessment of toxicity, using biochemical parameters including hepatic and non-hepatic enzymes. Antioxidant properties were examined in animals bearing EAC treated with daily oral administration of 100-300 mg/kg extract suspension. Results: Antitumor effects of EEST in EAC bearing mice was observed with LD50 1815 mg/kg. Parameters like body weight, tumor volume, packed cell volume, tumor cell count, mean survival time and increase in life span in animals in the EAC bearing animals treated with EEST 300 mg/kg was comparable with control group. Significant differences were also seen with changes in total protein content, hepatic enzymes contents, MDA level, and free radical scavenging enzymes in untreated vs. EEST treated group animals. Conclusions: Evaluation of antioxidant enzymes and hepatic enzymes in the EAC animal model treated with EEST exhibited similar effects as the positive control drug 5-flurouracil. S. tenerrimum extracts contain effective antioxidants with significant antitumor activity.

• KSBB Journal
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• 제28권1호
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• pp.54-59
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• 2013

Three isolates, E. faecium P1, P2 and P3, from intestinal drugs of three phamaceutical companies, four clinical vancomycin resistant isolates, E. faecium V1, V2, V3 and E. faecalis V4, and three isolates, E. faecalis DW01, DW07 and DW14, from infant feces were tested for the presence of virulence genes, ace, agg, esp, efaA, gelE, sprE, vanA and vanB as well as fsrABC, regulatory genes of gelE and sprE, cylMBA, cytolysin activation genes and cpd, cob and ccf, pheromone genes by PCR and for their phenotype activities such as protease, biofilm formation, cell clumping and hemolysis. The genes encoding cell surface adherence proteins, ace, agg, esp and efaA, were predominantly amplified from the vancomycin resistant strain V4 and the fecal isolates DW01, DW07 and DW14. Both protease and biofilm formation activity were detected only from E. faecalis V4 from which the PCR products of gelE and spreE as well as fsrABC were amplified. The pheromone genes were amplified from the V4, DW01, DW07 and DW14 strains and these strains showed clumping activity. Biofilm formation was observed from the strains DW01, DW07 and DW14, all of which produced PCR products of pheromone, and V4 as well. Whole cytolysin regulator genes were amplified from DW01, DW07 and DW14 and ${\beta}$-hemolysis activity was detected from these strains. Any virulence genes or activities except the pheomone gene ccf were not detected from the pharmaceutical isolates, E. faecium P1, P2 and P3.

• Radiation Oncology Journal
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• 제15권3호
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• pp.197-206
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• 1997

목적 : Phospholipase D (PLD)는 phosphatidylcholine을 phosphatidic acid (PA)와 choline으로 가수분해 시키는 효소이다. 최근 이 효소는 다른 phospholipase들과 유사하게 세포 신호전달과정에 관여하는 것으로 알려져 많은 관심의 대상이 되고 있으며, 아울러 발암과정에 관여하리라는 추측을 하게 하고 있다. 이 실험에서는 쥐를 방사선 조사하여 각 조직에서 올레산-PLO에 미치는 영향을 관찰하였다. 방법 : PLD assay를 위한 반응 혼합물에는 $0.1\;\muCi$의 $1,2-di[1^{14}C]palmitoyl$ phosphatidylcholine, 0.5mM phosphatidylcholine, 5mM sodium oleate, $0.2\%$ taurodeoxycholate, 50mM HEPES buffer(pH 6.5), 10mM $CaCl_2$와 25mM KF 를 함께 넣어주었다. 생성된 PA는 TLC로 분리하여 그 방사능을 측정하였다. 사용된 동물은 암컷 Wistar 쥐로서 코발트 60 원격치료 기기를 이용, 조사범위를 $10cm\times10cm$로하여 분당 선량율 2.7Gy로 방사선 조사선랸 l0Gy와 25Gy를 조사 하였다. 결과 : PLD 활성은 폐조직에서 가장 높았으며 신장, 근육, 리, 비장, 골수, 흥선. 간의 순으로 나타났다. 방사선 조사결과 PLD 활성에 변동을 보인 조직은 흥선, 비장, 폐와 골수이며, 특히 흉선과 비장은 PLD의 할성이 각각 2배 이상 증가한 것으로 관찰되었다. 이와는 반대고 골수의 PLD는 $30\%$ 이상 감소한 것으로 나타났다. 한편 PLD 활성값이 가장 낮은 판은 방사선 영향을 거의 받지 않는 것처럼 보였다. 결론 : 동물전신에 방사선 조사시 PLD가 가장 민감한 영향을 받는 조직은 림프양 기관과 조혈 세포인 것으로 보여 PLD가 이들 조직의 생리기능과 밀접한 관계가 있음을 암시해 주고있다. 더 나아가 방사선 긴장 (radiation stress)이 이들 조직의 세포증식내지 괴사현상연구에 중요한 수단을 제공해 줄 수 있을 것이다.

• 한국가축번식학회지
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• 제19권4호
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• pp.307-322
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• 1996

The corpus luteum (CL) is formed by the action of a surge of luteinizing hormone (LH) on the pre-ovulatory follicle. Luteal cells derived from granulosa and theca interna cells continue to secrete progesterone for about two weeks. LH in domestic animals is essential for the normal secretion of progesterone at all stages of the luteal phase. For this process in the rodents, 20$\alpha$-hydroxysteroid dehydrogenase (20$\alpha$-HSD) is indispensable. 20$\alpha$-HSD is an enzyme to be a biologically inactive steroid. This enzyme plays a critical role in the regulation of the rat luteal function and reported to be present in steroid-producing tissues such as the testis and adrenal gland. We have purified 20$\alpha$-HSD and found two distinct 20$\alpha$-HSD molecules (HSD-1 and HSD-2). Their molecular weights are both estimated to be 33kd.The amino acid compositions of HSD-1 and HSD-2 are mostly similar, but there is a slight difference in the content of lysine. We demonstrated that 1) CL of previous generations contribute more to whole ovarian 20$\alpha$-HSD activity, 2) newly formed corpora lutea contain only 20$\alpha$-HSD-1 activity, and 3) old CL express activities of each HSD isozyme as shown in the luteal tissue of cycling rats on the day of diestrus where only degenerating old CL exist. The increase in 20$\alpha$-HSD activity identified seems to be related to the increase in the numbers of 20$\alpha$-HSD-positive cells. Interestingly, 20$\alpha$-HSD-1 activities were strongly found in the follicle fluids and theca interna cells by immunohistochemical study. Thus, the activity of 20$\alpha$-HSD may be related to a survival mechanism of those luteal cells and follicles remaining in the ovaries. Luteal cells arise from two sources. The small luteal cells are all of theca cell origin, while the large luteal cells are mainly of granulosa cell origin. CL of Korean Native Cattle, as those of other animal species, contains two morphologycally and functionally distinct luteal cell populations, such as small and large luteal cells as well as nonluteal cells. In all reproductive states except in the late luteal phase, the bovine CL also contained more small luteal cells than large luteal cells. Luteal tissue secretes a variety of growth factors (proteins) and the pattern of secretion changes during all stages of the luteal phase. These growth factors could be important in regulating the function of the bovine corpus luteum and may act in a potential endocrine autocrine and paracrine mechanisms. Therefore, further work has to be done to elucidate the role of growth factors in the ovary, especially in the corpus luterum. Interest should be focussed on interaction of these growth factors in the regulation of luteal cell and the localization of cytokine synthesis in differnet luteal cells.

• 한국잡초학회지
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• 제15권2호
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• pp.148-153
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• 1995

Glyphosate (N-[phosphonomethyl]glycine)를 토마토(Lycopersicon esculentum Mil)의 동화부위(同化部位)에 부분처리(部分處理)하거나 전(全) 식물체(植物體)에 분무처리(噴霧處理)하였을 때 EPSP-synthase의 활성(活性) 감소(減少)가 나타났다. EPSP-synthase의 활성(活性)은 처리된 식물체(植物體)의 엽록소(葉綠素)의 감소보다 시기적으로 먼저 나타나는 현상이었다. EPSP-synthase의 활성(活性)은 glyphosate처리(處理)에 민감한 효소(酵素)로서 토마토의 세포현탁배양조직(細胞顯濁培養組織)과 분열조직(分裂組織)간에는 활성(活性)의 차이가 없없다. EPSP-synthase의 활성은 4-6 nkat/mg protein 정도이었다. EPSP-synthase의 활성(活性)억제는 glyphosate 처리(處理) 36시간 후 부터 나타나기 시작하였고, 엽록소(葉綠素)의 감소는 처리 48 시간 후 부터 나타나기 시작하였다. 세포현탁배양(細胞顯濁培養)에서 치사농도(致死濃度) 이하(以下)에서 glyphosate는 생체중(生體重)을 저하(低下)시켰으며 생육단계(生育段階) 중 lag-phase를 연장(延長)시켜 생육(生育)이 더디도록 하였다. Glyphosate 존재(存在)하에서 생체중(生體重)은 계대(繼代)배양 후 14일이 지난 뒤에 생체중(生體重)이 최고(最高)에 달하였다. EPSP-synthase에 대한 gly-phosate의 억제(抑制)효과는 lag-phase에서 심하게 나타났다.

• Journal of Periodontal and Implant Science
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• 제24권1호
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• pp.144-154
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• 1994

Final goal of periodontal treatment is to reconstruct the destructed periodontal tissue as well as to remove the necrotic pathologic elements. The purpose of this study is to investigate on the effect of Zizyphi extract to the inhibitory ability on collagenolytic activity of P gingivalis, biologic activity of gingival fibroblasts, and on the collagen and protein synthesis of gingival fibroblasts. Gingival fibroblast from giniva of first bicuspids from patient for orthodontic treatment were used and cultured. For the measurement of inhibitory ability of collagenolytic activity, crude enzyme was extracted and used on the basis of modified Ono's method. On the inhibition of collagenolytic enzyme from herbal extracts, collagenokit CLN-100 were used. The cellular activity of gingival fibroblast, were studied using MTT solution and measured optical density on 570mm by ELISA reader. To measure the effects on the ability of whole protein and collagen synthesis, cell membrane was destructed with ultrasonic grinder after culturing, centrifuged and counted by liquid scintilation counter. The inhibitory effects on producing of $IL-l{\beta}$ by monocyte, after promotion of producing $IL-l{\beta}$ by LPS, were compared with the mixture of herbal extracts and other drugs using thymocyte stimulation assay. About inhibitory effects of $PGF_2$. by gingival fibroblasts, herbal extract was compared with the addition of the other control groups using enzyme imunoassay. On the inhibition of collagenolytic activity by P. gingivalis, benzene extracts showed the most efficient inhibitory effects among the $19{\mu}g/ml$ of the compared extracts and 40.5% by Tetracycline. On the cellular activity promoting effects, compared extracts showed a bit of more effects than PDGF of $100{\mu}g/ml$ concentration and IGF of $20{\mu}g/ml$ concentration. All of the PDGF, IGF, Zizyphi Fructus extract should increase in collagen synthesis, but especially 70% ethylalcohol extracts of Zizyphi Fructus showed comparably high effects among the compared extracts. Effects on whole protein synthesis were slightly increased on every extract but especially 70% ethylalcohol extract showed significantly effective than any other estract. On the inhibitory effects of Zizyphi Fructus $IL-l{\beta}$ production by monocyte, compared extracts showed 70% of highly inhibitory effect than that of 60% inhibition effects on controlled group and each extracts showed no significant difference. In $PGF_2$ production inhibitroy effect of Zizyphi Fructus gingival fibroblasts, Herbal extracts showed 70% of inhibition comparing with tat of 90.2% of controlled group, but each extracts showed similar effects excluding the $H_2O$ extracts. These results suggested that Zizyphi Fructus might be useful medicine for inhibition of inflammatory mediator including $IL-l{\beta}$ and $PGF_2$.

• Journal of Microbiology and Biotechnology
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• 제20권9호
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• pp.1300-1306
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• 2010

Ethyl (R, S)-4-chloro-3-hydroxybutanoate (ECHB) is a useful chiral building block for the synthesis of L-carnitine and hypercholesterolemia drugs. The yeast reductase, YOL151W (GenBank locus tag), exhibits an enantioselective reduction activity, converting ethyl-4-chlorooxobutanoate (ECOB) exclusively into (R)-ECHB. YOL151W was generated in Escherichia coli cells and purified via Ni-NTA and desalting column chromatography. It evidenced an optimum temperature of $45^{\circ}C$ and an optimum pH of 6.5-7.5. Bacillus subtilis glucose dehydrogenase (GDH) was also expressed in Escherichia coli, and was used for the recycling of NADPH, required for the reduction reaction. Thereafter, Escherichia coli cells co-expressing YOL151W and GDH were constructed. After permeablization treatment, the Escherichia coli whole cells were utilized for ECHB synthesis. Through the use of this system, the 30 mM ECOB substrate could be converted to (R)-ECHB.

• Journal of Animal Science and Technology
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• 제48권6호
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• pp.907-920
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• 2006

본 실험은 Lactobacillus spp.와 Bacillus spp. 의 Salmonella typhimurium, E. coli 및 Listeria monocytogenes에 대한 억제활성과 송아지 분변 분리균주의 생장 및 억제활성의 특성을 알아보기 위해 실시되었다.Lactobacillus spp.와 Bacillus spp.의 병원성 세균 Salmonella typhimurium에 대한 억제활성은 Lactobacillus helveticus CU631이 가장 높았고, Bacillus spp.는 활성이 약하였다. 송아지 분변 분리균주를 동정한 결과 Lactobacillus pentosus CU13과 CU05, Pediococcus pentosaceus CUR02, Lactococcus lactis ssp lactis CUM14로 확인되었다. Lactobacillus rhamnosus CU02와 Lactobacillus pentosus CU13의 Listeria monocytogenes에 대한 억제활성에 영향을 미치는 배지성분과 첨가수준은 Tween 80 1.0%, peptone 3.0%, yeast extract 3.0%, glucose 3.0% beef extract 3.0%, NaCl 1.0～3.0%이며, whole cell과 세포벽 물질은 Listeria monocytogenes에 대하여 억제활성을 나타내었다. 억제 성향을 강하게 보인 균체 배양액을 80℃로 열처리한 결과 억제력은 나타나지 않았으나 catalase 및 Proteinase-K 처리는 억제활성에 영향을 미치지 않았다. 이 결과 억제활성 물질은 유기산에 의한 것으로 사료된다. Lactobacillus pentosus CU13과 Lactobacillus rhamnosus CU02의 21균주에 대한 억제 능력을 측정한 결과 병원성균주를 포함한 16균주에서 억제활성을 보였으나 5균주에서는 억제성향을 나타내지 않았다. Escherichia coli O157:H7을 감염시킨 mouse 중 Lactobacillus pentosus CU13을 접종한 경우 다소 체중 회복 현상이 나타났으나, Lactobacillus rhamnosus CU02을 접종한 경우 체중 회복이 빠르게 나타났다.

• 생약학회지
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• 제49권1호
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• pp.47-54
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• 2018

Astilbe rubra (AR) is a perennial, belongs to the Saxifragaceae family, it contains tannin and triterpene. AR has been used in republic korea to improve toxication, fever, pain and convulsion. Recently, number of natural products have been analyzed for potential pharmacological activities including anti-cancer, anti-obesity and anti-diabetic medication. Consequently, we investigated the growth inhibition effect of Astilbe rubra water extract (WAC), ethanol extract (EAC) and bergenin on Hela cell (human adenocarcinoma cell). From whole plant of A. rubra, bergenin was isolated by column chromatography and its structures were identified by $^1H$, $^{13}C$ NMR and IT TOF-ESI MS. High extraction efficiency of bergenin was shown at 0.95% under 60 min reflux extraction with 50% MeOH. The MTS assay showed that EAC (ethanol extract) treatment increased cell death in a dose-dependent manner. Moreover, EAC treatment on Hela cell increased apoptotic cell death and caspase-3 activity. Results suggest that EAC has growth inhibition effect on Hela cells, but not WAC and bergenin. $500{\mu}g/mL$ EAC treatment inhibited Hela cell at $60.2{\pm}1.5%$.

• Animal Bioscience
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• 제36권8호
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• pp.1285-1292
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• 2023

Objective: The objective of this study was to develop a novel endolysin (PanLys.1) for the specific killing of the ruminal hyper-ammonia-producing bacterium Peptostreptococcus anaerobius (P. anaerobius). Methods: Whole genome sequences of P. anaerobius strains and related bacteriophages were collected from the National Center for Biotechnology Information database, and the candidate gene for PanLys.1 was isolated based on amino acid sequences and conserved domain database (CDD) analysis. The gene was overexpressed using a pET system in Escherichia coli BL21 (DE3). The lytic activity of PanLys.1 was evaluated under various conditions (dosage, pH, temperature, NaCl, and metal ions) to determine the optimal lytic activity conditions. Finally, the killing activity of PanLys.1 against P. anaerobius was confirmed using an in vitro rumen fermentation system. Results: CDD analysis showed that PanLys.1 has a modular design with a catalytic domain, amidase-2, at the N-terminal, and a cell wall binding domain, from the CW-7 superfamily, at the C-terminal. The lytic activity of PanLys.1 against P. anaerobius was the highest at pH 8.0 (p<0.05) and was maintained at 37℃ to 45℃, and 0 to 250 mM NaCl. The activity of PanLys.1 significantly decreased (p<0.05) after Mn²⁺ or Zn²⁺ treatment. The relative abundance of P. anaerobius did not decrease after administration PanLys.1 under in vitro rumen conditions. Conclusion: The application of PanLys.1 to modulate P. anaerobius in the rumen might not be feasible because its lytic activity was not observed in in vitro rumen system.