• Title/Summary/Keyword: vero cell

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Evaluation of Cosmeceutical Properties of Fish Skin By-product Hydrolysates Collected During Surimi Manufacturing Process (연육 제조과정에서 발생하는 어피 부산물의 활용을 위한 Cosmeceutical 활성 평가)

  • Oh, Jae-Young;Lee, Hyo-Geun;Je, Jun-Geon;Wang, Lei;Kim, Hyun-Soo;Jeon, You-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.3
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    • pp.297-307
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    • 2020
  • This study investigated the cosmetic effects of enzymatic hydrolytes of an aquatic by-product, fish skin. The skins of olive flounder Paralichthys olivaceus (PO) and Alaska pollock Gadus chalcogrammus (AP) were hydrolyzed using pepsin, Alcalase, and Protemax. Three enzymatic hydrolytes were obtained and the inhibitory effects of these hydrolytes on the aging-related enzymes tyrosinase, elastase, and collagenase were determined. The results indicated that the pepsin hydrolytes of PO and PA had stronger activities than the other hydrolytes. PO and PA also significantly reduced the intracellular reactive oxygen species levels in and improved the viability of H2O2-treated Vero cells; decreased nitric oxide production by and increased the cell viability of lipopolysaccharide-treated RAW 264.7 cells; and reduced intracellular reactive oxygen species levels and improved the viability of ultraviolet B irradiated HaCaT cells and human dermal fibroblasts. Furthermore, PO and PA remarkably reduced the intra- and extracellular melanin contents of alpha-melanocyte-stimulating hormone-stimulated B16F10 cells. These results demonstrate that PO and PA have potential for use in the cosmetic industry.

Passage and Adaptation of Maaji Virus in Hamster (Maaji Virus의 Hamster 계대 및 적응)

  • Kim, Yun-Cheol;Paik, Woo-Hyun;Lee, Pyung-Woo
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.67-76
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    • 1996
  • The methods that make Hantavirus grow consist of inoculation into the experimental animals and cultured cells. The cultured cells, such as Vero-E6 and A549 cells, have been usually used for isolation of the virus and the animals, such as mice and rats, are used for large scale preparation of the virus so far. Furthermore, the cell can be used to maintain the virus and assay the infectivity and the animals can be used for the experiment of viral pathogenicity and challenge for assessment of vaccine. Apodemus mice, the own natural host of the virus, has been used for challenge test of Hantaan virus. However it has been pointed out to difficult handling and breeding the animal in laboratory. Therefore, we attempted to establish a new animal model for challenge test at the time of isolation of Maaji virus which is a new hantavirus similar but distinct to Hantaan virus. In suckling hamster, the titer of Maaji virus and the lethality to mice of the virus were increased gradually in the titer and lethality through passage by intracerebral (IC) inoculation. We tried to re-adapt this brain virus to lung of weanling hamster. The brain passaged virus was inoculated into weanling hamster intramuscularly. Again, the titer of the virus in lung was also increased by continuous passage of this virus. This facts could regarded as adaptation to new environment in which the virus proliferates. To identity the virus passaged in hamster with Maaji virus, both of the virus passaged in hamster brain and lung were compared with Maaji virus (MAA-I) and Hantaan virus (HTN 76-118) by means of restriction fragment length polymorphism (RFLP) and slingle strand conformation polymophism (SSCP). As a result, we conclude that Maaji virus could be adapted successfully to weanling hamster through this passage strategy. Utilizing this adapted Maaji virus strain, hamster model is able to be used for challenge test in hantaviral vaccinology and further experiments utilizing hamster system as a rather available and convenient lab animal are expected.

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Gallic Acid Enhancement of Gold Nanoparticle Anticancer Activity in Cervical Cancer Cells

  • Daduang, Jureerut;Palasap, Adisak;Daduang, Sakda;Boonsiri, Patcharee;Suwannalert, Prasit;Limpaiboon, Temduang
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.1
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    • pp.169-174
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    • 2015
  • Cervical cancer (CxCa) is the most common cancer in women and a prominent cause of cancer mortality worldwide. The primary cause of CxCa is human papillomavirus (HPV). Radiation therapy and chemotherapy have been used as standard treatments, but they have undesirable side effects for patients. It was reported that gallic acid has antioxidant, antimicrobial, and anticancer activities. Gold nanoparticles are currently being used in medicine as biosensors and drug delivery agents. This study aimed to develop a drug delivery agent using gold nanoparticles conjugated with gallic acid. The study was performed in uninfected (C33A) cervical cancer cells, cervical cancer cells infected with HPV type 16 (CaSki) or 18 (HeLa), and normal Vero kidney cells. The results showed that GA inhibited the proliferation of cancer cells by inducing apoptosis. To enhance the efficacy of this anticancer activity, 15-nm spherical gold nanoparticles (GNPs) were used to deliver GA to cancer cells. The GNPs-GA complex had a reduced ability compared to unmodified GA to inhibit the growth of CxCa cells. It was interesting that high-concentration ($150{\mu}M$) GNPs-GA was not toxic to normal cells, whereas GA alone was cytotoxic. In conclusion, GNPs-GA could inhibit CxCa cell proliferation less efficiently than GA, but it was not cytotoxic to normal cells. Thus, gold nanoparticles have the potential to be used as phytochemical delivery agents for alternative cancer treatment to reduce the side effects of radiotherapy and chemotherapy.

Antioxidant Activity of Pepsin Hydrolysate Derived from Edible Hippocampus abdominalis in vitro and in Zebrafish Models (빅벨리 해마(Hippocampus abdominalis) 유래 펩신 가수분해물의 In vitro와 In vivo에서의 항산화 효능)

  • Kim, Hyun-Soo;Shin, Byeung-Ok;Kim, Seo-Young;Wang, Lei;Lee, WonWoo;Kim, Yoon Taek;Rho, Sum;Cho, Moonjae;Jeon, You-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.4
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    • pp.445-453
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    • 2016
  • Seahorse Hippocampus abdominalis a marine teleost fish, has long been used as one of the essential materials in traditional Chinese medicine. However, the uses of seahorse have been limited due to its high cost, despite its beneficial biological activities. Seahorse has not been widely explored for its biofunctional properties and active components. In the present study, the enzymatic hydrolysates of seahorse were prepared by using two digestive enzymes (trypsin and pepsin) and five food grade enzymes (neutrase, protamex, alcalase, kojizyme, and flavourzyme). The enzymatic hydrolysates indicated higher hydrolysis yields than its water extract. Among them, the distilled water-pepsin hydrolysate (DP) which was obtained by distilled water extraction followed by pepsin hydrolysis, showed the highest yield and protein content as well as the highest alkyl radical scavenging activity. Also, it provided protective effects against oxidative stress induced by AAPH in vero cell and zebrafish. Further fractionation based on the molecular weight was carried out to identify it’s active components, and < 5 kDa (less than 5 kDa) molecular weight fraction was confirmed to have the highest antioxidant activity. In conclusion, this study suggests that DP of seahorse has antioxidant properties, and might be a novel and useful material from the marine origin for healthy functional foods and cosmetics.

Cytoprotective and Antioxidative Effects of Crude Drug Preparation (E-kong-san) (이공산(異功散)의 세포보호 및 항산화 작용)

  • Lee, Kyung-Tae;Choi, Jung-Hye;Rho, Young-Soo;Ahn, Kyoo-Seok;Chang, Sung-Goo;Oh, Soo-Myung;Jung, Jee-Chang
    • Korean Journal of Pharmacognosy
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    • v.30 no.3
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    • pp.255-260
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    • 1999
  • In the previous report, E-kong-san, which is usually used for recovering health in traditional medicine, has been shown to decrease cisplatin induced nephrotoxicity in vivo and in vitro. The significant reduction of E-kong-san on the cisplatin induced nephrotoxicity led us to investigate whether the effect of this water extract was a result of triggering antioxidation. In monkey kidney Vero cells, E-kong-san at $5{\sim}10\;mg/ml$ was able to attenuate 2mM cisplatin-stimulated cell death by 46.8% and 31.8%, respectively. E-kong-san showed strong free radical scavengering activities on 1,1-diphenyl-2-picrylhydrazil (DPPH) radical and xanthine/xanthine oxidase (XOD) generated superoxide anion radical $(O_2^{-.})$. We further studied the effects of E-kong-san on lipid peroxidation in rat liver microsomes induced by enzymatic and nonenzymatic methods. Moreover, E-kong-san exhibited significant inhibition on both ascorbic $acid/Fe^{2+}$ and $ADP/NADPH/Fe^{3+}$ induced lipid peroxidation in rat liver microsomes. Based on these results, we suggest that E-kong-san protects the cisplatin induced cytotoxicity by its antioxidative mechanism.

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Production of expressed protein from cloned ShigatoxinG 2e gene and Receptor Binding Affinity of the toxin (재조합 Shigatoxin 2e 유전자의 발현단백 생산 및 독소의 수용체 결합 친화성 확인)

  • Dong, Bun-youn;Kim, Sang-Hyun;Kim, Yeong-Il;Cho, Hyun-Ho;Lee, Woo-won;Kim, Kon-Sup;Kang, Ho-Jo;Kim, Yong-Hwan
    • Korean Journal of Veterinary Research
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    • v.44 no.2
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    • pp.251-257
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    • 2004
  • This study was designed to determine optimal condition for expression of cloned Shigatoxin2e(Stx2e) gene from transformed E. coli PED18, to compare the cytotoxicity titer between cloned Stx2e and Stx2e from original strain, and to confirm of receptor binding affinity of Stx2e for use of development of receptor binding ELISA to detect of Stx2e. The optimum composition of medium for expression of Stx2e gene in E.coli host-vector system was definded as the medium containing 0.5% glucose and 0.5 mM IPTG. The cytotoxicity titer of expressed Stx2e for Vero cell was 1000 fold higher than that of Stx2e from original strain AY93258. The binding affinity of Stx2e to receptor globotetraosyl ceramide($Gb_4$) was confirmed by immunobloting.

Construction and Characterization of the Vibrio parahaemolyticus Collagenase Inactivated Mutant (Vibrio parahaemolyticus collagenase 불활성화 돌연변이체의 제조 및 특성)

  • 이재원;전인준;강호영;차재호
    • Journal of Life Science
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    • v.14 no.2
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    • pp.362-367
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    • 2004
  • For better understanding of the host infection mechanism of Vibrio, a Vibrio parahaemolyticus collagenase mutant was generated by insertional inactivation of a vppC gene encoding extracellular collagenase. A recombinant DNA containing vppC::nptII was cloned into a suicide plasmid pDMS197, resulted in pVCM03. The recombinant suicide plasmid pVCM03 contained in E. coli $\chi$7213 was transferred to a wild-type V. parahaemolyticus 04 through conjugation. The recombinant vppC::nptII DNA in pVCM03 was exchanged with wild-type allele by homologous recombination resulting vppC mutant, V. parahaemolyticus CM. The mutant was selected and screened on TCBS media containing 10% sucrose and kanamycin. The mutation by allele exchange was confirmed with the comparison of the size of DNAs amplified by PCR. V. parahaemolyticus CM showed at least 4-fold less collagen-degrading activity than those of wild-type, and the mutant exhibited less cytotoxicity than that of wild-type in MTT assay.

Development of a multiplex-PCR for the rapid detection of Escherichia coli O157:H7 from raw beef (쇠고기중 Escherichia coli O157:H7 신속검출을 위한 multiplex - PCR 기법 개발)

  • Jung, Suk-chan;Jung, Byeong-yeal;Yoon, Jang-won;Cho, Yun-sang;Kim, Jong-yeom;Park, Yong-ho
    • Korean Journal of Veterinary Research
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    • v.38 no.1
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    • pp.173-181
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    • 1998
  • Esherichia coli O157 : H7의 slt I, slt II, uid A 및 eaeA 4종 유전자를 동시에 검출하기 위한 multiplex PCR 기법을 확립하고 쇠고기중 직접 E coli O157 : H7 검출시험을 실시하였다. 4 set의 primers를 이용한 multiplex PCR 기법으로 31종의 장내세균에 대한 특이성을 조사한 결과 E coli O157 : H7 에서 1,087bp (eae A), 584bp (slt II), 348bp (slt I) 또는 252bp (uid A)크기의 DNA를 동시에 특이적으로 검출할 수 있었다. E coli O157 : H7 15주는 모두 uid A 및 eae A 유전자가 동시에 검출되었고, 다른 장내세균에서는 검출되지 않았다. slt I 또는 slt II 유전자를 가지고 있는 E coli 표준균주 24종을 이용하여 multiplex PCR 기법과 Vero cell cytotoxicity assay을 비교검사한 결과 베로톡신 산생능과 PCR법의 결과는 일치하였다. mutiplex PCR 기법의 쇠고기중 검출한계는 modified EC(mEC)에서 증균없이는 E coli O157 : H7균 $10^4cells/g$ 이상에서 검출이 가능하였으나 mEC에 1차 증균후 modified TSB 증균하였을 경우에는 10cells/g이하까지도 검출이 가능하였다. 개발된 multiplex PCR 기법을 쇠고기 40종에 직접 적용한 결과 E coli O157 : H7은 검출되지 않았으나 slt I 및 slt II유전자를 가지고 있는 E coli 4종이 검출되었으며, 이들의 혈청형은 O6, O112, O115 및 O139 였다. 이 연구에서 개발된 multiplex PCR은 쇠고기중 E coli O157 : H7을 신속하고 특이적으로 검출하는데 사용할 수 있을 것으로 사료된다.

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Evaluation of the Antioxidant Efficacy of Padina boryana Extract from Maldivian Seaweed from Laccadive Sea Area (라카디브 해역 몰디브 자생 해조류 Padina boryana 추출물의 항산화 효능 평가)

  • Kim, Hyun-Soo;Wang, Lei;Jayawardena, Thilina U.;Lee, Jeong Min;Yim, Mi-Jin;Ko, Seok-Chun;Lee, Hyo-Geun;Je, Jun-Geon;Jeon, You-Jin;Lee, Dae-Sung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.54 no.2
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    • pp.162-169
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    • 2021
  • Global warming has affected the distribution of organisms for decades and has displayed rapid ascent recently. Research into the effects on tropical organisms are vital. Padina boryana is a resourceful marine microalgae in the Maldives Sea in the Laccadive region. A 70% ethanol extraction (PBE) of this seaweed was used to investigate its antioxidant potential. Both in vitro and in vivo models were implemented. PBE exhibited protective potential against H2O2 induced apoptosis. ROS levels were suppressed due to PBE. PBE expressed a cytoprotective nature. In vivo experiments involving the zebra fish model conformed its validity. The antioxidant efficacy of PBE was dose dependent. Study outcomes suggest PBE has potential as a novel and valuable marine resource to aid the functional food and cosmeceutical industries.

Effects of Bifidobacteria on the Growth and Caco-2 Cell Adherence of E. coli O157:H7 (Bifidobacteria 가 E. cold O157:H7의 생육 및 Caco-2 세포 정착에 미치는 영향)

  • 김응률;정후일;전석락;유제현
    • Microbiology and Biotechnology Letters
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    • v.29 no.3
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    • pp.169-175
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    • 2001
  • This study was conducted to investigate the effects of bifidobacteria on the growth and Caco-2 cell-adherence of Escherichia coli O157:H7 .Dur-ing momo-culture of E. coli O157:H7 and mixed culture with Bifidobacterium infantis K9, pH viable cell count, and ammonia concentration were measured Co-cultivation of E. coli O157:H7 with bifidobacteria. producing acidic metabolites rapidly decreased the viable cell count of E. coli O157:H7 In addition rapid decrease of ammo- nia concentration was observed during mixed culture after 8 hrs incubation compared to single culture of E. coli O157:H7 Therefore it is likely that bifidobacteria assimilate ammonia produced by E. coli O157:H7 P4 B, infantis K9 showed quite similar adherence on the Caco-2 cells in either case. On the other hand adherence of E. coli O157:H7 decreased from 2.6% to 1.86% when B infantis K9 was adhered to Caco-2 cell 2 hrs prior to the application of E. coli O157:H7 In conclusion in adherence of E coli O157:H7 to Caco -2 cell was inhibited by competition of its binding to the adherence site with bifidobacteria. In addition inhibitory effects of bifidobacteria on E coli O157:H7 appeared to be much higher with increae of the number of bifidobacteria and its ability of adherence to Caco-2 cells.

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