• Title/Summary/Keyword: Lysis

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Detection of Irradiated Fruits Using the DNA Comet Assay (DNA Comet Assay를 이용한 과일의 방사선 조사 확인)

  • Oh, Kyong-Nam;Park, Jun-Young;Kim, Kyeung-Eun;Yang, Jae-Seung
    • Korean Journal of Food Science and Technology
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    • v.32 no.3
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    • pp.531-537
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    • 2000
  • The simple microgel electrophoresis of single cells, a 'comet assay', on fruit seeds enabled the rapid identification of irradiated fruits by comparing the intact non-irradiated cells and the damaged cells of irradiated fruits. Grapes and plums were irradiated with 0.1, 0.5, 0.7, 1.0 kGy and strawberries, peaches, apples, and nectarines were irradiated with only 1.0 kGy. Seeds were isolated, crushed, and the suspended cells were embedded in an agarose layer. After lysis of the cells, they were subjected to microgel electrophoresis for 2 minutes, and then stained. The DNA radiation-induced fragmentation of all the fruits stretched and migrated out of the cells forming a tail toward the anode giving the appearance of a comet, while the undamaged cells appeared as intact nuclei without tails. Grape and plum seeds irradiated at 0.5 kGy and higher showed significant increases in tail length. With increasing the irradiation doses, longer extention of the DNA from the nucleus toward the anode was observed. Strawberry, peach, apple, and nectarine seeds irradiated with 1.0 kGy also showed the longer tails than non-irradiated ones. DNA comet assay as a rapid and inexpensive screening technique could be an officially validated method for the detection of irradiated fruits.

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Production of Vitamin $B_{12}$ by Using Protoplast Fusion between Bacillus natto and Bacillus megaterium (Bacillus natto 및 Bacillus megaterium의 원형질체 융합에 의한 Vitamin $B_{12}$의 생산)

  • Jin, Sung-Hyun;Park, Bub-Gyu;Roh, Myung-Hoon;Kim, Dong-Gyu;Ryu, Beung-Ho
    • Korean Journal of Food Science and Technology
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    • v.22 no.6
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    • pp.611-617
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    • 1990
  • This study was conducted to breed a high vitamin $B_{12}$ producer by the fusion of protoplasts between Bacillus natto and Bacillus megaterium. Auxotrophic mutants of Bacillus natto SH-34 ($thr^-try^-rif^r$) and Bacillus megaterium BK-13 ($arg^-ade^-lys^-str^r$) which showed high protease activity and production of vitamin $B_{12}$, respectively, were isolated for the fusion experiment. Protoplasts were induced by incubating the cells with lysis solution containing $500{\mu}/ml$ lysozyme, and the ratio of protoplast and regeneration formation were ranged from 99% and 67%, respectively. Fusion frequencies of fusants between Bacillus natto SH-34 and Bacillus megaterium BK-13 were appeared in the ranges of $1.0{\times}10^{-5}$ under the treatment of 30% PEG 6000 containing 3% PVP. The fusant, MNF-72 showed the highest product yield of $7.85{\mu}g/g-cell\;vitamin\;B_{12}$ in production medium. For the improvement of productivity, the immobilization of fusants with sodium alginate was carried out. In batch and continuous fermentation systems, the productivity were determined to be $0.58{\mu}g/ml.hr\;and\;0.80{\mu}g/ml.hr\;vitamin\;B_{12}$ under optimum condition, respectivity.

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Synthesis and Characterization of Porous Poly(ε-caprolactone)/Silica Nanocomposites (다공성 폴리(ε-카프로락톤)/실리카 복합체의 제조 및 특성평가)

  • Son, Siwon;Choi, Ji-Eun;Cho, Hun;Kang, DaeJun;Lee, Deuk Yong;Kim, Jin-Tae;Jang, Ju-Woong
    • Polymer(Korea)
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    • v.39 no.2
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    • pp.323-328
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    • 2015
  • Poly(${\varepsilon}$-caprolactone) (PCL) nanofibers and PCL/silica membranes were synthesized by sol-gel derived electrospinning and casting, respectively. Smooth PCL nanofibers were obtained from the precursor containing N,N-dimethylformamide (DMF). PCL/silica membranes were prepared by varying the tetraethyl orthosilicate (TEOS) contents from 0 to 40 vol% to investigate the effect of silica addition on mechanical properties and cytotoxicity of the membranes. Although the strength of the membranes decreased from 12 to 8 MPa with increasing the silica content, the strength remained almost constant 7 weeks after dipping in phosphate buffered saline solution (PBS). The strength reduction was attributed to the presence of a patterned surface pores and micro-pores present in the walls between pores. The crystal structure of the membranes was orthorhombic and the crystallite size decreased from 57 to 18 nm with increasing the silica content. From the agar overlay test, the PCL/silica membranes exhibited neither deformation and discoloration nor lysis of L-929 fibroblast cells.

Combined Therapy with Carboplatin and Meloxicam for Oral Squamous Cell Carcinoma in a Dog (구강 편평 세포암 이환견에서 Carboplatin과 Meloxicam의 병용투여 증례)

  • Jang, Hwan-Soo;Kim, Jun-Il;Kim, Jae-Hoon;Jang, Kwang-Ho
    • Journal of Veterinary Clinics
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    • v.28 no.3
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    • pp.314-317
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    • 2011
  • A 16-year-old, spayed female, Maltese dog weighing 4.3 kg was presented with a 2-week history of yellowish nose-and oral-discharge, anorexia and lethargy. In physical examination, indefinite ulcerative mass about $3{\times}3$ cm in size in the left sublingual region, severe gingivoperiodontitis, halitosis, moderate dental calculus, fever and swelling of mandible soft tissue were noted. In radiographic view, periodontitis and bone lysis findings on the left rostral part of maxilla and mandible were observed. In complete blood count and blood chemistry test, thrombocytosis, increased levels of serum $NH_3$, AST and ALP were obtained. Urinalysis revealed hematuria and proteinuria. In histological examination, squamous cell carcinoma was diagnosed. No surgery was performed, but combined treatment with carboplatin and piroxicam was initiated. Five days after initiation of medication, because of insistent vomiting, piroxicam was substituted to meloxicam. Although the therapeutic efficacy of meloxicam could not be known; administration of meloxicam might be less potential to gastrointestinal side-effects than piroxicam. The dog totally received 3 times of carboplatin infusion. Five days after third infusion of carboplatin, the dog was hospitalized with severe lethargy, vomiting and bloody diarrhea. In examination, severe renal failure signs were noted, and the dog was euthanized.

A Cytotoxicity of Carrier Oil and Essential Oils on Cells by Using of Aromatherapy (향기요법에 사용하는 캐리어 오일과 에션셜 오일의 세포에 대한 독성)

  • Yu, Byong-Soo;Kim, Sha-Sha;Yun, Young-Han;Kim, Ki-Young
    • Korean Journal of Human Ecology
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    • v.17 no.5
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    • pp.1027-1035
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    • 2008
  • Essentail oils and carrier oils are generally used for Aromatherapy. Therefore the toxicity, possibilities of irritations and sensitive reactions and injury of essential oils must be considered for clients and therapists. So that, in this studies a toxicity of jojoba and 4 species essential oils (fennel, mandarine, tea tree and cedarwood) were investigated by the measurement of MTT-assay and sirius red staining. Liver, kidney and brain tell were chosen for the cell viability assay and observation of morphological change. In the result, no cytotoxicity was observed on live., kidney and brain cell at concentration of 0.01 $\mu\el/m\el$ jojoba oil. And lysis and nucleus breaking were not observed at same concentration of jojoba oil on live., kidney and brain cell. fennel oil was showed 50% of cell viability and inhibited cell growth on liver, kidney and brain cell at relatively high concentration compared with the other oils. 50% of liver, kidney and brain cell viability and delayed cell growth of tea tree and mandarine oil were revealed at lower concentration than fennel oil. In cedarwood oil, 50% of liver cell viability at concentration of 0.00067 $\mu\el/m\el$ was showed, but cell viability and cell growth of kidney and beam cell were effected at the lowest concentration compared with other oils. So that, jojoba oil as using of carrier oil may be not harmful. And 3 essential oils from the fennel, tea tree and mamdarine may have very low toxicity, but cedarwood may be used carefully for inhalation. And over dosage of concentrated cedarwood oil should be not directly touched and exposured, and absolute essential oils must be diluted with carrier oils for topical and systematic massage.

Immuno-chromatographic Analysis for HPV-16 and 18 E7 Proteins as a Biomarker of Cervical Cancer Caused by Human Papillomavirus

  • Kim, Joo-Ho;Cho, Il-Hoon;Seo, Sung-Min;Kim, Ji-Sook;Oh, Kyu-Ha;Kang, Heun-Soo;Kim, In-Gyu;Paek, Se-Hwan
    • Bulletin of the Korean Chemical Society
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    • v.30 no.12
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    • pp.2999-3005
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    • 2009
  • Among the more than 120 different types of human papillomavirus (HPV), types 16 and 18 have been known to be high risk agents that cause cervical cancer. We examined, in an immuno-chromatographic analysis, the potential of using the early gene product, E7 protein, as a diagnostic marker of cervical cancer caused by HPV. We developed monoclonal antibodies specific to HPV-16 and 18 E7 proteins that were produced from bacterial cells using gene recombinant technology. For each E7 protein, the optimal antibody pair was selected using the immuno-chromatographic sandwichtype binding system based on the lateral flow through membrane pores. Under these conditions, this rapid testing assay had a detection capability as low as 2 ng/mL of E7 protein. Furthermore, since viral analysis required the host cell to be lysed using chemicals such as detergents, it was possible that the E7 protein was structurally damaged during this process, which would result in a decrease in detection sensitivity. Therefore, we examined the detrimental effects caused by different detergents on the E7 protein using HeLa cells as the host. In these experiments, we found that the damage caused by the detergent, nonylphenylpolyethylene glycol (NP-40), was minimal relative to Triton X-100 commonly used for the cell lysis. Temperature also affected the stability of the E7 protein, and we found that the E7 protein was stabilized at 4$^{\circ}C$ for about 2 h, which was 4 times longer than at room temperature. Finally, a HPV-infected cervical cancer cell line, which was used as a real sample model, was treated using the optimized conditions and the presence of E7 proteins were analyzed by immuno-chromatography. The results of this experiment demonstrated that this rapid test could specifically detect HPV-infected samples.

Antimicrobial & Physiological Characteristics of Ethanol Extract from Pinus rigida Miller Leaves (리기다소나무 잎 에탄올 추출물의 항균 및 생리특성)

  • Oh, Byung-Tae;Choi, Sung-Gil;Cho, Sung-Hwan
    • Food Science and Preservation
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    • v.13 no.5
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    • pp.629-633
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    • 2006
  • Pinus rigida Miller leaf extract (PRLE) showed antimicrobial activity remarkably against food pathogenic and spoilage bacteria at concentrations of $100{\sim}250{\mu}g/mL$. Alcohol-soluble PRLE had higher antimicrobial activity against Staphylococcus aureus and E-coli than any other-soluble PRLE such as butanol, ethyl acetate, ether and water. As PRLE concentration increased alcohol-soluble PRLE increased the remarkable inhibitory zone of microbial growth on the microbial media. PRLE showed good stability against temperature and pH in the range of $40{\sim}150^{\circ}C$ and $4{\sim}11$, respectively. This may indicate that PRLE can be a potential anti-microbial agent for industrial application. In addition, SEM of Listeria monocytogenes suggested that it antimicrobial component would perturb the functions of microbial cell membranes synergistically. In the feeding experiment the formaldehyde content in the serum of formalin-fed and PRLE-treated me decreased remarkably due to the lysis of formaldehyde and the rate of hemoglobin biosynthesis was recovered to the orignal state within a short breeding time.

Properties of Soil Suppressiveness to Cucumber Wilt, caused by Fusarium oxysporum f. sp. cucumerinum Owen (오이 덩굴쪼김병의 발병(發病)을 억제(抑制)하는 토양(土壤)의 특성(特性)에 관(關)하여)

  • Park, Chang-Seuk;Cho, Yong-Sup
    • Korean journal of applied entomology
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    • v.24 no.2 s.63
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    • pp.85-95
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    • 1985
  • It has been tried to find effective biological control measures involved in nature of soil suppressiveness to fusarium wilt of cncumber caused by Fusarium oxysporum f. sp. cucumerinum Owen. Total 28 soil samples were obtained from Jinju, Haman, Namji, Milyang and Suncheon vinyl house area. The disease response of test soil was quantified in terms of DI50 value which caculated from log-probit transformation of diseases response curves. Soils designated 5 from Jinju, 7 from Suncheon, 22 from Namji were recognized as suppressive to fusarium wilt of cucumber. This suppressiveness was completely nullified after autoclave. The disease suppressiveness of tested soil did not indicate any consistency according to either chemical property or texture of soil. Conidial germination, induction and germination of chlamydospore were markedly inhibited in supprerssive soil compared to those in intermediate or conducive soils, however, mycelial lysis did not appear to have direct relationship with disease suppressiveness of given soil. Population density of fluorescent Pseudomonads and Bacillus spp. in the soil originated from different degree of suppressiveness were not different significantly but the number of lytic bacterial plaques measured by triple layer agar method was remarkably higher in suppressive soil than that in intermediate or conducive soil.

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Optimized Condition of Genomic DNA Extraction and PCR Methods for GMO Detection in Potato (유전자재조합 감자의 검정을 위한 DNA분리 및 PCR검출의 최적조건 탐색)

  • Shin, Weon-Sun;Kim, Myung-Hee
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.591-597
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    • 2003
  • To compare the quality of genomic DNA extracted from potato for PCR detection, four different methods, such as silica-based membrane method, silica-coated bead method, STE solution treatment, and CTAB-phenol/chloroform method, were evaluated. Also, to remove an excessive carbohydrate from the potato, ${\alpha}$- and ${\beta}$-amylase were used individually and in combination. When used both silica-based membrane method and silica-coated bead method combined with enzymes, the genomic DNAs were extracted from the raw potato with high purity for PCR. However, the silica-coated head method combined with enzyme treatment was the most efficient for extraction of the genomic DNA from the frozen fried potatoes. When applied with STE solution, the highly purified DNA was extracted from the raw potatoes without enzyme treatment in adequate yield for PCR. In cases of processed potatoes, such as frozen-fried potato and fabricated potato chips, CTAB-phenol/chloroform method is mostly feasible for DNA extraction and PCR efficacy at high sensitivity. As the results of PCR amplification, 216bp of PCR product was detected on 2% agarose gel electrophoresis, but any amplicons derived from New leaf and New leaf Y gene was not detected in any sample.

Evaluation of Optimal Condition for Recombinant Bacterial Ghost Vaccine Production with Four Different Antigens of Streptococcus iniae-enolase, GAPDH, sagA, piaA (연쇄구균증 항원-enolase, GAPDH, sagA, piaA에 대한 재조합 고스트 박테리아 백신의 생산 최적화)

  • Ra, Chae-Hun;Kim, Yeong-Jin;Son, Chang-Woo;Jung, Dae-Young;Kim, Sung-Koo
    • Journal of Life Science
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    • v.19 no.7
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    • pp.845-851
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    • 2009
  • A vector harboring double cassettes; a heterologous gene expression cassette of pHCE-InaN-antigen and a ghost formation cassette of pAPR-cI-E lysis 37 SDM was constructed and introduced to E. coli DH5a. For the production of a bacterial ghost vaccine, bacterial ghosts from E. coli / Streptococcus iniae with four different types of antigens - enolase, GAPDH, sagA and piaA - were produced by the optimization of fermentation parameters such as a glucose concentration of 1 g/l, agitation of 300 rpm and aeration of 1 vvm. Efficiency of ghost bacteria formation was evaluated with cultures of OD$_{600}$=1.0, 2.0 and 3.0. The efficiency of the ghost bacteria formation was 99.54, 99.67, 99.99 and 99.99% with inductions at OD$_{600}$=3.0, 1.0, 2.0 and 1.0 for E. coli/S. iniae antigens enolase, piaA, GAPDH and sagA, respectively. Ghost bacteria as a vaccine was harvested by centrifugation. The antigen protein expressions were analyzed by SDS-PAGE and western blot analysis, and the molecular weights of the enolase, piaA, GAPDH and sagA were 78, 26, 67 and 26 kDa, respectively. The molecular weights of the expressed antigens were consistent with theoretical sizes obtained from the amino acid sequences.