• 제목/요약/키워드: E-Isolation

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Optimization of Culture Medium for Novel Cell-Associated Tannase Production from Bacillus massiliensis Using Response Surface Methodology

  • Belur, Prasanna D.;Goud, Rakesh;Goudar, Dinesh C.
    • Journal of Microbiology and Biotechnology
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    • 제22권2호
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    • pp.199-206
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    • 2012
  • Naturally immobilized tannase (tannin acyl hydrolase, E.C. 3.1.1.20) has many advantages, as it avoids the expensive and laborious operation of isolation, purification, and immobilization, plus it is highly stable in adverse pH and temperature. However, in the case of cell-associated enzymes, since the enzyme is associated with the biomass, separation of the pure biomass is necessary. However, tannic acid, a known inducer of tannase, forms insoluble complexes with media proteins, making it difficult to separate pure biomass. Therefore, this study optimizes the production of cell-associated tannase using a "protein-tannin complex" free media. An exploratory study was first conducted in shake-flasks to select the inducer, carbon source, and nitrogen sources. As a result it was found that gallic acid induces tannase synthesis, a tryptose broth gives higher biomass, and lactose supplementation is beneficial. The medium was then optimized using response surface methodology based on the full factorial central composite design in a 3 l bioreactor. A $2^3$ factorial design augmented by 7 axial points (${\alpha}$ = 1.682) and 2 replicates at the center point was implemented in 17 experiments. A mathematical model was also developed to show the effect of each medium component and their interactions on the production of cell-associated tannase. The validity of the proposed model was verified, and the optimized medium was shown to produce maximum cell-associated tannase activity of 9.65 U/l, which is 93.8% higher than the activity in the basal medium, after 12 h at pH 5.0, $30^{\circ}C$. The optimum medium consists of 38 g/l lactose, 50 g/l tryptose, and 2.8 g/l gallic acid.

Salmonella enteritidis의 편모항원에 대한 난황항체의 ,생산 x Production of Egg Yolk Antibodies against Flagella Antigen of Salmonella enteritidis

  • 김정우
    • 한국가금학회지
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    • 제25권4호
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    • pp.161-167
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    • 1998
  • 가금 및 가축의 살모넬라중을 일으키는 에서 편모항원을 순수분리하고 이를 면역원으로 산란계에 면역하여 난황 중에 형성된 항체와 혈청간에 면역 반응을 조사하고 생산된 에 대한 특이난황 항체의 특이성 검사와 항체회수율 검사를 실시한 결과S. enteritidis는 다음과 같다. 1. S. enteritidis에서 순수 분리한 flagella protein 의 분자량은 54.6 kDa으로 나타났다. 2. 산란계의 혈청중 항체가 형성은 면역 후 2주경부터 급격히 증가하기 시작하여 6주경에 최고의 항체가 수준에 도달하였다. 반면에 난황중 항체가는 면역 후 4주경부터 급격히 증가하기 시작하여 6주경에 혈청항체가 수준과 동일하게 도달되며, 10주 이후에는 혈청항체가 수준보다 높게 형성되어 일정한 수준을 유지하였다. 3. S. enteritidis의 flagella protein을 면역원으로 산란계에 면역하여 생산된 난황항체는 주로 S. enteritidis 균주에서만 특이적으로 반응하며 E. coli (Kl2:K99, K88+, 987P) 균체에서는 반응이 전혀 없는 것으로 나타났다. 4. 한개의 난으로부터 분리한 IgY의 함량은 106 mg이었으며 회수율은 88.3%였다.

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질경이로부터 항균성 화합물의 분리 및 동정 (Isolation and Identification of Antimicrobial Compound from Plantain (Plantago asiatica L.))

  • 김건희;김순임;한영실
    • 한국식품조리과학회지
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    • 제15권4호
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    • pp.410-417
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    • 1999
  • 우리나라 야산에서 쉽게 구할 수 있는 구황 식물인 질경이를 메탄올과 여러 용매로 추출하여 식품 부패 미생물에 대한 항균력을 실험하고 그 항균활성 물질을 분석하여 다음과 같은 결과를 얻었다. 질경이의 메탄올 추출물은 2000$\mu\textrm{g}$/ml 농도에서 B. subtilis 및 V. parahaemolyticus의 증식을 100% 억제하였다. 그리고 질경이의 ethylacetate 분획물은 1000$\mu\textrm{g}$/disc 농도에서 5종의 모든 실험 균주에 대하여 8.5~11mm 크기의 clear zone을 형성하여 가장 높은 항균력을 보였다. 질경이의 ethylacetate 분획물을 silica gel column chromatography와 TLC로 분리하여 얻은 fraction을 5가지 실험 균주에 대하여 항균 실험을 한 결과 3rd 분획물 중 4번째 분획물이 250$\mu\textrm{g}$/disc 농도에서 모든 균주에 대하여 10 mm 이상의 clear zone을 나타내었다. 항균력을 보인 질경이의 ethylacetate 3rd fraction No. 4를 HPLC로 단일 분리하여 얻은 peak IV로 부터 $^1$H-NMR 및 GC-MSD로 동정한 결과 hexadecanoic acid로 동정되었다.

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서해안 수산생물에서 분리한 대장균(Escherichia coli)의 항생제 내성 및 다제 내성 양상 비교 (Comparison of Antimicrobial Resistance and Multi-Drug Resistance Patterns of Escherichia coli Isolated from Aquatic Organisms Off the West Coast of South Korea)

  • 정연겸;박보미;김민주;박진일;정연중;오은경
    • 한국수산과학회지
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    • 제54권4호
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    • pp.388-396
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    • 2021
  • Antimicrobial resistance patterns of Escherichia coli were investigated. Strains were isolated from 310 shellfish, 36 crustaceans, and 12 fish collected off the West Coast of Korea from April 2019 to October 2020. Two hundred and ninety-five E. coli strains were isolated from shellfish, 100 from crustaceans, and 54 from fish. Strains isolated from shellfish showed the highest resistance to ampicillin (27.5%), whereas those from crustaceans were resistant to sulfisoxazole (30.0%) and those from fish were resistant to ampicillin (59.3%) and sulfisoxazole (59.3%). Ceftazidime resistance was observed in strains isolated from short neck and hard clams, whereas gentamicin resistance was observed in strains from fish. Multi-drug resistance was observed in 56 strains (48.7%) isolated from shellfish, 11 (28.2%) from crustaceans and 27 (73.0%) from fish. Depending on the source of isolation, the strains showed specific antimicrobial resistance tendency. Strains isolated from shellfish showed 12 different multi-drug resistance patterns, whereas those from crustaceans showed high resistance (59%) to a single antimicrobial agent and those from fish showed a broad trend of multi-drug resistance to more than eight antimicrobials.

Potential of polylactic-co-glycolic acid (PLGA) for delivery Jembrana disease DNA vaccine Model (pEGFP-C1-tat)

  • Unsunnidhal, Lalu;Wasito, Raden;Setyawan, Erif Maha Nugraha;Warsani, Ziana;Kusumawati, Asmarani
    • Journal of Veterinary Science
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    • 제22권6호
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    • pp.76.1-76.15
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    • 2021
  • Background: The development of a vaccine for Jembrana disease is needed to prevent losses in Indonesia's Bali cattle industry. A DNA vaccine model (pEGFP-C1-tat) that requires a functional delivery system will be developed. Polylactic-co-glycolic acid (PLGA) may have potential as a delivery system for the vaccine model. Objectives: This study aims to evaluate the in vitro potential of PLGA as a delivery system for pEGFP-C1-tat. Methods: Consensus and codon optimization for the tat gene was completed using a bioinformatic method, and the product was inserted into a pEGFP-C1 vector. Cloning of the pEGFP-C1-tat was successfully performed, and polymerase chain reaction (PCR) and restriction analysis confirmed DNA isolation. PLGA-pEGFP-C1-tat solutions were prepared for encapsulated formulation testing, physicochemical characterization, stability testing with DNase I, and cytotoxicity testing. The PLGA-pEGFP-C1-tat solutions were transfected in HeLa cells, and gene expression was observed by fluorescent microscopy and real-time PCR. Results: The successful acquisition of transformant bacteria was confirmed by PCR. The PLGA:DNA:polyvinyl alcohol ratio formulation with optimal encapsulation was 4%:0.5%:2%, physicochemical characterization of PLGA revealed a polydispersity index value of 0.246, a particle size of 925 nm, and a zeta potential value of -2.31 mV. PLGA succeeded in protecting pEGFP-C1-tat from enzymatic degradation, and the percentage viability from the cytotoxicity test of PLGA-pEGFP-C1-tat was 98.03%. The PLGA-pEGFP-C1-tat demonstrated luminescence of the EGFP-tat fusion protein and mRNA transcription was detected. Conclusions: PLGA has good potential as a delivery system for pEGFP-C1-tat.

A CAD/CAM-based strategy for concurrent endodontic and restorative treatment

  • Escobar, Patricia Maria;Kishen, Anil;Lopes, Fabiane Carneiro;Borges, Caroline Cristina;Kegler, Eugenio Gabriel;Sousa-Neto, Manoel Damiao
    • Restorative Dentistry and Endodontics
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    • 제44권3호
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    • pp.27.1-27.12
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    • 2019
  • This case report describes a technique in which endodontic treatment and permanent indirect restoration were completed in the same clinical appointment with the aid of a computer-aided design/computer-aided manufacturing (CAD/CAM) system. Two patients were diagnosed with irreversible pulpitis of the mandibular first molar. After access preparation, root canals were located, irrigation was performed until bleeding ceased, and the coronal tooth structure was prepared for indirect restoration. Then, utilizing an interim 3-mm build-up of the endodontic access cavity, a hemi-arch digital scan was performed with an intraoral scanner. Subsequent to digital scanning, restoration design was performed simultaneously with the endodontic procedure. The root canals were shaped using the Race system under irrigation with 2.5% sodium hypochlorite followed by root canal filling. The pulp chamber was subsequently filled with a 3-mm-thick composite resin restoration mimicking the interim build-up previously utilized to facilitate block milling in the CAD/CAM system. Clinical try-in of the permanent onlay restoration was followed by acid etching, application of a 5th generation adhesive, and cementation of the indirect restoration. Once the restoration was cemented, rubber dam isolation was removed, followed by occlusal adjustment and polishing. After 2 years of follow-up, the restorations were esthetically and functionally satisfactory, without complications.

Isolation, Identification and Enzymatic Activity of Halotolerant and Halophilic Fungi from the Great Sebkha of Oran in Northwestern of Algeria

  • Chamekh, Rajaa;Deniel, Franck;Donot, Christelle;Jany, Jean-Luc;Nodet, Patrice;Belabid, Lakhder
    • Mycobiology
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    • 제47권2호
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    • pp.230-241
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    • 2019
  • The Great Sebkha of Oran is a closed depression located in northwestern of Algeria. Despite the ranking of this sebkha among the wetlands of global importance by Ramsar Convention in 2002, no studies on the fungal community in this area have been carried out. In our study, samples were collected from two different regions. The first region is characterized by halophilic vegetation and cereal crops and the second by a total absence of vegetation. The isolated strains were identified morphologically then by molecular analysis. The biotechnological interest of the strains was evaluated by testing their ability to grow at different concentration of NaCl and to produce extracellular enzymes (i.e., lipase, amylase, protease, and cellulase) on solid medium. The results showed that the soil of sebkha is alkaline, with the exception of the soil of cereal crops that is neutral, and extremely saline. In this work, the species Gymnoascus halophilus, Trichoderma gamsii, the two phytopathogenic fungi, Fusarium brachygibbosum and Penicillium allii, and the teleomorphic form of P. longicatenatum observed for the first time in this species, were isolated for the first time in Algeria. The halotolerance test revealed that the majority of the isolated are halotolerant. Wallemia sp. and two strains of G. halophilus are the only obligate halophilic strains. All strains are capable to secrete at least one of the four tested enzymes. The most interesting species presenting the highest enzymatic index were Aspergillus sp. strain A4, Chaetomium sp. strain H1, P. vinaceum, G. halophilus, Wallemia sp. and Ustilago cynodontis.

Neuraminidase Inhibitors from the Fruiting Body of Glaziella splendens

  • Kim, Ji-Yul;Woo, E-Eum;Ha, Lee Su;Ki, Dae-Won;Lee, In-Kyoung;Yun, Bong-Sik
    • Mycobiology
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    • 제47권2호
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    • pp.256-260
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    • 2019
  • Neuraminidase (NA) cleaves the glycosidic bond linkages of sialic acids to release the mature virions from infected cells and has been an attractive therapeutic target for anti-influenza agents. In our ongoing investigation of NA inhibitors in mushroom extracts, we found that the extract the fruiting body of Glaziella splendens potently inhibited neuraminidase. The fruiting bodies of G. splendens were extracted and partitioned successively with hexane, ethyl acetate, and butanol. The ethyl acetate soluble-layer was subjected to silica gel and Sephadex LH-20 column chromatographies, and MPLC to obtain five compounds (1-5). Their structures were determined by spectroscopic methods. NA inhibitory activity of these compounds was evaluated using NAs from recombinant rvH1N1, H3N2, and H5N1 influenza A viruses. One compound (1) was elucidated as a new azaphilone derivative, and four compounds (2-5) were identified as entonaemin A, comazaphilone D, rubiginosin A, and entonaemin B, respectively. Compounds 3 and 4 showed considerable inhibitory activity against three types of neuraminidases with the $IC_{50}$ values of 30.9, 41.8, and $35.7{\mu}M$ for 3 and 46.5, 50.4, and $29.9{\mu}M$ for 4, respectively. This study reveals that the fruiting bodies of G. splendens possess azaphilone derivatives with the NA inhibitory activity. This is the first report on the isolation of neuraminidase inhibitors from the fruiting bodies of G. splendens.

언어와 감정-셸리의 『프랑켄슈타인』과 루소의『언어의 기원론』 (Shelley's Frankenstein and Rousseau's Essay on the Origin of Languages)

  • 김상욱
    • 영어영문학
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    • 제54권4호
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    • pp.483-509
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    • 2008
  • For the last decades, criticism on Frankenstein has tried to make a link between Victor's Creature and Rousseaurean "man in a state of nature." Like the Rousseaurean savage in a state of animal, the monster has only basic instincts least needed for his survival, i.e. self-preservation, but turns into a civilized man after learning language. Most critics argue that, despite the monster's acquisition of language, his failure in entry into a cultural and linguistic community is the outcome of a lack of sympathy for him by others, which displays the stark existence of epistemological barriers between them. That is to say, the monster imagines his being the same as others in the pre-linguistic stage but, in the linguistic stage, he realizes that he is different from others. Interpreting the Rousseaurean idea of language, which appears in his writings, as much more focused on emotion than many critics think, I read the dispute between Victor and his Creature as a variation of parent-offspring conflict. Shelley criticizes Rousseau's parental negligence in putting his children into a foundling hospital and leaving them dying there. The monster's revenge on uncaring Victor parallels the likely retaliation Rousseau's displaced children would perform against Rousseau, which Shelley imaginatively reproduces in her novel. The conflict between the monster and Victor is due to a disrupted attachment between parent and child in terms of Darwinian developmental psychology. Affective asynchrony between parent and child, which refers to a state of lack of mutual favorable feelings, accounts for numerous dysfunctional families. This paper shifts a focus from a semiotics-oriented perspective on the monster's social isolation to a Darwinian perspective, drawing attention to emotional problems transpiring in familial interactions. In doing so, it finds that language is a means of communicating one's internal emotions to others along with other means such as facial expressions and body movements. It also demonstrates that how to promote emotional well-being in either familial or social relationships entirely depends on the way in which one employs language that can entail either pleasure or anger on hearers' part.

Coping with large litters: management effects on welfare and nursing capacity of the sow

  • Peltoniemi, Olli;Han, Taehee;Yun, Jinhyeon
    • Journal of Animal Science and Technology
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    • 제63권2호
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    • pp.199-210
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    • 2021
  • A number of management issues can be used as drivers for change in order to improve animal welfare and nursing capacity of the hyperprolific sow. Group housing of sows during gestation is a recommended practice from the perspective of animal welfare. Related health issues include reproductive health and the locomotor system. It appears that management of pregnant sows in groups is challenging for a producer and considerable skill is required. We explored the benefits and challenges of group housing, including feeding issues. Increasing litter size requires additional attention to the mammary gland and its ability to provide sufficient nursing for the growing litter. We discuss the fundamentals of mammary development and the specific challenges related to the hyperprolific sow. We also address challenges with the farrowing environment. It appears that the old-fashioned farrowing crate is not only outdated in terms of welfare from the public's perspective, but also fails to provide the environment that the sow needs to support her physiology of farrowing, nursing, and maternal behaviour. Studies from our group and others indicate that providing the sow with a loose housing system adequate in space and nesting material, along with reasonable chance for isolation, can be considered as fundamental for successful farrowing of the hyperprolific sow. It has also been shown that management strategies, such as split suckling and cross fostering, are necessary to ensure proper colostrum intake for all piglets born alive in a large litter. We thus conclude that welfare and nursing capacity of the sow can be improved by management. However, current megatrends such as the climate change may change sow management and force the industry to rethink goals of breeding and, for instance, breeding for better resilience may need to be included as goals for the future.