• Title/Summary/Keyword: Culture method

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Comparison of Direct RT-PCR, Cell Culture RT-PCR and Cell IFA for Viability and Infectivity Assay of Cryptosporidium (크립토스포리디움 활성 및 감염성 판정을 위한 direct RT-PCR, cell culture RT-PCR 및 cell culture IFA의 비교)

  • Park, Sang-Jung;Yu, Jae-Ran;Kim, Jong-Min;Rim, Yeon-Taek;Jin, Ing-Nyol;Chung, Hyen-Mi
    • Journal of Life Science
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    • v.16 no.5
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    • pp.729-733
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    • 2006
  • Cryptosporidium is a waterborne pathogenic parasite which causes diarrhea. Immunomagnetic separation-immunofluorescent assay (IMS-IFA) has been a widely adopted for Cryptosporidium detection as standard method. However, this method does not provide information about viability or infectivity of Cryptosporidium. Therefore, many researchers have studied viability or infectivity analyses of Cryptosporidium with various methods such as vital staining, in vitro excystation, RT-PCR, cell culture, and mouse infection assay. In this study, two direct RT-PCR methods, cell culture RT-PCR and cell culture IFA were compared for sensitivity and other characteristics. The results showed that direct RT-PCR method with HSP70 genes had the highest sensitivity with detection up to 1 viable cell of Cryptosporidium. The infectious Cryptosporidium were detected up to 10 to 25 cells by cell culture methods in combination with RT-PCR and IFA. The infectious Cryptosporidium were apt to be quantified by cell culture IFA.

The Hatching Rate of Resting Eggs of the Rotifer Brachionus plicatilis according to Preservation Method (보관 방법에 따른 Rotifer Brachionus plicatilis 내구란의 부화)

  • Youn, Joo-Yeon;Hur, Sung-Bum
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.6
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    • pp.665-670
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    • 2011
  • The rotifer Brachionus plicatilis is one of the most important food organisms in aquaculture. The resting eggs produced by mictic female rotifers are easily stored and hatched, making them useful as the starter for the mass culture of rotifers in marine larval culture. This study examined the optimum preservation method for resting eggs to ensure a high hatching rate. To produce resting eggs, the marine rotifer B. plicatilis was cultured with Nannochloris oculata (KMMCC 16). The resting eggs were harvested and cryopreserved using 5% and 10% methanol (MeOH), dimethylsulfoxide (DMSO), and glycerol as cryoprotectant agents (CPAs). The cryopreservation comprised slow or rapid freezing and the resting eggs were stored for one month in liquid nitrogen ($-196^{\circ}C$). The resting eggs were also dried at different temperatures (30, 40, and $50^{\circ}C$) and for different times (1, 2, and 3 h). In general, the hatching rates of the resting eggs preserved with CPA were higher than those without CPA and the slow freezing method was better than the rapid freezing method. However, the optimum CPA concentration for the hatching rate of the resting eggs varied with the freezing method and kind of CPA, and the CPA also affected the viability of the resting eggs. Dried resting eggs had a high, rapid hatching rate over 80%. The moisture content of the resting eggs cryopreserved in liquid nitrogen affected the hatching rate. Drying at $30^{\circ}C$ for 1 hour resulted in a high hatching rate of the resting eggs. In conclusion, drying at $30^{\circ}C$ for 1 hour and preservation in liquid nitrogen with the slow freezing method, without CPA, is recommended for a high hatching rate (ca. 95%) of rotifer resting eggs.

Production of Lovastatin in Solid Culture (고체 배양법에 의한 Lovastatin생산)

  • 김현수;박지현
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.3
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    • pp.566-570
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    • 2004
  • Cultivation conditions for overproduction of lovastatins were investigated from the lovastatin producing strain N-03 which was obtained with NTG (N-methyl-N'-nitro-nitrosoguanidine) treatment from Aspergiliu ferrous ATCC 20542. Produced lactone and acid form of lovastatin were detected, and analyzed by HPLC method. In liquid culture, medium No. 2 containing soy protein produced higher amounts of the lovastatins than medium No. 1 (contained rapeseed oil). In solid culture, maximum production was obtained at 28$^{\circ}C$ for 15 days cultivation using cooked wheat bran. For the overproduction of lovastatin from this strain, solid culture method using plastic bag is more superior than liquid culture.

Development of Loading Machine of Culture Medium for Oyster Mushroom Production - Performance Test and Economic Analysis of Loading System - (느타리버섯 재배용 배지 입상 장치 개발(2) - 시작기 성능시험 및 경제성 평가 -)

  • Lee, Kyung-Jin;Lim, Hak-Kyu;Kim, Tae-Han
    • Journal of Biosystems Engineering
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    • v.34 no.4
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    • pp.220-227
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    • 2009
  • In the process of oyster mushroom production, loading work of culture medium needs the most intensive labor power. Therefore, development of culture medium machine causes to reduce the manpower and cost. The main objective of this study is to develop the culture medium loading machine and investigate the optimal operation conditions and to evaluate the economic value of the machine. The results are summarized as follows: 1. Optimum transporting velocity of the conveyor was 0.61 m/s 2. Optimum speed of blower was 3183 rpm at the transporting velocity of 0.61 m/s with the loading quantity of 3.41 t/hr 3. Recommendable opening area ratio of pressure controller was 1/2 at the blower speed of 3183 rpm and the transporting velocity of 0.61 m/s 4. The break even point resulted in $240\;m^2$ of cultivating area compared to the method of with portable workbench, and $350\;m^2$ of cultivating area compared to the method of with a tractor and a truck.

Antialgal Effect of a Novel Polysaccharolytic Sinorhizobium kostiense AFK-13 on Anabaena flos-aquae Causing Water Bloom

  • Kim, Jeong-Dong;Lee, Choul-Gyun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1613-1621
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    • 2006
  • Isolation and identification of algal lytic bacteria were carried out. Nine strains of algal lytic bacteria were isolated by the double-layer method using Anabaena flos-aquae as a sole nutrient. The isolate, AFK-13, showing the highest algal lytic activity was identified as Sinorhizobium kostiense based on the l6S rDNA sequence. The algal lytic experiments of the culture supernatants of AFK-13 demonstrated that the bacterial cell growth reached a maximum at 36-h culture, but the supernatant of 72-h culture exhibited the highest activity. Components among the extracellular products in the crude enzyme of the supernatant from S. kostiense AFK-13 culture were responsible for degradation of cell walls of Anabaena flos-aquae. Algal lytic assay tests of the culture supernatants suggest that the main substances for algal lytic activity could be proteinaceous. The activity of glucosidase was observed highly by polysaccharolytic analysis using the crude enzyme from S. kostiense AFK-13, whereas activities of galactosidase, mannosidase, rhamnosidase, and arabinosidase were also detected in low levels. The molecular weights (MW) of ${\alpha}-\;and\;{\beta}$-glucosidases were estimated to be approximately 50-100 kDa by the ultrafiltration method.

Analysis of S-glutathionylated proteins during adipocyte differentiation using eosin-glutathione and glutaredoxin 1

  • Hwang, Sungwon;Iram, Sana;Jin, Juno;Choi, Inho;Kim, Jihoe
    • BMB Reports
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    • v.55 no.3
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    • pp.154-159
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    • 2022
  • Protein S-glutathionylation is a reversible post-translational modification on cysteine residues forming a mixed disulfide with glutathione. S-glutathionylation, not only protects proteins from oxidation but also regulates the functions of proteins involved in various cellular signaling pathways. In this study, we developed a method for the detection of S-glutathionylated proteins (ProSSG) using eosin-glutathione (E-GSH) and mouse glutaredoxin 1 (mGrx1). ProSSG was efficiently and specifically labeled with E-GSH to form ProSSG-E via thiol-disulfide exchange. ProSSG-E was readily luminescent allowing the detection of ProSSG with semi-quantitative determination. In addition, a deglutathionylation enzyme mGrx1 specifically released E-GSH from ProSSG-E, which increased fluorescence allowing a sensitive determination of ProSSG levels. Application of the method to the adipocyte differentiation of 3T3-L1 cells showed specific detection of ProSSG and its increase upon differentiation induction, which was consistent with the result obtained by conventional immunoblot analysis, but with greater specificity and sensitivity.

A Design and Implementation of Control and Management System for Water Culture Device using Solar Tracking Method (광원 트래킹 기법을 이용한 수경재배기 제어 관리 시스템 설계 및 구현)

  • Park, Sung-Kyun;Jung, Se-Hoon;Oh, Min-Joo;Sim, Chun-Bo;Park, Dong-Gook;You, Kang-Soo
    • The Journal of the Korea institute of electronic communication sciences
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    • v.9 no.2
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    • pp.231-242
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    • 2014
  • It is throwing the spotlight on the cultivation crops about high quality crops and productivity improvement per unit area because of rapid climate change caused by global warming. Therefore, we propose a water culture management of circulation nutrient method control system applies to solar tracking method not using traditional method of deep flow technique and artificial light source. We design it in the form of the circulation nutrient method in waterway of a certain amount of nutrient solution and water flowed into the way of circular. In addition, we design a multistage structure in pyramid shape which be possible continuous photosynthesis action to crops of water culture bottom part. Also, solar tracking method is designed five sensor method of center hole sensor method for tracking shadow of solar light not using traditional two hole, four hole sensor method. Finally, through the water culture device applies to solar light tracking method was not introduced in existing study yet, we can reduce growth speed of crops which be possible continuous photosynthesis action to crops. Moreover, We can expect high productivity of per unit area which be possible all crops can be offered growth environment of same type by using form of pyramid shape of multistage structure without top or bottom part.

A Study on the Plan Photogrammetry for Clothing Design (피복구성학적 인체계측방법에 관한 연구 - 평면사진계측방법을 중심으로 -)

  • 박찬미;서미아
    • The Research Journal of the Costume Culture
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    • v.5 no.1
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    • pp.151-164
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    • 1997
  • This study pursues the problems of plan photogrammetry which is widely used in somatotyping at present, and find out a method which can improve accuracy of measurement on the basis of principles and mechanisms of photography-the basic foundation of the photographic analysis methods. As a result, this study proposes a new method which is based on the reference point method and perspective coordinate system. And the test measurement was operated to compare the measurement accuracy of the proposed method and the method based on reference grid screen method and perpendicular coordinate system which is commonly used at present. The result of this test measurement showed that the proposed method has higher accuracy. Two reasons can be pointed out for the improvement of measuring accuracy. The first reason is that the proposed perspective coordinate system reduces the perspective distortion of photography. And second reason is that measuring points can be closely placed to the scale and coordinate reference plan of measurement by the proposed reference point method which make possible to place measuring object (or person) at the center of scale and coordinate reference plan by utilizing reference points of measurement in the three dimensional space not on screen.

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TaqMan Probe Real-Time PCR for Quantitative Detection of Mycoplasma during Manufacture of Biologics (생물의약품 제조공정에서 마이코플라스마 정량 검출을 위한 TaqMan Probe Real-Time PCR)

  • Lee, Jae Il;Kim, In Seop
    • KSBB Journal
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    • v.29 no.5
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    • pp.361-371
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    • 2014
  • Mycoplasma is well recognized as one of the most prevalent and serious microbial contaminants of biologic manufacturing processes. Conventional methods for mycoplasma testing, direct culture method and indirect indicator cell culture method, are lengthy, costly and less sensitive to noncultivable species. In this report, we describe a new TaqMan probe-based real-time PCR method for rapid and quantitative detection of mycoplasma contamination during manufacture of biologics. Universal mycoplasma primers were used for mycoplasma PCR and mycoplasma DNA was quantified by use of a specific TaqMan probe. Specificity, sensitivity, and robustness of the real-time PCR method was validated according to the European Pharmacopoeia. The validation results met required criteria to justify its use as a replacement for the culture method. The established real-time PCR assay was successfully applied to the detection of mycoplasma from human keratinocyte and mesenchymal stem cell as well as Vero cell lines artificially infected with mycoplasma. The overall results indicated that this rapid, specific, sensitive, and robust assay can be reliably used for quantitative detection of mycoplasma contamination during manufacture of biologics.

Research of a new tie-dyeing tool based on 3D printing technology

  • Tu, Dan Dan;Kim, Sohyun
    • The Research Journal of the Costume Culture
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    • v.30 no.1
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    • pp.161-171
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    • 2022
  • Traditional tie-dyeing is widely implemented in the clothing handicraft culture in China, South Korea, and Japan. Since it was developed 2,000 years ago, it has become a popular method of fabric making in the world and is highly respected by fashion designers. Based on the existing traditional tie-dyeing methods, this study conducted specific research on the 3D printing technology of the SLS laser method and the micro tool design application method of the clamp-dyeing process. Through the experimental methods of this study, it proposes to use the "7000 Nylon" material, which is commonly used in 3D printing, to develop a new clamp-dyeing tool. This new tool can be widely used in the clamp-dyeing of fabrics, such as cotton, hemp, silk, and some chemical fibers. The applied method and principle can be consistent with the traditional clamp-dyeing method. Therefore, the innovation of tie-dyeing technology is the best protection measure for the development and inheritance of traditional fabric making. The continuation of artistic life needs originality, which is also the best response to market competition. At the same time, this new design of the clamp-dyeing tool has the characteristics of novelty, innovation, and rich changes, which aligns with the new fashion demands of current fabric design.