• Title/Summary/Keyword: Laboratory Culture

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Development of a tide-simulating apparatus for macroalgae

  • Kim, Jang-K.;Yarish, Charles
    • ALGAE
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    • v.25 no.1
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    • pp.37-44
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    • 2010
  • A tide-simulating apparatus was developed for culturing marine macroalgae. The objective of this study was to introduce a novel tide-simulating apparatus that can simulate a diurnal or semi-diurnal tidal cycle in the laboratory. In this apparatus, the seaweeds are move up and down and the water level remains the same during the simulated tidal cycle. The apparatus consists of 18 cylindrical culture tanks (3 blocks $\times$ 6 culture tanks) with 12 cm diameter and 24.5 cm long containing up to 2.5 L of seawater. There is a horizontal plate which covered all 18 culture tanks, and it is raised and lowered by a programmable motor that can regulate exposure time. In one application, seaweeds are attached to braided twine hung on Plexiglas air-tubing. The air-tubing is attached to a lid that is set on a horizontal plate. This apparatus is made of colorless Plexiglas to maximize light transmittance. This apparatus is easily disassembled and transportable to any indoor laboratory, wet laboratory, greenhouse, etc. This apparatus also offers considerable flexibility in terms of design. The size of culture tank can be redesigned by either increasing the height of cylinder or/and using a different diameter of cylindrical Plexiglas, therefore, larger/taller thalli can be cultivated. Growth rates of three eulittoral Porphyra species from different tidal elevations have been compared using this device.

The Rapid Drug Susceptibility Testing of Mycobacterium tuberculosis by GenoType$^{(R)}$ MTBDRplus in Contaminated Specimen

  • Heo, Reun;Kim, Yoon-Sik
    • Biomedical Science Letters
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    • v.19 no.4
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    • pp.330-337
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    • 2013
  • There are several methods currently being used to diagnose tuberculosis in patients, such as smear, PCR, tuberculosis culture and X-ray. For a proper medical treatment, antimicrobial susceptibility test and rapid drug susceptibility testing have been operated. Tuberculosis bacilli usually need 3~8 weeks of culture period because of delay in RNA synthesis and require 15~22 hours for generation. After a germ raises in culture, we initiated antimicrobial susceptibility test for a proper treatment. It has some difficulties to give a proper prescription for a tuberculosis patient because antimicrobial susceptibility test requires 4 weeks. To supplement this, we are practicing drug susceptibility testing which allow us to know the sensibility of RMP and INH after 2 or 3 days. But this is only possible when more than 2 positive germ. Therefore, we should practice rapid drug susceptibility testing with culture test. But if media is contaminated by other germs except Mycobacterium tuberculosis, it's hard to interpret result about culture test and to practice antimicrobial susceptibility test and rapid drug susceptibility testing. Because we have to practice again smear, culture test after extracting specimen from the patient, time is consumed and proper patient treatment is postponed. To address these problems and quick patient treatment, rapid drug susceptibility testing is practiced by using GenoType$^{(R)}$ MTDRplus method. As a result of this method we detected sensibility 10 and 7 cases and resistance 0 and 3 cases using RIM and INH respectively with other 1 case toward medicals out of the total 11 test. In conclusion rapid drug susceptibility testing can be used from the contaminated specimen after elimination of contaminated source from culture and proved that it can be practiced for rapid examination of a tuberculosis patient.

Embryogenic cell suspension culture and plant regeneration in zoysiagrass (Zoysia japonica Steud) (한국들잔디 배아세포의 부유배양과 식물체 재생)

  • Fang, Wenjuan;Han, Liebao;Qi, Chunhui;Li, Deying;Park, Tae-Yun
    • Asian Journal of Turfgrass Science
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    • v.23 no.2
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    • pp.345-352
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    • 2009
  • Zoysiagrass (Zoysia japonica Steud) is a warm season turfgrass species widely used for sports field and golf courses. Many cultivars are propagated through vegetative methods. This study was conducted to develop an optimum culture medium and culture conditions for embryogenic callus induction and plant regeneration, and to establish a cell suspension culture system for use in zoysiagrass breeding and propagation. The results indicated that adding $Cu^{++}$ at 2.5 mg $L^{-1}$ to the induction medium was optimum for callus induction. Increasing the numbers of sub-culture cycles improved the quality of calli. The optimum dosage for cell suspension culture ranged from 2.5 to 10 mL. The embryogenic callus suspension used in this study had a plant regeneration rate of 58%.

Induced Differentiation of Embryonic Stem Cells to Insulin Secreting Cells (배아줄기세표의 인슐린 분비세포로의 유도 분화에 대한 연구)

  • Sung, Ji-Hye;Lim, Chun-Kyu;Choi, Hye-Won;Lee, Hyoung-Song;Shin, Hyeon-Sang;Jun, Jin-Hyun;Yoon, Hyun-Soo;Koong, Mi-Kyoung
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.4
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    • pp.209-215
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    • 2004
  • Objective: Embryonic stem (ES) cells could be differentiated into the specific cell types by alternation of culture condition and modification of gene expression. This study was performed to evaluate the differentiation protocol for mouse and human ES cells to insulin secreting cells. Methods: Undifferentiated mouse (JH-I) and human (Miz-hESI) ES cells were cultured on STO feeder layer, and embryoid bodies (EBs) were formed by suspension culture. For the differentiation, EBs were cultured by sequential system with three stage protocol. The differentiating ES cells were collected and marker gene expressions were analyzed by seIni-quantitative RT-PCR in each stage. Amount of secreted insulin levels in culture media of human ES cells were measured by human insulin specific RIA kit. Results: During the differentiation process of human ES cells, GATA-4, a-fetoprotein, glucose transporter-2 and Ngn-3 expression were increased whereas OctA was decreased progressively. Insulin and albuInin mRNAs were expressed from stage IT in mouse ES cells and from stage III in human ES cells. We detected 3.0~7.9 IlU/rnl secretion of insulin from differentiated human ES cells by in vitro culture for 36 days. Conclusion: The sequential culture system could induce the differentiation of mouse and human ES cells into insulin secreting cells. This is the fIrst report of differentiation of human ES cells into insulin secreting cells by in vitro culture with serum and insulin free medium.

Gene Expression and Secretion of the Anticoagulant Hirudin in Saccharomyces cerevisiae

  • Sohn, Jung-Hoon;Lee, Sang-Kwon;Choi, Eui-Sung;Rhee, Sang-Ki
    • Journal of Microbiology and Biotechnology
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    • v.1 no.4
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    • pp.266-273
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    • 1991
  • Hirudin, a 65-amino acid protein isolated from the salivary gland of the bloodsucking leech, Hirudo medicinalis, is a potent thrombin-specific inhibitor and blocks the thrombin-mediated conversion of fibrinogen to fibrin in clot formation. We have studied the gene expression and secretion of hirudin in yeast. Saccharomyces cerevisiae. A gene coding for hirudin was synthesized based on the amino acid sequence and cloned into a yeast expression vector $YEG{\alpha}-1$ containing the ${\alpha}-mating$ factor pre-pro leader sequence and galactose-inducible promoter, GALl0. Recombinant S. cerevisiae was found to secrete biologically active hirudin into the extracellular medium. The secreted recombinant hirudin was recovered from the culture medium and purified with ultrafiltration and reverse phase high performance liquid chromatography. Approximately 1 mg of hirudin per liter was produced under suboptimal culture conditions and brought to about 90% purity in two steps of purification.

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Laboratory, Field and Deep Seawater Culture of Eucheuma serra-a High Lectin Yielding Red Alga

  • Dinabandhu Sahoo;Masao Ohno;Masanori Hiraoka
    • ALGAE
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    • v.17 no.2
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    • pp.127-133
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    • 2002
  • The red seaweed Eucheuma serra is a high yielding source of lectins. The plants were collected from a depth of 5-6 meters and cultured in the laboratory, field and deep seawater. A Daily Growht Rate (DGR) of 3.5% was observed at 18℃ with a low light of 30μmol photon $ m^{-2} · s^{_1}$ in the laboratory. When the plants were cultured in the field at different depths during winter onths of December and January, best growth was observed at 1 m depth and a DGR of 2.14±0.04% was recorded. The plants grown in the tank with a continuous supply of deep seawater showed a DGR of 8.2% The results indicate that E. serra can be cultivated in large scale both in deep seawater in the tank and in the field for the extraction of lectins at a commercial scale.

Improving Mycoplasma ovipneumoniae culture medium by a comparative transcriptome method

  • Wang, Xiaohui;Zhang, Wenguang;Hao, Yongqing
    • Journal of Veterinary Science
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    • v.21 no.2
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    • pp.30.1-30.11
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    • 2020
  • Mycoplasma ovipneumoniae (Mo) is difficult to culture, resulting in many difficulties in related research and application. Since nucleotide metabolism is a basic metabolism affects growth, this study conducted a "point-to-point" comparison of the corresponding growth phases between the Mo NM151 strain and the Mycoplasma mycoides subsp. capri (Mmc) PG3 strain. The results showed that the largest difference in nucleotide metabolism was found in the stationary phase. Nucleotide synthesis in PG3 was mostly de novo, while nucleotide synthesis in NM151 was primarily based on salvage synthesis. Compared with PG3, the missing reactions of NM151 referred to the synthesis of deoxythymine monophosphate. We proposed and validated a culture medium with added serine to fill this gap and prolong the stationary phase of NM151. This solved the problem of the fast death of Mo, which is significant for related research and application.