• Title/Summary/Keyword: agar-free

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Plant Regeneration from Protoplasts Isolated through Embryogenic Cell suspension Culture in Rice (벼 현탁배양을 통하여 분리된 원형질체로부터 식물체 재분화)

  • 정병균
    • Journal of Plant Biology
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    • v.36 no.3
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    • pp.211-218
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    • 1993
  • Plant regeneration was accomplished from protoplast culture of rice (Oryza sativa L. cv. Taebaeg). Embryogenic callus was induced from mature seed on MS medium containing 5 mM proline, 2.5 mg/L 2,4-D, 30 g/L sucrose in the dark at 28$^{\circ}C$ and used to establish embryogenic cell suspension culture. Suspension cells were subcultured every one week in N6 medium supplemented with 5 mM proline, 200 mg/L casein hydrolysate, 2.5 mg/L 2,4-D and amino acids of AA medium. Suspension cultures were composed of cells that were densely cytoplasmic, potentially embryogenic and were at least maintained for more than 6 months in liquid medium. Protoplasts were isolated from fast-growing suspension culture cells and cultured in a slightly modified KpR medium by mixed nurse culture. Isolated protoplasts began to divide within 5~7 days and thereafter, protoplast-derived calli were sequentially transferred to callus proliferating medium that soft agar MS medium contained 2 mg/L 2,4-D and produced distinct embryogenic cells. Microcolonies were then transferred to solid medium which consisted of MS medium containing 5 mg/L kinetin, 1 mg/L NAA, 1 mg/L ABA, 30 g/L sucrose and 10 g/L sorbitol under fluorescent light. Mulitple shoots of 4~5 per callus emerged and were transferred to hormone-free MS medium for root initiation. Thereafter, The plantlets were transferred to pots of soil to mature in the culture room.

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A Study of the Quality Characteristics of Yanggaeng Supplemented with Codonopsis lanceolata Traut (Benth et Hook) (더덕을 첨가한 양갱의 품질 특성에 관한 연구)

  • Kim, Myung-Hee;Chae, Hyun-Seok
    • Journal of the East Asian Society of Dietary Life
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    • v.21 no.2
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    • pp.228-234
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    • 2011
  • This study investigated the anti-oxidation activities of fresh Codonopsis lanceolata Traut (Benth et Hook) and red beans, and also the quality properties of fresh Codonopsis lanceolata Yanggaeng to which raw Codonopsis lanceolata Traut (Benth et Hook) was added. The DPPH free radical scavenging activities of fresh Codonopsis lanceolata Traut (Benth et Hook) and red beans were 31.42% and 37.98%, respectively. Thus, that of red beans was rather the total polyphenol contents of fresh Codonopsis lanceolata Traut (Benth et Hook) and red beans were 5.74 mg/g and 4.14 mg/g respectively, and the total flavonoid contents of fresh Codonopsis lanceolata Traut (Benth et Hook) and red beans were 5.07 mg/g and 1.29 mg/g respectively. As higher amounts of Codonopsis lanceolata Traut (Benth et Hook) were added to Yanggaeng, the water content in the Yanggaeng continued to increase. Preferences were in the following order: 21%>12%>28%>7%>0%, and it was found that the optimal level for addition of fresh Codonopsis lanceolata Traut (Benth et Hook) was between 12% and 21%. It was also shown that adjusting the amount of ingredients or agar would improve the product quality.

Factors influencing efficiency of somatic embryogenesis of Gentiana kurroo (Royle) cell suspension

  • Fiuk, Agnieszka;Rybczynski, Jan J.
    • Plant Biotechnology Reports
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    • v.2 no.1
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    • pp.33-39
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    • 2008
  • In this paper, we would like to show unexpected morphogenic potential of cell suspensions derived from seedling explants of Gentiana kurroo (Royle). Suspension cultures were established with the use of embryogenic callus derived from seedling explants (root, hypocotyl and cotyledons). Proembryogenic mass proliferated in liquid MS medium supplemented with $0.5mg\;l^{-1}$ 2,4-D and $1.0mg\;l^{-1}$ Kin. The highest growth coefficient was achieved for root derived cell suspensions. The microscopic analysis showed differences in aggregate structure depending on their size. To assess the embryogenic capability of the particular culture, 100 mg of cell aggregates was implanted on MS agar medium supplemented with Kin ($0.0-2.0mg\;l^{-1}$), $GA_3$ ($0.0-2.0mg\;l^{-1}$) and AS ($80.0mg\;l^{-1}$). The highest number of somatic embryos was obtained for cotyledon-derived cell suspension on $GA_3$-free medium, but the best morphological quality of embryos was observed in the presence of $0.5-1.0mg\;l^{-1}$ Kin, $0.5mg\;l^{-1}$ $GA_3$ and $80.0mg\;l^{-1}$ AS. The morphogenic competence of cultures also depended on the size of the aggregate fraction and was lower when size of aggregates decreased. Flow cytometry analysis reveled luck of uniformity of regenerants derived from hypocotyl suspension and 100% of uniformity for cotyledon suspension.

The First Acanthamoeba keratitis Case of Non-Contact Lens Wearer with HIV Infection in Thailand

  • Tananuvat, Napaporn;Techajongjintana, Natnaree;Somboon, Pradya;Wannasan, Anchalee
    • Parasites, Hosts and Diseases
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    • v.57 no.5
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    • pp.505-511
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    • 2019
  • Acanthamoeba keratitis (AK) is a rare sight-threatening corneal infection, often reporting from contact lens wearers. An asymptomatic human immunodeficiency virus (HIV)-infected Thai male without history of contact lens use complained foreign body sensation at his left eye during motorbike riding. He had neither specific keratitis symptoms nor common drugs responding, which contributed to delayed diagnosis. By corneal re-scraping, Acanthamoeba-like cysts were detected by calcofluor white staining and agar culture. The etiological agent obtained from the culture was molecularly confirmed by Acanthamoeba spp.-specific PCR, followed by DNA sequencing. The results from BLAST and phylogenetic analysis based on the DNA sequences, revealed that the pathogen was Acanthamoeba T4, the major genotype most frequently reported from clinical isolates. The infection was successfully treated with polyhexamethylene biguanide resulting in corneal scar. This appears the first reported AK case from a non-contact lens wearer with HIV infection in Thailand. Although AK is sporadic in developing countries, a role of free-living Acanthamoeba as an opportunistic pathogen should not be neglected. The report would increase awareness of AK, especially in the case presenting unspecific keratitis symptoms without clinical response to empirical antimicrobial therapy.

Studies on Bovine Serum Protein Fractions (I, II) (혈청단백질분획(血淸蛋白質分劃)에 대한 연구(硏究) (I, II))

  • Rim, Bong Ho
    • Korean Journal of Veterinary Research
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    • v.5 no.1
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    • pp.1-16
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    • 1965
  • I. A Comparison of Sodium Sulfate Precipitation and Zone(Paper, Agar) Electrophoresis; Many kinds of techniques have been used for fractionating serum proteins. In the present study, using bovine serum, the fractions obtained with sodium sulfate were compared with those determined by zone electrophoresis. 1. Fibrinogen was precipitated with 4 to 10 percent of sodium sulfate. 2. ${\gamma}$-globulin required 10 to 16 percent of the salt for precipitation. 3. ${\beta}$-globulin began to precipitate at 12 percent sodium sulfate, and completed precipitation at approximately 26 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. 4. ${\alpha}$-globulin completed precipitation at 13 to 28 percent sodium sulfate in paper electrophoresis and at 22 percent in agar electrophoresis. 5. Albumin began to precipitate at 14 percent of the salt, and was free from the mixture of globulins approximately at 28 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. The results of comparing fractions by the two methods were as follows: 1. Euglobulin (15%) was equal to the sum of the most ${\gamma}$-globulin and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. 2. Pseudoglobulin I (15-17.5%) corresponded to the most ${\alpha}$-, ${\beta}$-globulins and a small quantity of albumin. 3. Pseudoglobulin II(18-22%) was a mixture of the ${\alpha}$-, ${\beta}$-globulins and albumin fraction. 4. Albumin (above 22%) contained the most albumin fraction separated by zone electrophoresis and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. As mentioned above the fractions obtained with sodium sulfate were a mixture of the various proportion of the fractions determined by zone electrophoresis. The solubility of serum fractions to sodium sulfate coincided with the mobility of those by zone electrophoresis. (By percent of sodium sulfate we mean gram of sodium sulfate contained in $100m{\ell}$ of solution). II. Immunological Studies on Serum Protein Fractions with Sodium Sulfate; In the previous report the fractions of bovine serum protein with sodium sulfate compared with those obtained by zone electrophoresis, and the findings were that the former contained various proportion components of the latter. In this study the author studied whether or not the fractions with sodium sulfate are simple component antigenically by immunoelectrophoresis and micro double diffusion test (Immuno-precipitation), using rabbit antiserum to bovine serum. In immunoelectrophoresis, normal bovine serum developed with rabbit antibovine serum showed about ten distinct precipitin arcs. The distribution of these arcs was as follows: 1 albumin, 2 ${\alpha}_1$-, 3 ${\alpha}_2$-, 2 ${\beta}_1$-, ${\beta}_2$-, and 1 ${\gamma}$-globulin (Fig. 7, 9). In micro double diffusion test, five to six precipitation bands could be seen between antigens and antibody, the order of the precipitation bands location is albumin, ${\alpha}$-, ${\beta}$-, and ${\gamma}$-globulin from the side of antiserum well (Fig.19). Frequently the ${\alpha}$-, and ${\beta}$-precipitation bands were separated into two or three precipitation bands, which indicated that these globuline are not a pure component antigenically as shown in immuno-electrophoresis. In both Immunological methods, the two ${\alpha}$-, ${\beta}$-precipitin arcs and bands appeared clear and strong, indicating that the two globulins reacted as strong antigens. The precipitate reaction of ${\gamma}$-globulin was shown at 12 to 16 percent sodium sulfate; ${\beta}$-globulin at 12 to 20 percent; ${\alpha}$-globulin at 12 to 22 percent (immuno-electrophoresis), at 12 to 26 percent (Diffusion); and albumin at above 22 percent. Antigenically euglobulin contained ${\gamma}$-, ${\beta}$-, and ${\alpha}$-globulins, Pseudoglobulin I and Pseudoglobulin II were composed of ${\alpha}$-, and ${\beta}$-globulins, and albumin was a mixture of ${\alpha}$-globulin and albumin determined by zone electrophoresis. The results indicated that the fractions of serum protein obtained by either method were constituents of various proteins antigenically except ${\gamma}$-globulin and albumin by Zone electrophoresis.

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Neutral Electrolyzed Water for Prevention of Dental Caries (기존 구강청결제를 대체할 수 있는 치아우식 예방을 위한 전기분해수)

  • Lee, Kyam
    • Journal of the korean academy of Pediatric Dentistry
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    • v.43 no.3
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    • pp.306-312
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    • 2016
  • Gargle solution has typically been used for the prevention of oral infectious disease such as dental caries and periodontitis. However, the use of most gargle solutions is controversial in application for children because some gargle solutions have harmful side effects. Electrolyzed water is generated by passed an electric current and has antimicrobial activity. The purpose of this study was to investigate and compare the efficacy of electrolyzed water in various conditions for eliminating cariogenic bacteria. Electrolyzed water was generated by a platinum electrode in the presence of sodium chloride at various concentrations. Streptococcus mutans and Streptococcus sobrinus were cultivated into a brain heart infusion broth. After harvesting planktonic bacteria, the pellets were treated with the electrolyzed water and commercial gargle solutions and plated on a mitis-salivarius agar plate. Also, the anti-biofilm activity of the electrolyzed water and commercial gargle solutions was investigated after biofilm formation of S. mutans and S. sobrinus. The bacteria in the biofilm were plated onto a mitis-salivarius agar plate. The plates were incubated, and the colony forming unit was measured. The electrolyzed water containing sodium chloride showed significant antibacterial activity against S. mutans and S. sobrinus as well as some gargle solutions. Furthermore, the electrolyzed water had more disruptive effect on the biofilm of S. mutans and S. sobrinus and killed more bacteria in the biofilm than commercial gargle solutions. The results demonstrate that electrolyzed water may be a useful gargle solution for prevention of dental caries.

Production of Nucleotide by Immobilized Cell (고정화 미생물에 의한 뉴크레오타이드 생산)

  • CHO Jung-Il;JUNG Sung-Won
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.24 no.2
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    • pp.111-116
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    • 1991
  • The effective p.eduction of 5'-GMP(5'-Guanylic acid) by enzymatic conversion of 5'-XMP(5'-Xanthyic acid) was investigated. The Iyophilized Brevibacterium ammoniagenes ATCC 19216 which were used as the XHP aminase source, was immobilized by entrapping in K-carrageenan, agar, polyacrylamide or Ca-alginate. $3\%$ K-carrageenan was selected as the most suitable matrix. In the production of 5'-GMP using the free cells of 3. ammoniagenes ATCC 19216, the optimum conditions were $42^{\circ}C$, PH 7.0, 100mg/ml glucose, 120mg/ml cell ,8mg/ml $MgSO_4\cdot7H_2O$, 5mg/ml POESA, 5mg/ml phytic acid. Under the conditions, $94.5\%$ of 5'-GMP was converted within 8 hours. In the production of 5'-GMP using the immobilized whole cells of B. ammoniagenes ATCC 19216, the optimum conditions were $37^{\circ}C$, pH 7.5, 50mg/ml glucose, 1mg/ml $KH_2PO_4$, 10mg/ml phytic acid, 60mg/ml cell, 8mg/ml $MgSO_4\;\cdot\;7H_2O$, 5mg/ml POESA. Under the conditions, $64.7\%$ of 5'-GMP was converted within 40 hours.

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Immobilization of Nitrifier Consortium for the Removal of Ammonium Ion in the Recirculating Aquaculture System (양어장수내의 암모니아성 질소제거를 위한 질화세균군의 고정화)

  • KIM Sung-Koo;SEO Jae-Koan;LEE Jong-Seok;KONG In-Soo;SUH Keun-Hack
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.5
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    • pp.816-822
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    • 1997
  • The immobilization of a microorganism has been rapidly progressed with the development of biotechnology in recent years. Although it has been used as a tool to isolate products from biological media in various areas, it has not yet been practiced in the treatment of waste water. In this paper, we suggest a possibility to apply the immobilization technique In the recirculating aquaculture system. We examined the ability of $NH_4^+$ removal by nitrifier consortium immobilized in $Ba^{++}-alginate$, k-carrageenan and agar bead at the concentration of 50 g/L, respectively. In order to use the immobilized nitrifier consortium as media in the fludized bed reactor, the strength of bead was measured. $Ba^{++}-alginate$ as a support material showed higher strength of bead. Also, the nitrifier consortium immobilized in $Ba^{++}-alginate$ showed higher nitrification activity that could remove 20 mg/L ammonium ion than those immobilized in other two support materials, carrageenan and agar. The immobilized nitrifier consortium showed better nitrification activity than free nitrifier consortium.

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Processing of Fermented Squeezed-type Pen Shell By-product by Proteolytic Enzyme (효소분해법에 의한 키조개부산물 젓갈의 제조)

  • KANG Hoon-I;KANG Tae-Joong;BAE Tae-Jin;KIM Hyun-Ju
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.5
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    • pp.509-514
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    • 1994
  • A processing method for fermented paste of favorable flavor and texture using pen shell by-product was investigated. The pen shell by-product was homogenized with the addition of water and hydrolyzed with $5\%$ of Protin P(105 PU/g) at $55^{\circ}C$ for 1 hour. Flavor of the hydrolysate could be improved by thermal treatment at $100^{\circ}C$ for 40 minutes with $10\%$ of invert sugar. $2\%$ of agar-agar and $6\%$ of starch added to hydrolysate emulsified by $8\%$ of polyglycerol condensed ricinoleate(PGDR) were significantly effective for the improvement of rheological properties such as hardness, springiness and chewiness of the fermented paste. $15\%$ of table salt was finally added to the product of fermented pen shell paste. The contents of moisture, protein, lipid, carbohydrate and salinity of the product were $62.7\%,\;3.2\%,\;4.4\%,\;10.6\%\;and\;15.6\%$, respectively. The major free amino acids were glutamic acid, arginine, aspartic acid, leucine, lysine, glycine and alanine. The product was stable for the storage of 60 days at $23{\pm}3^{\circ}C$ on bacterial growth.

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EFFECTS OF SIGNAL TRANSDUCTION PATHWAY IN THE RAS-INDUCED CELLULAR TRANSFORMATION OF HUMAN EPITHELIAL CELLS IN CULTURE (인체 상피세포에서 ras-종양유전자의 발암화가 신호 전달 기작에 미치는 영향)

  • Jang, Do-Geun;Byeon, Ki-Jeong;Kim, Chin-Soo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.3
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    • pp.254-261
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    • 2000
  • The present study has attempted to look into the mechanism of ras-induced carcinogenesis in a human epithelial cell system. Human epithelial cells immortalized with Ad12-SV40 hybrid virus were used to assess carcinogenic potential of the ras-oncogene. Cells transfected with pSV2-ras showed characteristics of cellular transformation. The transformation parameters such as cell density, soft-agar colony formation, and cell aggregation were significantly increased in the cells expressing ras oncoprotein. In addition, the duration required for the appearance of foci was shortened in the ras-transfected cells. Consistent with other reports, our results demonstrated an evidence that the ras-oncogene induced the cellular transformation of human epithelial cell system. When a high concentration of glucocorticoid was added into the media, transformation process was accelerated. It is speculated that glucocorticoid may provide an advantageous environment for the proliferation of the transformed cells. The induction of the intracellular free calcium concentrations following agonist treatment was significantly lower in the transformed cells than in the control cells. These effects were more manifested in the presence of extracellular cacium, indicating that the transformation process may alter the influx pathway of extracellular calcium. The induction of $IP_3$ following agonist treatment was also lower in the transformed cells than in the control cells. Thus, it is suggested that phospholipase C-coupled pathway was down-regulated in the process of the ras-induced transformation. While the levels of $TGF-{\beta}_1$ and PAI-2 mRNAs were decreased, the level of fibronectin mRNA was increased. The results indicate that mechanism of the ras-induced transformation may be associated with the altered expressions of growth regulatory factors. The present study demonstrates an evidence that the ras-induced cellular transformation may be associated with alteration of signal transduction and growth regulatory factors. The study will contribute to improve the understanding of molecular mechanism of epithelium-derived cancers including oral cancer.

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