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The Selection of Domestically Bred Cultivars for Spray-type Chrysanthemum Transformation (스프레이 국화 형질전환을 위한 국내 육성 품종 선발)

  • Suh, Eun-Jung;Han, Bong Hee;Lee, Yeon-Hee;Lee, Seong-Kon;Hong, Joon Ki;Kim, Kyung Hwan
    • Horticultural Science & Technology
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    • v.33 no.6
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    • pp.947-954
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    • 2015
  • To select suitable spray chrysanthemum cultivars for Agrobacterium-mediated transformation, thirty-nine (39) spray cultivars bred in the National Institutes of Korea and a standard cultivar Jinba from Japan were collected and tested for regeneration rate and Agrobacterium infection assays. MS medium with $0.5mg{\cdot}L^{-1}$ IAA and $1.0mg{\cdot}L^{-1}$ BAP was used for shoot regeneration from leaf disks and internodes. The shoot regeneration rate in leaf disks was the highest in cultivar BRM, followed by cultivars VS, WW and YTM. The cultivar JB (Jinba) used as a transformation material in previous reports ranked similarly to cultivars PK and SPP. In shoot regeneration from internodes, the shoot regeneration rate was the highest for cultivar PA, followed by cultivar WW. The infection rate of leaves and internodes of 40 chrysanthemum cultivars with agrobacterium was investigated. Cultivars WPP, YNW, VS, PP, WW, FA, PA and YMN showed the highest infection levels in leaves, whereas cultivars WPP, PA, PK and YNW had the highest infection levels in internodes. Considering all of these results, cultivars VS and WW were the most appropriate for gene transformation of chrysanthemum using leaves, while cultivar PA was for internodes.

THE EFFECT OF THE GENISTEIN ON THE PROLIFERATION OF HT1080 AND EXPRESSION OF MEMBRANE TYPE 1-MATRIX METALLOPROTEINASE (MT1-MMP) mRNA (Genistein이 사람 섬유육종 세포주 증식 및 Membrane Type 1-Matrix Metalloproteinase (MT1-MMP) mRNA 발현에 미치는 영향)

  • Kang, Jin-Han;Myoung, Hoon;Kim, Myung-Jin
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.4
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    • pp.314-320
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    • 2001
  • Matrix metalloproteinases have long been viewed as ideal candidates for proteinases that enables tumor cells to permeated basement membrane defenses and invade surrounding tissue. There is growing evidence that the MMPs have an expanded role, as they are important for the creation and maintenance of a microenvironment that facilitates growth and angiogenesis of tumors at primary and metastatic sites. MT-MMPs are not secreted but instead remaining attached to cell surfaces. Although not all of the MT-MMPs are fully characterized, MT-MMPs have important role in localizing and activating secreted MMPs. The MMP genes are transcriptionally responsive to a wide variety of oncogene, growth factors, cytokine, and hormones. Currently, a number of MMP inhibitors are being developed and some have reached clinical trials as anti-metastatic or anti-cancer therapies. MT1-MMP is involved in the activation of proMMP-2. MT1-MMP is significant not only as a tumor marker but as a new target for chemotherapy against cancer. The purpose of this study was to evaluate the effects of protein kinase C inhibitor(genistein) on the proliferation of HT1080 and expression of MT1-MMP mRNA. Human fibrosarcoma cell line HT1080 was cultured and divided 2 groups. The experimental group was treated with $100{\mu}M$ genistein and incubated 12h, 24h for $[3^H]-thymidine$ uptake assay and northern hybridization individually. And the control group was treated with same amount of PBS for the above procedures. $[3^H]-thymidine$ incorporation was measured with ${\beta}$ ray detector. And RT-PCR and northern blotting for MT1-MMP mRNA was performed. The results were as follows 1. $[3^H]-thymidine$ uptake was reduced in experimental group with statistical significance. 2. MT1-MMP mRNA expression was significantly reduced in experimental group. These results showed that protein kinase C inhibitor (genistein) inhibited proliferation of HT1080 and almost completely blocked transcription of MT1-MMP mRNA. So, it is possible to use the protein kinase inhibitor (genistein) as anti-metastatic and anti-proliferative agent.

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Serological and Molecular Detection of Toxoplasma gondii and Babesia microti in the Blood of Rescued Wild Animals in Gangwon-do (Province), Korea

  • Hong, Sung-Hee;Kim, Hee-Jong;Jeong, Young-Il;Cho, Shin-Hyeong;Lee, Won-Ja;Kim, Jong-Tak;Lee, Sang-Eun
    • Parasites, Hosts and Diseases
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    • v.55 no.2
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    • pp.207-212
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    • 2017
  • Infections of Toxoplasma gondii and Babesia microti are reported in many wild animals worldwide, but information on their incidence and molecular detection in Korean wild fields is limited. In this study, the prevalence of T. gondii and B. microti infection in blood samples of 5 animal species (37 Chinese water deer, 23 raccoon dogs, 6 roe deer, 1 wild boar, and 3 Eurasian badgers) was examined during 2008-2009 in Gangwon-do (Province), the Republic of Korea (=Korea) by using serological and molecular tests. The overall seropositivity of T. gondii was 8.6% (6/70); 10.8% in Chinese water deer, 4.3% in raccoon dogs, and 16.7% in roe deer. PCR revealed only 1 case of T. gondii infection in Chinese water deer, and phylogenic analysis showed that the positive isolate was practically identical to the highly pathogenetic strain type I. In B. microti PCR, the positive rate was 5.7% (4/70), including 2 Chinese water deer and 2 Eurasian badgers. Phylogenetic analysis results of 18S rRNA and the ${\beta}$-tubulin gene showed that all positive isolates were US-type B. microti. To our knowledge, this is the first report of B. microti detected in Chinese water deer and Eurasian badger from Korea. These results indicate a potentially high prevalence of T. gondii and B. microti in wild animals of Gangwon-do, Korea. Furthermore, Chinese water deer might act as a reservoir for parasite infections of domestic animals.

Effects of Microbacterium laevaniformans Levans Molecular Weight on Cytotoxicity

  • Oh, Im-Kyung;Yoo, Sang-Ho;Bae, In-Young;Cha, Jae-Ho;Lee, Hyeon-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.985-990
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    • 2004
  • Levans produced from Microbacterium laevaniformans were isolated, characterized, and fractionated by molecular weight. TLC, HPLC, and GC-MS analyses of the exopolysaccharide showed that it was a fructan-type polymer and was composed of (2,6)- and (2,1)-glycosidic linkages. $^{13}C$-NMR analysis proved that the polysaccharide was mainly a $\beta$-(2,6)-linked levan-type polysaccharide. To investigate the cytotoxicity of the acetone-precipitated levan fractions such as M1, M2, and M3, HepG2, P388D1, U937, SNU-1, and SNUC2A cell lines were screened. Among the cell lines tested, the cytotoxicity of M1- M3 fractions were detected from only SNU-1 and HepG2 cells at the dosage level of $100-800\mu\textrm{g}ml$. The M2 fraction M_r$, 80,000) at 400 $mu{g/ml}$ had the greatest cell growth inhibition (84.6%) on SNU-1, while the M1 $(M_r$, 50,000) at $800\mu\textrm{g}ml$ showed the greatest (46.32%) on HepG2. To obtain more uniform M_r$ fractions of levan, the levan was further fractionated from S1 $(M_r$ 1,000,000) to S5 $(M_r$ 10,000) using gel permeation chromatography. Again, the S1-S5 fractions had strong cytotoxicity on SNU-1 and HepG2 cell lines. The greatest inhibition effects of S4 $(M_r$ 80,000) on SNU-1 and S5 $(M_r$ 10,000) on HepG2 were shown to be 49.5% and 73.0%, respectively. The cytotoxicity of the levan fractions was more effective on SNU-1 than on HepG2. Although the relationship between the Mw and the cytotoxicity was not clear, smaller $M_r$, fractions of levan showed greater growth inhibition effect on the cancer cell lines in general. Therefore, it was indicated that a specific Mw class of levan is responsible for the effective cytotoxicity.

Plasma Levels of Hormones and Metabolites as Affected by the Forages Type in Two Different Types of Crossbred Holstein Cattle

  • Chaiyabutr, N.;Preuksagorn, S.;Komolvanich, S.;Chanpongsang, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.10
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    • pp.1359-1366
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    • 2000
  • An experiment was carried out to study plasma levels of hormones and metabolites of crossbred Holstein cattle during late pregnancy (28 days pre partum), early lactation (30 days post partum), mid-lactation (120 days post partum) and late lactation (210 days post partum). Two breed types of Holstein $Friesian{\times}Red$ Sindhi (50:50 = 50%HF) and Holstein $Friesian{\times}Red$ Sindhi (87.5:12.5 = 87.5%HF) were divided into four groups of four animals each. Two groups of each breed were fed with either rice straw treated with 5% urea or pangola hay (Digitaria decumbens) as the source of roughage throughout the experiments. There were a substantial increases in the mean levels of total triiodothyronine ($T_3$), insulin and glucagon at the onset of lactation, and maintained in a high levels during lactation advance for all groups of experiments. The mean levels of prolactin and thyroxine ($T_4$) were not significantly different among groups of animals, but the plasma cortisol concentration was slightly higher in both groups of 50%HF in comparison with those of 87.5%HF animals. The mean levels of plasma growth hormone (GH) of both groups of 87.5%HF animals feeding on either hay or urea treated rice straw markedly rose in the early period of lactation and markedly reduced in mid- and late lactation. These changes were accompanied with changes of milk yield. In contrast to 50%HF animals, plasma GH levels were considerably higher in the late pregnant period than in the early period of lactation and it remained constant as its value at the early lactation throughout the experimental period. The high levels of both plasma progesterone and estradiol concentration significantly declined after parturition and remained low through lactating period. The plasma glucose level in the 50%HF animals feeding on either hay or urea treated rice straw was higher than the 87.5%HF animals in all periods of experiments. Changes in plasma FFA levels of both types of crossbred animals were depended on the endocrine status during late pregnancy and lactation. The levels of plasma FFA of 50%HF animals were significantly higher (p<0.05) than those of 87.5%HF animals during late pregnancy. Both plasma ${\beta}$-hydroxybutyrate and lactate concentrations were not affected by feeding on either hay or urea treated rice straw during late pregnancy and lactation. These data demonstrate that there were no differences in the physiological performances in the same crossbred animals fed either hay or urea treated rice straw. The 87.5%HF animal has the genetic potential for a high milk yield and homeorhetic adaptation for mammary function differed from 50%HF animals during periods of lactation. Altering lactation persistency in 87.5%HF is regulated mainly by chronically acting growth hormones through the period of lactation.

Safety evaluation of type B transport container for tritium storage vessel (B형 삼중수소 운반용기 안정성 평가)

  • Lee, Min-Soo;Paek, Seung-Woo;Kim, Kwang-Rag;Ahn, Do-Hee;Yim, Sung-Paal;Chung, Hong-Suk;Choi, Heui-Joo;Choi, Jeong-Won;Son, Soon-Hwan;Song, Kyu-Min
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.5 no.2
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    • pp.155-169
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    • 2007
  • A transport container for a 500 kCi tritium storage vessel was developed, which could be used for the transport of metal tritide from Wolsong TRF facility to a disposal site. The structural, thermal, shielding, and confinement analyses were performed for the container in a view of Type B. As a result of structural analysis, the developed container sustained its integrity under normal and accidental conditions. The maximum temperature increase of the inner storage vessel by radiation was evaluated at $134.8^{\circ}C at room temperature. In $800^{\circ}C$ fire test, The thermal barrier of container sustained the inner vessel at $405^{\circ}C after 30 min, which temperature was allowable for the container integrity since maximum design temperature of inner vessel was $550^{\circ}C. In the evaluation of the shielding, the activity of radiation was nearly zero on the outer surface of inner vessel. Consequently the transport container for a 500 kCi tritium was evaluated to pass all the safety tests including accidental condition, so it was concluded that the designed transport container is proper to be used.

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Effect of Enamel Matrix Drivatives application on the expression of PDLs17, PDLs22 of cultured human periodontal ligament cells in vitro (Enamel Matrix Derivatives가 사람 치주인대 세포의 특이유전자인 PDLs17, PDLs22의 발현에 끼치는 효과)

  • Han, Geun-A;Jang, Hyun-seon;Kok, Jung-Ki;Park, Ju-Chol;Kim, Heoung-Jung;Kim, Jung-Gwan;Kim, Byung-Ock
    • Journal of Periodontal and Implant Science
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    • v.34 no.2
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    • pp.333-344
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    • 2004
  • The enamel matrix derivative (EMD) has been recently used in the periodontal regenerative techniques. The present study was established to investigate the influence of EMD on human periodontal ligament cells using expression of mRNA of periodontal ligament specific gene (PDLs)17, PDLs22, type I collagen when EMD applied to periodontal ligament cells. Periodontal ligament cells were obtained from a healthy periodontium and cultured in Dulbecco's modified Eagle's medium (DMEM) plus 10% fetal bovine serum and ${\beta}-glycerophosphate$ with ascorbic acid. Test groups were two; One adds EMD in culture media and another added EMD and Dexamethasone (DEX) in culture media. Positive control group added DEX in culture media, and negative control group adds niether of EMD nor DEX. $Emdogain^{(R)}$ (Biora, Sweden, 30 mg/ml) was diluted by 75 ${\mu}g/ml$ concentration to culture media. For reverse transcription-polymerase chain reaction (RT-PCR), total RNA isolated on days 0, 7, 14 and 21. mRNA of PDLs17 was expressed on days 14 and 21 in EMD or DEX group, and expressed on days 7, 14 and 21 in EMD plus DEX group, the other side, expressed on days 21 in negative control group. mRNA of PDLs22 expressed on days 7, 14 and 21 in EMD group, and expressed on days 14 and 21 in DEX group, and expressed on days 7, 14 and 21 in EMD plus DEX group. Negative control group expressed on days 14 and 21. Type I collagen was expressed on all days and all groups. These results indicate that EMD promotes differentiation of periodontal ligament cells, and this is considered to offer basis that can apply EMD to periodontal tissue regeneration technique.

Effect of Electron-beam Irradiaton on the Artificial Bone Substitutes Composed of Hydroxyapatite and Tricalcium Phosphate Mixtures with Type I Collagen (수산화인회석과 인산삼칼슘 및 1형 콜라젠 혼합골의 전자빔 조사 효과)

  • Park, Jung Min;Kim, Soung Min;Kim, Min Keun;Park, Young Wook;Myoung, Hoon;Lee, Byung Cheol;Lee, Jong Ho;Lee, Suk Keun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.35 no.1
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    • pp.38-50
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    • 2013
  • Purpose: The aim of this study is to evaluate the effect and potential of electron beam (E-beam) irradiation treatment to the synthetic bony mixtures composed of hydroxyapatite (HA; Bongros$^{(R)}$, Bio@ Co., Korea) and tricalcium phosphate (${\beta}$-TCP, Sigma-Aldrich Co., USA), mixed at various ratios and of type I collagen (Rat tail, BD Biosciences Co., Sweden) as an organic matrix. Methods: We used 1.0~2.0 MeV linear accelerator and 2.0 MeV superconductive linear accelerator (power 100 KW, pressure 115 kPa, temperature $-30{\sim}120^{\circ}C$, sensor sensitivity 0.1~1.2 mV/kPa, generating power sensitivity 44.75 mV/kPa, supply voltage $5{\pm}0.25$ V) with different irradiation dose, such as 1, 30 and 60 kGy. Structural changes in this synthetic bone material were studied in vitro, by scanning electron microscopy (SEM), elementary analysis and field emission scanning electron microscope (FE-SEM), attenuated total reflection (ATR), and electron spectroscopy for chemical analysis (ESCA). Results: The large particular size of HA was changed after E-beam irradiation, to which small particle of TCP was engaged with organic collagen components in SEM findings. Conclusion: The important new in vitro data to be applicable as the substitutes of artificial bone materials in dental and medical fields will be able to be summarized.

Effects of Dietary Fructose and Glucose on Hepatic Steatosis and NLRP3 Inflammasome in a Rodent Model of Obesity and Type 2 Diabetes (비만 및 제2형 당뇨병 쥐 모델에서 과당과 포도당의 섭취가 지방간과 NLRP3 염증조절결합체에 미치는 영향)

  • Lee, Hee Jae;Yang, Soo Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.10
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    • pp.1576-1584
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    • 2013
  • This study is carried out to assess the relative effects of different doses of dietary glucose or fructose on non-alcoholic fatty liver disease (NAFLD) and hepatic metaflammation in a rodent model of type 2 diabetes. KK/HlJ male mice were fed experimental diets as follows: 1) control (CON), 2) moderate glucose (MG, 30% of total calories as glucose), 3) high glucose (HG, 60% of total calories as glucose), 4) moderate fructose (MF, 30% of total calories as fructose), and 5) high fructose (HF, 60% of total calories as fructose) for three weeks. Food intake was not affected by treatments. Compared with HF, HG not only increased serum fasting glucose and area under the curve during oral glucose tolerance test, but also decreased the levels of serum insulin and adiponectin. It indicated that glucose control was complicated via high glucose intake. High fructose treatment led to increased triglyceride in the serum and liver. In comparison to HG, high fructose diet activated NOD-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome consisting of apoptosis-associated speck-like protein containing a CARD (ASC), NLRP3 and caspase 1, which increases interleukin (IL)-$1{\beta}$ maturation and secretion. The activation of NLRP3 inflammasome was accompanied by increased levels of tumor necrosis factor alpha (TNF-${\alpha}$) and IL-6. However, the expression of NLRP3 inflammasome components and pro-inflammatory cytokines did not differ between CON and HG. These data suggested that dietary fructose triggers hepatic metaflammation accompanied by NLRP3 inflammasome activation and has deleterious effects on NAFLD.

Establishment of Human Embryonic Stem Cells Derived from Frozen-Thawed Blastocysts (동결-융해된 인간 배반포기 배 유래의 배아 간(幹) 세포 배양)

  • Kim, Eun-Young;Nam, Hwa-Kyung;Lee, Keum-Sil;Park, Sae-Young;Park, Eun-Mi;Yoon, Ji-Yeon;Heo, Young-Tae;Cho, Hyun-Jung;Park, Se-Pill;Chung, Kil-Saeng;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.28 no.1
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    • pp.33-40
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    • 2001
  • Objective: This study was to establish the human embryonic stem (ES) cells derived from frozen-thawed blastocyst stage embryo that were destined to be discarded after five years in routine human IVF-ET program. Methods: Frozen-thawed and survived human blastocysts were treated by immunosurgery, and recovered ICM cells were cultured onto STO feeder cell layer and ICM colony was subcultured by mechanical dissociation into clumps. To identify ES cell, alkaline phosphatase staining and expression of Oct4 in replated ICM colonies were examined. Also, to examine the possibility of ES cell differentiation, retinoic acid (RA), basic fibroblast growth factor (b-FGF), nerve growth factor (NGF) were added in culture medium. In addition, to classify the specific cell type, differentiated cells were stained by indirect immunocytochemistry. Results: One ICM colony recovered from frozen-thawed six blastocysts was subcultured, continuously replated during 40 passage culture duration without differentiation. Subcultured colonies were strong positively stained by alkaline phophatase. When the expression of Oct4 in cultured ES colony was examined, Oct4b type is more clearly indicated than Oct4a one although there was not detected in embryoid body or differentiated cells. In differentiated cardiomyocytes from ES colony, cells were beaten regularly (60 times/min). In differentiated neural cells from ES colony, neurofilament (NF) 200 kDa protein, microtubule associated protein (MAP) 2 and ${\beta}$-tubulin of specific marker in neurons, glial fibrillary acidic protein (GFAP) of specific marker in astrocytes and galactocelebrocide (GalC) of specific marker in oligodendrocytes were confirmed by indirect immunocytochemistry. Also, muscle cells were detected by indirect immunocytochemistry. In addition, ES colonies can be successfully cryopreserved. Conclusion: This study suggested that establishment of human ES cells can be successfully derived from frozen-thawed blastocysts that were destined to be discarded, and obtained specific cell types (cardiomyocytes, neurons and muscle cells) through the in vitro differentiation procedures of ES cells.

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