• Title/Summary/Keyword: $\beta$-

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Beta-4 Integrin Transfection, Cloning and Functional Assay in Squamous Cell Carcinoma (Beta-4 Integrin 유전자 주입, 클로닝과 편평상피암에서의 Beta-4 Integrin 기능에 관한 연구)

  • Kim Young-Min;Carey Tomas E.
    • Korean Journal of Head & Neck Oncology
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    • v.13 no.2
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    • pp.169-179
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    • 1997
  • 서론 : Laminin의 수용기로 알려진 Integrin $\alpha6\beta4$의 세포내 표현 정도는 편평상피암을 위시한 여러 악성종양의 전이능력 및 예후와 밀접한 상관관계가 없다고 알려져 있다. 이 Integrin은 Laminin과 같은 세포와 리간드와 결합하면 상피세포의 기저막 지주 구조물인 hemidesmosome의 세포체질 요소(cytoskeletal element)와 연관되어 그 결과 세포의 기저막과 세포내 케라틴을 연결하는 역할을 한다. Integrin $\alpha6\beta4$는 구조적으로 다른 많은 integrin들과 달리 $\beta$4의 세포질내 영역(cytoplasmic domain)이 특징적으로 크다. 이 세포질내 영역 $\beta$4 integrin의 기능은 아직 밝혀지지 않고 있으나 아마 세포 성장의 신호전달 및 악성종양의 특징인 침윤 전이에 관련할 것으로 보아지고 있다. 재료 및 방법: 저자들은 우선 $\beta$4 integrin의 wild type s-DNA와 $\beta$4 세포질내 영역(cytoplasmic domain) 및 $\beta$4의 tyrosine 인산화 반응 부위가 각각 결손된 c-DNA를 PCR을 통하여 합성하여 pRc/CMV 벡터에 삽입한 후 원래 $\beta$4 integrin의 발현이 결집된 인간 방광암 세포에 Calcium phosphate precipitation 방법으로 주입(transfection)시켜 형질변환된 세포를 면역형광법, Flow cytometry 및 Immunoprecipitation 방법으로 클로닝하여 wild type $\beta$4-full length(Clone FL), truncated $\beta$4-cytoplasmic domain(C1one CD), 및 mutated $\beta$4-tyrosine phosphorylation site (Clone M)을 얻었다. 암 세포의 부착 및 침투 능력의 기능적 연구로 모노 클로날 항체와 fibronectin, laminin, Matrigel을 단백질 기질로 사용하였으며 결과 비교를 위하여 pRc/CMV 벡터만 주입시켰던 클로운과 방광암 세포주를 $\beta$4 integrin 음성 대조군으로 또한 이 Integrin의 높은 발현을 보이는 두경부 편평상피암 세포주를 양성 대조군으로 이용하였다. 결과 : 세포부착능력에 있어서 온전한 $\beta$4 cytoplasmic domain이 존재하는 클로운이 laminin에 강한 부착능력을 보였으나 fibronectin의 부착정도는 $\beta$4 integrin의 표현정도와 관계없이 모든 클로운에서 비슷하였다. Matrigel을 투과하는 암세포 침윤 능력에서는 $\beta$4 integrin의 표현이 존재하는 클로운들이 투과 능력이 높았으나 세포외 리간드가 없는 control membrane을 사용하였을 때와 비교하여 투과능력의 차이를 보이지 않았다. 결론 : 유전자 주입(transfection) 방법으로 integrin의 다양한 클로운의 합성이 가능하여 이 Integrin의 암 세포의 부착 및 침투 능력에서의 기능을 규명 할 수 있게 한다. $\beta$4 integrin은 편평상피 암세포의 부착에 있어서 세포외 리간드 laminin과 특이 결합하여 부착 능력을 높이는 중요한 역할을 하며 편평상피 암세포의 침투에 있어서는 $\beta$4 integrin의 표현이 침투 능력을 높이는 역할을 하나 이때에는 laminin과 같은 리간드와의 특이 결합에 의존하지는 않는 것으로 사료된다.

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Effect of ${\beta}-Estradiol$ on the Growth of Primary Rabbit Proximal Tubule Cells in Serum-free Medium (${\beta}-Estradiol$이 토끼 근위 세뇨관 상피세포의 성장에 미치는 영향)

  • Park, Sang-Ho;Chung, Joo-Ho;Ko, Kye-Chang;Jung, Jee-Chang
    • The Korean Journal of Pharmacology
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    • v.29 no.1
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    • pp.73-83
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    • 1993
  • In order to examine the effect of ${\beta}-estradiol$ on the cell growth, using a primary rabbit kidney poximal tubule cell culture system. We investigated the effect of ${\beta}-estradiol$ on alpha 1 (IV) collagen and ${\beta}-actin$ mRNA levels from primary rabbit kidney cell cultures, and also the effects of 3 growth factors and ${\beta}-estradiol$ supplementation on the growth of primary rabbit kidney proximal tubule cells in the serum-free medium. 1 nM of ${\beta}-estradiol$ showed a sizable potentiation effect on the growth of the proximal tubule cell in serum-free medium, but higher concentration (> 10 nM) of estradiol indeed inhibited the growth. In the absence of hydrocortisone as a growth supplement in serum-free medium, ${\beta}-estradiol$ caused to potentiate the growth of the cell. In the presence of hydrocortisone, ${\beta}-estradiol$ also potentiated the growth of the proximal tubule cells. According to the Northern analysis, ${\beta}-estradiol$ increased the level of ${\beta}-actin$ mRNA, although mRNA level of the alpha I(IV) collagen was not changed significantly.

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A BIOACTIVITY STUDY OF PORTLAND CEMENT MIXED WITH β-GLYCEROPHOSPHATE ON HUMAN PULP CELL (β-glycerophosphate 혼합시 인간 치수 세포에 대한 Portland cement의 생활성에 관한 연구)

  • Oh, Young-Hwan;Jang, Young-Joo;Cho, Yong-Bum
    • Restorative Dentistry and Endodontics
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    • v.34 no.5
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    • pp.415-423
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    • 2009
  • The purpose of this study is to investigate the response of human pulp cell on Portland cement mixed with $\beta$-glycerophosphate. To investigate the effect of $\beta$-glycerophosphate and/or dexamethasone on human pulp cell, ALP activity on various concentration of $\beta$-glycerophosphate and dexamethasone was measured and mineral nodule of human pulp cell was stained with Alizarin red S. MTS assay and ALP activity of human pulp cell on Portland cement mixed with various concentration of $\beta$-glycerophosphate (10 mM, 100mM, 1M) was measured and the specimens were examined under SEM. Addition of $\beta$-glycerophosphate or dexamethasone alone had no effect however, the addition of 5 mM $\beta$-glycerophosphate and 100 nM dexamethasone had the largest increasement in ALP activity. There was no toxicity in all samples and the data showed that Portland cement mixed with 10 mM $\beta$-glycerophosphate had more increase in ALP activity compared with control. In conclusion, Portland cement mixed with $\beta$-glycerophosphate has no toxicity and promotes differentiation and mineralization of pulp cell compared with additive-free Portland cement. This implicated that application of Portland cement mixed with $\beta$-glycerophosphate might form more reparative dentin and in turn it would bring direct pulp capping to success.

Chlorination of Phenyl Derivatives : Chlorination of ethyl -${\alpha},{\beta}-dichloro-{\beta}$-phenyl propionate under gamma ray irradiation (芳香族 誘導體의 염素化反應 Ethyl-${\alpha},{\beta}-dichloro-{\beta}$-phenyl propionate의 gamma 線 鹽素化反應)

  • Kim, You-Sun;Kim, Ki-Soo
    • Journal of the Korean Chemical Society
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    • v.12 no.2
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    • pp.55-60
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    • 1968
  • Chlorination of aromatic derivatives under UV light and ${\gamma}$-ray irradiation was studied. Ethyl ${\alpha} ,{\beta} -dichloro-{\beta}$-phenyl propionate gave the p-chlorophenyl derivatives when chlorination was done under UV light. The same type of the product was obtained in the reaction where the mole ratio of the ester and chlorine was 1 to 2 and the chlorination was done under ${\gamma}$-ray irradiation. When the mole ratio of the ester and chlorine was 1 to 8, the chlorination reaction under ${\gamma}$-ray irradiation gave a poly-chlorinated derivatives which was identified as a side chain chlorinated p-chlorophenyl derivatives. Ethyl ${\alpha} ,{\beta} -dichloro-{\beta}$-(p-chlorophenyl) propionate gave the same type of the side chain chlorinated p-chloro derivatives by the chlorination under ${\gamma}$-ray irradiation, whereas ethyl ${\alpha} ,{\beta} -dichloro-{\beta}$-(o-chlorophenyl) derivatives gave o,p-dichlorophenyl derivatives. The identifications of the products were based on a radio thin layer chromatography and activation analysis of chlorine contents of product. The chlorination reaction was discussed in regards to the effect of phenyl substituents to the formation of reaction product and the procedures were described.

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A Study on the Binding Characteristics of $\beta$-Cyclodextrin with Benzene and Its Application on the Bioremediation ($\beta$-시클로덱스트린($\beta$-Cyclodextrin)의 결합 특성과 벤젠의 생물학적 분해에의 적용에 대한 연구)

  • 최종규;손현석;조경덕
    • Journal of Environmental Health Sciences
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    • v.28 no.5
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    • pp.65-70
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    • 2002
  • Recently, surfactants were frequently used in order to desorb the hydrophobic organic compounds (HOCs) from soil and to enhance the bioavailability. Among them, -cyclodextrin ($\beta$-CD) is one of those. This study was performed to investigate the binding characteristics between benzene and $\beta$-CD and to examine the bioavailability of benzene. First, we investigated binding characteristics between benzene and $\beta$-CD in water and water/soil system. Then, we examined the effect of $\beta$-CD on the biodegradation of benzene in water and water/soil system. Experimental results on the binding characteristics showed that $\beta$-CD resulted in an efficient complex formation with benzene. As -CD concentration increased, the benzene concentration complexed with $\beta$-CD rapidly increased to 30-40% initial benzene added, and reached the equilibrium. We also investigated the effect of $\beta$-CD on the desorption of benzene from soil in the water/soil system. As $\beta$-CD concentration increased, benzene concentration desorbed into water increased up to 90%. How-ever, in its application to biodegradation of benzene in water and water/soil system, the biodegradation rate of benzene did not improved in the presence of $\beta$-CD compared with in the absense of $\beta$-CD. This result indicated that $\beta$-CD was more preferentially used as a carbon source than benzene. Therefore, for remediation of benzene contaminated soils, $\beta$-CD can be used as a surfactant to desert benzene from soil, and then ex-situ chemical treatment can be applied for the remediation.

The Relationship Between the Expression of Estrogen Receptor ${\beta}$ and Recurrence in Breast Cancer (에스트로겐 수용체 ${\beta}$ 발현과 유방암 재발과의 관련성)

  • Kang, Su-Hwan;Choi, Jung-Eun;Lee, Soo-Jung
    • Journal of Yeungnam Medical Science
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    • v.28 no.2
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    • pp.153-164
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    • 2011
  • Background: It has been reported that estrogen receptor beta ($ER{\beta}$) mRNA expression was down-regulated during carcinogenesis and was inversely related to estrogen receptor alpha ($ER{\alpha}$) expression in breast cancer. The association of $ER{\beta}$ mRNA expression to tamoxifen resistance has also been reported. In this study, the expression of $ER{\alpha}$ and $ER{\beta}$ via immunohistochemistry (IHC) and reverse transcription-polymerase chain reaction (RT-PCR) was prompted, and an attempt was made to find out the relationship between $ER{\beta}$ expression and recurrence in the hormonal therapy group, and between $ER{\beta}$ expression and known prognostic factors. Methods: Tumor specimens were obtained at surgery from 67 female breast cancer patients during the period of September 1995 to December 2000. All the specimens were frozen in liquid nitrogen and kept at $-70^{\circ}C$ until they were used. The medical records were analyzed retrospectively. The expressions of ER were analyzed using IHC and RT-PCR methods. Results: The median follow-up was at 93.0 months (range: 14-157 months). The percentage of $ER{\alpha}+/ER{\beta}+$, $ER{\alpha}+/ER{\beta}-$, $ER{\alpha}-/ER{\beta}+$, and $ER{\alpha}-/ER{\beta}$ group were 35.9% 9.4%, 47.2%, and 7.5%, respectively, in 53 patients with hormonal therapy. $ER{\beta}$ was positive in 42 (82.3%) of 51 ER-positive patients. In the hormonal therapy group, the recurrence rates of each group was 15.8%, 0%, 40.0%, and 0%, respectively. In this group, the $ER{\beta}$ expression tended to recur, but there was no clinical significance (p=0.084). Conclusion: The $ER{\beta}$ expression may be a predictive marker of a poor response to endocrine therapy in breast cancer patients, although this needs to be confirmed in additional studies.

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Increase of Epigallocatechin in Green Tea Extract by Lactic Acid Bacteria Fermentation (젖산균 발효를 통한 녹차 추출물의 Epigallocatechin 함량의 증대)

  • Choi, Chan-Yeong;Park, Eun-Hee;Ju, Yoong-Woon;Kim, Myoung-Dong
    • Microbiology and Biotechnology Letters
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    • v.44 no.1
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    • pp.62-67
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    • 2016
  • Hydrolytic enzyme activities, including those of ${\beta}$-glucosidase, ${\beta}$-glucuronidase, ${\beta}$-xylosidase, ${\beta}$-galactosidase, ${\beta}$-arabinofuranosidase, ${\beta}$-arabinosidase, and ${\beta}$-arabinopyranosidase, which are useful for bioconversion, were explored in lactic acid bacteria isolated from Korean traditional fermented foods. Nine bacterial strains were selected for the fermentation of green tea extract prepared by supercritical fluid extraction. Changes in the concentrations of catechin, epicatechin, epicatechin gallate, epigallocatechin, and epigallocatechin-3-gallate in green tea extract were investigated after fermentation by the selected lactic acid bacteria strains. The strain Leuconostoc mesenteroides MBE1424, which showed the highest ${\beta}$-glucuronidase enzyme activity among the tested bacterial strains, increased the epigallocatechin content of the green tea extract by 60%. In addition, L. mesenteroides MBE1424 was more resistant than the control strain at high temperature and showed a maximum specific growth rate at $40^{\circ}C$. L. mesenteroides MBE1424 was presumed to have an enzyme system containing ${\beta}$-glucuronidase with utility in the bioconversion of green tea extract.

Cloning and Molecular Characterization of ${\beta}$-1,3-Glucan Synthase from Sparassis crispa

  • Yang, Yun Hui;Kang, Hyeon-Woo;Ro, Hyeon-Su
    • Mycobiology
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    • v.42 no.2
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    • pp.167-173
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    • 2014
  • A ${\beta}$-glucan synthase gene was isolated from the genomic DNA of polypore mushroom Sparassis crispa, which reportedly produces unusually high amount of soluble ${\beta}$-1,3-glucan (${\beta}$-glucan). Sequencing and subsequent open reading frame analysis of the isolated gene revealed that the gene (5,502 bp) consisted of 10 exons separated by nine introns. The predicted mRNA encoded a ${\beta}$-glucan synthase protein, consisting of 1,576 amino acid residues. Comparison of the predicted protein sequence with multiple fungal ${\beta}$-glucan synthases estimated that the isolated gene contained a complete N-terminus but was lacking approximately 70 amino acid residues in the C-terminus. Fungal ${\beta}$-glucan synthases are integral membrane proteins, containing the two catalytic and two transmembrane domains. The lacking C-terminal part of S. crispa ${\beta}$-glucan synthase was estimated to include catalytically insignificant transmembrane ${\alpha}$-helices and loops. Sequence analysis of 101 fungal ${\beta}$-glucan synthases, obtained from public databases, revealed that the ${\beta}$-glucan synthases with various fungal origins were categorized into corresponding fungal groups in the classification system. Interestingly, mushrooms belonging to the class Agaricomycetes were found to contain two distinct types (Type I and II) of ${\beta}$-glucan synthases with the type-specific sequence signatures in the loop regions. S. crispa ${\beta}$-glucan synthase in this study belonged to Type II family, meaning Type I ${\beta}$-glucan synthase is expected to be discovered in S. crispa. The high productivity of soluble ${\beta}$-glucan was not explained but detailed biochemical studies on the catalytic loop domain in the S. crispa ${\beta}$-glucan synthase will provide better explanations.

Varietal and Annual Variations of β-Glucan Contents in Korean Barley (Hordeum vulgare L.) and Oat (Avena sativa L.) Cultivars

  • Lee, Mi-Ja;Yoo, Jae-Soo;Kim, Yang-Kil;Park, Jong-Chul;Kim, Tae-Soo;Choi, Jae-Seong;Kim, Kee-Jong;Kim, Hyung-Soon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.3
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    • pp.284-291
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    • 2011
  • Varietal and annual variations in the contents of ${\beta}$-glucan fractions per weight grain samples were examined in sixteen covered and eighteen naked barley and five oat cultivars developed in Korea. Also, the effect of pearling on ${\beta}$-glucan content was investigated. Average contents of total, soluble and insoluble ${\beta}$-glucan fractions were 5.25, 3.72, and 1.53%, respectively, in covered barley, and 5.86, 3.51, and 2.35%, respectively, in naked barley. Soluble ${\beta}$-glucan content was higher in covered barley, though total ${\beta}$-glucan content higher in naked barley. The total and insoluble ${\beta}$-glucan contents were higher in pearled grains. Total ${\beta}$-glucan content was higher in waxy barley than in non-waxy barley. Duwonchapssalbori, a two-rowed and waxy naked barley cultivar, was highest in total, soluble and insoluble ${\beta}$-glucan contents. Highly significant positive correlations were observed between total ${\beta}$-glucan and soluble ${\beta}$-glucan contents both in covered and naked barley. There were significant annual variations in total ${\beta}$-glucan content in barley. Average contents of total, soluble and insoluble ${\beta}$-glucans of oat cultivars were 4.33, 3.44, and 0.89%, respectively. Contents of all fractions of ${\beta}$-glucans were higher in barley than in oat. These results would be useful for the breeding of high ${\beta}$-glucan variety and also for the use barley and oat as valueadded food ingredients.

Linkage Structure Analysis of Barley and Oat $\beta$-Glucans by High Performance Anion Exchange Chromatography

  • Ryu, Je-Hoon;Yoo, Dong-Hyung;Lee, Byung-Hoo;Lee, Su-Yong;Joo, Mi-Hyun;Yoo, Sang-Ho
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.271-274
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    • 2009
  • Cereal $\beta$-glucans, linked essentially by mixed $\beta$-(1,4/1,3) glycosidic bonds, were extracted, purified, and structurally identified. Previously chemical structure of barley $\beta$-glucans was characterized from 3 varieties of 'Gang', 'Ohl', and 'Gwangan', and the (1,4)/(1,3) linkage ratio of the $\beta$-glucans was identical. In this study, $\beta$-glucans from 1 barley ('Chal') and 3 oat ('Ohl', 'Samhan', and 'Donghan') varieties were structurally scrutinized, and the linkage pattern of total 7 cereal $\beta$-glucans was compared. The amount of 2 major 3-O-$\beta$-cellobiosyl-D-glucose (DP3) and 3-O-$\beta$-cellotriosyl-D-glucose (DP4) from barley and oat accounted for only 66.6-73.3 and 68.12-81.89% of water-extractable $\beta$-glucan fractions, and the (1,4)/(1,3) linkage ratios of both barley and oat $\beta$-glucans were within very narrow range of 2.27-2.31 and 2.38-2.39, respectively, among the cultivars tested. Structural difference in the cereal $\beta$-glucans was evident when DP3:DP4 ratio in the $\beta$-glucan structure was compared. As a result, this ratio was significantly greater for barley $\beta$-glucan (2.26-2.74) than for oat (1.54-1.66). Chal-B had the greatest DP3 to DP4 ratio among the samples, which in turn reflected the least amount of (1,4)-linkages.