• Title/Summary/Keyword: BAC

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Expression of Polyhistidine-Containing Fusion Human HepG2 Type Glucose Transport Protein in Spodoptera Cells and Its Purification Using a Metal Affinity Chromatography

  • Lee, Chong-Kee
    • Biomedical Science Letters
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    • v.16 no.3
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    • pp.201-206
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    • 2010
  • In order to develop procedures for the rapid isolation of recombinant sugar transporter in functional form from away from the endogenous insect cell transporter, gene fusion techniques were exploited. Briefly, BamH1-digested human HepG2 type glucose transport protein cDNA was first cloned into a transfer vector pBlueBacHis, containing a tract of six histidine residues. Recombinant baculoviruses including the human cDNA were then generated by allelic exchange following transfection of insect cells with wild-type BaculoGold virus DNA and the recombinant transfer vector. Plaque assay was then performed to obtain and purify recombinant viruses expressing the human transport protein. All the cell samples that had been infected with viruses from the several blue plaques exhibited a positive reaction in the immnuassay, demonstrating expression of the glucose transport protein. In contrast, no color development in the immunoassay was observed for cells infected with the wild-type virus or no virus. Immunoblot analysis showed that a major immunoreactive band of apparent Mr 43,000~44,000 was evident in the lysate from cells infected with the recombinant baculovirus. Following expression of the recombinant fusion protein with the metal-binding domain and enterokinase cleavage site, the fusion protein was recovered by competition with imidizole using immobilized metal charged resin. The leader peptide was then removed from the fusion protein by cleavage with porcine enterokinase. Final separation of the recombinant protein of the interest was achieved by passage over $Ni^{2+}$-charged resin under binding conditions. The expressed transport protein bound cytochalasin B and demonstrated a functional similarity to its human counterpart.

Isolation and Some Properties of Bitter Taste Compounds from Cultured Oyster, Crassostrea gigas (양식산 굴로부터 쓴맛 성분의 분리 및 성질)

  • LEE Jong-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.1
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    • pp.98-104
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    • 1995
  • Five bitter taste compounds (OY-22, OY-23, OY-24, OY-25 and OY-26) were firstly isolated from cultured oyster (Crassostrea gigas) at Gamak Bay, Sourthern coast of Korea, between November, 1989 and January, 1990, and smoked-canned oyster, which were produced by the same oysters. They were presumed as cyclic peptides composed with 6 or 7 amino acids, including sulfur on the basis of NMR and MS spectra. Val and Leu in OY-24, leu and lie in OY-25 and tow leucines in OY-26 were detected from those each compounds, seperately, by the amino acid analysis. Another amino acids were thought as non-constitutional amino acids. They showed non-toxic to mice $(100{\mu}g/20g\;mice i.p.)$ and non actibacterial artivities to Asp. niger and B. subtilis $(10{\mu}g/disk)$. The chemical structures and other biological activities of them are now in studying.

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Production of the BmCecB1 antimicrobial peptide in transgenic silkworm

  • Kim, Seong Wan;Kim, Seong Ryul;Park, Seung Won;Choi, Kwang Ho;Goo, Tae Won
    • International Journal of Industrial Entomology and Biomaterials
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    • v.31 no.2
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    • pp.85-89
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    • 2015
  • This peptide has antibacterial activity against several Gram-positive and Gram-negative bacteria. Bombyx mori cecropinB1(BmCecB1) is antimicrobial peptides from Bombyx mori and belongs to cecropin family. Antimicrobial peptides are important components of the innate immune systems in all living organism. To produce the BmCecB1 antimicrobial peptide, we constructed transgenic silkworm that expressed BmCecB1 gene under the control BmA3 promoter using piggyBac vector. The use of the 3xP3-driven EGFP cDNA as a marker allowed us to rapidly distinguish transgenic silkworm. Mixtures of the donor vector and helper vector were micro-injected into 600 eggs of bivoltin silkworms, Baegokjam. In total, 49 larvae (G0) were hatched and allowed to develop into moths. The resulting G1 generation consisted of 22 broods, and we selected 2 broods containing at least 1 EGFP-positive embryo. The rate of successful transgenesis for the G1 broods was 9%. We identified 9 EGFP-positive G1 moths and these were backcrossed with wild-type moths. With the aim of identifying a BmCecB1 as antimicrobial peptide, we investigated the Radical diffusion Assay (RDA) and then demonstrated that BmCecB1 possesses high antibacterial activities against Gram-negative bacteria.

Well Differentiated Adenosquamous Carcinoma of Lung Mimicking Benign Lesions in Fine Needle Aspiration Cytology - Report of a Case - (세침흡인 세포검사에서 양성 질환을 닮은 폐의 고분화 샘편평 암종 - 1예 보고 -)

  • Bae, Jong-Yup;Oh, Hoon-Kyu;Park, Jae-Bok
    • The Korean Journal of Cytopathology
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    • v.15 no.2
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    • pp.101-105
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    • 2004
  • Fine needle aspiration (FNA) cytological examination is an appropriate method for the evaluation of pulmonary nodules. In major types of lung cancer, Its diagnostic accuracy is quite high. However, it is sometimes difficult, using this technique, to differentiate between some unusual phenotypes including adenosquamous carcinoma, bronchioloalveolar carcinoma (BAC), neuroendocrine tumor, mucoepidermoid carcinoma, and sclerosing hemangioma. Here, we present a case involving extremely well differentiated adenosquamous carcinoma, mimicking benign lesions, such as pulmonary scar and adenomatoid malformation with squamous metaplasia. The patient was a 68-year-old man presenting with a solitary pulmonary nodule$(1.6\times1.6cm)$, which was incidentally found at the periphery of the right lower lobe. FNA revealed some clusters of glandular cells with minimal atypia, in addition to squamous cells at a nearly full maturational state. Histological examination verified the cytological diagnosis on a lobectomy specimen. The tumor exhibited a well differentiated adenocarcinoma component, mimicking the bronchioles in scarred lung tissue. and a well differentiated squamous cell carcinoma component, mimucking the squamous cell nests of adenoacanthoma, in the other organs. In the present case, the possibility of adenosquamous carcinoma should have been considered if squamous cells were seen in the FNA from the peripheral pulmonary nodule, even though they appeared to be benign.

Effect of Distribution System Materials and Water Quality on Heterotrophic Plate Counts and Biofilm Proliferation

  • CHANG , YOUNG-CHEOL;JUNG, KWEON
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1114-1119
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    • 2004
  • The biofilms on pipe walls in water distribution systems are of interest since they can lead to chlorine demand, coliform growth, pipe corrosion, and water taste and odor problems. As such, the study described in this paper is part of an AWWARF and Tampa Bay Water tailored collaboration project to determine the effect of blending different source waters on the water quality in various distribution systems. The project was based on 18 independent pilot distribution systems (PDS), each being fed by a different water blend (7 finished waters blended in different proportions). The source waters compared were groundwater, surface water, and brackish water, which were treated in a variety of pilot distribution systems, including reverse osmosis (RO) (desalination), both membrane and chemical softening, and ozonation-biological activated carbon (BAC), resulting in a total of 7 different finished waters. The observations from this study consistently demonstrated that unlined ductile iron was more heavily colonized by a biomass than galvanized steel, lined ductile iron, and PVC (in that order) and that the fixed biomass accumulation was more influenced by the nature of the supporting material than by the water quality (including the secondary residual levels). However, although the bulk liquid water cultivable bacterial counts (i.e. heterotrophic plate counts or HPCs) did not increase with a greater biofilm accumulation, the results also suggested that high HPCs corresponded to a low disinfectant residual more than a high biofilm inventory. Furthermore, temperature was found to affect the biofilms, plus the AOC was important when the residual was between 0.6 and 2.0 mg $Cl_2/l$. An additional aspect of the current study was that the potential of the exoproteolytic activity (PEPA) technique was used along with a traditional so-called destructive technique in which the biofilm was scrapped off the coupon surface, resuspended, and cultivated on an R2A agar. Both techniques indicated similar trends and relative comparisons among the PDSs, yet the culturable biofilm values for the traditional method were several orders of magnitude lower than the PEPA values.

Bombyx mori Nucleopolyhedrovirus Bacmid Enabling Rapid Generation of Recombinant Virus by In Vitro Transposition

  • Tao, Xue Ying;Choi, Jae Young;Kim, Yang-Su;Lee, Seok Hee;An, Saes Byeol;Pang, Ying;Kim, Jong Hoon;Kim, Woo Jin;Je, Yeon Ho
    • Journal of Microbiology and Biotechnology
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    • v.25 no.3
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    • pp.386-392
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    • 2015
  • A novel recombinant bacmid, bEasyBm, that enables the easy and fast generation of pure recombinant baculovirus without any purification step was constructed. In bEasyBm, attR recombination sites were introduced to facilitate the generation of a recombinant viral genome by in vitro transposition. Moreover, the extracellular RNase gene from Bacillus amyloliquefaciens, barnase, was expressed under the control of the Cotesia plutellae bracovirus early promoter to negatively select against the nonrecombinant background. The bEasyBm bacmid could only replicate in host insect cells when the barnase gene was replaced with the gene of interest by in vitro transposition. When bEasyBm was transposed with pDualBac-EGFP, the resulting recombinant virus, EasyBm-EGFP, showed high levels of EGFP expression efficiency compared with that of non-purified recombinant virus BmGOZA-EGFP, which was constructed using the bBmGOZA system. In addition, nonrecombinant backgrounds were not detected in unpurified EasyBm-EGFP stocks. Based on these results, a high-throughput system for the generation of multiple recombinant viruses at a time was established.

Effects of Hyperbaric Oxygen Intervention on Alcohol-Induced Oxidative Stress and Hangover in Hyperbaric Oxygen Chamber (고압산소챔버 중재가 알코올에 의한 산화적 스트레스 및 숙취 해소에 미치는 영향)

  • Kwon, Hyeok Chan;Park, Soon Won;Cho, Hyun Jeong;Kang, Ji Hyun;Lee, Hyun Ju;Tae, Ki Sik
    • Journal of Biomedical Engineering Research
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    • v.39 no.5
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    • pp.175-182
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    • 2018
  • The purpose of this study is to evaluate the reliability and effectiveness of hyperbaric oxygen chamber on alcohol-induced oxidative stress and hangover. In order to evaluate them, 20 healthy adults were tested for blood and hangover stress tests. When an exponential change was examined after 3 interventions (hangover drink, hyperbaric oxygen chamber, and normal chamber as placebo effect) of 1 hour, the heart rate of subjects experiencing a hyperbaric oxygen chamber showed a statistically significant decrease (p < 0.001). The tests of blood alcohol concentration (BAC) as for exquisite hematology analysis, glucose, creatinine, and AST (aspartate aminotransferase) as well as ALT (alanine aminotransferase) used as liver-damage indicator show that the hyperbaric chamber has the effective effect. In the test of lactic acid, CRP (c-reactive protein), cortisol, and creatinine, the hyperbaric chamber shows much more excellent effect than the hangover drink and normal chamber as control groups. Further studies on hyperbaric oxygen chamber intervention including this study could be very helpful for improving lives of both the patients and healthy people.

Quantitative Analysis of Human- and Cow-Specific 16S rRNA Gene Markers for Assessment of Fecal Pollution in River Waters by Real-Time PCR

  • Jeong, Ju-Yong;Park, Hee-Deung;Lee, Kyong-Hee;Hwang, Jae-Hong;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.245-253
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    • 2010
  • The base sequences representing human- and cow-specific 168 rRNA gene markers identified in a T-RFLP analysis were recovered from clone libraries. The human- and cow-specific primers were designed from these sequences and their specificities were analyzed with fecal DNAs from human, cow, and pig. The AllBac primer set showed positive results for all human, cow, and pig samples, whereas the human-specific primer set showed positive result only for the human sample but not for the cow or pig samples. Likewise, the cow-specific primer set showed positive results only for the cow sample but not for the human or pig samples. Real-time PCR assay with these primers was developed for the identification and quantification of fecal pollution in the river water. The human- and cow-specific markers were detected in the order of 9 $\log_{10}$ copies per gram wet feces, which were two orders of magnitude lower than those of total Bacteroidales. For the river water samples, the human-specific marker was detected in $1.7-6.2\;\log_{10}$ copies/100 ml water, which was 2.4-4.9 orders of magnitude lower than those of total Bacteroidales. There was no significant correlation between total Bacteroidales and conventional fecal indicators, but there was a high correlation between Bacteroidales and the human-specific marker. This assay could reliably identify and quantify the fecal pollution sources, enabling effective measures in the watersheds and facilitating water quality management.

Construction of Deletion Map of 16q by LOH Analysis from HCC Patients and Physical Map on 16q 23.3 - 24.1 Region

  • Chung, Jiyeol;Choi, Nae Yun;Shim, Myoung Sup;Choi, Dong Wook;Kang, Hyen Sam;Kim, Chang Min;Kim, Ung Jin;Park, Sun Hwa;Kim, Hyeon;Lee, Byeong Jae
    • Genomics & Informatics
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    • v.1 no.2
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    • pp.101-107
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    • 2003
  • Loss of heterozygosity (LOH) has been used to detect deleted regions of a specific chromosome in cancer cells. LOH on chromosome 16q has been reported to occur frequently in progressed hepatocellular carcinoma (HCC). Liver tissues from 37 Korean HCC patients were analyzed for LOH by using 25 polymorphic microsatellite markers distributed along 16q. Out of the 37 HCC patients studied, 21 patients (56.8%) showed LOH in various regions of 16q with at least one polymorphic marker. Puring the analysis of these 21 LOH cases, 6 patients showed interstitial LOHs in which the boundary of the LOH region was defined. With two rounds of LOH analysis, five commonly occurring interstitial LOH regions were identified; 16q21-22.1, 16q22.2 - 22.3, 16q22.3, 16q23.2 and 16q23.3 - 24.1. Among the five LOH regions the 16q23.3 - 24.1 region has been reported to be related with chromosome instability. A complete physical map, which covers the 3.2 Mb region of 16q23.3 - 24.1 (D16S402 and D16S486), was constructed to identify novel candidate tumor suppressor genes. We provide the minimally tiling path map consisting of 28 BAC clones. There was one gap between NT_10422.11 and NT_019609.9 of the human genome sequence contig (NCBI sequence build 33, April 29, 2003). This gap can be filled by sequencing the R-1425M20 clone which bridges these sequence contigs.

Aspect of Blood Pressure after Oral Administration of Alcohol : Effect of Dissolved Oxygen on Blood Pressure (알코올 투여에 따른 혈압의 변화 양상 : 용존산소량에 따른 혈압 변화)

  • Song, Byung-Jeong;Lee, Byung-Yo;Chae, Jung-Woo;An, Jung-Hwa;Kwon, Kwang-Il
    • Korean Journal of Clinical Pharmacy
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    • v.22 no.3
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    • pp.228-233
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    • 2012
  • Purpose: The purpose of this study was to elucidate the effect of dissolved oxygen in alcohol to blood pressure of healthy persons. Methods: Subjects (n=30) were randomized in a double blind crossover study to receive 120 mL, 240 mL, 360 mL of alcohol (Korean spirit, 19.59 v/v%, dissolved oxygen is 8 ppm and 20 ppm). Blood alcohol concentration (BAC) and blood pressure were measured applying Lion SD-400 Alcolmeter$^{(R)}$ Breathalyser and Tensoval duo control. Pharmacokinetic parameters ($C_{max}$, $T_{max}$, $AUC_{last}$, $K_{el}$, $V_d$, Clearance) were calculated using Winnonlin$^{(R)}$ program. The difference of parameters and values were analysed by student t-test using Microsoft$^{(R)}$ Excel program. Results: The $AUC_{last}$ values of 8 ppm group and 20 ppm group in 240 ml administration were $6.15{\pm}2.60cg{\cdot}min/ml$, $5.33{\pm}1.84cg{\cdot}min/ml$ (p<0.05) and those in 360 mL were $11.93{\pm}5.70cg{\cdot}min/ml$, $10.33{\pm}4.60cg{\cdot}min/ml$ (p < 0.01), respectively. Thus, the $AUC_{last}$ was significantly decreased. On the other hands, there was a significant change in systolic blood pressure (SBP) after alcohol administration. All measured value after 360 mL of alcohol administration was significantly decreased (p < 0.01). Conclusions: The dissolved oxygen in alcoholic beverage has no effect on blood pressures but the alcohol administration has an effect on blood pressure. Thus, SBP can be used as a biomarker of alcohol administration and utilized in PK/PD modeling of alcohol.