• Title/Summary/Keyword: Molecular Characteristics

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Proteomic Analysis of the Triglyceride-Rich Lipoprotein-Laden Foam Cells

  • Lu, Yanjun;Guo, Jianli;Di, Yong;Zong, Yiqiang;Qu, Shen;Tian, Jun
    • Molecules and Cells
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    • v.28 no.3
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    • pp.175-181
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    • 2009
  • In hypertriglyceridaemic individuals, atherosclerogenesis is associated with the increased concentrations of very low density lipoprotein (VLDL) and VLDL-associated remnant particles. In vitro studies have suggested that VLDL induces foam cells formation. To reveal the changes of the proteins expression in the process of foam cells formation induced by VLDL, we performed a proteomic analysis of the foam cells based on the stimulation of differentiated THP-1 cells with VLDL. Using two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis, 14 differentially expressed proteins, containing 8 up-regulated proteins and 6 down-regulated proteins were identified. The proteins are involved in energy metabolism, oxidative stress, cell growth, differentiation and apoptosis, such as adipose differentiation-related protein (ADRP), enolase, S100A11, heat shock protein 27 and so on. In addition, the expression of some selected proteins was confirmed by Western blot and RT-PCR analysis. The results suggest that VLDL not only induces lipid accumulation, but also brings about foam cells diverse characteristics by altering the expression of various proteins.

Identification of strain harboring both aac(6')-Ib and aac(6')-Ib-cr variant simultaneously in Escherichia coli and Klebsiella pneumoniae

  • Kim, Yun-Tae;Jang, Ji-Hyun;Kim, Hyun-Chul;Kim, Hyo-Gyeong;Lee, Kyoung-Ryul;Park, Kyung-Sun;Lee, Hee-Joo;Kim, Young-Jin
    • BMB Reports
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    • v.44 no.4
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    • pp.262-266
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    • 2011
  • The aac(6')-Ib gene is the most prevalent gene that encodes aminoglycoside-modifying enzymes and confers resistance to tobramycin, kanamycin, and amikacin. The aac(6')-Ib-cr variant gene can induce resistance against aminoglycoside and fluoroquinolone simultaneously. Two main methods, sequence analysis and the restriction enzyme method, can detect the aac(6')-Ib-cr variant in clinical strains. We collected the 85 strains that were believed to be aac(6')-Ib positive from clinical isolates. Among them, 38 strains were the wild-type; the remaining 47 strains were the aac(6')-Ib-cr variant. Of these 47 strains, 19 simultaneously harbored aac(6')-Ib and aac(6')-Ib-cr. Our study aims to report the characteristics of the 19 strains that simultaneously harbored both genes. This study is the first investigation published in Korea of strains that included both aac(6')-Ib and aac(6')-Ib-cr variant.

Bacterial Fruit Rot of Apricot Caused by Burkholderia cepacia in China

  • Fang, Yuan;Li, Bin;Wang, Fang;Liu, Baoping;Wu, Zhiyi;Su, Ting;Qiu, Wen;Xie, Guanlin
    • The Plant Pathology Journal
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    • v.25 no.4
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    • pp.429-432
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    • 2009
  • An unreported disease of apricot was observed in orchards in Zhejiang province, China. Symptoms started as water soaked lesions on the fruit surface. Later, water-soaked areas developed and spread to the entire fruit, resulting in soft rot of the whole fruit. The causal organism isolated from symptomatic fruits was identified as Burkholderia cepacia based on its biochemical and physiological characteristics and confirmed by the cellular fatty acid composition and Biolog data as well as 16S rRNA gene sequence analysis. The bacterial isolates caused similar symptoms when inoculated onto fruits of apricot. In addition, European plum, Japanese plum, nectarine and kiwifruit were susceptible to the B. cepacia pathogen. However, the B. cepacia pathogen failed to cause any visible symptoms when it was inoculated onto 16 other fruits. This is the first report of a bacterial disease of apricot caused by B. cepacia in China.

Isolation, identification, and culture conditions of the strain producing antibacterial antibiotic (항세균성 항생물질을 생산하는 미생물의 분리, 동정 및 배양조건)

  • Yoo, Jae-Hong;Yoon, Sang-Hong;Koo, Bon-Sung;Yeo, Yun-Soo;Park, In-Cheol;Lee, Byung-Moo;Ryu, Jin-Chang
    • The Korean Journal of Pesticide Science
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    • v.3 no.2
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    • pp.1-7
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    • 1999
  • The strain with antibacterial activity was isolated among soil samples collected in Suwon area. The isolated strain was identified as Bacillus sp. YR-1 with respect to its morphological, cultural, and physiological characteristics. Optimal medium for the highest production of antibiotic was composed of sucrose 2.0%(w/v), peptone 2.0%(w/v) and NaCl 0.1%(w/v). The maximum production of antibiotic was shown at $35^{\circ}C$ for 48 hours with the initial pH 7.0.

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Molecular Characterization of Protease Producing Idiomarina Species Isolated from Peruvian Saline Environments

  • Flores-Fernandez, Carol N.;Chavez-Hidalgo, Elizabeth;Santos, Marco;Zavaleta, Amparo I.;Arahal, David R.
    • Microbiology and Biotechnology Letters
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    • v.47 no.3
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    • pp.401-411
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    • 2019
  • All Idiomarina species are isolated from saline environments; microorganisms in such extreme habitats develop metabolic adaptations and can produce compounds such as proteases with an industrial potential. ARDRA and 16S rRNA gene sequencing are established methods for performing phylogenetic analysis and taxonomic identification. However, 16S-23S ITS is more variable than the 16S rRNA gene within a genus, and is therefore, used as a marker to achieve a more precise identification. In this study, ten protease producing Idiomarina strains isolated from the Peruvian salterns were characterized using biochemical and molecular methods to determine their bacterial diversity and industrial potential. In addition, comparison between the length and nucleotide sequences of a 16S-23S ITS region allowed us to assess the inter and intraspecies variability. Based on the 16S rRNA gene, two species of Idiomarina were identified (I. zobellii and I. fontislapidosi). However, biochemical tests revealed that there were differences between the strains of the same species. Moreover, it was found that the ITS contains two tRNA genes, $tRNA^{Ile(GAT)}$ and $tRNA^{Ala(TGC)}$, which are separated by an ISR of a variable size between strains of I. zobellii. In one strain of I. zobellii (PM21), we found nonconserved nucleotides that were previously not reported in the $tRNA^{Ala}$ gene sequences of Idiomarina spp. Thus, based on the biochemical and molecular characteristics, we can conclude that protease producing Idiomarina strains have industrial potential; only two I. zobellii strains (PM48 and PM72) exhibited the same properties. The differences between the other strains could be explained by the presence of subspecies.

Characterization of Dissolved Organics Based on Their Origins (상수 원수에 따른 용존 유기물의 특성 평가)

  • 허준무;박종안;장봉기;이종화
    • Journal of Environmental Science International
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    • v.8 no.3
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    • pp.337-347
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    • 1999
  • This study was carried out to evaluate the characteristics of dissolved organics based on their origins, which were divided into two categories. The first group consisted of river, lake and secondary sewage treatment effluent, which were chosen as representative of their origins. The second group were artificial samples which were made of AHA(Aldrich humic acids) and WHA(Wako humic acids). Physicochemical characteristics, biological degradability and THMEP(trihalomethane formation potential) of the samples were analysed based on the AMWD(apparent molecular weight distribution). Large portion of dissolved organic carbon(DOC) in the river and lake samples was comprised of LMW(low molecular weight), which that of AHA and WHA was HMW(high molecular weight). The DOC of the lake was evenly distributed in the all range of molecular weight. The river, lake and secondary treated effluent have lower ultraviolet(UV) absorbance at 254nm, and have a higher amount of humic acids. Higher absorbance of humic acids means that aliphatic bond and benzenoid type components that absorb UV light were contained in these kind of humic acids. It was expected that lake sample was the most biodegradable in the different samples investigated, and in order of secondary sewage treatment effluent, river, WHA and AHA based on the result of determination of specific ultraviolet absorbance(SUVA). Biodegradability showed similar result except for AHA, while dissolved organics in the range of LMW decreased during the biodegradability test, and on the contrary those of HMW increased. Production of the SMPs(soluble micobial products) was observed during humicfication of dissolved organics and the SMPs were higher production of the SMPs. THM formation was high in the samples containing HMW and similar tendency was shown in the THMEP(trihalomethane formation potential), except for WHA.

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Isolation from Gloydius blomhoffii siniticus Venom of a Fibrin(ogen)olytic Enzyme Consisting of Two Heterogenous Polypeptides

  • Choi, Suk-Ho;Lee, Seung-Bae
    • Journal of Pharmacopuncture
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    • v.16 no.2
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    • pp.46-54
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    • 2013
  • Objective: This study was undertaken to isolate a fibrin(ogen)olytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate the enzymatic characteristics and hemorrhagic activity of the isolated enzyme as a potential pharmacopuncture agent. Methods: The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were determined by using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined. Results: The fibrin(ogen)olytic enzyme with the molecular weight of 27 kDa (FE-27kDa) isolated from G. b. siniticus venom consisted of two heterogenous disulfide bond-linked polypeptides with the molecular weights of 15 kDa and 18 kDa. When more than $20{\mu}g$ of FE-27kDa was applied on the fibrin plate, fibrinolysis zone was formed as indicating its fibrinolytic activity. The fibrinolytic activity was inhibited completely by phenylmethanesulfonylfluoride (PMSF) and ethylenediaminetetraacetic acid (EDTA) and partially by thiothreitol and cysteine. Metal ions such as $Hg^{2+}$ and $Fe^{2+}$ inhibited the fibrinolytic activity completely, but $Mn^{2+}$ did not. FE-27kDa preferentially hydrolyzed ${\alpha}$-chain of fibrinogen and slowly hydrolyzed ${\beta}$-chain, but did not hydrolyze ${\gamma}$-chain. High-molecular-weight polypeptides of gelatin were hydrolyzed partially into polypeptides with molecular weights of more than 45 kDa. A dosage of more than $10{\mu}g$ of FE-27kDa per mouse was required to induce hemorrhage beneath the skin. Conclusion: FE-27kDa was a serine proteinase consisting of two heterogeneous polypeptides, hydrolyzed fibrin, fibrinogen, and gelatin, and caused hemorrhage beneath the skin of mouse. This study suggests that the potential of FE-27kDa as pharmacopuncture agent should be limited due to low fibrinolytic activity and a possible side effect of hemorrhage.

Relationship between Telomerase Activity and Expression of Caspase-3 in Colorectal Cancer

  • Lee Kyung Eun;Kim Na Young;Hong Young Seoub;Park Ki Jae;Choi Hong Jo;Roh Young Hoon;Roh Mee Sook
    • Biomedical Science Letters
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    • v.11 no.4
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    • pp.481-486
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    • 2005
  • This study was performed to define roles of telomerase and apoptosis and their relationships with clinicopathologic characteristics in colorectal cancers. We performed TRAP (Telomeric Repeat Amplification Protoco1)-ELISA assay for telomerase activity and immunohistochemistry of active caspase-3 expression for apoptosis in 35 colorectal cancers. Increased telomerase activity was detected in $71.4\%$ (25/35) and average apoptotic index was 14.6 per 1000 tumor cells. Telomerase activity and caspase 3 expression had no significant association with clinicopathological characteristics, however, increased telomerase activity was more frequently found in progressed colorectal cancers. Although there is no definitive relation, low apoptotic index group was more frequent in cases with increased telomerase activity. These date indicate that telomerase might be involved in progression of colorectal cancers. We suggest that there is a need for further study to define the relationship between telomerase and apoptosis in colorectal cancers.

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A Novel Glycosyl Hydrolase Family 16 β-Agarase from the Agar-Utilizing Marine Bacterium Gilvimarinus agarilyticus JEA5: the First Molecular and Biochemical Characterization of Agarase in Genus Gilvimarinus

  • Lee, Youngdeuk;Jo, Eunyoung;Lee, Yeon-Ju;Hettiarachchi, Sachithra Amarin;Park, Gun-Hoo;Lee, Su-Jin;Heo, Soo-Jin;Kang, Do-Hyung;Oh, Chulhong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.776-783
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    • 2018
  • The agarase gene gaa16a was identified from a draft genome sequence of Gilvimarinus agarilyticus JEA5, an agar-utilizing marine bacterium. Recently, three agarase-producing bacteria, G. chinensis, G. polysaccharolyticus, and G. agarilyticus, in the genus Gilvimarinus were reported. However, there have been no reports of the molecular characteristics and biochemical properties of these agarases. In this study, we analyzed the molecular characteristics and biochemical properties of agarases in Gilvimarinus. Gaa16A comprised a 1,323-bp open reading frame encoding 441 amino acids. The predicted molecular mass and isoelectric point were 49 kDa and 4.9, respectively. The amino acid sequence of Gaa16A showed features typical of glycosyl hydrolase family 16 (GH16) ${\beta}$-agarases, including a GH16 domain, carbohydrate-binding region (RICIN domain), and signal peptide. Recombinant Gaa16A (excluding the signal peptide and carbohydrate-binding region, rGaa16A) was expressed as a fused protein with maltose-binding protein at its N-terminus in Escherichia coli. rGaa16A had maximum activity at $55^{\circ}C$ and pH 7.0 and 103 U/mg of specific activity in the presence of 2.5 mM $CaCl_2$. The enzyme hydrolyzed agarose to yield neoagarotetraose as the main product. This enzyme may be useful for industrial production of functional neoagaro-oligosaccharides.

Taxonomic status of three taxa of Elsholtzia (E. hallasanensis, E. springia, and E. splendens var. fasciflora) (Lamiaceae) based on molecular data

  • Lee, Chang Shook;Hwang, Kung Ae;Kim, Jin Ok;Suh, Hyoung Min;Lee, Nam Sook
    • Korean Journal of Plant Taxonomy
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    • v.41 no.3
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    • pp.259-266
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    • 2011
  • Elsholtzia hallasanensis, E. springia, and E. splendens var. fasciflora (Lamiaceae) were reported recently as new species or new varieties of E. splendens according to their morphological characteristics. To reappraise the taxonomic status of these additional taxa and to determine the relationships between all Korean Elsholtzia taxa except E. saxatilis, which is distributed in North Korea, molecular studies based on the nrDNA (ITS) and cpDNA (rpl16, and trnH-psbA) sequences of seven taxa of Elsholtzia and one outgroup were carried out. The molecular data support that E. angustifolia and E. minima are distinct species from E. splendens and E. ciliata, respectively, because they have several private marker genes and show monophyly. The molecular data also support that E. splendens has a very close taxonomic relationship with both E. hallasanensis and E. springia. We found that E. splendens var. fasciflora, with multiple inflorescence, was based on several private marker genes and on the monophyly of its trees, suggesting that it can be considered as a variety. Elsholtzia springia, with the same sequences and the same morphological characteristics with E. hallasanensis after transplanting, should be treated as a synonym of E. hallasanensis. Moreover, we consider the taxonomic status of E. hallasanensis as E. splendens var. hallasanensis (Y. Lee) N.S. Lee & C.S. Lee, stat. nov.