• Title/Summary/Keyword: isoenzymes

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Effects of Enzyme Inducers and Glutathione on the Embryotoxicity of Cyclophosphamide in Cultured Rat Embryos (효소유도제 및 glutathione이 전배자배양된 랫드태자에서 cyclophosphamide의 독성에 미치는 영향)

  • 한순영;신재호;권석철;강명옥;이유미;김판기;양미라;박귀례;장성재
    • Toxicological Research
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    • v.11 no.1
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    • pp.31-36
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    • 1995
  • Cyclophosphamide (CP) must be enzymatically activated by cytochrome P450(CYP)-linked mixed-function oxidation pathway to be either mutagenic or teratogenic. Influences of alterations in hepatic mixed-function oxidase acitivity and glutathione (GSH) content on the embryotoxicity of CP were studied in rat whole embryo culture system. The embryotoxicity of CP was compared using rat S-9 fraction (S-9) pretreated with chemicals inducing different CYP isozymes, acetone (ACE), Aroclor 1254 (ARO), $\beta$-naphthoflavone (NAF) and phenobarbital (PHE). When 10.5 day embryos were cultured in the immediately centrifuged rat serum for 48 hrs using general gas char{ging schedule, CP$(40{\mu}g/ml)$ with S-9 induced by either NAF or PHE increased the incidence of realformations and significantly decreased embryonic growth compared with the non-induced S-9 group. ACE or ARO induced S-9 group showed no significant difference in embryonic growth. These data suggest that PB and/or NAF inducible CYP isoenzymes are mainly involved in the activation of CP. To examine the effect of GSH on the embryotoxicity of CP, 10.5 day embryos were exposed to CP and S-9 after preincubation with 10 mM of GSH for 3 hrs. In the GSH pretreated group the growth of embryos increased significantly compared with that of the untreated group, suggesting that GSH may protect embryos in culture from some toxic effects of CP.

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Extra-LDH Isoenzyme($LD_6$) in Myocardial Infarction with Serious Course - Two cases report - (예후가 불량한 심근경색환자에 나타난 혈청 LD Isoenzyme의 Extra Band($LD_6$) - 2 증례 보고 -)

  • Kim, Kyung-Dong;Choi, Myung-Sook;Lee, Chae-Hoon;Jeon, Chang-Ho;Kim, Chung-Sook;Kim, Young-Jo
    • Journal of Yeungnam Medical Science
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    • v.5 no.2
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    • pp.247-254
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    • 1988
  • Serum lactate dehydrogenase isoenzymes are separated 5 fractions($LD_1-LD_5$) in normal serum by agrose electrophoresis. An unusual band on the cathodic side of $LD_5$ isoenzyme has been found on lactate dehydrogenase electrophores is of serum, called $LD_6$, and the presence of which signifies a poor prognosis for the patients of myocardial infarction. In recent, we have experienced $LD_6$ in two patients with myocardial infarction. Cardiac arrest was noted within 24 hours after exhibition of $LD_6$ extraband and they did not recover from their illness.

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Diurnal Changes of Chlorophyll Fluorescence and Antioxidative Enzyme Activity of the Leaves from Four Subtropical Plants (아열대성 식물 4종의 엽록소형광과 항산화효소 활성의 일주기적 변화)

  • Oh, Soon-Ja;Goh, Chang-Hyo;Koh, Seok-Chan
    • Journal of Environmental Science International
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    • v.16 no.5
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    • pp.633-640
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    • 2007
  • The diurnal changes of chlorophyll fluorescence and antioxidative enzyme activity were investigated in the leaves from four subtropical plant species (Crinum asiaticum var. japonicum Bak., Osmanthus insularis Koidz., Asplenium antiquum Makino and Chloranthus glaber Makino) under the natural habitats in summer and winter. The intensity of chlorophyll fluorescence was lower in O-, I-, J-, P-steps of O-J-I-P transient in winter than summer, and prominent diurnal change was not found in the fluorescence intensity of four subtropical plant species in winter. The activity and isoenzyme pattern of SOD and catalase did irregularly change seasonally and diurnally in four subtropical plant species. In contrast, the peroxidase activity and isoenzyme pattern was different depending on plant species and growth seasons; The activity increased slightly more in winter than in summer in four subtropical plant species, and several isoenzymes appeared in the leaves from C. asiaticum var japonicum, O. insularis and A. antiquum in winter.

Analysis of Differentially Expressed Proteins in Bovine Longissimus Dorsi and Biceps Femoris Muscles

  • Kim, S.M.;Park, M.Y.;Seo, K.S.;Yoon, D.H.;Lee, H.-G.;Choi, Y.J.;Kim, S.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.10
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    • pp.1496-1502
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    • 2006
  • Skeletal muscle contains slow and fast twitch fibers. These skeletal muscle fibers express type I and type II myosin, respectively, and these myosin isoenzymes have different ATPase activity. The aim of this study was to investigate protein profiles of bovine skeletal muscles by proteomic analysis. Fifty seven spots of distinct proteins were excised and characterized. The expression of sixteen spots was differed in longissimus dorsi muscle with a minimal 2-fold change compared to biceps femoris muscle. The majority of differentially expressed proteins belonged to metabolic regulation-related proteins such as glyceraldehyde 3-phosphate dehydrogenase, triosephosphate isomerase and carbonic anhydrase 3. The real time-PCR assay confirmed an increase or induction of specific genes: RGS12TS isoform, GAPDH, triosephosphate isomerase and carbonic anhydrase. These results suggest that the expression of metabolic proteins is under a specific control system in different bovine skeletal muscle. These observations could have significant implications for understanding the physiological regulation of bovine skeletal muscles.

Covalent Interactions of Reactive Pentachlorophenol Metabolites with Cellular Macromolecules (Pentachlorophenol 대사물과 세포내 거대분자물의 반응에 관한 연구)

  • 정요찬;윤병수;이영순;조명행
    • Toxicological Research
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    • v.13 no.3
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    • pp.257-263
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    • 1997
  • Pentachlorophenol(PCP) which ks widely used in wood preservation, pulp and paper mills, has led to a substantial envirortmental contamination. To get the reliable data for the effective health risk assessment with PCP, covalent binding potential of PCP to cellular macromolecules and glutathione(GSH) was investigated after intraperitoneal administration of $^{14}C-PCP$ to rats. PCP metabolites were able to bind covalently to serum albumin and hepatic protein in a dose- and time-dependent manner. Hepatic protein adducts of PCP metabolites were increased as a function of cytochrome P-450 activities, whereas, albumin adducts significantly decreased. Covalent binding of PCP metabolites with DNA or hemoglobin was not observed. GSH levels in liver tissue decreased over 12hrs, however, the level was recovered after 48hrs. Tetrachloro-1,4-benzoquinone (1,4-TCBQ), one of the most reactive PCP metabolites, conjugated with GSH very rapidly. Base on our results, we could conclude that PCP metabolized to reactive electrophilic metabolites by cytochrome P-450 isoenzymes and conjugated rapidly with neighboring protein or nonprotein sulfhydryl before reacting with DNA or hemoglobin. We propose that albumin adducts and mercapturic acids of PCP metabolites can be used good biomarker of recent PCP exposure.

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Biochemical and molecular characterization of a strain KA/S2 of Acnnthamoebc castellanii isolated from Korean soil (카스텔란니가시아메바(Acanthamoeba castellanii) 한국 토양분리주 KA/S2의 생화학적 및 분자생물학적 특성)

  • 정동일;공현희
    • Parasites, Hosts and Diseases
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    • v.34 no.1
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    • pp.79-86
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    • 1996
  • A strain, KA/S2, isolated from Korean soil and morphologically assigned to Acanthcmoebc cQsteLlcnii, was characterized by isoenzyme analysis , and total proteins profile, End mitochondrial (Mt) DNA restriction fragment length polymorphism (RFLP) , and compared with four reference strains assigned to the species (the authenitic Castellani, Neff, Ma, and Chang strains). It was found that four isoenzyme, total proteins, and Mt DNA RFLP patterns by eight restriction endonucleases of the strain KA/S2 were identical with those of the Neff strain, isolated from soil of California, USA. The Chang strain was unique in its morphology and total protein patterns. Interstrain polymorphisms of isoensyme profiles and Mt DNA RFLP patterns were observed among the Castellani, Neff, Ma, and Chang strains. Mt DNA RFLP was confirmed to be highly appropriate for the strain characterization and identification of Acnnthamoeba spry.

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Dynamic Gene Expression Profiling of Escherichia coli in Carbon Source Transition from Glucose to Acetate

  • Oh Min-Kyu;Cha Mee-Jeong;Lee Sun-Gu;Rohlin Lars;Liao James C.
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.543-549
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    • 2006
  • DNA microarray was used to study the transcription profiling of Escherichia coli adapting to acetate as a sole carbon source. Bacteria grown in glucose minimal media were used as a reference. The dynamic expression levels of 3,497 genes were monitored at seven time points during this adaptation. Among the central metabolic genes, the glycolytic and glucose phosphotransferase genes were repressed as the bacteria entered stationary phase, whereas the glyoxylate pathway, TCA cycle, and gluconeogenic genes were induced. Distinct induction or repression patterns were recognized among different pathway genes. For example, the repression of glycolytic genes and the induction of gluconeogenic ones started immediately after glucose was depleted. On the other hand, the regulation of the pentose phosphate pathway genes and glyoxylate genes gradually responded to the glucose depletion or was more related to growth in acetate. When the whole genome was considered, many of the CRP, FadR, and Cra regulons were immediately responsive to the glucose depletion, whereas the $\sigma^s$, Lrp, and IHF regulons were gradually responsive to the glucose depletion. The expression profiling also provided differential regulations between isoenzymes; for example, malic enzymes A (sfcA) and B (maeB). The expression profiles of three genes were confirmed with RT-PCR.

Purification and characterization of catalase-3 of deinococcus radiophilus

  • Lee, In-Jeong;Lee, young-Nam
    • Journal of Microbiology
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    • v.33 no.3
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    • pp.239-243
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    • 1995
  • Deinococcus radiophilus, an UV resistant bacterium seemed to contain three issoenzymes of catalase. Among them, the samllest and most abundant species in cell-free extract, catalase-3 which also exhibited peroxidase activity was purified to electrophoretic homogeneity (145-fold purification) by chromatographic procedures. Its molecular weight was 155 kDa composed of four 38 kDa subunits. The $K_{m}$ value of catalase-3 for H$\_$2/O$\_$2/ was approximately 0.5 mM. This enzyme showed a typical ferric heme spectrum with maximum absorption at 405 nm. Upon binding to cyanide, the 405 nm peak shifted to 420 nm. Catalase-3 was very sensitive to inhibitors of heme proteins, such as cyanide, azide and hydroxylamine. A ratio of A$\_$405/A$\_$28O/ was 0.5 Catalase-3 was active over a wide range of pH, between pH 7 and 10. The enzyme was rather heat-labile and partially sensitive to edthanol-chloroform treatment, but resistant to 3-amino-1, 2, 4-triazole. Catalase-3 of D. radiophilus, which is a bifunction catalatic peroxidatic enzyme seemed to share certain molecular properties with the typical catalase and the catalase-[roxidase along with its own unique features.

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Acanthamoeba sohi, n. sp., a pathogenic Korean isolate YM-4 from a freshwater fish

  • Im, Kyung-Il;Shin, Ho-Joon
    • Parasites, Hosts and Diseases
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    • v.41 no.4
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    • pp.181-188
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    • 2003
  • A new species of Acanthamoeba was isolated from a freshwater fish in Korea and tentatively named Acanthamoeba sp. YM-4 (Korean isolate YM-4). The trophozoites were $11.0-23.0{\;}{\mu\textrm{m}}$ in length and had hyaline filamentous projections. Cysts were similar to those of A. culbertsoni and A. royreba, which were previously designated as Acanthamoeba group III. Acanthamoeba YM-4 can survive at $40^{\circ}C$, and its generation time was 19.6 hr, which was longer than that of A. culbertsoni. In terms of the in vitro cytotoxicity of lysates, Acanthamoeba YM-4 was weaker than A. culbertsoni, but stronger than A. polyphaga. On the basis of the mortality of experimentally infected mice, Acanthamoeba YM-4 was found to be highly virulent. The isoenzymes profile of Acanthamoeba YM-4 was similar to that of A. royreba. An anti-Acanthamoeba YM-4 monoclonal antibody, McAY7, was found to react only with Acanthamoeba YM-4, and not with A. culbertsoni. Random amplified polymorphic DNA marker analysis and RFLP analysis of mitochondrial DNA and of 18S small subunit ribosomal RNA, placed Acanthamoeba YM-4 in a separate cluster on the basis of phylogenetic distances. Thus the Acanthamoeba Korean isolate YM-4 was identified as a new species, and assigned as Acanthamoeba sohi.

Genetic Variation of Several Isoenzymes in Pinus densiflora for. multicaulis (반송(Pinus densiflora for. multicaulis)의 몇 가지 동위효소(同位酵素)의 유전변이(遺傳變異))

  • Hwang, Jae Woo;Lee, Seok Woo
    • Journal of Korean Society of Forest Science
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    • v.85 no.3
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    • pp.409-415
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    • 1996
  • Isozyme variations in 4 enzyme systems in Pinus densiflora for. multicaulis in 31 individual trees collected throughout the country were studied by starch gel electrophoresis using haploid megagametophyte tissue to compare with those of Pinus densiflora. A minimum of 7 loci were found to code for isozymes of the enzyme systems. No variation was found at locus GOT-A; at the remaining 6 variable loci(GDH-A, GOT-B, GOT-C, IDH-A, LAP-,A, LAP-B) and 2 to 4 alleles were identified. We could not find any marker alleles to distinguish Pinus densiflora for. multicaulis from Pinus densiflora at the isozymes studied here. Allele frequency distributions at each loci were almost all the same as those of P. densiflora. The percentage of polymorphic loci(99% level), the number of alleles per locus, the observed and expected heterozygosities were 85.7, 2.3, 0.165 and 0.186%, respectively. The level of genetic diversity in Pinus densiflora for. multicaulis seemed to be less than that of P. densiflora.

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