• Title/Summary/Keyword: Isoenzyme

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Bacterial endosymbiosis within the cytoplasm of Acanthamoeba Lwnunensis isolated from a contact lens storage case (콘택트렌즈 보존 용기 유래 Acnnthamoebc lugdunensis을 KA/LS주의 내공생세균)

  • 정동일;공현희
    • Parasites, Hosts and Diseases
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    • v.35 no.2
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    • pp.127-134
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    • 1997
  • Transmission electron microscopy of an ArGnthnmoebo isolate (KA/LS) from a contact lens case revealed bacterial endosymbionts within cytoplasm of the amoebae. The Acnnthamoebn isolate belonged to the morphological group ll. Based on the polymerase chain reaction (PCR) - restriction fragment leilgth polymorphism (RFLP) of 185 ribosomal RNA coding DNA (rDNA) , the isolate was identified as A. Iwnunensis. Strain typing by isoenzyme analysis using isorlectric focusing (IEF) and mitochondrial (Ent) DNA RFLP revealed that the isolate was closely related with KA/Ll , the most predominant type of isolates from contact lens storage casas, KA/E2, a clinical isolate, KA/W4, previou:fly reported to host endosymbionts. and L3a strains of A. Iwnunensis. The endosymbionts were similar to those of KA/W4 in a.jpects that they were randomly distributed in both trophozoites and cysts, and were rod-shaped bacteri3 measuring approximately 1.38 x 0.50 ㎛. But the number of endosymbionts per amoeba was significantly lower than that of KA/W4. They were neither limited by phagosomal membranes nor included in lacunae- like stnlcture.

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Establishment and Characterization of the Fibroblast Line from Silkie Bantam

  • Li, L.F.;Guan, W.J.;Li, H.;Bai, X.J.;Ma, Y.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.4
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    • pp.492-499
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    • 2009
  • A Silkie Bantam embryo fibroblast line (named SBF59 line) was successfully established by using direct explant culture and cryopreservation techniques. Cell morphology, viability, dynamic growth and contamination were tested and the karyotype and levels of isoenzymes of lactic dehydrogenase and malic dehydrogenase were analyzed. Four kinds of fluorescent protein extrogenes, including $pEGFP-N_3$, $pECFP-N_1$, $pEYFP-N_1$ and $pDsRed1-N_1$ were transfected into the cells. The results showed that the cells were healthy and possessed a fibrous structure without a change in morphology. The average viability of the cells was 96% before freezing and 90.5% after thawing. The growth curve appeared as typical "S" shape and the cell growth passed through a detention phase, a logarithmic phase and a platform phase; the estimated population doubling time (PDT) was 38.5 h; assays for the presence of bacteria, fungi, viruses and mycoplasmas were negative; the cell line showed no cross contamination when assessed by isoenzyme analysis; the chromosome number was 2n = 78 on more than 88% of occasions; four kinds of fluorescent protein extro-genes appeared to be expressed effectively with a high transfection efficiency between 18.3% and 42.3%. The cell line met the required quality control standard. It not only preserves the genetic resources of the important Silkie Bantam at the cellular level but also provides valuable materials for genomic, post-genomic, somatic cell cloning research and other applications.

Studies on the Total Creatine Phosphokinase(CPK) Activities and CPK Isoenzymes Fractions of Organ Tissues in Chicken (닭의 장기조직의 Creatine phosphokinase(CPK) 총활성 및 CPK Isoenzyme 분획에 관한 연구)

  • Kim, Bong-Sik;Hurh, In;Park, Gyan-Myung;Yoo, Seog-Ho;Kim, Won-Sun
    • Korean Journal of Veterinary Service
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    • v.17 no.2
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    • pp.114-121
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    • 1994
  • Clinically healthy 12 female Manina breed chicken (6 of 75 days old : group 46 of 145 days old: group B, female) were examined to establish physiological basic data on organ tissues total Creatine phosphokinase (CPK) activities and CPK isoenzymes fractions. The tissues examined were the Lung, Heart, Liver, Proventriculus, Gizzard, Duodenum, Colon and Muscle. The results obtained were summarized as follows : 1. Total CPK activities were high with decreasing order of the Muscle > Proventriculus >Giz-zard >Heart >Duodenum> Colon >Lung >Liver in group A and Muscle >Proventriculus >Giz-zard >Heart > Colon >Duodenum > Lung > Liver in group B. Significance of total CPK activities in group difference was only found Colon, group B showed higher values than that of group A (p< 0.01). 2. In the pattern of CPK isoenzymes fractions, Lung, Heart, Liver, Proventriculus, Gizzard, Duodenum and Muscle were high with decreasing order of CK2 >CK3, Colon showed the pattern with decreasing order of CK3 >CK2. Significance of CPK isoenzymes fractions in group difference was only found Liver, CK2 in group B(P<0.01) and CK3 in group A(P<0.01) were higher than that of the other group.

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Identification of three pathways for p-cresol catabolism and their gene expression in Pseudomonas alkylphenolica KL28 (Pseudomonas alkylphenolica KL28에 존재하는 3종류의 p-cresol 분해 경로 및 유전자 발현)

  • Sung, Jin Il;Lee, Kyoung
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.298-305
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    • 2016
  • Previously our laboratory showed that Pseudomonas alkylphenolica KL28 possesses two different lap and pcu gene clusters for p-cresol catabolism. In this study, additional gene cluster (pchACXF-pcaHG-orf4-pcaBC) has been identified to encode enzymes necessary for catabolism of p-cresol to ${\beta}$-carboxy-cis,cis-muconate. This gene cluster showed almost identical nucleotide sequence homologies to those in the plasmid of Pseudomonas putida NCIMB 9866 and 9869, British origins, indicating the possibility of a horizontal gene transfer. Through mutagenesis of each gene cluster and gfp-based promoter reporter assays, it has been shown that the three gene clusters are functionally operated and pch genes are induced by p-cresol. Furthermore, the pcu gene cluster of the three was shown to be dominantly expressed in utilization of p-cresol. Mutation of the pcu gene was defective in aerial structure formation under p-cresol vapor, indicating the utilization rate of carbon source is one of key elements for the multicellular development of this strain.

Bronchiolitis Obliterans Organizing Pneumonia as the First Manifestation of Polymyositis (다발성근염의 선행증상으로 나타난 폐쇄성 세기관지염 기질화 폐렴)

  • Lee, Jong-Hoon;Son, Choon-Hee;Jeong, Won-Tae;Lee, Ki-Nam;Lee, Young-Hoon;Choi, Pil-Jo;Jeong, Jin-Sook;Lee, Chang-Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.1
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    • pp.89-95
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    • 1999
  • Bronchiolitis obliterans organizing pneumonia (BOOP) preceding polymyositis is rare. In this report, a 40-year-old patient with fever, chillness, generalized myalgia and progressive exertional dyspnea, had bilateral interstitial infiltrates on chest radiograph. High-Resolution CT showed subpleural and peribronchial distribution of airspace consolidation. Open lung biopsy was consistent with BOOP. Prednisolone therapy led to improvement, but during tapering of prednisolone for 3 months to 30 mg, he complained of weakness of both lower legs. One month later, prednisolone was tapered to 15 mg a day, fever. chillness and generalized myalgia were recurred. He complained of weakness of both arms. The creatine kinase (CK) with MM isoenzyme, lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) were elevated. Anti-Jo1 antibody was positive. Vastus lateralis muscle biopsy was compatible with polymyositis. After injection of methylprednisolone for 1 week, the patient became afebrile, the dyspnea resolved, the pulmonary infiltrates decreased, and the muscle strength improved. The serum CK, LDH, AST levels declined significantly. Patients with idiopathic BOOP should have follow-up for the possible development of connective tissue disorders including polymyositis.

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A Study on the Production of Aromatic Amino Acids by Escherichia coli. (Escherichia coli에 의한 방향족 아미노산 생산에 관한 연구)

  • Park, Young-Jin
    • Microbiology and Biotechnology Letters
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    • v.13 no.2
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    • pp.119-127
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    • 1985
  • A series of Escherichia coli mutants were exmined for ability to convert glucose and ammonium salts into phenylalanine. This enabled the biochemical changes having major. effects on phenylaianine yield, and interactions between mutations, to be identified. Changes to the common pathway of aromatic biosynthesis having a major effects include desensitization of the first enzyme (3-deoxy-D-arabinoheptulosonate synthase) to end-product inhibition, and removal of repression of enzyme synthesis. It is suggested that the 3-deoxy-D-arabino-heptulosonate synthase Phe isoenzyme has a more important effect on yield. Similarly, removal of repression and end-product inhibition on the phenylalanine terminal pathway increased yield, and changes to both common and branch pathways were synergistic. Blockage of the typrosine and tryptophan pathways had minor effects on phenylalanine yield, and a mutation affecting aramatic amino acid transport (aroP) decreased yield. With multiple-mutation strains hish specific rates of product formation (ie 0.1-0.17g phenylalanine/g cells/h) were obtained.

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Comparison of Two Laccases from Trametes versicolor for Application in the Decolorization of Dyes

  • Li, Qi;Ge, Lin;Cai, Junli;Pei, Jianjun;Xie, Jingcong;Zhao, Linguo
    • Journal of Microbiology and Biotechnology
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    • v.24 no.4
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    • pp.545-555
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    • 2014
  • It has been previously demonstrated that laccases exhibit great potential for use in several industrial and environmental applications. In this paper, two laccase isoenzyme genes, lccB and lccC, were cloned and expressed in Pichia pastoris GS115. The sequence analysis indicated that the lccB and lccC genes consisted of 1,563 and 1,584 bp, and their open reading frames encoded 520 and 527 amino acids, respectively. They had 72.7% degree of identity in nucleotides and 86.7% in amino acids. The expression levels of LccB and LccC were up to 32,479 and 34,231 U/l, respectively. The recombinant laccases were purified by ultrafiltration and $(NH_4)_2SO_4$ precipitation, showing a single band on SDS-PAGE, which had a molecular mass of 58 kDa. The optimal pH and temperature for LccB were 2.0 and $55^{\circ}C$ with 2,2'-azinobis-[ 3-ethylbenzthiazolinesulfonic acid (ABTS) as a substrate, whereas LccC exhibited optimal pH and temperature at 3.0 and $60^{\circ}C$. The apparent kinetic parameters of LccB were 0.43 mM for ABTS with a $V_{max}$ value of 51.28 U/mg, and the Km and $V_{max}$ values for LccC were 0.29 mM and 62.89 U/mg. The recombinant laccases were able to decolorize five types of dyes. Acid Violet 43 (100 g/ml) was completely decolorized by LccB or LccC (2 U/ml), and the decolorization of Reactive Blue KN-R (100 g/ml) was 91.6% by LccC (2 U/ml). Thus, the study characterizes useful laccase isoenzymes from T. versicolor that have the capability of being incorporated into the treatment of similar azo and anthraquinone dyes from dyeing industries.

A Study of Clinical Effect in Camikegungtang and Gamikuyibitang (가미계궁탕(加味桂芎湯) 및 가미귀비탕(加味歸脾湯)의 임상효능(臨床效能)에 관(關)한 연구(硏究))

  • Ryu Yeong-Su;Ryu Hui-Yeong
    • Journal of Oriental Neuropsychiatry
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    • v.3 no.2
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    • pp.1-33
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    • 1992
  • In order to observe clinical effect of Gamikeguntang(加味柱芎湯) and Gamikuyibitang(加味歸脾湯), I reached following conclusion through the clinical pathology and physicohemical investigation, The following results were obtained. 1) The amplirude on AVL lead which is observed in ECG was increased significantly both in medication group of Gamikegungtang (加味柱芎湯) and of Gamikegungtang(加味歸脾湯). 2) In Hematology, the values of RBC, Hb and Hct are totally increased in the medication group of Gamikuyibitang(加味歸脾湯). 3) In serum Lipid, Total cholesterol and Tryglyceride are significantly decreased and HDL-cholesteral is significantly increased both in medication group of Gamikegungtang(加味柱芎湯) and Gamikuyibitang(加味歸脾湯). Especiay, HDL-cholesterol is remarkably increased in the medication group of Gamikegungtang(加味柱芎湯). 4) The serum Bile acid is remarkably decreased in the medication group of Gamikegungtang(加味柱芎湯), but is not in the medication group of Gamikuyibitang(加味歸脾湯). 5) The value of Cardiac enzyme is remarkably decreased both in the medication group of Gamikegungtang(加味柱芎湯) and Gamikuyibitnag(加味歸脾湯). especially, in the group of Gamikuybitang(加味歸脾湯). 6) In determination of LDH isoenzyme, $LD_4$ is remarkably decreased in the medication group of Gamikegungtang(加味柱芎湯) and $LD_1\;LD_2$ are remarkably decreased in the medication group of Gamikuyibitang(加味歸脾湯). In view of the result so far achieved, I knew that Gamikegungtang(加味柱芎湯) had an indirect effect on Palpitation which is called mental disorder by means of treatment of liver and gall bladder disease, and that Gamiluyibitang(加味歸脾湯) make increase activation of heart whose blood is deficient( 心血不足). In conclusion, I think that we need comparative studies between the main prescription of Kegungtang(柱芎湯). Kuyibitnag(歸脾湯) and the additional prescription of them from now on.

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Chronic cold stress-induced myocardial injury: effects on oxidative stress, inflammation and pyroptosis

  • Hongming Lv;Yvxi He;Jingjing Wu; Li Zhen ;Yvwei Zheng
    • Journal of Veterinary Science
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    • v.24 no.1
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    • pp.2.1-2.14
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    • 2023
  • Background: Hypothermia is a crucial environmental factor that elevates the risk of cardiovascular disease, but the underlying effect is unclear. Objectives: This study examined the role of cold stress (CS) in cardiac injury and its underlying mechanisms. Methods: In this study, a chronic CS-induced myocardial injury model was used; mice were subjected to chronic CS (4℃) for three hours per day for three weeks. Results: CS could result in myocardial injury by inducing the levels of heat shock proteins 70 (HSP70), enhancing the generation of creatine phosphokinase-isoenzyme (CKMB) and malondialdehyde (MDA), increasing the contents of tumor necrosis factor-α (TNF-α), high mobility group box 1 (HMGB1) interleukin1b (IL-1β), IL-18, IL-6, and triggering the depletion of superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH). Multiple signaling pathways were activated by cold exposure, including pyroptosis-associated NOD-like receptor 3 (NLRP3)-regulated caspase-1-dependent/Gasdermin D (GSDMD), inflammation-related toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)-mediated nuclear factor kappa B (NF-κB), and mitogen-activated protein kinase (MAPK), as well as oxidative stressinvolved thioredoxin-1/thioredoxin-interacting protein (Txnip) signaling pathways, which play a pivotal role in myocardial injury resulting from hypothermia. Conclusions: These findings provide new insights into the increased risk of cardiovascular disease at extremely low temperatures.

Protein Expression in Pig Species Longissimus dorsi Muscles among Different Breeds and Growth Stages (돼지의 품종 및 성장 단계에 따른 등심조직의 단백질 발현 양상 비교, 분석)

  • Kim, Byung-Uk;Kim, Sam-Woong;Hong, Yeon-Hee;Jeong, Mi-Ae;Ryu, Yeon-Sun;Park, Hwa-Chun;Jung, Jong-Hyun;Kwon, Young-Min;Choi, In-Soon;Lee, Sang-Suk;Kim, Chul-Wook;Cho, Kwang-Keun
    • Journal of Life Science
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    • v.22 no.6
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    • pp.713-722
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    • 2012
  • When proteins extracted from longissimus dorsi muscles of Landrace and Berkshire at the finishing stage were compared by 2-DE, the Landrace demonstrated a quantitative increase in proteins related to slow skeletal muscle function, such as serum albumin precursor, troponin T (slow skeletal muscle; sTnT) and myoglobin. In contrast, the Berkshire exhibited comparatively elevated enzymes involved in metabolic pathways, fast skeletal muscle function, and energy production, such as heat shock 27-kDa protein (HSP27)-1, TnT (fast skeletal muscle; fTnT), muscle creatine kinase, phosphoglucomutase 1 (PGM1), triosephosphate isomerase (Tpi1) and adenylate kinase isoenzyme 1 (AK1). When compared to growing Berkshire, finishing Berkshire showed increased levels of aldehyde dehydrogenase 1 family, member L1 (ALDHL1), and muscle creatine kinase. In contrast, the growing Berkshire muscle had elevated levels of HSP27-1, sTnT, fTnT, serum albumin precursor, PGM1, AK1, and Tpi 1 as compared to the finishing Berkshire. The Landrace longissimus dorsi muscle may be composed of slower skeletal muscle, whereas Berkshire is composed of a faster skeletal muscle. The uniquely elevated quantities of proteins involved in skeletal muscle function, energy metabolism, and cytoskeleton function in the growing Berkshire indicate that these factors support growth and maintenance during the growing stage when compared with the finishing Berkshire.