• Title/Summary/Keyword: light-chain

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Altered Gene Expression in Cerulein-Stimulated Pancreatic Acinar Cells: Pathologic Mechanism of Acute Pancreatitis

  • Yu, Ji-Hoon;Lim, Joo-Weon;Kim, Hye-Young
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.6
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    • pp.409-416
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    • 2009
  • Acute pancreatitis is a multifactorial disease associated with the premature activation of digestive enzymes. The genes expressed in pancreatic acinar cells determine the severity of the disease. The present study determined the differentially expressed genes in pancreatic acinar cells treated with cerulein as an in vitro model of acute pancreatitis. Pancreatic acinar AR42J cells were stimulated with $10^{-8}$ M cerulein for 4 h, and genes with altered expression were identified using a cDNA microarray for 4,000 rat genes and validated by real-time PCR. These genes showed a 2.5-fold or higher increase with cerulein: lithostatin, guanylate cyclase, myosin light chain kinase 2, cathepsin C, progestin-induced protein, and pancreatic trypsin 2. Stathin 1 and ribosomal protein S13 showed a 2.5-fold or higher decreases in expression. Real-time PCR analysis showed time-dependent alterations of these genes. Using commercially available antibodies specific for guanylate cyclase, myosin light chain kinase 2, and cathepsin C, a time-dependent increase in these proteins were observed by Western blotting. Thus, disturbances in proliferation, differentiation, cytoskeleton arrangement, enzyme activity, and secretion may be underlying mechanisms of acute pancreatitis.

Proteome Analysis of Bovine Longissimus dorsi Muscle Associated with the Marbling Score

  • Shen, Y.N.;Kim, S.H.;Yoon, D.H.;Lee, H.G.;Kang, H.S.;Seo, K.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.8
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    • pp.1083-1088
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    • 2012
  • The breeding value of marbling score in skeletal muscle is an important factor for evaluating beef quality. In the present study, we investigated proteins associated with the breeding value of the marbling score for bovine sirloin to select potential biomarkers to improve meat quality through comparative proteomic analysis. Proteins isolated from muscle were separated by two-dimensional gel electrophoresis. After analyzing images of the stained gel, seven protein spots for the high marbling score group were identified corresponding to changes in expression that were at least two-fold compared to the low marbling score group. Four spots with increased intensities in the high marbling score group were identified as phosphoglycerate kinase 1, triosephophate isomerase, acidic ribosomal phosphoprotein PO, and capping protein (actin filament) Z-line alpha 2. Spots with decreased intensities in the high marbling score group compared to the low score group were identified as 14-3-3 epsilon, carbonic anhydrase II, and myosin light chain 1. Expression of myosin light chain 1 and carbonic anhydrase 2 was confirmed by Western blotting. Taken together, these data could help improve the economic performance of cattle and provide useful information about the underlying the function of bovine skeletal muscle.

Effects of Minor Arginyl tRNA and Isoleucyl tRNA on the Expression of Clostridium botulinum Neurotoxin Light Chain in Escherichia coli

  • Kim, Jin-Sook;Seong, Hye-Young;Kim, Mi-Wha;Ku, Jong-Seo;Choi, Soon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.13 no.2
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    • pp.287-291
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    • 2003
  • Botulinum neurotoxin type A (BONT/A) is an extremely potent toxin, which is produced by Clostridium botulinum. The light chain of this protein (BONT/A LC), which is known as a zinc endopeptidase, cleaves SNAP-25 involved in the exocytosis process. In this work, the expression of recombinant BoNT/A LC in E. coli is described. The BONT/A LC gene of C. botulinum contains a high frequency of the arginine AGA and isoleucine ATA codons that are rarely used in genes of E. coli, hampering the translation of recombinant protein. The argD and ilex tRNA genes were cloned into pACYC184 vector, resulting in pAAD131X plasmid. The translational stress of the toxin gene related to codon bias was reversed by fupplernentation of the AGA arginyl tRNA of T4 phage and AUA isoleucyl tRNA of E. coli. This system may be applicable for the expression of a variety of AT-rich heterologous genes in E. coli.

Rapid Progression of Solitary Plasmacytoma to Multiple Myeloma in Lumbar Vertebra

  • Yang, Jin Seo;Cho, Yong Jun;Kang, Suk Hyung;Choi, Hyuk Jai
    • Journal of Korean Neurosurgical Society
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    • v.54 no.5
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    • pp.426-430
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    • 2013
  • The prognosis of solitary plasmacytoma varies greatly, with some patients recovering after surgical removal or local fractional radiation therapy, and others progressing to multiple myeloma years later. Primary detection of progression to multiple myeloma is important in the treatment of solitary plasmacytoma. There have been several analyses of the risk factors involved in the early progression to multiple myeloma. We describe one case of solitary plasmacytoma of the lumbar vertebra that was treated with surgical decompression with stabilization and additional radiotherapy. The patient had no factors associated with rapid progression to multiple myeloma such as age, size, immunologic results, pathological findings, and serum free light chain ratio at the time of diagnosis. However, his condition progressed to multiple myeloma less than two months after the initial diagnosis of solitary plasmacytoma. We suggest that surgeons should be vigilant in watching for rapid progression to multiple myeloma even in case that the patient with solitary plasmacytoma has no risk factors for rapid progression to multiple myeloma.

Gene Expression in the Muscles of young and Mature Channel Catfish (Ictalurus punctatus) as Analyzed by Expressed Sequence Tags and Gene Filters

  • Soon-Hag Kim
    • Journal of Aquaculture
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    • v.16 no.1
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    • pp.8-14
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    • 2003
  • To generate expressed sequence tags for genomics research involving genetic linkage analysis, to examine gene expression profiles in muscles of channel catfish in a non-normalized muscle cDNA library, to compare gene expression in young and mature channel catfish muscles using the EST reagents and gene filters to demonstrate the feasibility of functional genomics research in small laboratories. 102 randomly picked cDNA clones were analyzed from the catfish muscle cDNA library. Of the sequences generated, 90.2% of ESTs was identified as known genes by identity comparisons. These 92 clones of known gene products represent transcriptional products of 24 genes. The 10 clones of unknown gene products represent 8 genes. The major transcripts (70.1% of the analyzed ESTs) in the catfish muscle are from many major genes involved in muscle contraction, relaxation, energy metabolism and calcium binding such as alpha actin, creatine kinase, parvalbumin, myosin, troponins, and tropomyosins. Gene expression of the unique ESTs was comparatively studied in the young and adult catfish muscles. Significant differences were observed for aldolase, myostatin, myosin light chain, parvalbumin, and an unknown gene. While myosin light chain and an unknown gene (CM 192) are down-regulated in the mature fish muscle, the aldolase, myostatin, and parvalbumin are significantly up-regulated in the mature fish muscle. Although the physiological significance of the changes in expression levels needs to be further addressed, this research demonstrates the feasibility and power of functional genomics in channel catfish. Channel catfish muscle gene expression profiles provide a valuable molecular muscle physiology blueprint for functional comparative genomics.

Effects of inverse lighting and extreme heat diet on short chain fatty acid and blood lipid profile in extreme heat stress-exposed broilers (폭염 브로일러 닭의 혈액지질 및 짧은 사슬지방산에 대한 폭염사료와 역전점등 효과)

  • Park, Sang-Oh;Hwangbo, Jong;Park, Byung-Sung;Choi, Hee-Chul
    • Journal of the Korean Applied Science and Technology
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    • v.30 no.3
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    • pp.400-410
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    • 2013
  • This study was carried out to investigate the effects of feeding the broilers that are exposed to extreme heat stress by control of inverse lighting times with night restricted feeding of extreme heat diet(EHD1, 2: extreme heat diet) containing different amount of soy oil, molasses, amino acids and vitamin C on short chain fatty acid and blood lipid profile. 300 broiler chickens(Abaica strain) were randomized into four dietary treatment groups according to a randomized block design on the day they were hatched. The four dietary treatment groups were: T1(EHD 1, 10:00~19:00 Dark, 19:00~10:00 Light), T2(EHD 2, 10:00~19:00 Dark, 19:00~10:00 Light), T3(EHD 1, 09:00~18:00 Dark, 18:00~09:00 Light), T4(EHD 2, 09:00~18:00 Dark, 18:00~09:00 Light). The body weight gain of the broilers was highest in T2, and high in order T1, T4, T3(p<0.05). Weights of the lymphoid organ, thymus and bursa of Fabricius were high in T1, T2 as compared to T3, T4 but spleen was lower in T4 than T1, T2, T3(p<0.05). Blood triglyceride, total cholesterol and glucose were higher in T1, T2 than T3, T4(p<0.05). LDL-C was high in orderT4, T3, T2, T1 but HDL-C showed the opposite trend(p<0.05). Blood concentrations of IgG, IgG and IgM were higher in T1, T2 than inT3, T4, but the corticosterone concentration decreased significantly in them. In T1 and T2, Lactobacillus in the feces increased, but total aerobic bacteria, E.coli, coliform bacteria was decreased rather significantly, compared with those in T3 and T4(p<0.05). Concentrations of acetic acid, propionic acid and total SCFA in cecum were high in order T2, T1, T3, T4, but butyric acid, isobutyric acid, valeric acid, isovaleric acid were lower in T1, T2 than in T3, T4 (p<0.05).

Photosynthetic Characteristics of Polyvinylalcohol-Immobilized Spinach Chloroplasts (Polyvinylalcohol에 고정화한 시금치 엽록체의 광합성특성에 대한 연구)

  • 박인호
    • Journal of Plant Biology
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    • v.34 no.3
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    • pp.215-221
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    • 1991
  • Photoxynthetic properties of polyvinylalcohol (PVA)-immobilized chloroplast especially regarded to stability of photosynthetic electron transport and the fluorescence induction pattern were studied. When isolated spinach chloroplasts were immobilized with PVA, it showed good preservation of photosynthetic electron transport activity, especially PS II activity, during storage at -15$^{\circ}C$, 4$^{\circ}C$ and 2$0^{\circ}C$. And immobilized chloroplasts revealed similar thermostability of whole chain electron transport to free chloroplsts. And the absorption peak of red band of chloroplasts showed the blue-shift of 2-4 nm after immobilization. Fv/Fm ratio of chlorophyll fluorescence slightly decreased after immobilization. White light pulse after continuous light do not induced the additional fluorescence rise. This means chlorophyll fluorescence at room temperature reached to Fmax under continuous light in the immobilized chloroplasts. It seems that PVA may be a good candidate for immobilization matrix for the preservation of photosynthetic function of thylakoids and for the continuous use of chloroplast membranes of higher plants for solar energy storage and conversion.

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A Study on the Dynamics Behavior of Mixed Organic Monolayers (혼합 유기단분자의 동적 거동에 관한 연구)

  • Kim, Sung-Jin;Lee, Kyung-Sup
    • Proceedings of the KIEE Conference
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    • 2000.11c
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    • pp.470-472
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    • 2000
  • In this paper generation from of displacement current was compared and measured with light stimulus induce monolayers which 8A5H with azobenzene and arac.acid mixed. Light response of two monolayers which dynamics behavior are different was compared and measured though they are the same isomer The experimental results are as following; In the case of light stimulus mixed monolayers reacted less than 845H about 5[fA]. This is the mason molecule dynamic behaviour and trans was not activated due to its very chain length.

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Single-Chain Fv Fragment of Catalytic Antibody 4f4f with Glycosidase Activity: Design, Expression, and Purification

  • Jang, Chang-Hwan;Chung, Hyun-Ho;Yu, Jae-Hoon;Chang, Yung-Jin;Kim, Hyong-Bai;Paek, Se-Hwan;Shin, Dong-Hoon;Kim, Kyung-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.376-380
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    • 1999
  • Constructs, encoding a single-chain variable fragment of a catalytic antibody 4f4f (scFv-4f4f) with glycosidase activity, were made by combining the coding sequences for the heavy and light chain variable domains with a sequence encoding a linker (GGGGS). Using three different plasmid systems, single-chain antibodies were expressed separately in Escherichia coli, demonstrating significant differences in the expression level and amounts in soluble form of the recombinant protein. The protein expression from pET3a-scFv-4f4f was up to 20% of the total soluble proteins and, more importantly, the proteins were mostly found in a soluble form. An SDS-PAGE analysis of the purified single-chain proteins, yielding higher than 5mg from a 1-1 culture, showed a single band corresponding to its molecular weight of 29,100. A preliminary study shows that the expressed scFv-4f4f is catalytically active. The catalytic parameters for the hydrolysis of p-nitrophenyl-$\beta$-D-glucopyranoside by scFv-4f4f are being investigated.

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Isolation of Human scFv Antibodies Specific for House Dust Mite Antigens from an Asthma Patient by Using a Phage Display Library

  • Jung, Wang-lim;Lee, Hee-kyung;Yong, Tae-soon;Cha, Sang-hoon
    • IMMUNE NETWORK
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    • v.2 no.2
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    • pp.91-95
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    • 2002
  • Background: In order to characterize human antibodies with specificity for mite allergens at the molecular level, a scFv phage display library was constructed using peripheral blood mononuclear lymphocytes from an asthma patient allergic to mite as Ig gene sources. Methods: Immunoglobulin $V_H$ and V gene fragments were obtained by polymerase chain reaction, and randomly combined in pCANTAB-5E vector. The resulting human scFv phage display library had $3{\times}10^4$ independent clones, and biopanning was performed with house dust mite extracts. Results: Four scFv clones specific for house dust mite extract were isolated. Immunoblot assay showed that our clones reacted to 25 kDa and 50~60 kDa proteins with unknown identity in mite extracts. Sequence analysis indicated that two clones (b7 and c15) are identical, and all clones belong to human $V_H3$ subgroup. On the other hand, light chain usage was different in that two clones (a2 and b7 / c15) belonging to V ${\kappa}4$ subgroup, but a4 used V ${\kappa}1$ light chain gene. Conclusion: Our approach should facilitate provision of useful information on the antibody responses against allergens at the molecular level in humans.