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Stimulation of Nitric Oxide Production in RAW 264.7 Macrophages by the Peptides Derived from Silk Fibroin. (실크 피브로인 유래 펩타이드에 의한 RAW 264.7 Macrophage의 Nitric Oxide 생성 촉진)

  • 박금주;현창기
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.39-45
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    • 2002
  • It was found that the peptides originated from the hydrolysates of silk fibroin have in vitro immunostimulating effects in murine macrophage RAW264.7 cells. The stimulation effects on nitric oxide (NO) production resulted from treatments of acid or enzymatic hydrolysates were measured. The silk fibroin preparation isolated from cocoon was most efficiently digested by acid hydrolysis. Even though the sole treatment of acid hydrolysate stimulated the NO production in dose-dependent pattern, a part of its activity was found to be caused by the contaminated endotoxin, LPS. When each endotoxin-free hydrolysates obtained by filtering it through an ultrafiltration membrane of molecular weight (MW) cut-off 10,000 to eliminate LPS was used, the peptic hydrolysate with lowest degree of hydrolysis showed the highest activity. The fractions of peptic hydrolysate with MW ranges of 1,000∼10,000, 500∼1,000 and below 500 also showed a higher MW-higher activity correlation. From the analyses of amino acid composition of each hydrolysate, it was found that the contents of arginine, lysine, alanine and glycine residues affected the activity level of hydrolysate. The results of this study showed a possibility of utilizing fibroin as a source for immunostimulating (chemopreventive) functional peptides.

Accumulation of mtDNA Deletion (${\Delta}mtDNA^{4977}$) showing Tissue-Specific and Age-Related Variation (조직별 및 나이에 따른 마이토콘드리아 DNA 결손 (${\Delta}mtDNA^{4977}$)의 축적)

  • Jeong, Hye-Jin;Chung, Hyung-Min;Cho, Sung-Won;Kim, Hyun-Ah;Lee, Kyung-Sool;Kwon, Hwang;Choi, Dong-Hee;Kwak, In-Pyung;Yoon, Tae-Ki;Lee, Sook-Hwan
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.3
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    • pp.203-206
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    • 2003
  • Objectives: Controversial arguments exists on both the case for and against on the accumulation of mitochondrial DNA (mtDNA) deletion in association to tissue and age. The debate continues as to whether this mutation is a major contributor to the phenotypic expression of aging and common degenerative diseases or simply a clinical insignificant epiphenomenon. The objective of this study was to determine whether the accumulation of mtDNA deletion is correlated with age-related and tissue-specific variation. Materials and Methods: One hundred and fifty-seven tissues from blood, ovary, uterine muscle, and abdominal muscle were obtained from patients ranging in age from 31$\sim$60 years. After reviewing the clinical reports, patients with mitochondrial disorder were excluded from this study. The tissues were obtained at gynecological surgeries with the consent of the patient. Total DNA isolated from blood, ovary, uterine muscle, and abdominal muscle was amplified by two rounds of PCR using two pairs of primers corresponding to positions 8225-8247 (sense), 13551-13574 (antisense) for the area around deleted mtDNA and 8421-8440 (sense), 13520-13501 (antisense) for nested PCR product. A statistical analysis was performed by $x^2$-test. Results: About 0% of blood, 94.8% of ovary, 71.4% of uterine muscle, and 86.1% abdominal muscle harbored mtDNA deletion. When we examined the proportion of deleted mtDNA according to age deletion rate was 90% of ovary, 63.6% of uterine muscle, 77.7% of abdominal muscle in thirties and 100% of all tissue in fifties. Conclusion: The findings of this study suggest that the mtDNA deletion is varied in tissue-specific pattern and increases with aging.

Physicochemical Properties of Waxy Rice Starches Prepared from Three Different Cultivars (다수계 찰벼와 일반찰벼 전분의 이화학적 성질)

  • Kim, Hyong-Soo;Kang, Ock-Joo;Yoon, Gae-Soon
    • Applied Biological Chemistry
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    • v.26 no.4
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    • pp.211-216
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    • 1983
  • The physicochemical properties of starches isolated from three kinds of waxy ricer were investigated. The average diammeter of Orchal starch granule was 3.6u, and those of Hangang and Baegoon starch granules were 4.8u and 5.6u, respectively. X-ray diffraction patterns of all samples were A types. Optical transmittance of starch suspensions was increased rapidly $60{\sim}62^{\circ}C$, $50{\sim}55^{\circ}C$ and $50{\sim}60^{\circ}C$ in Olchal, Hangang and Baegoon, respectively, and all of them exhibited maximum transmittance $75{\sim}80^{\circ}C$ in range. Initial gelationization temperature by means of amylogram pattern was $60{\sim}61^{\circ}C$, then, Olchal starch had a little higher temperature than others. Raising power of them was $260\sim220$, and then, Olchal starch has a little higher raising power. But, water binding capacity of Baegoon and Hangang were somewhat higher than that of Olchal. Blue values of Olchal and Hangang were similar with 0.13, and that of Baegoon was 0. 14. Alkari numbers of Olchal, Hangang and Baegoon were 4.2, 4.9 and 5.1. The degree of retrogradation of Hangang and Baegoon starch gels were somewhat higher than that of Olchal starch gel.

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Effects of Extracellular $Ca^{2+}$ and $Ca^{2+}$-Antagonists on Endothelium-Dependent Relaxation in Rabbit Aorta (토끼 대동맥 평활근의 내피세포 의존성 이완에 미치는 $Ca^{2+}$$Ca^{2+}$ 길항제의 효과)

  • Suh, Suk-Hyo;Goo, Yong-Sook;Park, Choon-Ok;Hwang, Sang-Ik;Kim, Ki-Whan
    • The Korean Journal of Physiology
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    • v.24 no.1
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    • pp.91-102
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    • 1990
  • The effects of extracellular $Ca^{2+}$ and various $Ca^{2+}$ antagonists on endothelium-dependent relaxation to acetylcholine were studied in the isolated rabbit thoracic aorta in order to elucidate the control mechanism of endothelium derived relaxing factor (EDRF) release. Endothelium was removed from aortic strips by gentle rubbing with cotton ball. The effect of hemoglobin on basal tension was also observed with hemolysate. The results obtained were as follows: 1) Endothelium-dependent relaxation (EDR) to acetylcholine (ACh) showed biphasic pattern; the initial rapid relaxation phase and the late slow relaxation phase. 2) With the depletion of the extracellular $Ca^{2+}$, EDR was gradually suppressed, especially the late slow relaxation. 3) Verapamil, nifedipine, $Mn^{2+}$ and $Cd^{2+}$ had not any effect on EDR, while $La^{3+}$ and $Co^{2+}$ suppressed EDR completely. 4) The resting tension of the strips with rubbed endothelium was not altered by the addition of hemoglobin. That of the strips with intact endothelium, however, was enhanced and EDR to ACh was completely blocked From these results, we suggest that extracellular $Ca^{2+}$ is necessary for ACh-induced slow relaxation while $Ca^{2+}$ antagonists have not any effect on EDR.

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Epidemiological properties of salmonellosis of poultry farms in Kyongbuk province at recent year (최근 경북지역의 양계농장에서 발생한 salmonellosis의 역학적 특성)

  • 오강희;박노찬;김영환;조민희;이준규;신상희;손재권;정종식
    • Korean Journal of Veterinary Service
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    • v.23 no.1
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    • pp.45-59
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    • 2000
  • The present study was conducted to investigate the epidemiological properties of salmonellosis of poultry farms in Kyongbuk province during the relied from November 1998 to November 1999 And antibiotic susceptibility and biochemical characteristics of 120 Salmonella cultures isolated from chicken samples were also investigated. The results obtained through this study were summarized as follows, 1. Among 667,200 chickens of 31 flocks in 17 layer farms and 9 broiler farms, 61,350 chickens of 31 flocks were died with salmonellosis. The death rate of 25 farms varied from 0.1% to 75.0%, and the average death rate was 9.2%. 2. According to etiological agents, fowl typhoid was shown the most predominant outbreak among the salmonellosis during a year, which accounted for 8R.0%(22/25) of the total case 3. The serotypes of 120 Salmonella isolates were identified as 7 strains(5.8%) of S pullorum, 10 strains(8.4%) of S typhimurium and 103 strains(85.8%) of S gallinarum. 4. Most outbreak of fowl typhoid were prevalent on the layer chicken farms(77.1%), and the summer season(45.5%) also appeared the most hazardous season during the year. 5. It seemed that the Hyline breed(70.6%) was the most susceptible among the layer chicken breeds, and followed by Isabrown(23.5%), Tetra(5.9%) in order. 6. In layers, 76.4% of fowl typhoid occurred commonly from 14 to 40 weeks including the early laying peroid, but in broiler farms, all cases was outbreak within first second weeks. 7 All the strains of S pullorum were resistant to lincomycin(Lm), penicillin(Pm), and steptomycin(Sm), but sensitive to amikacin(Ak), ampicillin(Am), cephalothin(Ce), ciprofloxacin (CiP), chloramphenicol(Cm), colistin(Co), enrofloxacin(Enr), furazolidone(Fu), gentamicin(Gm), kanamycin(Km), neomycin(Nm), polymyxin(Po), and teracycline(Tc). All the strains of S typhimurium were resistant to Lm(100%), Pm(100%), Po(90%), and Sm(90%), but were sensitive to Ak, Am, Ce, CiP, Cm, Co, Enr, Fu, Gm, Km, Nm, and Tc. 8. Minimum inhibitory concentration(MIC) of 103 strains of S gallinarum were also evaluated and their patterns were much more variable than others. All the strains of S gallinarum were sensitive to Ak, Am, Ce, Cip, Cm, Enr, Fu, Km, and Nm, but resistant to Lm(100%), and Sm(100%), 99(96.1%) to Co, 83(80.6%) to Pm, and 83(80.6%) to Po, 55(53.4%) to Gm, and 33(32.0%) to Tc. 9. The multiple drug resistance patterns of 120 Salmonella strains were CoLmPmPo Sm pattern(34.2%), CoGmLmPmPoSmTc(20.8%), CoGmLmPmPoSm(13.3%), CoGmLmSm(7.5%), LmPmSm(7.5%), LmPm(6.7%), CoLmSmTc(3.3%),, CoGmLmSmTc(1.7%), GmLmSmTc(1.7%), CoGmLmPoSm(0.1%), LmPmPo(0.1%), CoLm Sm(0.1%), and LmSm(0.1%), in order.

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IDIOPATHIC GINGIVAL FIBROMATOSIS AT BIRTH (출생 시 발생한 특발성 치은섬유종증)

  • Lee, Hyo-Seol;Choi, Hyung-Jun;Choi, Byung-Jai;Sohn, Hyung-Kyu;Lee, Jae-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.35 no.4
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    • pp.766-770
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    • 2008
  • Gingival fibromatosis is a rare condition characterized by varying degrees of gingival hyperplasia. Gingival fibromatosis usually occurs as an isolated disorder or can be associated with a variety of other syndromes. It usually appears at the time of eruption of permanent dentition but, can develop at the time of eruption of the primary dentition and rarely at birth. It may deform palatal contour and subsequently restrict the tongue movement, resulting in interference during speech and mastication. In addition, it incapacitates maintenance of normal lip closure. A 14-month-old girl visited the department of pediatric dentistry, Yonsei University Dental Hospital, for the congenital gingival overgrowth. There was no one in the family, who showed similar pattern of gingival growth. The intraoral clinical examination revealed generalized severe gingival enlargement throughout the maxillary and the mandibular arches. Enlarged gingival tissue was pink and had firm consistency. She was referred for chromosomal analysis, which confirmed absence of any known syndrome. Under local anesthesia, "Punch-biopsy" was performed on the labial area, and the specimen was histologically diagnosed as gingival fibromatosis. For she did not have any medical problem nor familiar history, she was diagnosed as having idiopathic gingival fibromatosis. Regarding her age and behavior, close follow-up was decided.

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Bitterness and Solubility of Soy Protein, Casein, Gluten, and Gelatin Hydrolysates Treated with Various Enzymes (효소종류에 따른 대두단백, 카제인, 글루텐, 젤라틴 단백질 가수분해물의 쓴맛과 용해도 특성)

  • Kim, Mi-Ryung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.4
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    • pp.587-594
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    • 2010
  • To develop commercially available food protein hydrolysates, the effects of different types of enzymes and substrates on bitterness and solubility of partially hydrolyzed food proteins were investigated. Four types of proteins (casein, isolated soy protein (ISP), wheat gluten, and gelatin) and five types of proteolytic enzymes (a microbial alkaline protease (alcalase), a microbial neutral protease (neutrase), papain, bromelain, trypsin) were used. To profile the pattern of hydrolysis, the degree of hydrolysis (DH) were monitored during 180 min of reaction time by pH-stat method. Casein showed the highest susceptibility to hydrolysis for all five proteases compared to those of ISP, gluten, and gelatin. In addition, the bitter intensity and solubility (nitrogen soluble index, NSI) of each protein hydrolysate were compared at DH 10%. Bitterness and solubility of protein hydrolysates were highly affected by DH and the types of enzymes and substrates. At DH=10%, casein hydrolysate by trypsin, ISP and gluten hydrolysates by either bromelain or neutrase, and gelatin hydrolysates by the five proteases tested in this study were highly soluble and less bitter.

A STUDY ON THE RESPONSES OF OSTEOBLASTS TO VARIOUS SURFACE-TREATED TITANIUM

  • Lee Joung-Min;Kim Yung-Soo;Kim Chang-Whe;Jang Kyung-Soo;Lim Young-Jun
    • The Journal of Korean Academy of Prosthodontics
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    • v.42 no.3
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    • pp.307-326
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    • 2004
  • Statement of problem. The long-term success of implants is the development of a stable direct connection between bone and implant surface, which must be structural and functional. To improve a direct implant fixation to the bone, various strategies have been developed focusing on the surface of materials. Among them, altering the surface properties can modify cellular responses such as cell adhesion, cell motility and bone deposition. Purpose. This study was to evaluate the cellular behaviors on the surface-modified titanium by morphological observation, cellular proliferation and differentiation. Material and methods. Specimens were divided into five groups, depending on their surface treatment: electropolishing(EP) anoclizing(AN), machining(MA), blasting with hydroxyapatite particle(RBM) and electrical discharge machining(EDM). Physicochemical properties and microstructures of the specimens were examined and the responses of osteoblast-like cells were investigated. The microtopography of specimens was observed by scanning electron microscopy(SEM). Surface roughness was measured by a three-dimensional roughness measuring system. The microstructure was analyzed by X-ray diffractometer(XRD) and scanning auger electron microscopy(AES). To evaluate cellular responses to modified titanium surfaces, osteoblasts isolated from neonatal rat were cultured. The cellular morphology and total protein amounts of osteoblast-like cell were taken as the marker for cellular proliferation, while the expression of alkaline phosphatase was used as the early differentiation marker for osteoblast. In addition, the type I collagen production was determined to be a reliable indicator of bone matrix synthesis. Results. 1. Each prepared specimen showed specific microtopography at SEM examination. The RBM group had a rough and irregular pattern with reticulated appearance. The EDM-treated surface had evident cracks and was heterogeneous consisting of broad sheet or plate with smooth edges and clusters of small grains, deep pores or craters. 2. Surface roughness values were, from the lowest to the highest, electropolished group, anodized group, machined group, RBM group and EDM group. 3. All groups showed amorphous structures. Especially anodized group was found to have increased surface oxide thickness and EDM group had titaniumcarbide(TiC) structure. 4. Cells on electropolished, anodized and machined surfaces developed flattened cell shape and cells on RBM appeared spherical and EDM showed both. After 14 days, the cells cultured from all groups were formed to be confluent and exhibited multilayer proliferation, often overlapped or stratified. 5. Total protein amounts were formed to be quite similar among all the group at 48 hours. At 14 days, the electropolished group and the anodized group induced more total protein amount than the RBM group(P<.05). 6. There was no significant difference among five groups for alkaline phosphatase(ALP) activity at 48 hours. The AN group showed significantly higher ALP activity than any other groups at 14 days(P<.05). 7. All the groups showed similar collagen synthesis except the EDM group. The amount of collagen on the electropolished and anodized surfaces were higher than that on the EDM surface(P<.05).

Characteristics and Action Pattern of Pretense from Bacillus subtilis CCKS-111 in Korean Traditional Soy Sauce (한국재래간장으로 부터 분리한 Bacillus subtilis CCKS-111이 생성하는 Protease의 특성 및 작용양상)

  • Choi, Cheong;Choi, Kwang-Soo;Cho, Young-Je;Lim, Sung-il;Kim, Sung;Son, Jun-Ho;Lee, Hee-Duck;Kim, Young-Hwal
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.25 no.6
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    • pp.915-921
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    • 1996
  • An alkaline pretense Producing microorganism was isolated from Korean traditional soy sauce and identified as Bacillus subtilis CCKS-111. The optimum culture condition of Bacillus subtilis CCKS-111 for the production of alkaline pretense was as follow: 2% soluble starch, 0.2% peptone, 0.1% (NB$_4$)$_2$S$_2$O$_{8}$ , 0.2% MgSO$_4$, pH 7.0, 35$^{\circ}C$ and 24hrs. The optimum pH and temperature for the enzyme activity of alkaline pretense producing Bacillus subtilis CCKS-111 were pH 9.0 and 5$0^{\circ}C$, respectively. The enzyme was relatively stable at pH 6.0~11.0 and at temperature below 4$0^{\circ}C$. The activity of the enzyme was inhibited by $K^{+}$ and Hg$^{2+}$, whereas Cu$^{2+}$ exhibited rather activating effects on the enzyme activity. Ethylenediaminetetraacetic acid and phenylmethanesulfonyl fluoride inhibited the enzyme activity. This indicates that this is serine pretense which requires metal ion group for the enzyme activity. Km value was 2.313$\times$10$^{-4}$ M/L, V$_{max}$ value was 39.216$\mu\textrm{g}$/min. This enzyme hydrolyzed casein more rapidly than the hemoglobin.lobin.

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GENE EXPRESSION CHARACTERISTICS OF PUTATIVE PROINFLAMMATORY CYTOKINES AND RECEPTOR MOLECULE CLONING (Putative proinflammatory cytokine유전자의 발현양상과 수용체 분자의 cloing)

  • Oh, Kwi-Ok;Song, Yo-Han;Seo, Young-Seok;Lee, Dong-Whan;Moon, Dae-Hee;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
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    • v.24 no.3
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    • pp.472-482
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    • 1994
  • Cytokines expressed specifically in leukocytes subsets and in activated cells, which are involved in chemotaxis and activation of leukocytes, are recently defined as chemokines. Macrophage inflammatory $protein-1{\alpha}(MIP-1{\alpha})$ and $MIP-1{\beta}$ are members of C-C chemokine subfamily which produces wide immunomodulatory, proinflammatory, and hematopoietic modulatory actions. We have studied their gene expression by using Northern blot analysis in various blood cells such as cytolytic T lymphocyte(CTL), helper T lymphocyte(HTL), macrophage, and B lymphocyte. Resting CTL line CTLL-R8 expressed $MIP-1{\alpha}$ mRNA which was downregulated by ConA stimulation. Both of resting and ConA stimulated HTL line Hut78 and Jurkat did not express $MIP-1{\alpha}$ mRNA. There was detectable $MIP-1{\alpha}$ transcript in HTL hybridoma 2B4.11 which was a little upstimulated by ConA stimulation. B cell line 230, and macrophage cell line RAW264.7 and WR19M.1 showed distinct $MIP-1{\alpha}$ message which were induced after LPS stimulation. Expression pattern of $MIP-1{\beta}$ in all cell lines or cell were almost identical to that of $MIP-1{\alpha}$. Also strategies employed to identify and characterize the biological functions was preceded by receptor cloning to trace the shorcut to the final goal of cytokine research. For the cloning of $MIP-1{\alpha}$ receptor(R), we used synthetic oligonucleotides of transmembrane(T) conserved sequences of already cloned human(h) IL-8-R, and performed reverse transcription-polymerase chain reaction(RT-PCR) amplification using murine(m) macrophage cell line mRNA. Among 5RT-PCR products, we isolated a homologous cDNA with hIL-8-R which were shown to be putative mIL-8-R cDNA.

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