• Title/Summary/Keyword: Enzyme I

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분자유전학적인 기술을 이용한 육 감별법

  • Kim, Tae-Heon
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2000.11a
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    • pp.59-75
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    • 2000
  • This study was carried out to develop a DNA marker for identifying between Korean cattle (Hanwoo) and other breeds. First experiment was performed to isolate Hanwoo specific DNA marker at sequence characterized amplified regions (SCARs). Five breeds of cattle including Hanwoo, Holstein, Hereford, Angus and Charolais were represented with the from 8 to 20 individuals. Fourteen primers of 300 arbitrary primers of 10 nucleotides showed reproducible polymorphism across the breeds. An amplified band of 0.9 kb in the primer MG-3 showed the specificity to Holstein breed. And MG-6 and MG-12 detected the Hereford and Hanwoo specific markers at the size of 2.0 kb and 1.0 kb, respectively. A 1.0 kb band of MG-12 was cloned and sequenced. A SCAR primer was designed based on the obtained sequences. It was possible to identify the Hanwoo from Holstein breed. Second experiment was carried out to observe the genotype frequencies of MC1R in 1,044 samples of imported beef and eight different cattle breeds including Hanwoo, Holstein, Angus, Brown-Swiss, Charolais, Limousin, Simmental and Hereford. The primers for the amplification of bovine MC1R gene were designed based on a bovine MC1R gene sequence (GenBank accession no.Y19103). A size of 350 bp was amplified by polymerase chain reaction(PCR), digested with two different restriction enzyme, BsrFI and MspA II, and electrophoresed in 2.5% Metaphore agarose gel for determination of genotypes. Genotype frequencies of Hanwoo were 0.10 in E+e and 0.90 in ee. Allele ED was shown in all of Holstein and Angus breeds tested which have black coat color phenotypes. We suggested that SCAR marker and the bovine MC1R gene could be used as a DNA marker for distinguishing beef between Hanwoo and Holstein.

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Quality characteristics of fermented soybean products produced by lactic acid bacteria isolated from traditional soybean paste (전통 장류 유래 유산균을 이용한 콩 발효물의 품질특성)

  • Lee, Sun Young;Seo, Bo Young;Eom, Jeong Seon;Choi, Hye Sun
    • Food Science and Preservation
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    • v.24 no.2
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    • pp.187-195
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    • 2017
  • This study evaluated quality characteristics of soybean fermented by selected lactic acid bacteria, which were the enzyme strains with high antimicrobial activities isolated from traditionally prepared soybean paste. We determined total aerobic and lactic acid bacteria counts, protease and amylase activities, reducing sugar and amino-type nitrogen contents, and the amounts of amino acids, organic acids, and aroma-compounds. The total aerobic bacteria counts in soybean fermented with strain I13 ($7.75{\times}10^9CFU/mL$) were the highest among all the strains analyzed. Lactic acid bacteria numbers were $2.85{\times}10^9$ to $4.35{\times}10^9CFU/mL$ in soybean fermented with isolates. Amylase and protease activities of the JSB22 sample were the highest among all sample. Reducing sugar and amino-type nitrogen contents of soybean fermented with JSB22 (1.23%, 94.52 mg%) were highest. Total amino acid content of the samples was 15.88-17.62%, and glutamic acid, aspartic acid, leucine, lysine, and arginine were the major amino acids. Lactic acid (0.82-3.65 g/100 g), oxalic acid (22.74-63.57 mg/100 g), and fumaric acid (2.88-6.33 mg/100 g) were predominant organic acids. A total of 39 volatile aroma-compounds were identified, including 2 esters, 5 ketones, 7 alcohols, 14 hydrocarbons, 2 heterocyclic compounds, 4 acids, and 5 miscellaneous compounds. These results represent useful information for the development of a starter (single or complex) and will be used for production of functional fermented soybean foods.

A Study on Bone Formation & Osteoporosis by Taeyoungion-Jahage Extracts

  • Kim, Yi-Geun;Seong, Jun-Ho;Kim, Dong-Il;Lee, Tae-Kyun;Kim, Jun-Ki;Park, Young-Duck
    • The Journal of Korean Obstetrics and Gynecology
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    • v.15 no.4
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    • pp.45-60
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    • 2002
  • Mouse calvarial osteoblast cells were isolated and cultured. To examine whether the cells produce active gelatinases in culture medium or not,the cells were analyzed using by zymograsphic analysis on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). We show that mouse calvarial osteoblasts in culture constitutively synthesize progelatinase- A. Then, mouse osteoblasts, which were stimulated by PTH, $1,25(OH)_2D_3$, mononuclear cell conditioned medium (MCM) and IL-1 as bone resorption agent's, showed increased collagenolysis by producing the active gelatinase. However, treatment of indomethacin and dexamethasone significantly decreased those effects of collagenolysis in mouse osteoblastic cells. On the other hand, IL-1 in stimulating bone resorption was examined using fetal mouse long bone organ culture. IL-1 stimulated bone resorption and produced marked resorption when present simultaneously. Furthermore, when it was examined the effects of indomethacin and dexamethasone on the dose dependent responses of $IL-1{\alpha}$, indomethacin and dexametasone produced a rightward shift in the IL-1 dose response curve. The results of in vitro cytotoxicities showed that Taeyoungjon-Jahage water extracts(T.Y.J-J.H.G extracts) have no any cytotoxicities in concentrations of $1-200\;{\mu}g/ml$ and furthermore there is no any cytotoxicity even in concentration of $300\;{\mu}g/ml$ on mouse calvarial bone cells. T.Y.J.-J.H.G. extracts had protective activity against PTH (2 units/mI), or MCM (5%, v/v), or $rhIL-1{\alpha}$ (1 ng/mI) or $1,25(OH)_2D3$ (10 ng/ml) , $IL-1{\alpha}$ and $IL-1{\beta}-induced$ collagenolysis in the mouse calvarial cells. Pretreatment of the T.Y.J.-J.H.G.extracts for 1 h, which by itself had little effect on cell survival, did not enhance the collagenolysis, nor significantly reduced the collagenolysis by pretreatment. Furthermore. the medicinal extracts were shown to have the protective effects against collagenolysis induced by $IL-1{\alpha}$ and $IL-1{\beta}$. Pretreatment of the extracts for 1 h significantly reduced the collagenolysis. Interestingly, the T.Y.J.-J.H.G. extracts were shown to have the inhibiting effects against gelatinase enzyme and processing activity induced by the bone resortion agents of PTH, $1,25(OH)_2D_3$, $IL-1{\beta}$ and $IL-1{\alpha}$, with strong protective effect in pretreatment with the extracts. T.Y.J.-J.H.G. extracts were shown to have the inhibiting effects against $IL-1{\alpha}-$ and $IL-1{\beta}-stimulated$ bone resorption and the effect of the pretreatment with a various concentrations of the medicinal extracts were significant. The inhibition extent and phenomena of IL-1 stimulated bone resorption by nonsteroidal anti-inflammatory agents of indomethacin and dexamethasone were similar to those obtained by T.Y.J.-J.H.G. extracts treatment in the mouse calvarial tissue culture system. These results indicated that the T.Y.J.-J.H.G.-water extracts are highly stable and applicable to clinical uses in osteoporosis.

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Incorporation of RAPD linkage Map Into RFLP Map in Glycine max (L, ) Merr (콩의 RAPD 연관지도를 RFLP 연관지도와 합병)

  • Choi, In-Soo;Kim, Yong-Chul
    • Journal of Life Science
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    • v.13 no.3
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    • pp.280-290
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    • 2003
  • The incorporation of RAPD markers into the previous classical and RFLP genetic linkage maps will facilitate the generation of a detailed genetic map by compensating for the lack of one type of marker in the region of interest. The objective of this paper was to present features we observed when we associated RAPD map from an intraspecific cross of a Glycine max$\times$G. max, 'Essex'$\times$PI 437654 with the public RFLP map developed from an interspecific cross of G. max$\times$G. soja. Among 27 linkage groups of RAPD map, eight linkage groups contained probe/enzyme combination RFLP markers, which allowed us the incorporation of RAPD markers into the public RFLP map. Map position rearrangement was observed. In incorporating L.G.C-3 into the public RFLP linkage group a1 and a2, both pSAC3 and pA136 region, and pA170/EcoRV and pB170/HindIII region were in opposite order, respectively. And, pk400 was localized 1.8 cM from pA96-1 and 8.4 cM from pB172 in the public RFLP map, but was localized 9.9 cM from i locus and 18.9 cM from pA85 in our study. A noticeable expansion of the map distances in the intraspecific cross of Essex and PI 437654 was also observed. Map distance between probes pA890 and pK493 in L.G.C-1 was 48.6 cM, but it was only 13.3 cM in the public RFLP map. The distances from the probe pB32-2 to pA670 and from pA670 to pA668 in L.G. C-2 were 50.9 cM and 31.7 cM, but they were 35.9 cM and 13.5 cM in the public RFLP map. The detection of duplicate loci from the same probe that were mapped on the same or/and different linkage group was another feature we observed.

Extract of Rubus coreanus Fruits Increases Expression and Activity of Endothelial Nitric Oxide Synthase in the Human Umbilical Vein Endothelial Cells (복분자 추출물에 의한 내피세포 NO 합성효소의 활성과 발현 증가)

  • Yoon, Hyun-Joong;Park, Soo-Young;Oh, Sung-Tack;Lee, Kee-Young;Yang, Sung-Yeul
    • Journal of Life Science
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    • v.21 no.1
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    • pp.44-55
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    • 2011
  • This study aimed to investigate the effects of water extract of Rubus coreanus (RCE) on the expression and activity of endothelial nitric oxide synthase (eNOS), as well as its signal transduction pathways in human umbilical vein endothelial cells (HUVECs). The specific inhibitors of NOS show RCE treatment increases NO production in HUVECs due to the up-regulation of eNOS rather than iNOS. The real-time expression level of eNOS mRNA was also increased upon RCE treatment in HUVECs. While a PKC-specific inhibitor, RO-317549, did not alter RCE-induced NO production in HUVECs, tamoxifen (estrogen receptor-specific inhibitor), PD98059 (ERK-specific inhibitor) and LY-294002 (PI3K/Akt-specific inhibitor) did have suppressive effects. Increased NO production by RCE seems to result from a higher level of active eNOS (pSer1177). Specifically, inhibition of ERK not only decreased the level of active eNOS, but also increased the inactive form of the enzyme (pThr495) in HUVECs. This study suggests that RCE treatment increases NO production in HUVECs due to the increased expression and activity of eNOS. It is also shown that RCE-induced eNOS activation occurs partly through the binding of RCE to the estrogen receptor, along with ERK and PI3K/Akt-dependent signal transduction pathways. In addition, the regulatory binding proteins of eNOS including Hsp90 and caveolin-1 were related to these effects of RCE on eNOS activity in HUVECs.

Effects of the Deer Antler Extract on Scopolamine-induced Memory Impairment and Its Related Enzyme Activities (녹용 추출물이 치매 동물모델의 기억력 개선과 관련효소 활성에 미치는 효과)

  • Lee, Mi-Ra;Sun, Bai-Shen;Gu, Li-Juan;Wang, Chun-Yan;Fang, Zhe-Ming;Wang, Zhen;Mo, Eun-Kyoung;Ly, Sun-Young;Sung, Chang-Keun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.4
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    • pp.409-414
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    • 2009
  • The aim of this study was to investigate the ameliorating effects of deer antler extract on the learning and memory impairments induced by the administration of scopolamine (2 mg/kg, i.p.) in rats. Tacrine was used as a positive control agent for evaluating the cognition enhancing activity of deer antler extract in scopolamine-induced amnesia models. The results showed that the deer antler extract-treated group (200 mg/kg, p.o.) and the tacrine-treated group (10 mg/kg, p.o.) significantly ameliorated scopolamine-induced amnesia based on the Morris water maze test. Although there was no statistical significance of brain ACh contents among the experimental groups, the brain ACh contents of the deer antler extract-treated group was slightly higher than that of the scopolamine-treated group. The inhibitory effect of deer antler extract on the acetylcholinesterase activity in the brain was significantly lower than that of scopolamine-treated group. The tacrine- and the deer antler-treated groups reduced the MAO-B activity compared to the scopolamine-treated group, but not significantly. These results suggest that the deer antler extract could be an effective agent for the prevention of the cognitive impairment induced by cholinergic dysfunction.

An Analysis of 94 Percutaneous Renal Biopsies (경피적 신생검 94례에 대한 분석)

  • Kang, Ho-Jung;Lim, Sang-Woo;Do, Joo-Yeung;Yoon, Kyung-Woo
    • Journal of Yeungnam Medical Science
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    • v.12 no.1
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    • pp.84-95
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    • 1995
  • A clinical and histopathological study was performed on ninety-four patients with nephrotic syndrome (91 idiopathic and 3 secondary) who were admitted to Department of Internal Medicine, Yeungnam University Hospital during the period of nine years, from January 1985 to May 1994. The results were as following. 1. the ratio of male to female was 1.76:1. In young age group, minimal change was the most predominant type. In old age group, membranous glomerulonephritis and focal glomerulosclerosis were predominant types. 2. The primary nephrotic syndromes were 96.8% and secondary nephrotic syndromes were 3.2%. Histopathologic findings of 94 renal biopsy tissue were classified into minimal change (43.6%), mesangial proliferative glomerulonephritis (29.8%), membranous glomerulonephritis (12.8%), TypeI membranous proliferative glomerulonephritis (4.3%), focal glomerulosclerosis (3.2%) and others (6.4%). 3. The response of eighty-six patients treated with steroid showed complete remission in 51.2%, partial remission in 20.9%, steroid dependent in 2.3%, and no effect in 25.6% of cases respectively. The response to steroid therapy was most effective in the patients with minimal change lesion. 4. In the patient with membranous proliferative glomerulonephlitis, long-term angiotensin converting enzyme inhibitor treatment showed less deterioration of renal function.

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Anti-inflammatory and Antioxidative Effects of Lotus Root Extract in LPS-PG-Stimulated Human Gingival Fibroblast-1 Cells (치주염 원인균 LPS-PG로 유도된 인체 치은섬유아세포에서 연뿌리 추출물에 대한 항염증 및 항산화 효과)

  • Lee, Young-Kyung;Kim, Chul Hwan;Jeong, Dae Won;Lee, Ki Won;Oh, Young Taek;Kim, Jeong Il;Jeong, Jin-Woo
    • Korean Journal of Plant Resources
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    • v.35 no.5
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    • pp.565-573
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    • 2022
  • Gingival inflammation is one of the main causes that can be related to various periodontal diseases. Human gingival fibroblast (HGF) is the major constituent in periodontal connective tissue and secretes various inflammatory mediators, such as nitric oxide (NO) and prostaglandin E2 (PGE2), upon lipopolysaccharide stimulation. This study is aimed at investigating the anti-inflammatory and antioxidative activities of Lotus Root extract (LRE) in Porphyromonas gingivalis derived lipopolysaccharide (LPS-PG)-stimulated HGF-1 cells. The concentration of NO and PGE2, as well as their responsible enzymes, inducible NO synthase (iNOS), and cyclooxygenase-2 (COX-2), was analyzed by Griess reaction, ELISA, and western blot analysis. LPS-PG sharply elevated the production and protein expression of inflammatory mediators, which were significantly attenuated by LRE treatment in a dose-dependent manner. LRE treatment also suppressed activation of Toll-like receptor 4 (TLR4)/myeloid differentiation primary response gene 88 (MyD88) and nuclear factor-κB (NF-κB) in LPS-PG-stimulated HGF-1 cells. In addition, one of phase II enzyme, NAD(P)H quinone dehydrogenase (NQO)-1, and its transcription factor, Nuclear factor erythroid 2-related factor 2 (Nrf2), were significantly induced by LRE treatment. Consequently, these results suggest that LRE ameliorates LPS-PG-induced inflammatory responses by attenuating TLR4/MyD88-mediated NF-κB, and activating NQO-1/Nrf2 antioxidant response element signaling pathways in HGF-1 cells.

Studies on Microbial and Enzymatic Actions during the Ripening Process of Salted Alaska Pollack Tripe (창난 젓갈의 숙성 과정 중 미생물 및 자기소화효소 작용에 관한 연구)

  • Chae, Soo-Kyu
    • The Korean Journal of Food And Nutrition
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    • v.24 no.3
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    • pp.340-349
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    • 2011
  • This study examined the roles of autolytic enzymes and microorganisms in the ripening process of salted Alaska pollack tripe made with various concentrations of salt i.e, 7.5% and 20% by weight. Salted Alaska pollack tripe treated with antibiotic agents for the inhibition of microbial growth and a control were prepared experimentally, and changes in chemical composition and viable cell counts were investigated, individually, during the ripening process. Just after the preparation of the low salt Alaska pollack tripe made with 7.5% salt, viable bacterial cells occurred at a level of $10^5$ CFU/g. In the control, bacterial counts increased rapidly to $10^7$ CFU/g by the 14th day of ripening. However, in the sample treated with antibiotic agents, counts were decreased to a level of $10^4$ CFU/g by the 3rd day of ripening and increased gradually to $10^6$ CFU/g by the 5th day of ripening, and then the same value was maintained there-after. Just after the preparation of the high salt Alaska pollack tripe made with 20% salt, viable bacterial cells occurred at a level of $10^3$ CFU/g. In both the samples treated with antibiotic agents and the control, bacterial counts decreased rapidly to $10^0$ CFU/g by the 45th day of ripening and increased gradually there-after. The content of amino type nitrogen was 76.3 mg% just after the preparation of the low salt Alaska pollack tripe made with 7.5% salt. Amino type nitrogen content was increased to 283.5 mg% by the 5th day of proper ripening in the control, but it was increased to 208.0 mg% in the sample treated with antibiotic agents. The difference in amino type nitrogen content was 75.5 mg/100 g. The content of amino type nitrogen was 57.2 mg% just after the preparation of the high salt Alaska pollack tripe made with 20% salt. Amino type nitrogen content was increased to 198.3 mg by the 60th day of proper ripening in the control, but it was increased to 162.0 mg% in the sample treated with the antibiotic agents. The difference in amino type nitrogen content was 36.3 mg/100 g. The contents of VBN and TMA-N were 102.1 mg% and 20.5 mg%, respectively, at the 7th day of ripening in the low salt Alaska pollack tripe made with 7.5% salt. The content of VBN was 60.0 mg% and TMA-N was not detected at the 21st day of ripening in the sample treated with antibiotic agents. The control sample was spoiled by the 7th day of ripening but the sample treated with antibiotic agents was not spoiled by the 21st day of ripening. On the other hand, VBN content was 37.2 mg% and TMA-N was not detected at the 90th day of ripening in the high salt Alaska pollack tripe made with 20% salt, and the control sample was not spoiled.

Angiotensin Converting Enzyme Gene Polymorphism in Alport Syndrome (알포트증후군 환자에서 안지오텐신전환효소 유전자 다형성의 의의)

  • Kim Ji-Hong;Lee Jae-Seung;Kim Pyung-Kil
    • Childhood Kidney Diseases
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    • v.8 no.1
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    • pp.18-25
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    • 2004
  • Purpose : Alport syndrome is clinically characterized by hereditary progressive nephritis causing ESRD with irregular thickening of the GBM and sensory neural hearing loss. The mutations of type IV collagen gene(COL4A5) located on the long arm of X chromosome is considered responsible for most of the structural abnormalities in the GBM of Alport patients. Since no definite clinical prognostic predictor has been reported in the disease yet, we designed this study to evaluate the significance of genetic polymorphism of the angiotensin converting enzyme in children with Alport syndrome as a prognostic factor for disease progression. Methods : ACE I/D genotype were examined by PCR amplification of the genomic DNA in 12 patients with Alport syndrome and 12 of their family members. Alport patients were divided into two groups; the conservative group, those who had preserved renal function for more than 10 years of age, the early CRF group, those who had progressed to CRF within 10 years of age. Results : The mean age of onset was $3.45{\pm}2.4$ years in the conservative group, $4.4{\pm}1.2$ years in the early CRF group. Sex ratios were 5:3 and 2:1 in each group. Among 12 cases of patients, 4 cases were in early CRF group and their mean duration of onset to CRF was 4.5 yews(8.9 years of age). Eight patients(67%) were in the conservative group and they had normal renal function for more than 10 years of age(mean duration of renal preservation was 10.6 years). The incidence of II type ACE gene were in 25.0%(3 cases), ID type in 41.7%(5 cases), DD type in 33.3%(4 cases). There was no significant difference between Alport patient and normal control(II type 44.3%, ID type 40.9%, DD type 14.8%). The incidence of DD type of early CRF group were higher than that of the conservative group(75% vs 12.5%)(p<0.05). There was no difference in ACE gene polymorphism between normal Alport family members and control group. Conclusion : Even though there was no significant difference of ACE polymorphism between Alport patients and the normal control group, the incidence of DD type is significantly increased in early CRF group which means DD type of ACE polymorphism has a possibility of being a predictor for early progression to CRF in Alport patients.

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