• Title/Summary/Keyword: 43 kD protein

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Identification and Functional Analysis of Escherichia coli RNase E Mutants (Escherichia coli 리보핵산 내부분해효소 RNase E의 돌연변이체 선별 및 특성분석)

  • Shin, Eun-Kyoung;Go, Ha-Young;Kim, Young-Min;Ju, Se-Jin;Lee, Kang-Seok
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.325-330
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    • 2007
  • RNase E is an essential Escherichia coli endoribonuclease that plays a major role in the decay and processing of a large fraction of RNAs in the cell and expression of N-terminal domain consisted of 1-498 amino acids (N-Rne) is sufficient to support normal cellular growth. By utilizing these properties of RNase E, we developed a genetic system to screen for amino acid substitutions in the catalytic domain of the protein (N-Rne) that lead to various phenotypes. Using this system, we identified three kinds of mutants. A mutant N-Rne containing amino acid substitution in the S1 domain (I6T) of the protein was not able to support survival of E. coli cells, and another mutant N-Rne with amino acid substitution at the position 488 (R488C) in the small domain enabled N-Rne to have an elevated ribonucleolytic activity, while amino acid substitution in the DNase I domain (N305D) only enabled N-Rne to support survival of E. roli cells when the mutant N-Rne was over-expressed. Analysis of copy number of ColEl-type plasmid revealed that effects of amino acid substitution on the ability of N-Rne to support cellular growth stemmed from their differential effects on the ribonucleolytic activity of N-Rne in the cell. These results imply that the genetic system developed in this study can be used to isolate mutant RNase E with various phenotypes, which would help to unveil a functional role of each subdomain of the protein in the regulation of RNA stability in E. coli.

Comparison of Radiation Adaptive Responses in Peripheral Lymphocytes of Patients Undergoing Tc-99m MDP and Tc-99m DTPA Scintigraphies (Tc-99m MDP와 Tc-99m DTPA 신티그라피를 시행한 환자의 말초혈액 림프구에서 유도되는 방사선 적응반응의 비교)

  • Bom, Hee-Seung;Li, Ming-Hao;Min, Jung-Jun;Kwon, An-Sung;Kim, Ji-Yeul
    • The Korean Journal of Nuclear Medicine
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    • v.34 no.3
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    • pp.252-259
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    • 2000
  • Purpose: The purpose of this study was to compare the radiation adaptive response (RAR) in peripheral lymphocytes (PL) of patients induced by Tc-99m MDP and Tc-99m DTPA scintigraphies. Materials and Methods: Lymphocytes from 45 patients (25 males, 20 females, mean age $44{\pm}18$ years) were collected before and after scintigraphies using 740 MBq Tc-99m MDP (n=22) or Tc-99m DTPA (n=23). Lymphocytes from 20 controls (12 males, 8 females, mean age $43{\pm}7$ years) were also collected. They were exposed to challenge dose of 2 Gy ${\gamma}-rays$ using a Cs-137 cell irradiator Number of ring-form (R) and dicentric (D) chromosomes was counted under the light microscope. From them a representative score, Ydr, was calculated as Ydr=(D+R)/cells. Adaptation index (AI) was defined as difference of Ydr between unconditioned and conditioned lymphocytes. Ydr was also measured after an administration of cycloheximide (CHM), a protein synthesis inhibitor, before challenge dose. Results: RAR was induced in both groups of patients. CHM abolished the adaptive response in both groups. AI of Tc-99m MDP group was significantly higher than that of Tc-99m BTPA group. Conclusion: Tc-99m MDP induced RAR was more prominent than those induced by Tc-99m DTPA.

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Effect of Nitrogen Application Level and Regrowth Period on Composition of Fatty Acid in Tall fescue and Bermudagrass (질소시비 수준 및 재생기간이 톨페스큐와 버뮤다그라스의 지방산조성에 미치는 영향)

  • Park, Hyung-Soo;Hwang, Kyung-Jun;Park, Nam-Gun;Kim, Young-Jun;Jeong, Ha-Yeon;Jo, Nam-Chul;Ko, Moon-Suck
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.29 no.1
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    • pp.43-50
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    • 2009
  • Two experiments were conducted to evaluate the effects of nitrogen (N) application level and regrowth period on the fatty acid (FA) concentration and forage production of tall fescue (Festuca arundinacea Schr.) and bermudagrass (Cynodon dactylon (L.) Pers.). N was applied at 0, 50 and 100 kg/ha, and swards were cut after regrowth periods (15d, 30d and 45d). Concentrations of Individual FA were determined by gas chromatography. FA composition of tall fescue contains a high proportion ($87{\sim}88%$) of total FA contents as palmitic (C16:0), linoleic (C18:2) and linolenic acid (C18:3). The FA composition was not affected by N application, but a longer regrowth period significantly (p<0.05) decreased the composition of linolenic acid (C18:3) and increased those of linoleic acid (C18:2). The FA composition of bermudagrass was not affected by N application, but a longer regrowth period significantly (p<0.001) decreased the composition of linoleic (C18:2) and linolenic acid (C18:3). Linoleic and linolenic acfd of tall fescue were higher than those of bermudagrass. Dry matter (DM) yield of bermudagrass was significantly (P<0.05) affected by N application level and regrowth period. DM yield was higher at high N application and was higher at longer regrowth period. The crude protein concentration was higher at high N application and was low at longer regrowth period. Application of nitrogen fertilizer Increases dry matter (DM) yield and crude protein (CP) content of bermudagrass. These studies demonstrate opportunities to affect the FA concentration and composition of FA in forage through management strategies, which could affect milk FA composition.

BIOLOGICAL RESPONSES OF OSTEOBLAST-LIKE CELLS TO DIFFERENT TITANIUM SURFACE BY ANODIZING MODIFICATION

  • Kim Myung-Joo;Kim Chang-Whe;Lim Young-Jun;Park Hyun-Joo
    • The Journal of Korean Academy of Prosthodontics
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    • v.43 no.6
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    • pp.751-763
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    • 2005
  • Statement of problem. To improve a direct implant fixation to the bone, various strategies have been developed focusing on the surface of materials. The surface quality of the implant depends on the chemical, physical, mechanical and topographical properties of the surface. The different properties will interact with each other and a change in thickness of the oxide layer may also result in a change in surface energy, the surface topography and surface, chemical composition. However, there is limited the comprehensive study with regard to changed surface and biologic behavior of osteoblast by anodization. Purpose of study. The aim of this study was to analyze the characteristics of an oxide layer formed and to evaluate the cellular biologic behaviors on titanium by anodic oxidation (anodization) by cellular proliferation, differentiation, ECM formation and gene expression. And the phospholipase activity was measured on the anodized surface as preliminary study to understand how surface properties of Ti implant are transduced into downstream cellular events. Methods and Materials. The surface of a commercially pure titanium(Grade 2) was modified by anodic oxidation. The group 1 samples had a machined surface and other three experimental specimens were anodized under a constant voltage of 270 V(Group 2), 350 V(Group 3), and 450 V(Group 4). The specimen characteristics were inspected using the following five categories; the surface morphology, the surface roughness, the thickness of oxide layer, the crystallinity, and the chemical composition of the oxide layer. Cell numbers were taken as a marker for cell proliferation. While the expression of alkaline phosphatase and Runx2 (Cbfa1) was used as early differentiation marker for osteoblast. The type I collagen production was determined, which constitutes the main structural protein of the extracellular matrix. Phospholipase $A_2$ and D activity were detected. Results. (1) The anodized titanium had a porous oxide layer, and there was increase in both the size and number of pores with increasing anodizing voltage. (2) With increasing voltage, the surface roughness and thickness of the oxide film increased significantly (p<0.01), the $TiO_2$phase changed from anatase to rutile. During the anodic oxidization, Ca and P ions were more incorporated into the oxide layer. (3) The in vitro cell responses of the specimen were also dependant on the oxidation conditions. With increasing voltage, the ALP activity, type I collagen production, and Cbfa 1 gene expression increased significantly (p<0.01), while the cell proliferation decreased. (4) In preliminary study on the relation of surface property and phospholipase, PLD activity was increased but $PLA_2$ activity did not changed according to applied voltage. Conclusion. The anodized titanium shows improved surface characteristics than the machined titanium. The surface properties acquired by anodization appear to give rise more mature osteoblast characteristics and might result in increased bone growth, and contribute to the achievement of a tight fixation. The precise mechanism of surface property signaling is not known, may be related to phospholipase D.

Chemical Composition and Nutritional Value of Algae Meal Produced from Dairy Cow Wastes as a Feedstuff (젖소폐기물에서 생산된 Algae Meal의 화학적조성 및 사료적가치)

  • Kim, Y.K.;Eun, J.S.;Kim, S.D.
    • Journal of the Korea Organic Resources Recycling Association
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    • v.6 no.1
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    • pp.75-85
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    • 1998
  • Algal meal (cell) was produced from the solution of dairy cow wastes by fermentation of ulothrix. sp. and chlorella sp. Raw wastes mainly feces were diluted with ground water to give dry matter concentration of 0.5 w/v of wastes in 20 l amounts of ten plastic containers. Each containers were covered with plastic nets and vinyl films to protect from the insects and rain. Algea cells were harvested every 3 to 5 days and dried by sunlight and artifitial heat. Dried cells were ground by a feed meal, and analyzed and tested for the chemical composition of dry cell, in vitro DM and protein digestibility and the safty of algae. Protein contents in algae meals, ulothrix (29.37%) and chlorella (29.24%) were similar. However, chlorella contained lower Neutral detergent fiber (5.92%) than ulothrix(20,76%), and higher ash (32.86%) and calcium (12.62%) than ulothrix (28.66% and 6.09%) (P<.01). Ulothrix protein had higher for essential amino acids; valine, isoleucine and phenylalanine, than chlorella (P<.05). Algal fats contained high saturated fatty acids, C16:0 and C18:0, for ulothrix and high unsaturated fatty acids, C18:1 and C18:2, for chlorella (P<.01). In vitro digestibility of. ulothrix tended to be higher for DM, but lower for protein than chlorella. The weight gain and survival percentage were higher for pond fishes (loaches, Misgurnus sp. ) fed diet added chlorella meal than diets added ulothrix meal and control diet (P<.05).

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Anti-aging Effects of L-Carnitine on Human Skin (L-카르니틴의 사람피부에 대한 항노화 효과)

  • Lee Bum-Chun;Choe Tae-Boo;Sim Gwan-Sub;Lee Geun-Soo;Park Sung-Min;Lee Chun-Il;Pyo Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.3 s.47
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    • pp.393-397
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    • 2004
  • L-Carnitine $({\beta}-hydroxy-{\gamma}-trimethyl-ammoniumbutyric{\;}acid)$ is a small water-soluble molecule important in mammalian fat metabolism. It is essential for the normal oxidation of fatty acids by the mitochondria, and is involved in the trans-esterification and excretion of acyl-CoA esters. In this paper, to investigate the relationship between aging and L-carnitine, we investigated the effects of in vitro matrix-metalloproteinase (MMP) inhibition and activity and expression of UYA-induced MMPs in human skin fibroblasts. Also, we studied to develop as anti-aging cosmetics with L-carnitine. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. ELISA (enzyme linked immune sorbent assay), gelatin-substrate zymography, RT-PCR ELISA techniques were used for the effects of L-carnitine on MMP expression, activity, and MMP mRNA expression in UVA irradiated fibroblast $(5\;J/cm^2)$, respectively. In addition, we performed clinical study with L-carnitine cream. L-carnitine inhibited the activities of MMP-1 in a dose-dependent manner and the $IC_{50}$ values calculated from semi-log plots were 2.45 mM, and L-carnitine showed strong inhibition on MMP-2 (gelatinase) activity in UVA irradiated fibroblast by zymography. Also, UVA induced MMP-1, 2 expression was reduced $43\%,\;53\%$ by treated with L-carnitine at 1.25 mM, and MMP-1 mRNA expression was reduced dose-dependent manner. Therefore L-carnitine was able to significantly inhibit the MMP activity, and regulate MMP expression in protein and mRNA level. The results of clinical study showed that $1.0\%$ L-carnitine treated group reduced wrinkle significantly compared with placebo treated group (P<0.05). All these results suggest that L-carnitine may be useful as new anti-aging cosmetics for protection against UVA induced Mm expression and activity.

Study on Characteristic of Pyropia Dentata Three Cultivar in Haenam Aquafarm, Jellanam-do (전남 해남에서 잇바디돌김 3종의 양식 특성 연구)

  • Han, M.K.;Jeong, D.S.;Kim, C.W.;Choi, S.J.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.23 no.2
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    • pp.43-50
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    • 2021
  • Growth tests on the Yuldo, Supum1 and Supum2 cultivars of Pyropia dentata were performed at the Eoran and Imha aquafarm, Haenam in Jeollanamdo, from October to December in 2017. The mean water temperature ranged from 5.4 to 26.4 ℃. In Eoran aquafarm (flating raft method), dissolved total nitrogen (DTN), dissolved total phosphorus (DTP), total nitrogen (TN), total phosphorus (TP) and chemical oxygen demand (COD) ranged from 0.091 to 0.181 mg/L, 0.007 to 0.019 mg/L, 0.114 to 0.187 mg/L, 0.008 to 0.033 mg/L and 0.200 to1.000 mg/L, respectively. In Imha aquafarm (fixed pold method), DTN, DTP, TN, TP and COD ranged from 0.118 to 0.276 mg/L, 0.005 to 0.024 mg/L, 0.155 to 0.305 mg/L, 0.009 to 0.042 mg/L and 0.300 to1.400 mg/L, respectively. In order to investigate the number of conchospores attached on the Pyropia net, which was cut into about 4cm long. The mean number of conchospores of Yuldo, Supum1 and Supum2 cultivars were 39.5, 26.5, 72.5, respectively. Young thalli were harvested two times at Eoran aquafarm, and three times at Imha aquafarm. In eoran aquafarm, the mean thallus length of Yuldo, Supum1 and Supum2 cultivars were 49.9, 46.0, 42.0 cm on October and 163, 126.0, 263.0 cm on November, respectively. The mean thallus width of Yuldo, Supum1 and Supum2 cultivars were 5.8, 4.6, 11.5 cm on October and 20.9, 11.5, 14.0 cm on November, respectively. In Imha aquafarm, the mean thallus length of Yuldo, Supum1 and Supum2 cultivars were 119.0, 60.9, 71.0 cm on October, 196.0, 132.0, 262.0 cm on November and 187.0, 281.0, 296.0 cm on December, respectively. The mean thallus width of Yuldo, Supum1 and Supum2 cultivars were 4.2, 3.4, 3.1 cm on October, 8.9, 6.2, 6.6 cm on November and 11.7, 11.6, 9.4 cm on December, respectively. In eoran aquafarm, contents of moisture, crude ash, crude lipid, crude protein and carbohydrate of three cultivars ranged from 11.64 to 20.15, 19.54 to 21.19, 0.00 to 0.18, 29.78 to 37.81, 29.16 to 29.71, respectively. In Imha aquafarm, contents of moisture, crude ash, crude lipid, crude protein and carbohydrate of three cultivars ranged from 8.43 to 9.15, 11.42 to 17.49, 0.00 to 0.00, 31.90 to 37.54, 36.30 to 42.24, respectively.

Antimicrobial and Antioxidant Activity of the Discorea alata L. (Discorea alata L.의 항균 및 항산화 활성)

  • Kwon, Jeong-Eun;Kwon, Jung-Bae;Kwun, In-Sook;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.283-288
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    • 2010
  • Yam (Dioscorea spp.) has been used as important edible and medicinal natural resource in worldwide and D. alata L. is most popular nourishment among the yam. In this study the composition, color, antioxidation and antimicrobial activity of D. alata Gyeongbuk No. 6 (GB-6), which was established in Gyeongbuk Agricultural Research & Extension Services, Andong, Korea, was compared to those of D. batatas Gyeongbuk No. 1 (GB-1), a major domestic cultivation species. Water content of GB-6 was $78.02{\pm}0.16%$, which is slightly lower than that of GB-1 ($82.6{\pm}0.07%$). The contents of crude protein, crude fat, crude fiber and ash of GB-6 were 0.95, 0.26, 0.85 and 0.70%, whereas those of GB-1 were 1.58, 0.15, 1.39 and 0.88%, respectively. Analysis of color using colormeter showed that the GB-6 is slight dark-yellow than GB-1, and total polyphenol content of GB-6 was 2-fold higher compared than that of GB-1. Sequential organic solvent fractions from methanol extract of GB-6 showed that the ethylacetate fraction has highest total polyphenol ($144.1{\pm}3.20\;mg/g$). Determination of antioxidation activity showed that the ethylacete fraction and water fraction has strong DPPH radical scavenging activity ($IC_{50}=78.32\;{\mu}g/mL$) and reducing power, respectively. In antimicrobial activity assay, the n-hexane and ethylacetate fraction showed antibacterial activity against B. subtilis, L.monocytogenes, S. epidermidis, S. aureus, P. vulgaris, and S. typhimurium. These results provide the possibility of domestic cultivated D. alata GB-6 as a healthy food.

Effects of Mulberry-Leaf Powder Tofu Consumption on Serum Lipid Profiles, Ca, Ca/P Ratio and Pb Status in Middle-Aged Women (뽕잎분말 첨가두부 섭취가 비만 중년여성의 혈청 지질, 칼슘, 칼슘/인 비율 및 납 수준변화에 미친 영향)

  • Kim, A.J.;Kim, M.H.;Chung, Kun-Sub
    • Korean Journal of Food Science and Technology
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    • v.38 no.3
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    • pp.432-437
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    • 2006
  • This study was carried out to investigate the effects of mulberry-leaf powder Tofu (MPT) on serum lipids profiles, Ca levels, Ca/P ratio and Pb levels in 30 middle aged obese women living in the Choongnam area. 100 g/day MPT was consumed for 4 weeks. The nutrient contents per 100 g MPT were 86.71 kcal (energy), 8.98 g protein, 0.53 mg fiber, 211.33 mg Ca and 1.59 g fat. Anthropometric measurements, 24 h recall dietary intakes, serum levels of protein, albumin, glucose, Ca and Pb, lipid profiles (cholesterol, triglyceride, HDL-cholesterol, LDL-cholesterol), and the Ca/P ratio were analyzed before and after consumption of MPT. After consumption of MPT, there were no significant differences in anthropometric measurements, the levels of serum protein, albumin, glucose, total cholesterol, HDL-cholesterol, lipase activity, HTR (HDL-cholesterol/total cholesterol), CRF (cardiac risk factor), Ca, Ca/P ratio and Pb. There were decreases in the levels of serum triglyceride, LDL-cholesterol, AI (atherogenic index) and LHR (LDL-cholesterol/HDL-cholesterol). Significantly increased dietary intakes of plant protein, total Ca, and plant Ca were observed.

Modeling net energy requirements of 2 to 3-week-old Cherry Valley ducks

  • Yang, Ting;Yu, Lexiao;Wen, Min;Zhao, Hua;Chen, Xiaoling;Liu, Guangmang;Tian, Gang;Cai, Jingyi;Jia, Gang
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.10
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    • pp.1624-1632
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    • 2020
  • Objective: A total of three hundred unsexed ducks were utilized to estimate net energy requirements of maintenance (NEm) and weight gain (NEg) for 2 to 3-week-old Cherry Valley ducks and to establish a model equation to predict NE requirements using the factorial method. Methods: To determine the apparent metabolizable energy (AME) of the diet, fifty 7-day-old ducks at approximately equal body weights (BWs) were randomly assigned into five groups that were fed at different levels (ad libitum, 85%, 75%, 65%, and 55% of ad libitum intake), and the endogenous acid-insoluble ash as indigestible marker. The two hundred and fifty 7-day-old ducks were used for a comparative slaughter experiment. At the beginning of the experiment, ten ducks were sacrificed to determine the initial body composition and energy content. The remaining ducks were randomly assigned into five groups (same as metabolic experiment). Ducks of the ad libitum group were slaughtered at 14 and 21-day-old. At the end of the experiment, two ducks were selected from each replicate and slaughtered to determine the body composition and energy content. Results: The results of the metabolizable experiment showed AME values of 13.43 to 13.77 MJ/kg for ducks at different feed intakes. The results of the comparative slaughter experiment showed the NEm value for 2 to 3-week-old Cherry Valley ducks was 549.54 kJ/kg of BW0.75/d, and the NEg value was 10.41 kJ/g. The deposition efficiency values of fat (Kf) and crude protein (Kp) were 0.96 and 0.60, respectively, and the values of efficiency of energy utilization (Kg) and maintenance efficiency (Km) were 0.75 and 0.88, respectively. Conclusion: The equation for the prediction of NE requirements for 2 to 3-week-old Cherry Valley ducks was the following: NE = 549.54 BW0.75+10.41 ΔW, where ΔW is the weight gain (g).