• 제목/요약/키워드: Mg and Pi

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Isolation and properties of protease Pi in escherichia coli (대장균 세포내 단백질 분해효소, protease Pi의 정제와 특성)

  • 이영섭;곽태환;임정빈;정진하
    • Korean Journal of Microbiology
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    • v.24 no.2
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    • pp.119-126
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    • 1986
  • A periplasmic endoprotease, named protease Pi, was purified to homogeneity from Escherkchia coli by conventional procedure with insulin as substrate. This enzyme degrades insulin and glucagon to trichloroacetic acid-soluble meterials, but shows little or no hydrolysis of bovine serum albumin, casein or globin. Its molecular weight was 110, 000 when determined by gel filtration on Sephacryl S-300 and was 105, 000 when estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Thus, it appears to be single polypeptide. This snzyme is metalloprotease, since it is completely inhibited by o-phenanthroline and can be activated by addition of divalent metal cations, such as $Mg^{2+}\;and\;Co^{2+}$. It is destinct from protease Ci, a cytoplasmic insulin degrading enzyme, since protease Pi is localized to the periplasm. Since protease Pi selectively degrades GTP cyclohydrolase I, it appears to play a role in the regulation of pteridine biosynthesis.

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Synergetic effect of soluble whey protein hydrolysate and Panax ginseng berry extract on muscle atrophy in hindlimb-immobilized C57BL/6 mice

  • Han, Min Ji;Shin, Ji Eun;Park, Seok Jun;Choung, Se-Young
    • Journal of Ginseng Research
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    • v.46 no.2
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    • pp.283-289
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    • 2022
  • Background: Sarcopenia, defined as loss of muscle mass and strength with age, becomes a public health concern as the elderly population increases. This study aimed to determine whether the mixture of soluble whey protein hydrolysate (WPH) and Panax ginseng berry extract (GBE) has a synergetic effect on sarcopenia and, if so, to identify the relevant mechanisms and optimal mixing ratio. Methods: In the first experiment, C57BL/6 mice were hindlimb immobilized for one-week and then administered WPH 800 mg/kg, GBE 100 mg/kg, WPH 800 mg/kg+ GBE 100 mg/kg mixture, and Fructus Schisandrae extract (SFE) 200 mg/kg for two weeks. In the second experiment, experimental design was same, but mice were administered three different doses of WPH and GBE mixture (WPH 800 mg/kg+ GBE 100 mg/kg, WPH 800 mg/kg+ GBE 90 mg/kg, WPH 1000 mg/kg+ GBE 75 mg/kg). Results: In the first experiment, we confirmed the synergetic effect of WPH and GBE on muscle mass and identified that GBE was more effective on the protein synthesis side, and WPH tended to be slightly more effective for protein degradation. In the second experiment, among three different ratios, the WPH 800 mg/kg+ GBE 100 mg/kg was most effective for muscle mass and strength. The mixtures activated muscle protein synthesis via PI3K/Akt/mTORc1 pathway and inhibited muscle protein degradation via suppressing ubiquitin-proteasome system (UPS) and autophagy-lysosome system (ALS), and these effects were more GBE dose-dependent than WPH. Conclusion: The WPH and GBE mixture having a synergetic effect is a potential agent to prevent sarcopenia.

Changes in the serum immunoglobulin levels and viral antibody titers of colostrum-conferred Korean native calves during the first 12 weeks postpartum (초유를 섭취한 한우 송아지의 출생후 12주 동안의 혈청 면역글로불린과 각종 바이러스 항체가의 변화)

  • Kim, Doo;Han, Hong-ryul
    • Korean Journal of Veterinary Research
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    • v.29 no.2
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    • pp.83-90
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    • 1989
  • The changes in serum total protein and immunoglobulin levels, and BVD, IBR and PI-3 viral neutralizing antibody titers in colostrum-conferred Korean native calves during the first 12 weeks postpartum were studied, and the results obtained were summerized as follows: The Mean concentration of total protein, total immunoglobulin, IgG, IgM and IgA in sera of 9 calves at birth were $3.8{\pm}0.5g/dl$, $0.27{\pm}0.15mg/ml$, $0.06{\pm}0.08mg/ml$, $0.21{\pm}0.11mg/ml$, and extremely low concentration, respectively. Serum total protein level reached a maximum at 20 hours after birth, total immunoglobulin, IgG, and IgM levels at 24 hours, and IgA level at 28 hours, respectively. Serum IgA level reached a minimum at 4 weeks old, IgM level at 5 weeks, total immunoglobulin level at 8 weeks, and IgG level at 10 weeks, respectively. After then those levels had begun to increase, but total protein level was still decreasing at 12 weeks old. The half-lives of IgG, IgM, and IgA were 21.1 days, 4.0 days, and 2.6 days-respectively. In 10 Korean native cows immediately after parturition, serum neutralizing antibody titers specific to BVD, IBR and PI-3 virus were $8.7{\pm}1.5{\log}_2$, $5.7{\pm}1.2{\log}_2$, and $6.8{\pm}1.01{\log}_2$, respectively. And colostral neutralizing antibody titers against BVD, IBR, and PI-3 virus were $10.1{\pm}1.4{\log}_2$, $6.8{\pm}1.3{\log}_2$ and $7.8{\pm}1.7{\log}_2$, respectively. Before suckling the colostrum, SN antibody titers against BVD, IBR, and PI-3 virus were undetectable from all of 9 Korean native calves. Nevertheless SN antibody titer against BVD virus reached a maximum level ($9.2{\pm}0.6{\log}_2$) at 24 hours after birth, that against IBR virus ($6.1{\pm}1.0{\log}_2$) at 20 hours after birth, and that against PI-3 virus ($6.8{\pm}0.9{\log}_2$) at 32 hours after birth, respectively. In 12 weeks old calves, the SN antibodies against BVD and IBR virus were still decreasing, but that against PI-3 virus reached a minimum at 10 weeks, and increased after 12 weeks of age. The half-lives of SN antibodies against BVD, PI-3 and IBR, virus were 16.0 days, 22.6 days, and 25.5 days, respectively.

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Anti-diabetic Effect of Wen-Pi-Tang-Hab-Wu-Ling-San Extract in Streptozotocin-induced Diabetic Rats (Streptozotocin으로 유도한 당뇨병 쥐에서 $WHW^{(R)}$의 항당뇨 효과에 대한 연구)

  • Bae, Hyo-Sang;Nam, Jung-Ki;Jung, Jun-Ki;Oh, Seung-Yeol;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.85-91
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    • 2008
  • Objectives : This study aimed to evaluate the anti-diabetic effect of Wen-Pi-Tang-Hab-Wu-Ling-San (WHW) extract in streptozotocin(STZ)-induced type-1 diabetic rats. Methods : Experimental diabetes were induced by intraperitoneal injection of streptozotocin (60 mg/kg). Two groups of STZ-induced diabetic rats were given the following treatments for 2 weeks by oral Administrations : (1) WHW 10 mg/kg, (2) WHW 100 mg/kg. In addition, vehicle-treated diabetic and nondiabetic controls were used in the experiment. The effects of WHW extract on STZ-induced diabetes were observed by measuring the changes of body weights and the levels of fasting blood glucose, insulin, urea nitrogen (BUN) and creatinine level in sera of rats, respectively. Results : In comparison control group, WHW-treated groups (100 mg/kg) were significantly decreased fasting blood glucose levels and increased serum insulin levels in STZ-induced diabetic rats. Moreover, WHW-treated groups (100 mg/kg) were reduced s-creatinie levels in STZ-induced diabetic rats. In addition, the changes related to diabetic nephropathy with body weight were significantly lower in WHW extract-dosing groups than in the diabetic control. Conclusions : The study thus showed that WHW extract enhanced the anti-diabetic effect in STZ-induced diabetic rats by improving the hypoglycemia. It also increased pancreatic insulin content in these rats.

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Characterization of Cordierite by XPS (Ⅰ) (XPS에 의한 코디에리트의 특성 연구 (연구Ⅰ))

  • Han, Byoung-Sung
    • Journal of the Korean Institute of Telematics and Electronics
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    • v.26 no.3
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    • pp.124-129
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    • 1989
  • The cordierite is of great interest for microelectronic packaging. Its main advantages are low dielectric constant and low thermal expansion. The cordierite precursor obtained by sol-gel synthesis whose sintering temperature is about $900^{circ}C$ is an amorphous and cristal white powder. Green and fired cordierite samples were studied by XPS for microscopic properties. At the surface the results of XPS show forte diminution of Mg in comparison with its value at volume and the deficit of Mg compensates by augmentation of Al and Si. $pi$-cordierite phase is present near the surface $<100{\AA}$ and small quantities of magnesium aluminate ($MgAl_2O_4) is present in the bulk. Sintering of the green cordierite introduces no chemical modification at the surface.

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Transfer and Expression of a Tomato Inhibitor II and $\beta$-Glucuronidae Fusion Gene in Flowering Cabbage, Brassica oleracea var. acephala DC. (꽃양배추로의 프로타제 저해제 II와 $\beta$-Glucuronidase 융합 유전자의 도입 및 발현)

  • 김창길;정재동;안진흥;김경민
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.1
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    • pp.37-43
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    • 1998
  • The highest transformation frequency was observed when cotyledonary and hypocotyl explants of flowering cabbage (Brassica oleracea var. acephala DC) 'Eunbae' were cultured on shoot induction medium without kanamycin for 1 day, then cocultured with Agrobacterium tumefaciens LBA4404;;pGA1036 harboring tomato inhibitor II promoter and $\beta$-glucuronidae (GUS) fusion gene for 3 days. These explants were transferred to MS medium containing 20 mg/L kanamycin, 500 mg/L carbenicillin, and 1 mg/L BA. The explants were subsequently subcultured every 2 weeks. Incorporation of the GUS gene into flowering cabbage was confirmed by PCR analysis of DNA. Southern blot analysis showed that ECL-labeled GUS gene was hybridized to the expected amplified genomic DNA fragment of about 366 bp from transgenic flowering cabbage. Histochemical analysis based on the enzymatic activity of the GUS protein indicated that PI-II promoter activity was sysmatically associated with vascular tissue in wonded as well as in non-wounded leaves, petioles and stems, but not in roots. Partial wounding with razor blade showed not systemic induction but partial induction.

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Properties and Thermostability of Gelatin-degrading Proteinases in the Fruit of Actinidia chinensis (Kiwifruit) (Kiwifruit 과육에 존재하는 단백질분해효소의 특성과 열안정성)

  • 오순자;김성철;고석찬
    • Journal of Life Science
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    • v.12 no.6
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    • pp.752-758
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    • 2002
  • This study was investigated on properties and thermostability of gelatin-degrading proteinases in the fruit of Actinidia chinensis (kiwifruit) for the industrial application. Three gelatin-degrading proteinases (PI, PII and PIII) were detected from the pulp of fruits. The molecular weights of these proteinases, PI, PII and PIII, were approximately 220 kD, 51 kD, and 26 kD respectively, on the basis of gelatin-containing SDS-PACE. The optimum pH of these proteinases ranged from 2.0 to 5.0 with a maximal activity at pH 4.0. These proteinases had a high sensitivity to E-64 and iodoacetate which are cysteine protease inhibitors, and required DTT, cysteine, and $\beta$-mercaptoethanol for their activities which are stimulators for cysteine proteases. These results indicate that these proteinases are cysteine proteinases and the proteinase PIII is actinidin (EC 3.4.22.14), based on the molecular weight and/or susceptibility against proteinase inhibitors. These proteinases were strongly activated by $Ca^{2+}$, $Mg^{2+}$ and $Mn^{2+}$, whereas strongly inhibited by Zn$^{2+}$ and Hg$^{2+}$. However, these proteinases have slightly different susceptibility against other cations ($Ca^{2+}$, $Cu^{2+}$, $Al^{3+}$, $Ca^{3+}$. The temperature stability of proteinase PIII was more stable than proteinases PI and PII. Moreover, proteinase PIII remained stable below $50^{\circ}C$ for 48hr, showing the residual activity above 75% of the enzyme activity.

The Effect of Gingko Biloba Extract on Energy Metabolic Status in C3H Mouse Fibrosarcoma : Evaluated by in vivo $^{31}P$ Magnetic Resonance Spectroscopy (Gingko biloba extract가 C3H 마우스 섬유육종의 에너지 대사 상태에 미치는 영향 : 생체내 $^{31}P$ 자기공명 분광법을 이용한 분석)

  • Ha Sung Whan;Kim Won Dong;Ahn Yong Chan;Park Chan Il;Lim Tae Hwan;Lee Tae Kuen
    • Radiation Oncology Journal
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    • v.20 no.2
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    • pp.147-154
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    • 2002
  • Purpose : Gingko biloba extract (GBE), a natural product extracted from Gingko leaves, is known to increase the radiosensitivity of tumors. This radiosensitization probably arises from the increase in the peripheral blood flow by decreasing the blood viscosity and relaxing the vasospasm. The influence of a GBE on the metabolic status in fibrosarcoma II (FSall) of a C3H mouse was investigated using $^{31}P$ magnetic resonance spectroscopy (MRS). Materials and Methods : Eighteen C3H mice with fibrosarcoma II $(from\;100\;mm^3\;to\;130\;mm^3)$ were prepared for this experiment. The mice were divided into 2 groups; one (9 mice) without a priming dose, and the other (9 mice) with a priming dose of GBE. The GBE priming dose (100 mg/kg) was administered by an intraperitoneal (i.p.) injection 24 hours prior to the measurement. First $^{31}P$ MRS spectra were measured in the mice from each group as a baseline and test dose of GBE (100 mg/kg) was then administered to each group. One hour later, the $^{31}P$ MRS spectra were measured again to evaluate the change in the energy metabolic status. Results : In the group without the priming dose, the mean pH, PCr/Pi, PME/ATP, Pi/ATP, PCr/(Pi+PME) values 1 hour after the test dose were not changed significantly compared to the values at the baseline. However, in the group with the priming dose, the mean PCr/Pi, Pi/ATP, PCr/(Pi+PME) values 1 hour after the test dose changed from the baseline values of 0.49, 0.77, 0.17 to 0.74, 0.57, 0.28 respectively. According to the paired t-test, the differences were statistically significant. Conclusion : The above findings suggest that the metabolic status is significantly improved after administering GBE if the priming dose is given 24 hours earlier. This shows that the radiosensitizing effect of GBE is based on the increase of tumor blood flow and the improvement in the metabolic status.

Analysis of Cholangitis Rates with Extended Perioperative Antibiotics and Adjuvant Corticosteroids in Biliary Atresia

  • Goh, Lynette;Phua, Kong Boo;Low, Yee;Chiang, Li Wei;Yong, Chen;Chiou, Fang Kuan
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.24 no.4
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    • pp.366-376
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    • 2021
  • Purpose: There is no consensus regarding adjuvant therapies following Kasai portoenterostomy (KP) for biliary atresia (BA). This study aimed to analyze the effect of extended perioperative intravenous antibiotics (PI-Abx) and adjuvant corticosteroid on cholangitis and jaundice clearance rates in the 3 years post-KP in children with BA. Methods: Data of patients who underwent KP between 1999-2018 at a single center were retrospectively analyzed. Group A (1999-2010) received PI-Abx for 5 days, Group B (2010-2012) received PI-Abx for 5 days plus low-dose prednisolone (2 mg/kg), and Group C (2012-2017) received PI-Abx for 14 days plus high-dose prednisolone (5 mg/kg). Results: Fifty-four patients were included with groups A, B, and C comprising 25, 9, and 20 patients, respectively. The number of episodes of cholangitis was 1.0, 1.6, and 1.3 per patient (p=NS) within the first year and 1.8, 2.3, and 1.7 (p=NS) over 3 years in Groups A, B, and C, respectively. The jaundice clearance rate at 6 months was 52%, 78%, and 50% (p=NS), and the 3-year native liver survival (NLS) rate was 76%, 100%, and 80% (p=NS) in Groups A, B, and C, respectively. A near-significant association was observed between the incidence of cholangitis within the first year and decompensated liver cirrhosis/death at 3 years post KP (p=0.09). Persistence of jaundice at 6 months was significantly associated with decompensated cirrhosis/death at 3 years (p<0.001). Conclusion: The extended duration of PI-Abx and adjuvant corticosteroids was not associated with improved rates of cholangitis, jaundice clearance, or NLS in patients with BA.

Development of Fibrinolytic Agents from Snake Venoms

  • 김영식;한범수;장일무
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.279-279
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    • 1994
  • Fibrinolytic proteases, piscivorase I (PI) and piscivorase II (PII), were isolated from Agkistrodon piscivorus piscivorus (eastern cotonmouth moccasin) venom using gel filtration on Bio-Gel P100 and ion-exchange chromatography on CM-Sepharose. The molecular welghts of two proteases were approximately 23400 and 29000. Their isoelectric points 6.6 and 8.5, respectively. The partial amino acid sequences of PI were characterized by tryptic digestion. PI readily cleaves the A${\alpha}$-and B${\beta}$-chaln of fibronogen, but PII rapidly cleaves A${\alpha}$-chain and more slowly the B${\beta}$-chain, They were activated by Ca$\^$2+/, Mg$\^$2+/ and Ba$\^$2+/, but inhibited by Zn$\^$2+/, Cu$\^$2+/ and Mn$\^$2+/. Two enzymes were also inhibited by cysten, ${\beta}$-mercapto -ethanol, and by metal chelators such as EDTA and EGTA, but not by benzamidine, PMSF, soybean trypsin inhibitor and aprotinin. They did not act like thrombin, plasmin and kallikrein, using specific chromogenllc substrates. Two protease did not induce platelet aggregation. PI showed low hemorrhagic activity at dosage of 50 $\mu\textrm{g}$.

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