• Journal of Microbiology and Biotechnology
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• 제11권5호
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• pp.838-844
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• 2001

The enzyme Fuc aldolase from Methanococcus jannaschii that catalyzes the aldol condensation of DHAP and L-lactaldehyde to give fuculose-1-phosphate was inactivated by DEP. The inactivation was pseudo first-order in the enzyme and DEP, which was biphasic. A pseudo second-order rate constant of 120$M^{-1}min^{-1}$ was obtained at pH 6.0 and $25{\circ}C$. Quantifying the increase in absorbance at 240nm showed that four histidine residues per subunit were modified during the nearly complete inactivation. The statistical analysis and the time course of the modification suggested that two or three histidine residues were essential for activity. The rate of inactivation was dependent on the pH, and the pH inactivation data implied the involvement of the amino acid residue with a $pK_a$ value of 5.7. Fuc aldolase was protected against DEP inactivation by DHAP, indicating that the histidine residues were located at the active site of Fuc aldolase. DL-Glyceraldehyde, as an alternative substrate to L-lactaldehyde, showed no specific protection for the Fuc aldolase.

• 한국미생물·생명공학회지
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• 제20권1호
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• pp.61-67
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• 1992

유기용매 이상계에서의 thermolysin을 사용하여 아스파탐 전구체 합성시 pH, 온도, 기질농도, 그리고 유기용매상에 대한 수용액상의 부비피 ($\alpha$)등의 변화에 따른 기질의 분해 반응, 효소의 안정성, 그리고 Z-APM 합성에 미치는 복합적인 영향을 조사함으로써 반응조건의 최적화를 도모하였다. 유기용매 이상계에서의 L-PM.HCL의 자연분해는 수용액에서보다 훨씬 느리게 일어나며, 또한 $\alpha$가 증가할 수록 분해속도가 감소하는 것을 알 수 있었다.

• BMB Reports
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• 제32권5호
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• pp.515-520
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• 1999

Incubation of two types of glutamate dehydrogenase isoproteins from bovine brain with the arginine-specific dicarbonyl reagent phenylglyoxal resulted in a biphasic loss of enzyme activity. Reaction of the glutamate dehydrogenase isoproteins with phenylglyoxal caused a rapid loss of 53~62% of the enzyme activities and modification of two residues of arginine per enzyme subunit. Prolonged incubation of the glutamate dehydrogenase isoproteins with phenylglyoxal resulted in the modification of an additional four residues of arginine per enzyme subunit without further loss of the residual activities. Partial protection against inactivation was provided by the coenzyme NADH or substrate 2-oxoglutarate. The most marked decrease in the rate of inactivation was observed by the combined addition of NADH and 2-oxoglutarate, suggesting that the first two modified arginine residues are in the vicinity of the catalytic site. However, inactivation of the glutamate dehydrogenase isoproteins by phenylglyoxal appears to be partial with approximately 40% activity remained after an extended reaction time with excess reagent, suggesting that the modified arginine residues may not be directly involved in catalysis. The lack of complete protection by substrates also suggest the possibility that the modified arginine residues are not directly involved at the active site, and the partial loss of activity by the modification of arginine residues may be due to a conformational change. There were no significant differences between the two glutamate dehydrogenase isoproteins in sensitivities to inactivation by phenylglyoxal, indicating that the microenvironmental structures of the glutamate dehydrogenase isoproteins are very similar to each other.

• Journal of Acupuncture Research
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• 제18권1호
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• pp.76-87
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• 2001

Objective : Zusanli(ST36) and Hoku(Li4) are analgesic acupuncture points frequently used for acupuncture in Oriental medicine. The present study was conducted to see the antinociceptive effects produced by electroacupuncture combined two frequencies(Low, High) and two different acupuncture points(LI4, ST36) in the rat tail flick test. Method : In this study the Rats (Sprague-Dawley, 250-300g) were partially anesthetized with thiopental sodium(40mg/kg, i.p.). The basal reaction time for the tail-flick was 3${\pm}$0.5 sec. Low frequency(3Hz, 5V, biphasic) and high frequency(100Hz, 5V, biphasic) were applied to the inserted needle for the period of insertion(twenty minutes). Experimental groups are divied as follow; a) electroacupuncture stimulation groups at Hoku with or high frequency(L-EA, H-EA), b) electroacupuncture stimulation groups at Zusanli with low or high frequency(1-EA, h-EA), c) low frequency at Hoku and Zusanli(LIEA), d) low frequency at Hoku and high frequency at Zusanli(LhEA), e) high frequency at Hoku and low frequency at Zusanli(HIEA), f) high frequency at Hoku and Zusanli(HhEA) Results : The individual stimulation at either Hoku or Zusanli with low frequency has stronger and longer analgesic effect than high frequency stimulation. In addition, the combined stimulation at Hoku and Zusanli with low frequency has superior effect to individual stimulation with low frequency. LhEA and LIEA have superior effect to other stimulation groups among the combined groups. In order to determine the involvement of opioid system on the different antinociceptive effects, Naloxone, an opioid antagonist, was used in the combined groups. LIEA is the most sensitive when naloxone was administrated among study groups. HhEA is the least sensitive in the administration of naloxone. Conclusion : From results, this study confirmed that the opioid system is involved in analgesic effect of low frequency stimulation of acupuncture point, and we also can suggest the stronger analgesic effect of combining stimulation points is due to the theory of spatial summation in the nervous system.

• Journal of Periodontal and Implant Science
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• 제50권1호
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• pp.14-27
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• 2020

Purpose: To overcome several drawbacks of chemically-crosslinked collagen membranes, modification processes such as ultraviolet (UV) crosslinking and the addition of biphasic calcium phosphate (BCP) to collagen membranes have been introduced. This study evaluated the efficacy and biocompatibility of BCP-supplemented UV-crosslinked collagen membrane for guided bone regeneration (GBR) in a rabbit calvarial model. Methods: Four circular bone defects (diameter, 8 mm) were created in the calvarium of 10 rabbits. Each defect was randomly allocated to one of the following groups: 1) the sham control group (spontaneous healing); 2) the M group (defect coverage with a BCP-supplemented UV-crosslinked collagen membrane and no graft material); 3) the BG (defects filled with BCP particles without membrane coverage); and 4) the BG+M group (defects filled with BCP particles and covered with a BCP-supplemented UV-crosslinked collagen membrane in a conventional GBR procedure). At 2 and 8 weeks, rabbits were sacrificed, and experimental defects were investigated histologically and by micro-computed tomography (micro-CT). Results: In both micro-CT and histometric analyses, the BG and BG+M groups at both 2 and 8 weeks showed significantly higher new bone formation than the control group. On micro-CT, the new bone volume of the BG+M group (48.39±5.47 ㎣) was larger than that of the BG group (38.71±2.24 ㎣, P=0.032) at 8 weeks. Histologically, greater new bone area was observed in the BG+M group than in the BG or M groups. BCP-supplemented UV-crosslinked collagen membrane did not cause an abnormal cellular reaction and was stable until 8 weeks. Conclusions: Enhanced new bone formation in GBR can be achieved by simultaneously using bone graft material and a BCP-supplemented UV-crosslinked collagen membrane, which showed high biocompatibility and resistance to degradation, making it a biocompatible alternative to chemically-crosslinked collagen membranes.

• The Korean Journal of Pain
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• 제14권1호
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• pp.1-6
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• 2001

Background: Subcutaneous injection of 5% formalin into the hind paw of the rat produces a biphasic nociceptive response. The second phase depends on changes in the dorsal horn cell function that occur shortly after an initial C-fiber discharge, spinal sensitization, or windup phenomenon. This study was performed to investigate the role of glutamate during spinal sensitization. Methods: Sprague-Dawley rats weighing 200 to 250 g were used for this study. Under light anesthesia (0.5% isoflurane) the rats were segregated in a specially designed cage and $50{\mu}l$ 0.5% formalin was injected subcutaneously in the foot dorsum of right hindlimb. Forty minutes after the formalin injection, the rat was quickly decapitated and spinal cord was removed. The spinal segments at the level of L3 (largest area) was collected and stored in a deep freezer ($-70^{\circ}C$). The mRNA gene expression of N-methyl-D-aspartate receptor (NMDAR) and the metabotropic glutamate receptor subtype 5 (mGluR5) were determined by the polymerase chain reaction. Results: The number of flinches was $19.8{\pm}2.3/min$. at one minute after formalin injection and decreased to zero after then. The second peak appeared at 35 and 40 minutes after formalin injection. The values were $17.8{\pm}2.2$ and $17.2{\pm}3.0/min$. The mRNA gene expressions of NMDAR and mGluR5 were increased by $459.0{\pm}46.8%$ (P < 0.01) and $111.1{\pm}4.8%$ (P > 0.05) respectively at 40 minutes after formalin injection. The increased rate of NMDAR was significantly higher than that of mGluR5 (P < 0.01). Conclusions: From these results it suggested that NMDAR partly contributed to the mechanism of central sensitization after the formalin test but mGluR5 did not.

• BMB Reports
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• 제28권1호
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• pp.72-78
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• 1995

Time-dependent inactivation of E. coli GTP cyclohydrolase I with a 2',3'-dialdehyde derivative of GTP (oGTP) was directed to the active site of the enzyme, and was dependent on the concentration of oGTP. The kinetics of inactivation were biphasic with a rapid reaction occurring immediately upon exposure of the enzyme to oGTP followed by a slow rate of inactivation. The $K_i$ value of oGTP for the enzyme was 0.25 mM. Inactivation was prevented by preincubation of the enzyme with GTP, the substrate of the enzyme. At 100% inactivation, 2.3 mol of [8.5'-$^3H$]oGTP were bound per each enzyme subunit, which consists of two identical polypeptides. The active site residue which reacted with the affinity label was lysine. oGTP interacted selectively with the ${\varepsilon}$-amino group of lysine in the GTP-binding site to form a morpholine-like structure which was stable without sodium borohydride treatment. However, triphosphate group was eliminated during the hydrolysis step. To identify the active site of the enzyme, [8.5'-$^3H$]oGTP-labeled enzyme was cleaved by endoproteinase Lys-C, and the $^3H$-labeled peptide was purified by HPLC. The amino acid sequence of the active site peptide was Pro-Ser-Leu-Ser-Lys, which corresponds to the aminoterminal sequence of the enzyme.

• The Korean Journal of Pain
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• 제30권2호
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• pp.98-103
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• 2017

Background: The root of Peucedanum japonicum Thunb., a perennial herb found in Japan, the Philippines, China, and Korea, is used as an analgesic. In a previous study, sec-O-glucosylhamaudol (SOG) showed an analgesic effect. This study was performed to examine the antinociceptive effect of intrathecal SOG in the formalin test. Methods: Male Sprague-Dawley rats were implanted with an intrathecal catheter. Rats were randomly treated with a vehicle and SOG ($10{\mu}g$, $30{\mu}g$, $60{\mu}g$, and $100{\mu}g$) before formalin injection. Five percent formalin was injected into the hind-paw, and a biphasic reaction followed, consisting of flinching and licking behaviors (phase 1, 0-10 min; phase 2, 10-60 min). Naloxone was injected 10 min before administration of SOG $100{\mu}g$ to evaluate the involvement of SOG with an opioid receptor. Dose-responsiveness and ED50 values were calculated. Results: Intrathecal SOG showed a significant reduction of the flinching responses at both phases in a dose-dependent manner. Significant effects were showed from the dose of $30{\mu}g$ and maximum effects were achieved at a dose of $100{\mu}g$ in both phases. The ED50 value (95% confidence intervals) of intrathecal SOG was 30.3 $(25.8-35.5){\mu}g$ during phase 1, and 48.0 (41.4-55.7) during phase 2. The antinociceptive effects of SOG ($100{\mu}g$) were significantly reverted at both phases of the formalin test by naloxone. Conclusions: These results demonstrate that intrathecal SOG has a very strong antinociceptive effect in the formalin test and it seems the effect is related to an opioid receptor.

• The Korean Journal of Physiology and Pharmacology
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• 제3권3호
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• pp.251-261
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• 1999

The effects of carnosine and related compounds (CRCs) including anserine, homocarnosine, histidine, and ${\beta}-alanine$ on monosaccharide autoxidation and $H_2O_2$ formation were investigated. The incubation of CRCs with D-glucose, D-glucosamine, and D, L-glyceraldehyde at $37^{\circ}C$ increased the absorption maxima at 285 nm, 273 nm, and $290{\sim}330$ nm, respectively. D, L-glyceraldehyde was the most reactive sugar with CRCs. The presence of copper strongly stimulated the reaction of carnosine and anserine with D-glucose or D-glucosamine. Carnosine and anserine stimulated $H_2O_2$ formation from D-glucose autoxidation in a dose-dependent manner in the presence of 10 ${\mu}M$ Cu (II). The presence of human serum albumin (HSA) decreased their effect on $H_2O_2$ formation. Carnosine and anserine has a biphasic effect on ${\alpha}-ketoaldehyde$ formation from glucose autoxidation. CRCs inhibited glycation of HSA as determined by hydroxymethyl furfural, lysine residue with free ${\varepsilon}-amino$ group, and fructosamine assay. These results suggest that CRCs may be protective against diabetic complications by reacting with sugars and protecting glycation of protein.

• Korean Chemical Engineering Research
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• 제48권5호
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• pp.643-648
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• 2010

본 연구에서는 이온성 액체인 염화콜린에 5가지의 금속염화물을 첨가하여 루이스산 이온성 액체 촉매를 제조하고, 이 촉매를 사용하여 대두유로부터 바이오디젤을 합성하였다. 먼저 단독의 금속염화물인 염화주석과 염화아연, 염화알루미늄, 염화철(III), 염화구리(I) 촉매에 대하여 363~423 K 온도 범위에서 에스터 교환 반응의 반응성을 조사하였다. 5가지의 금속염화물 중 염화주석이 우수한 촉매 활성을 나타내었고, 이러한 경향과 같이 5가지의 루이스산 이온성 액체 촉매 중 $[Me_3NC_2H_4OH]Cl-2SnCl_2$의 촉매가 403 K에서 14시간 동안 유지:메탄올:촉매의 몰 비율 1:12:0.9인 조건으로 최대 91.1%의 높은 반응수율을 나타내었다. 단독의 염화주석 촉매와는 달리, $[Me_3NC_2H_4OH]Cl-2SnCl_2$의 촉매는 반응 후 액체-액체 이상계를 형성하여 반응물과 생성물로부터 쉽게 분리할 수 있으며, 5회 이상 재사용 후에도 활성이 거의 감소하지 않았다. 이러한 결과는 촉매의 수분에 대해 안정성과 강한 루이스 산성도의 특성에 기인한 것으로 생각된다. 또한 촉매에 대한 반응시간과 촉매 및 메탄올 몰 비율 등의 반응변수들에 대한 영향이 조사되었다.