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Determination of Microquantities of Ammonia by Enzymatic Analysis (효소분석법에 의한 미량암모니아의 정량)

  • 성하진;양한철
    • Microbiology and Biotechnology Letters
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    • v.14 no.6
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    • pp.495-500
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    • 1986
  • Enzymatic micro-assay methods were studied those were capable of determining ammonia down to 10$^{-5}$M(0.01 $\mu$mole/ml) in the presence of other nitrogenous compounds such as protein and amino acid. Microquantities of ammonia (0.01-0.1 $\mu$mole) could be determined indirectly by measuring phosphorous, one of the products of the enzymatic reaction catalyzed by glutamine synthetase. In this reaction, L-glutamate, ATP and ammonium chloride were used as substrates, and phosphorous was formed in propotion to the concentration of ammonium chloride In the reaction mixture. Another procedure was examined in which glutamine synthetase reaction coupled with pyruvate kinase and lactate dehydrogenase reactions was used. One mililiter of the assay mixture contained; phosphoenol pyruvate, 3 mM, L-glutamate, 10 mM; ATP, 1mM: MgSO$_4$, 20 mM: KCl, 75mM: NADH, 0.2mM: Tris-HCl buffer(pH 7.0), 100mM; pyruvate kinase, 10 U: lactate dehydrogenase, 12 U and glutamine synthetase, 4 U. After preincubation for 20 min at 3$0^{\circ}C$, NH$_4$Cl was added and the rates of NADH oxidation were followed at 340nm. The effective range of this method was proved to be from 0.01 to 0.05 $\mu$mole/$m{\ell}$. Glutamine synthetase reaction coupled with glutamate synthase reaction could also be effectively used for determining microquantities of ammonia. The one mililiter assay mixture contained; ATP, 5mM: L-glutamate, 5mM; L-ketoglutarate, 5mM; MgCl$_2$, 15mM; NADPH, 0.15mM; Tris-HCl buffer(pH 7.0); 100mM; glutamine synthetase, 1U and glutamate synthase, 0.5U. After preincubation for 20min at 3$0^{\circ}C$ NH$_4$Cl was added and the rates of NADPH oxidation were followed at 340nm. The effective range of this procedure was appeared to be from 0.01 to 0.05$\mu$mole/$m{\ell}$.

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Marine Terrace of Daebo-Guryongpo-Gampo, SE Korea(II) (대보-구룡포-감포 지역의 해안단구(II))

  • 최성자
    • Economic and Environmental Geology
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    • v.37 no.2
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    • pp.245-253
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    • 2004
  • The 36km-long Daebo-Campo coast has a well-developed marine terraces divided to six steps by elevation of paleoshoreline : 0.5 m(T1), 10 m(T2), 30 m(T3), 40 m(T4), 60 m(T5) and 75 m(T6). The 2$^{nd}$ and 3$^{nd}$ platforms in Daebo to Guryongpo are wider and more distinctive than those of Guryongpo to Gampo. The 3$^{nd}$ terrace of 30 m high is subdivided to two flights as lower(T3b) and upper(T3a) by old sea cliff. Platform age is unclear because of coral fossil free. However, the terrace age could be determined with convergent OSL ages from beach sediments on 2$^{nd}$ step(T2). OSL ages of the terrace of 10 m high range in 60-70 ka. It reveals that the 2$^{nd}$ -step platform correlates to Oxygen Isotope Time scale, substage 5a(ca. 80 ka), and that uplift rate is ca. 0.19 m/ka for 2$^{nd}$ terrace at Daebo-Campo coast. If considering equivalent uplift rate for all terraces since the Late Pleistocene, the 3$^{rd}$ and 4$^{th}$ terraces would be 5e substage and 7 stage. The 30 m-high terrace provides a good indicator for uplift at Daebo-Gampo coast since 125,000 yrs(MIS 5e). It suggests that the local neotectonic deformation might cause an optional uplift rate of ca. 0.19 m/ka along the SE coast of Korea.

Determination of the Optimum Sampling Area for the Benthic Community Study of the Songdo Tidal Flat and Youngil Bay Subtidal Sediment (송도 갯벌과 영일만 조하대 저서동물의 군집조사를 위한 적정 채집면적의 결정)

  • Koh, Chul-Hwan;Kang, Seong-Gil;Lee, Chang-Bok
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.4 no.1
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    • pp.63-70
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    • 1999
  • The optimum sampling area which can be applied to the benthic community study is estimated from large survey data in the Songdo tidal flat and subtidal zone of Youngil Bay, Korea. A total of 250 samples by 0.02 $m^2$ box corer for the benthic fauna in Songdo tidal flat and 50 samples by 0.1 $m^2$ van Veen grab in Youngil Bay were taken from the total sampling area of 5 $m^2$. It was assumed that the sampling area could contain sufficient information on sediment fauna, if cumulative number of species, ecological indices, and similarity index by cluster analysis reflect the similarity level of 75% to those found at total sampling area (5 $m^2$). A total of 56 and 60 species occurred from Songdo tidal flat and Youngil Bay, respectively. The cumulative curve of the species number ($N_{sp}$) as a function of the sampling area (A in $m^2$ ) was fitted as $N_{sp}=37.379A^{0.257}$ ($r^2=0.99$) for intertidal fauna and $N_{sp}=40.895A^{0.257}$ ($r^2=0.98$) for subtidal fauna. Based on these curves and 75% of similarity to the total sampling area (5 $m^2$), the optimum sampling area was proposed as 1.6 $m^2$ for the intertidal and 1.5 $m^2$ for the subtidal fauna. Ecological indices (species diversity, richness, evenness and dominance indices) were again calculated on the basis of species composition in differently simulated sample sizes. Changes in ecological indices with these sample sizes indicated that samplings could be done by collecting fauna from < 0.5 $m^2$-1.5 $m^2$ on the Songdo tidal flat and from < 0.5 $m^2$-1.2 $m^2$ in Youngil Bay. Changes in similarity level of all units of each simulated sample size showed that sampling area of 0.3 $m^2$ (Songdo tidal flat) and 0.6 $m^2$ (Youngil Bay) should be taken to obtain a similarity level of 75%. In conclusion, sampling area which was determined by cumulative number of species, ecological indices and similarity index by cluster analysis could be determined as 1.5 $m^2$ (0.02 $m^2$ box corer, n=75) for Songdo tidal flat and 1.2 $m^2$ (0.1 $m^2$ van Veen grab, n=12) for Youngil Bay. If these sampling areas could be covered in the field survey, population densities of seven dominant species comprising 68% of the total faunal abundance occurring on Songdo tidal flat and six species comprising 90% in Youngil Bay can be estimated at the precision level of P=0.2.

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Antioxidative Activity of Rumex crispus L. Extract (소리쟁이 추출물의 항산화 효능)

  • Rhim, Tae-Jin;Choi, Moo-Young;Park, Hee-Juhn
    • Korean Journal of Plant Resources
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    • v.25 no.5
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    • pp.568-577
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    • 2012
  • The objective of this study was to investigate the antioxidative capacity of ethanol extracts from Rumex crispus L. The concentration of R. crispus L. extract at which DPPH radical scavenging activity was inhibited by 50% was 2.15 mg/mL, which was lower than that of ${\alpha}$-tocopherol (0.43 mg/mL), as compared to 100% by pyrogallol as a reference. Total antioxidant status was examined by total antioxidant capacity against ABTS radical reactions. Total antioxidant capacities of R. crispus L. extract at concentrations of 0.1 and 1 mg/mL were 0.47 and 2.33 mM Trolox equivalents, respectively, which were higher than those of ${\alpha}$-tocopherol. Superoxide scavenging activities of R. crispus L. extract at concentrations of 0.1 and 1 mg/mL were 21.5 and 78.9%, respectively, which were not significantly (p>0.05) different from those of catechin. Oxygen radical absorbance capacities of R. crispus L. extract at concentrations of 20 and 100 ${\mu}g/mL$ were 62.5 and 156.4 ${\mu}M$ Trolox equivalents, respectively, which were lower than those of ascorbic acid. Cupric reducing antioxidant capacities of R. crispus L. extract at concentrations of 0.1 and 1 mg/mL were 0.28 and 1.88 mM Trolox equivalents, which were similar or significantly (p<0.05) higher than those of ${\alpha}$-tocopherol, respectively. R. crispus L. extract prevented supercoiled DNA strand breakage induced by hydroxyl radical and peroxyl radical. Total phenolic contents of R. crispus L. extract at concentrations of 0.5 and 5 mg/mL were 0.58 and 3.85 mM gallic acid equivalents, respectively. R. crispus L. extract at concentration of 0.1 and 0.5 mg/mL inhibited 0.2 mM tert-butyl hydroperoxide-induced cytotoxicity by 38.5 and 63.5%, respectively, in HepG2 cell culture system. Thus, strong antioxidant and cytotoxicity-inhibiting effects of R. crispus L. extract seem to be due to, at least in part, the prevention from free radicals-induced oxidation as well as high levels in total phenolic contents.

Effects of Temperature, Diluents, and Plastic Tubes on the Motility and Acrosome Intactness of Fresh Rooster Semen (닭 신선 정자 운동성과 첨체안정성에 보존 온도, 희석제 및 용기가 미치는 영향)

  • Kim, Sung Woo;Lee, Jae-Yeong;Kim, Chan-Lan;Ko, Yeoung-Gyu;Lee, Sung Soo
    • Korean Journal of Poultry Science
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    • v.48 no.4
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    • pp.185-191
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    • 2021
  • Chicken spermatozoa have the ability to survive in low-temperature environments; however, the effects of low temperature on sperm motility and acrosome damage have not been studied in detail. The present study investigated semen longevity following dilution of rooster semen with Beltsville Poultry Semen Extender (BPSE) and Lake extender in preservation vessels (1.5 mL e-tube and 0.5 mL straw). Spermatozoa motility in the closed-type vessel (0.5 mL straw) was higher than that in the 1.5 mL e-tube on day 3 of preservation (68.6±3.1% vs. 22.1±5.7%). The motility of rooster semen diluted with BPSE in 0.5 mL straw was also higher than that of the Lake extender on day 3 of preservation (57.7±5.6% vs. 37.7±5.4%). Furthermore, acrosome intactness was higher in 0.5 mL straw than in the 1.5 mL e-tube, and the rate of acrosome cap damage increased with preservation days. The present study demonstrates that a closed 0.5-mL straw vessel could be used for low-temperature semen preservation, with an increased motility rate and acrosome integrity in fresh rooster semen.

Characterization of Mitochondrial NADH Dehydrogenase in Lentinus edodes (표고버섯의 미토콘드리아성 NADH 탈수소효소의 특성)

  • Kim, Eun-Mi;Min, Ji-Young;Min, Tae-Jin
    • The Korean Journal of Mycology
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    • v.26 no.1 s.84
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    • pp.119-126
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    • 1998
  • Mitochondria were isolated from Lentinus edodes and properties of the mitochondrial NADH dehydrogenase were studied. Optimal pH, temperature, and thermal stability of the enzyme were estimated to be 7.6, $33^{\circ}C$, and stable for one hour at $50^{\circ}C$. The apparent $K_m$ for the NADH was 0.33 mM. This enzyme catalyzed to transfer electrons from NADH to ferricyanide, decylubiquinone, and 2,6-dichloro-phenol-indophenol. 0.5 mM antimycin A and 0.01 mM dibromothymoquinone strongly inhibited 87.8% and 76.5% of the enzyme activities. 0.01 mM oligomycin known as an inhibitor of ATPase also strongly inhibited 79.2% of activities. 0.5 mM 5,5'-dithiobis-(2-nitrobenzoic acid) and 1.0 mM N-ethylmaleimide known as a modifier of SH group inhibited 50.4% and 36.7% of activities. 1 mM ethyl 2,4-dihydroxy-6-methyl benzoate and 10 mM orcinol, which had been known as an antibiotics isolated from Umbilicaria vellea according to our previous work, stimulated 68.4% and 48.1% of the enzyme activities.

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The Effect of Pressure and Hose Length on the Travelling Distance of Particles in Power Sprayer (토출압력(吐出壓力) 및 호오스길이가 도달성(到達性)에 미치는 영향(影響)에 관(關)한 연구(硏究))

  • Kwon, Soon Hong;Choi, Kyu Hong
    • Journal of Biosystems Engineering
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    • v.7 no.2
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    • pp.30-35
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    • 1983
  • To investigate the factors affecting the transportability of spray droplets, the maximum distance, the effective distance, the ratio of even distribution, and the diameters of particles were measured in accordance with the different pressure levels of power sprayer using 3 hole swath type nozzle, and the results are summerized as follows; 1. The distance of the most dense point from the nozzle was shortened by 0.5 meter with the 100-meter-long hose. The maximum reaching distances were reduced by 1.5 and 1.0 meters for the 13m/m and the 8.5 m/m hoses respectively, and the effective distance were reduced by 0.5 meter for both cases. 2. The effective distance can not be extended beyond 14 meters even if the length of hose is minimized at the rated pump pressure 28 kg/$cm^2$, it was 1 meter longer for 13m/m hose compared to the 8.5m/m one. 3. In case of 13m/m hose, the most dense point can be extended further by 0.5 meter increasing the pump pressure by 8 kg/$cm^2$, and the maximum distance and effective distance were increased by 2.0 and 0.5 meters respectively. There was no significant effect of pressure changes on the transportability in case of 8.5m/m hose. 4. Both the reduction of hose length and the increase of pump pressure influenced in large extent to the atomization effect of droplets. It was noticed that the diameter of droplet is related to the pump pressure and inside diameter of hose. 5. The pressure drop in 100-meter-long and l3m/m hose was 5~7kg/$cm^2$ at the pump pressure range of 25~33kg/$cm^2$, and it is an equivalent of 2% per 10 meter length of hose.

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Purification and some properties of polyphenol oxidase from Spuriopimpinella bracycarpa (참나물로부터 추출한 polyphenol oxidase의 부분정제 및 성질)

  • Ham, Seung-Shi;Hong, Eun-Hee;Lee, Sang-Young;Park, Gwi-Gun;Omura, Hirohisa
    • Applied Biological Chemistry
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    • v.34 no.1
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    • pp.49-53
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    • 1991
  • Three polyphenol oxidase(polyphenol oxidase I, II and III ) were isolated from the crude extract of a Spuriopimpinella bracycarpa by $(NH_4)_2SO_4$ precipitation and subsequent Sephadex G-150 chromatography. The final preparation thus obtained showed three peaks of enzyme activity. Optimum pH and temperature for the activity of polyphenol oxidase were 7.5 and $30^{\circ}C$, respectively. The enzyme was completely inactivated when i4 was treated at$70^{\circ}C$ for 30min and at $80^{\circ}C$ for 5min at pH 6.5. The enzyme was partially inactivated by ascorbic acid, glutathione and potassium cyanide (0.1mM), and was completely inhibited by L-cysteine, ascorbic acid, glutathione and potassium cyanide(0.5 and 1.0mM). The enzyme has good activity on catechol and 3,4-dihydroxytoluene but was strongly inactivated on pyrogallol, dopamine and DL-dopa. The Michaelis cons4ant of the enzyme was 86.5mM with catechol as a substrate.

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Modification of Endothelium on Contractile Response of Brain Vessels to Contracting Agents (혈관 수축제의 뇌혈관 수축반응에 대한 혈관근 내피세포의 역할)

  • Kook, Young-Johng;Baik, Yung-Hong;Kim, Jong-Keun;Choi, Bong-Kyu;Choi, Soo-Hyung;Kim, Yung-In
    • The Korean Journal of Pharmacology
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    • v.24 no.2
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    • pp.203-216
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    • 1988
  • To delineate the mechanisms of vasoconstriction and vasodilation in cerebral arteries the effects of some vasoconstrictors and calcium antagonists on the basilar artery (BA) and arterial circle of Willis (WC) were examined and also the role of endothelium in the action of these drugs was investigated in pigs, cats and rabbits. In pig cerebral arteries, dose-dependent contractile responses were elicited by KCI, histamine, 5-hydroxytryptamine (5-HT) and angiotensin, but norepinephrine (NE), phenylephrine (PE) and epinephrine (EP) elicited dose-dependent contractions only under pretreatment with propranolol 10-6 M. The magnitudes of maximal contractile effects of these drugs were different from each other, and 5-H~ was the largest and angiotensin the smallest. Some calcium antagonists dose-dependently inhibited KCI (35 mM)-induced contraction and the order of potency in inhibiting the contraction was nifedipine > > diltiazem > flunarizine > oxybutynin > isosorbide dinitrate (ISDN) > glyceryl trinitrate. 5-HT (10-6 M)-induced contraction was dosedependently inhibited by nifedipine but slightly inhibited by diltiazem and ISDN. In rings with intact endothelium, KCI (35 mM)-induced contraction was not affected by acetylcholine (ACh) but $PGF_{2{\alpha}}$ (lO-SM)-induced contraction was dose-dependently relaxed by ACh and adenosine. This endothelium-dependent relaxation was not affected by nifedipine (l0-6M)-pretreatment but markedly inhibited by methylene blue (50,uM)-pretreatment. In the porcine arterial rings without endothelium, ACh had no effect or even contracted the $PGF_{2{\alpha}}-induced$ contraction. However, the dosedependent relaxing effect of ACh appeared when the deendothelized porcine ring and rabbit thoracic aorta with intact endotheli urn were simultaneously suspended into a bath and this relaxing effect was also inhibited by methylene blue-pretreatment. In cat cerebral arteries, 5-HT and NE elicited dose-dependent contractile responses and ACh also produced dose-dependent contraction regardless of the existence of endothelium. ACh-induced contraction was most prominent. 5-HT (IO-SM)induced contraction was not relaxed but contracted additionally by ACh even in the intact endothelial ring. In rabbit cerebral arteries, 5-HT and NE elicited dose-dependent contractile responses and 5-HT-induced contraction was more prominent. In the intact endothelial preparations, 5-HT (lO-s M)-induced contraction was markedly relaxed by the addition of ACh( IO-SM) and this endothelium-dependent relaxing effect was inhibited by atropine (l0-7M)-pretreatment but notaffected by diltiazem (l0-6M)-pretreatment. These results suggest that ACh elicits endotheliumdependent relaxing effect mediated by muscarinic receptors in cerebral arteries of pig and rabbit, and that ACh acts as vasoconstrictor in cat cerebral artery.

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A Study on the Structural Standard of the Tube and Coupler Scaffold (단관비계의 구조규격에 관한 연구)

  • 이영섭
    • Journal of the Korean Society of Safety
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    • v.5 no.2
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    • pp.66-75
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    • 1990
  • This study is conducted to establish the structural standard of tube and coupler scaffold which is suitable for our present stuation through the comparison analysis for domestic and foreign standards as well as measurement of field survey. The results of this study are as follows : 1) The load is classified by three categories, light-duty(equal and lower than 150kg/m$^2$), medium-duty(150-250 kg/m$^2$), heavy-duty(250-350kg/m$^2$), and the equivalent horizontal length of side posts is each, 1.5-1.8m, 1.2-1.5m, equal and lower than 1.2m, and the equivalent horizontal length between front and rear posts is each 1.2-1.5m, 0.9-1.2m, equal and lower thatn 0.9m, in accordance with the load classification. 2) The height between upper and lower runner is equal and lower than 1.5m, and the brace across the width of scaffold should be installed within 15m in horizontal direction at 45 degree angle. 3) The entire scaffold should be securely tied to the wall of permanent structure with uslng anchor and bolt at intervals not to exceed 6m in case of non-connection and 4.5m in case of connection in both horizontal and vertical direction. 4) The post should be installed on the sound foundation tied to lumber footing with using base plate, and standard platform plank should be produced in the factory and widely used in the construction field.

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