• Title/Summary/Keyword: Chemiluminescence method

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The Effect of L-carnitine and Acetylcarnitine on Sperm Parameters in vitro (생체외 L-carnitine과 Acetylcarnitine의 정자지표 개선 효과)

  • Lee, Wan;Park, Nam-Cheol
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.2
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    • pp.149-157
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    • 2002
  • Objectives: To assess the scavenging effect of carnitine derivatives on oxidative damage to sperm during sperm processing, cryopreservation and thawing. Materials and Methods: Fresh semen samples from 20 normal healthy volunteers were collected by masturbation after at least 48 hours abstinence. After liquefaction of semen samples at room temperature, the specimens were diluted with sperm wash media (Ham's F-10, Life technologics) to a uniform density of $20{\times}10^6/ml$. L-carnitine or acetylcarnitine were added with various concentration of $0{\mu}M$, $10{\mu}M$, $30{\mu}M$ in semen sample or cryoprotectant. All specimens were cryopreservated at $-196^{circ}C$ $LN_2$ for 3 days. Sperm motility, vitality, fertilizing capacity, reactive oxygen species formation and the level of lipid peroxidation were analyzed by computer assisted semen analyzer, eosin-nigrosin stain, hypoosmotic swelling test, chemiluminescence and thiobarbituric acid method, respectively, during sperm processing, cryopreservation and thawing. Results: The sperm motility was only increased in proportion to the concentration of acetylcarnitine with no statistical significance (p>0.05). The sperm vitality was also significantly improved in proportion to the concentration of acetylcarnitine with statistical significance (p<0.05). The sperm fertilizing capacity was significantly increased in proportion to the concentration of L-carnitine and acetylcarnitine and reactive oxygen species generation and lipid peroxidation were significantly decreased with same fashion (p<0.05). On comparison of effects between L-carnitine and acetylcarnitine, acetylcarnitine was superior to L-carnitine on the improvement of sperm motility and vitality as well as the suppression of reactive oxygen species generation and lipid peroxidation. Conclusions: These results suggest that carnitine derivatives have a scavenging effect against oxidative damages during sperm processing, cryopreservation and thawing. Therefore, carnitine derivatives may be useful as an oral antioxidant in patients with male infertility due to increased ROS generation.

Relationship between exhaled nitric oxide and pulmonary function test in children with asthma (소아 천식에서 호기산화질소와 폐기능 검사의 관계)

  • Ko, Han-Seok;Chung, Sung-Hoon;Choi, Yong-Sung;Choi, Sun-Hee;Rha, Yeong-Ho
    • Clinical and Experimental Pediatrics
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    • v.51 no.2
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    • pp.181-187
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    • 2008
  • Purpose : Asthma is characterized by reversible airway obstruction and bronchial hyperresponsiveness result from airway inflammation. Fraction of nitric oxide in expired air (FeNO) has recently been investigated as a noninvasive measure of airway inflammation. FeNO has been reported to correlate with induced sputum eosinophilia and methacholine challenge test that it is represent severity of asthma. The purpose of this study was to analyze the relationship of FeNO with pulmonary function tests in patients with intermittent asthma. Methods : Eighty children included in this study were diagnosed as asthma from April through August, 2005 in Department of Pediatrics, College of Medicine, Kyunghee University. They aged from 4 to 15 years who were able to conduct spirometry and FeNO monitoring. They did not have upper respiratory tract infection and did not use an asthma controller which contain corticosteroids within 4 weeks. Pulmonary function test was done and FeNO was measured with online tidal breathing method using a chemiluminescence NO analyzer (CLD 88 sp, Eco Medics, Duernten, Switzerland). The correlations between pulmonary function test and FeNO were analyzed using Spearman correlation coefficient method. Results : The mean of FeNO of subject was 16.88 parts per billion (ppb). The mean of forced expiratory volume in 1 second ($FEV_1$) was $0.890{\pm}0.455L$ and forced vital capacity (FVC) was $1.071{\pm}0.630L$. The mean of predicted $FEV_1%$ ($FEV_1%pred$) was $98.39{\pm}34.27%$ and $FEV_1/FVC$ was $88.53{\pm}19.49$. FeNO was significantly correlate with $FEV_1$ (r=0.345, P<0.01) and FVC (r=0.244, P<0.05). FeNO did not correlate with $FEV_1%pred$ or $FEV_1/FVC$. Conclusion : The measurement of FeNO could be a useful marker in the management of childhood asthma and it is evolving to provide a complementary role alongside existing pulmonary function test. We propose that measuring technique and establishment of normal reference range are important area for future research.

Determination of aromatic amino acids by chemiluminometric assay with Luminol-H2O2-Cu(II) system (Luminol-H2O2-Cu(II) 시스템을 이용한 방향족 아미노산의 화학발광법적 정량)

  • Kim, Kyung-Min;Kim, Young-Ho;Lee, Sang-Hak
    • Analytical Science and Technology
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    • v.25 no.3
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    • pp.171-177
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    • 2012
  • A determination method of aromatic amino acids such as trytophan (Trp), tyrosine (Tyr), and phenylalanine (Phe) using luminol-$H_2O_2$-Cu(II) system has been presented. In the presence of an aromatic amino acid, the enhanced chemiluminescence (CL) intensity of luminol-$H_2O_2$-Cu(II) system was obtained by forming a complex between Cu(II) and the amino acid. Based on the above phenomenon, a sensitive and fast determination of three aromatic amino acids was performed using the CL method in batch-type detection system. To optimize determination conditions, the kinetic influence of an aromatic amino acid on the luminol-$H_2O_2$-Cu(II) system and the effects of $H_2O_2$ and Cu(II) concentration, pH, and buffers were investigated. Under the optimized conditions, the calibration curve was linear over the range from $1.0{\times}10^{-6}$ to $2.0{\times}10^{-5}\;M$ for Trp, $1.0{\times}10^{-6}$ to $2.0{\times}10^{-5}\;M$ for Try, and $2.0{\times}10^{-6}$ to $2.0{\times}10^{-5}\;M$ for Phe, respectively. In this range, reproducibility (RSD, n = 4) of Trp, Try, and Phe were 3.21%, 2.64%, and 2.48%, respectively. The limit of detection ($3{\sigma}/s$) was calculated to be $6.8{\times}10^{-7}\;M$ for Trp, $5.7{\times}10^{-7}\;M$ for Try, and $9.6{\times}10^{-7}\;M$ for Phe.

A Study of the characteristics of NOx measurement and analysis methods of the SCR system for ships (선박용 SCR 시스템 NOx 측정 및 분석 방식의 특성 연구)

  • Kim, Sung-Yoon;Lee, Young-Ho;Kim, Min;Park, Sam-Sik
    • Journal of Advanced Marine Engineering and Technology
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    • v.39 no.4
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    • pp.387-392
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    • 2015
  • A method for measuring and analyzing the $NO_x$ in ships is described in $NO_x$ Technical Code 2008. The analysis device, as required by the Code, has been to use a Chemi-luminescence detection method or Heated Chemi-luminescence detection. on the other hand, selective catalytic reduction using $NH_3$ as a reducing agent has an interference effect on the analyzer, and causes measurement error. In this study, the Chemi-luminescence detection method was examined according to how it affects the concentration of $O_2$, CO, $SO_2$, $NH_3$. Fourier transform infrared spectrometry analysis equipment and measurement methods were compared. In order to confirm the effect of the physical interference of the measuring device, it was confirmed by decomposing a measuring device. Consequently, white precipitate and moisture were generated inside the chemiluminescence detection system and I found that affecting interference. The influence of interference highlights the need to consider the minimized $NO_x$ measurement method.

Absorption, Excretion and Antioxidative Effect of Rebamipide on Reproductive Organ (Rebamipide의 생식기관 내 흡수, 배설 및 항산화제로서 불임치료효과)

  • Kim, Jong Il;Park, Hyun Jun;Park, Nam Cheol
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.4
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    • pp.301-314
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    • 2005
  • Objective: Rebamipide is a propionic acid derivative that has an action of the inhibition of superoxide production and removal of hydroxyl radical with the sperm incubation and cryopreservation. In the present study, to investigate whether rebamipide is useful to treat male infertility and sterility, the author observed the antioxidative effects in patient with male infertility and also examined its absorption and distribution in rat genital organ. Methods: To measure the distribution of rebamipide in reproductive organ in the rat, carbon indicated rebamipide, $^{14}C-OPC-12759$, was orally administered to 10 Spraque-Dawley rats and its organ concentration in serum, liver, kidney, stomach, duodenum, colon, urinary bladder, seminal vesicle, epididymis and testicle were measured each time after 0.5, 1, 2, 4, 8 and 24 hours by using HPLC fluorescent method. The concentrations in semen were measured by HPLC fluorescent method in a sample of 50 infertile males who took 900 mg of rebamipide daily for 3 months. To measure the antioxidative effect and fertility rate for 3 months, each month before and after the treatment, sperm motility, vitality, the oxygen free radical formation, level of peroxidation, fetilizing capacity of semen sample which were obtained from infertile male patients by masturbation after at least 48 hours abstinence were analyzed by computer assisted semen analyzer, eosin-nigrosin stain, chemiluminescence, thiobarbituric acid method and hypo-osmotic swelling test. Simultaneously in a sample that wanted baby, both pregnancy and delivery were researched. Results: The $^{14}C-OPC-12759$ concentration in the body of white rats was highest in gastrointestinal organ like stomach, smal intestine and duodenum and followed by genital organ like seminal vesicle, testis and epididymis. The rebamipide concentration in semen of infertile males was $220.77{\pm}327.84ng/mL$ (SD) which showed a large deviation but it was higher than serum which was $126{\pm}76ng/mL$ (SD). In the infertile males, after the treatment with rebamipide, the level of seminal reactive oxygen species (ROS) and lipid peroxidation have significantly decreased in duration of the treatment (p<0.05) and sperm vitality and fertilizing capacity except sperm motility significantly improved on post treatment of 2~3 months (p<0.05). Out of the 41 cases who hoped for pregnancy, 15 cases (36.6%) became pregnant and 12 cases had childbrith, 2 cases had miscarriage and one case is ongoing. The side effect was observed in 1 case (2%) which experienced diarrhea but it was lost spontaneously. Conclusions: We conclude from this study that rebamipide showed relatively high tendancy of absorption and excretion in the genital organ. In infertile males who had elevated ROS in semen, by specifically inhibiting the cell damage from the antioxidation, a way to preserve sperm motility, vitality and fertilizing capacity was confirmed.

Role of Mitochondria in Oxidative Damage of Post-Ischemic Reperfused Hearts (허혈/재관류 심장의 산화손상에서 미토콘드리아의 역할)

  • Park, Jong-Wan;Chun, Yang-Sook;Kim, Myung-Suk
    • The Korean Journal of Pharmacology
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    • v.32 no.2
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    • pp.201-209
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    • 1996
  • Restoration of the blood flow after a period of ischemia is accompanied by generation of toxic oxygen radicals. This phenomenon may account for the occurrence of reperfusion-mediated tissue injury in ischemic hearts. In in vitro studies, although oxygen radicals can be generated from a variety of sources, including xanthine oxidase system, activated leucocytes, mitochondria and others, the most important source and mechanism of oxygen radical production in the post-ischemic reperfused hearts is unclear. In the present study, we tested the hypothesis that the respiratory chain of mitochondria might be an important source of oxygen radicals which are responsible for the development of the reperfusion injury of ischemic hearts. Langendorff-perfused, isolated rat hearts were subjected to 30 min of global ischemia at $37^{\circ}C$, followed by reperfusion. Amytal, a reversible inhibitor of mitochondrial respiration, was employed to assess the mitochondrial contributions to the development of the reperfusion injury. Intact mitochonria were isolated from the control and the post-ischemic reperfused hearts. Mitochondrial oxygen radical generation was measured by chemiluminescence method and the oxidative tissue damage was estimated by measuring a lipid peroxidation product, malondialdehyde(MDA). To evaluate the extent of the reperfusion injury, post-ischemic functional recovery and lactate dehydrogenase(LDH) release were assessed and compared in Amytal-treated and -untreated hearts. Upon reperfusion of the ischemic hearts, MDA release into the coronary effluent was markedly increased. MDA content of mitochondria isolated from the post-ischemic reperfused hearts was increased to 152% of preischemic value, whereas minimal change was observed in extramitochondrial fraction. The generation of superoxide anion was increased about twice in mitochondria from the reperfused hearts than in those from the control hearts. Amytal inhibited the mitochondrial superoxide generation significantly and also suppressed MDA production in the reperfused hearts. Additionally, Amytal prevented the contractile dysfunction and the increased release of LDH observed in the reperfused hearts. In conclusion, these results indicate that the respiratory chain of mitochondria may be an important source of oxygen radical formation in post-ischemic reperfused hearts, and that oxygen radicals originating from the mitochondria may contribute to the development of myocardial reperfusion injury.

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The Effects of Puerariae Radix Catechins Administration on Liver Function in Carbon Tetrachloride-Treated Rats (사염화탄소를 투여한 흰쥐에 있어서의 간 기능에 미치는 칡 카테킨의 효과)

  • 이치호;한석현;김종배;민상기
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.5
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    • pp.713-719
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    • 1995
  • These studies were carried out to investigate the effects of Puerariae radix catechins(PRC) administration on the biochemical parameters of liver function in liver of carbon tetrachloride(CCl4)-treated rats. Thirty six healthy Sprague-Dawley rats weighing about 120g were used for this experiment and divided intot he following 3 groups : normal control group(NCON), $CCl_4$ control group(CCON), PRC treated group(PRC). Fifty percent $CCl_4$ in oil was administered(I.P.) by 2ml per kg body weight two times a week for 3 weeks. PRC treated groups were administered orally at the leaves of 1% per day in distilled water for 8 weeks. Lipid hydroperoxides were analyzed by using chemiluminescence-high performance liquid chromatography(CL-HPLC) method as a phosphatidylcholine hydroperoxide value(PCOOH) in liver tissues. $CCl_4$ treatment significantly(p<0.05) resulted in an increase in GPT & GOT activities and liver hydroperoxide values comparing with those of the untreated control, while administration of PRC to the $CCl_4-treated$ rats significantly(p<0.001) decreased GPT & GOT activities and liver hydroperoxide value. Their ultrastructual changes of hepatocellular organelles were shown to clarify the morphologic nature of protective effects of PRC on hepatocytic injuries. $CCl_4$ treatment observed to change the ultrastructual nature of outer membrane of hepatocytes. However, the hepatic changes on PRC treatment to $CCl_4$ group was not found. PRC administration may inhibit the formatiion of liver lipid hydroperoxides in vivo and were very effective in recovering the liver function in $CCl_4-treated$ rats.

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Protective Effects of Cellular Membrane and Component Analysis of Polygonum aviculare Extracts (마디풀 추출물의 세포 보호 효과 및 주성분 분석)

  • Park, Soo Nam;Kim, Min-Ji;Kim, Su Ji
    • Microbiology and Biotechnology Letters
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    • v.42 no.1
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    • pp.51-57
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    • 2014
  • In this study, the antioxidative effects and component analysis of Polygonum aviculare (P. aviculare) extracts were investigated. The ethyl acetate and the aglycone fraction from P. aviculare extracts were more active than (+)-${\alpha}$-tocopherol and $\small{L}$-ascorbic acid, which are known as strong antioxidants for their antioxidative activity by the DPPH method and chemiluminescence assay. The cellular protective effects of fractions of P. aviculare on the rose-bengal sensitized photohemolysis of human erythrocytes, increased in a concentration dependent manner ($1-10{\mu}l$). In particular, the ethyl acetate fraction at a concentration of $10{\mu}l$ showed the most prominent protective effect among all the extracts (${\tau}_{50}$, 314.70 min). TLC and HPLC chromatogram of the ethyl acetate fraction of P. aviculare extracts revealed 3 main bands (PA8, PA5, PA6) and peaks (peak 1, peak 2, peak 3), which were identified as myricetin-3-O-rhamnoside (myricitrin, PA8, peak 1), quercetin-3-${\alpha}$-rhamnoside (quercitrin, PA6, peak 3) by LC/ESI-MS/MS and $^1H$-NMR respectively. These results indicate that fractions from P. aviculare could be applicable to new functional cosmetics as antioxidants.

Measurement of Progesterone in Plasma and Milk by RIA and CIA (RIA 및 CIA에 의한 혈장과 우유내 Progesterone 측정)

  • 이경찬
    • Korean Journal of Animal Reproduction
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    • v.14 no.1
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    • pp.57-65
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    • 1990
  • These experiment were carried out ; (1) to investigate the changes of progesterone in plasma, whole milk and skim milk during oestrus cycle and pregnancy and ; (20) to evaluate a chemiluminescence Immunoassay(CIA) as an alternative method by measuring the progesterone concentration is skim milk by RIA and CIA. The results obtained in these experiments were summerized as follows ; 1. Plasma progesterone levels in non-pregnant Holstein during oestrus cycle were relatively low until day 8 after oestrus. And then, the progesterone level began to increase and reached a peak with 6.3ng/ml on day 14 and then declined repidly to 1.5ng/ml and 2.2ng/ml on day 18 and 20, respectively. 2. Whole milk progesterone level in pregnant Holstein increased from 1.0ng/ml on oestrus to 16.0ng/ml on day 8 and then remained from 11.0ng/ml on day 10 to 22.0ng/ml on day 22. 3. In non-pregnant Holstein, whole milk pregesterone lev디 was 1.5ng/ml on oestrus and began to increase rapidly from day 6 after oestrus and exhibited a ranged of levels, 17.8~20.0ng/ml from day 6 to day 16 after oestrus. 4. Skim milk progesterone levels in pregnant Holstein were a range of 130~490pg/ml at the time of estrus and began to increase continually till then showing constant levels ranging from 1300pg/ml on day 10 to 1650pg/ml on day 22. 5. In non-pregnant Holstein, skim milk progesterone level was 160pg/ml on oestrus and began to increase from 190pg/ml on day 2 after oestrus to day 8 and then keep constant levels ran ging from 1050 to 1300pg/ml from day 8 to day 16 and then decreased to 240~450pg/ml from day 18 to day 22 after oestrus. 6. The results obtained from CIA for the analysis of skim milk progesterone were in good agreement with the values derived by RIA.(r=0.914)

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Anti-oxidative Activities of Castanea crenata Leaf Extract/Fractions and Application on Cosmetics (밤나무 잎 추출물의 항노화 활성 및 화장품에의 응용( I ))

  • Kim, Jin-Young;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.4
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    • pp.259-268
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    • 2008
  • In this study, the antioxidative effects, inhibitory effects on tyrosinase and elastase and components of Castanea crenata leaf were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl radical, DPPH) scavenging activity ($FSC_{50}$) of extract / fractions of Castanea crenata left was in the order: 50% ethanol extract ($13.6{\mu}g/mL$) < ethyl acetate fraction (6.2) < aglycone fraction (2.1). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$ of extract / fractions from Castanea crenata leaf extract / fractions on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activity was in the order: aglycone fraction (0.8) < 50% ethanol extract (0.5) < ethyl acetate fraction (0.3). The scavenging activity ($IC_{50}$ for ${O_2}^{{\cdot}\;-}$ (superoxide anion radical) generated by NBT method was in the order: ethyl acetate fraction (145.5) < aglycone fraction (65.5). The protective effects on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethyl acetate fraction exhibited the most prominent cellular protective effect (${\tau}_{50}$, $191.9{\pm}12.2\;min$ at $10{\mu}g/mL$). The inhibitory effect of aglycone fraction ($9.1{\mu}g/mL$) on elastase was higher than oleanolic and ($13.7{\mu}g/mL$). And the inhibitory effect of aglycone fraction ($21.6{\mu}g/mL$) on tyrosinase was higher than arbutin ($226.2{\mu}g/mL$). But 50% ethanol extract rarely exhibited the inhibitory activity on tryosinase and elastase. Flavonoids were contained in Castanea crenata left (96.3 mg / 100 g dried Castanea crenata leaf). And flavonoids contained in ethyl acetate fraction were kaempferol, quercetin, quercitrin, and so on. Quercitrin is the most abundant component. These results indicate that extract / fractions of Castanea crenata can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging free radical and ROS, Castanea crenata leaf extract/ fractions could be used as new cosmeceutical for whitening and anti-wrinkle products.