• Title/Summary/Keyword: human plasma

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Synthetic Peptide-Based Enzyme-Linked Immunosorbent Assay for Human $\alpha$-Fetoprotein

  • Yoon, Mi-Chung;Lee, Hyun-Hee
    • Biomedical Science Letters
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    • v.7 no.3
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    • pp.103-110
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    • 2001
  • $\alpha$-Fetoprotein(AFP) is a good marker for the detection of several diseases such as hepatocellular carcinoma, gonadal germ cell tumor, gastric tumor, and Down's syndrome. In this study, we developed ELISA, using synthetic peptides corresponding to the epitopes of AFP. Five kinds of peptides were synthesized from AFP to produce antibodies in rats that recognize AFP in human plasma as well as amniotic fluid and do not cross-react with serum albumin. All five kinds of antibodies showed good reactivities with their peptide-keyhole limpet hemocyanin conjugates. Anti-synthetic peptide 1 (R-N-E-Y-G-I-A-S-I-L, 4-13) antibody, in particular, reacted well with AEP as well as synthetic peptide 1-KLH but not with human serum albumin. The binding affinity(Kd) was 2.7$\times$10$^{-9}$M for peptide 1 and 6.8$\times$10$^{-8}$M for AEP. The range for measurement of AFP was 10~1,000 ng/ml. The within-assay and between-assay coefficients of variance(CV) were 4.83% and 10.97%, respectively. In a sample of 31 sera and 33 amniotic fluids, there was a good correlation between AFP values determined in this assay and those in a commercial kit. These results indicate that the antibodies against synthetic peptides corresponding to the epitopes of AFP are highly specific to APP and synthetic peptide-based ELISA would be useful for the measurement of human AFP.

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Changes of Glycosidase Activity of Frozen-Thawed Spermatozoa in Human

  • Lee, Chae-Sik;Lee, Sang-Chan;Lee, Ji-Eun;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.35 no.2
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    • pp.185-190
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    • 2011
  • To evaluate the effect of spermatozoa culture on glycosidase activity of frozen-thawed spermatozoa in human, the spermatozoa were treated experimentally and assayed for activities of ${\alpha}$-L-fucosidase, ${\alpha}$-D-mannosidase, ${\beta}$-D-galactosidase and N-acetyl-${\beta}$-D-glucosaminidase (${\beta}$-GlcNAc'ase). The ${\beta}$-GlcNAc'ase activity was at least two-folds higher than other glycosidases regardless of spermatozoa incubation (p<0.05). The spermatozoa motility was decreased with incubation periods, but no effects by different glycosidases on the changes of spermatozoa motility during the various periods of incubation. In all glycosidases, the spermatozoa-zona binding rates in spermatozoa without incubation were higher than in spermatozoa incubated for 2 h (p<0.05). ${\beta}$-GlcNAc'ase is present mainly in the plasma membrane of spermatozoa frozen-thawed in human. It was also shown that the glycosidase activity was increased in all glycosidases in spite of lower sperm-zona binding by spermatozoa incubation.

Functional Properties of Modified Low Density Lipoprotein and Degradation of Modified LDL by Human Monocyte-Macrophages

  • Kim, Tae-Woong;Park, Jae-Hoon;Park, Young-June;Son, Heung-Soon;Yang, Ki-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.3
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    • pp.362-370
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    • 1995
  • Human plasma low density lipoprotein(LDL) is the main carrier for cholesterol, and recent studies suggest the normal LDL can be readily oxidized by free radical and not interact with LDL receptor. Lipoprotein pariticles are consisted of lipid andprotein, and fatty acids of lipoproteins are prone to oxidation. LDL particles readily undergo oxidative modification by copper. From the results, oxidized LDL altered its biological properties. A marked increase in the electrophoretic mobility of LDl on agarose gel indicated that negative surface charge of the LDL particles was increased. Also, the results from the HPLC showed that oxidized LDL was degraded into several polypeptides nonenzymatically. Degradation tests which measured the amount of 5-IAF labelled oxidized LDL were carried out by monocyte and hepatocyte cell culture. Hepatocyte cell culture of modified LDL did not show consistent pattern. However, binding rate of modified LDL with HMDM(human monocyte derived macrophage) was enhanced with oxidation, but was retarded by addition of antioxidants(hyaluronic acid, vitamin A, vitamin E). Also comparisons of oxidized-LDL, acetyl-LDL and MDA-LDL showed significant differences in the chemical properteis and binding affinity to HMDM. Thus, modificaition of normal LDL altered its biological properties.

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Fatty Acid Composition and Functional Properties of Low Density Lipoprotein and Oxidized LDL from Human Plasma (인체 혈장에서 분리한 LDL과 LDL의 지방산 조성과 기능성의 변화)

  • Jae-Hoon Choi;Hyun-Mi Cho;Heung-Soo Son;Tae-Woong Kim
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.3
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    • pp.402-408
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    • 1994
  • Human plasma low density lipoprotein (LDL) is the major factor of coronary heart disease.But recent studies suggest the normal LDL can be realdily oxidized by oxygen free radicals and not interact with LDL receptors.Lipoprotei particles consist of lipid and protein, and fatty acids are prone to oxidatioin.The fatty acid compositions of LDL from Koreans was compared with that of Westerners.From the results, the raio of unsaturated fatty acids of korean and Westerner approximately 30 and 70%, respectively.which means Westerners are more labile in the lipid oxidation of LDL than Koreams.Normal LDL was incubated with $CuSO_4$ in PBS to lead for the peroxidation of LDL, and it was tested by the detection of TBARS and free radicals.Then, ascorbate, ${\alpha}-tocopherol$ and hyaluronic acid were found to have effects of antioxidants on LDL oxidation.The amount of free radical increased as the extent of oxidation increased.The time course of free radical formation was similar to TBARS.Therefore, determination of free radical by Luminometer was much more convenient than that of TBARS.

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Effects of High Polyunsaturated Fat Diet on Human Plasma Cholesterol Esterification and Transfer (고 불포화 지방산 식이가 인체 혈청 Cholesterol의 Esterification과 Transfer에 미치는 영향)

  • Lee Myoung Sook
    • Journal of Nutrition and Health
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    • v.26 no.7
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    • pp.819-828
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    • 1993
  • The reaction of lecithin cholesterol acyltransferase(LCAT) and cholesteryl ester transfer protein (CETP) are important in cholesterol esterification and transfer for the reverse cholesterol transport(RCT) system. The purpose of this study were to assess the effects of fatty acid unsaturation on RCT system. After 12 female human subjects consumed a prescribed high saturated fat diet prior to the period, two groups of six subjects were provided with a high PUFA(corn oil) or a high SFA(butter) as major fats in a 40 en % fat diet. Butter feeding increased plasma total-(34%), esterified-(96%), HDL_3$-(23%), LDL-(20%), and VLDL plus LDL(35%) cholesterol while corn oil feeding decreased esterified-(25%), LDL(15%) cholesterol and TG(27%). There were significant differences of fats on total-(p=0.0001), esterified-(p=0.0001), total HDL-(p=0.005), $HDL_2$-(p=0.01)and LDL-(p=0.0001) cholesterol. LCAT activity did not change during the period but highly correlated to apo A-I in HDL which was increased in the corn oil group. The 2.5 fold increased CETP activity in the butter group during the period might be related to changes in plasma VLDL plus LDL cholesterol level which were increased in the butter group.

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Studies on Protein Contents and Enzyme Activities of Human Seminal Plasma (사람정장(精漿)의 단백질농도(蛋白質濃度)와 효소활성도(酵素活性度)에 관한 연구(硏究))

  • Park, Kyu-Hong;Lee, Hee-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.10 no.1
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    • pp.7-24
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    • 1983
  • On the basis of the semen analysis in 66 subjects, they were divided into six different groups: Group I consisted of 16 normal subjects with sperm counts of over 40 ${\times}10^6$/ml and motility of over 40 percent, Group II, 7 subjects with normal sperm counts, but motility of under 40 percent, Group III, 15 oligospermic patients with under 40 ${\times}10^6$/ml, Group IV 14 azoospermic patients, Group V, 10 patients with vasectomy and Group VI, 4 abnormal patients with 2 cases of hypoplastic testis, 1 case of Klinefelter's syndrome and 1 case of testis tumor. After seperation of semen into sperm and seminal plasma by centrifugation, the protein contents and the activities of hyaluronidase, ${\beta}$-N acetylglucosaminidase, ${\beta}$-glucuronidase, arylsulfatase, acrosin and azocoll proteinase in seminal plasma were measured. Vasectomy group has 30 percent less of total protein than normal group. For the comparison of enzyme activities of seminal plasma, it could be assumed that the enzymes in seminal plasma were not contaminated with the enzymes of spermatozoa by testing the enzymes of the seminal plasma from the vasectomy and azoospermic groups. It had been reported that hyaluronidase was only released from spermatozoa, however, the result obtained in this investigation showed that azoospermic and vasectomy group had high specific activities of hyaluronidase. The results indicated that hyaluronidase was not only from the testis but also from the male accessory sexual glands. Oligospermic group (Group III) showed the lowest total activity of hyaluronidase among them. The specific activities of ${\beta}$ -N-acetylglucosaminidase was high in oligospermic group (Group III) and low in vasectomy group (Group V). These results were contradictory with the pattern of hyaluronidase activities. This indicated that the spermatozoa which were stayed in epididymis would increase the activity of this enzyme. The specific activity of ${\beta}$ - glucuronidase was low in oligospermic and vasectomy groups. Group VI including testis tumor had remarkably high arylsulfatase activity. Arylsulfatase, a typical lysosomal enzyme, has been known to be released unusually large amounts from certain tumor cells. Arylsulfatase was also released with high activities from azoospermic and vascetomy group. This result indicated that this enzyme was also released from the sources other than testis. Acrosin, a proteolytic enzyme locating in the sperm acrosome, was not found throughout all the samples of seminal plasma. The activities of azocoll proteinase, a non-specific neutral proteinase was nearly identical in all the groups. This enzyme must have been released from the sources other than testis.

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Bioequivalence and Pharmacokinetics of 70 mg Alendronate Sodium Tablets by Measuring Alendronate in Plasma

  • Yun Min-Hyuk;Woo Jong-Su;Kwon Kwang-Il
    • Archives of Pharmacal Research
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    • v.29 no.4
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    • pp.328-332
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    • 2006
  • The bioequivalence and pharmacokinetics of alendronate sodium tablets were examined by determining the plasma concentration of alendronate. Two groups, consisting of 24 healthy volunteers, each received a 70 mg reference alendronate sodium tablet and a test tablet in a $2{\times}2$ crossover study. There was a 6-day washout period between doses. The plasma alendronate concentration was monitored for 7 h after the dose, using HPLC-Fluorescence Detector (FD). The area under the plasma concentration-time curve from time 0 to the last sampling time at 7 h $(AUC_{0-7h})$ was calculated using the linear-log trapezoidal rule. The maximum plasma drug concentration $(C_{max})$ and the time to reach $C_{max}(T_{max})$ were derived from the plasma concentration-time data. Analysis of variance was performed using logarithmically transformed $AUC_{0-7h}\;and\;C_{max}$, and untransformed $T_{max}$. For the test medication versus the reference medication, the $AUC_{0-7h}\;were\;87.63{\pm}29.27\;vs.\;102.44{\pm}69.96ng\;h\;mL^{-1}$ and the $C_{max}$ values were $34.29{\pm}13.77\;vs.\;38.47{\pm}24.39ng\;mL^{-1}$ respectively. The $90\%$ confidence intervals of the mean differences of the logarithmic transformed $AUC_{0-7h}$ and $C_{max}$ values were log 0.8234-log 1.1597 and log 0.8222-log 1.1409, respectively, satisfying the bioequivalence criteria guidelines of both the US Food and Drug Administration and the Korea Food and Drug Administration. The other pharmacokinetic parameters for the test drug versus reference drug, respectively, were: $t_{1/2},\;1.87{\pm}0.62\;vs.\;1.77{\pm}0.54\;h;\;V/F,\;2061.30{\pm}986.49\;vs.\;2576.45{\pm}1826.05\;L;\;CL/F,\;835.32{\pm}357.35\;vs.\;889.48{\pm}485.87\;L\;h^{-1}; K_{el},\;0.42{\pm}0.14\;vs.\;0.40{\pm}0.18\;h^{-1};\;Ka,\;4.46{\pm}3.63\;vs.\;3.80{\pm}3.64\;h^{-1};\;and\;T_{lag},\;0.19{\pm}0.09\;vs.\;0.18{\pm}0.06\;h$. These results indicated that two alendronate formulations(70-mg alendronate sodium) were biologically equivalent and can be prescribed interchangeably.

Experiment on the Sterilization Performance of Airborne Bacteria in Indoor Spaces using the Variation of Ozone Concentration Generated According to the Discharge Time of a Plasma Module with a Dielectric Barrier Discharge Technology (유전체 장벽방전 플라즈마 방전시간에 따른 오존 발생 농도변화의 값을 통한 실내 공간 내 부유세균 살균성능에 대한 실험)

  • Su Yeon Lee;Chang Soo Kim;Gyu Ri Kim;Jong Eon Im
    • Journal of the Society of Disaster Information
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    • v.19 no.2
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    • pp.344-351
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    • 2023
  • Purpose: This study aimed to evaluate the effectiveness of a dielectric barrier discharge (DBD) plasma module for sterilizing airborne bacteria in indoor spaces and measure the concentration of ozone generated during plasma discharge. Method: The DBD plasma module was installed in a 76m3 space, and air samples were collected under various discharge times to compare the reduction of airborne bacteria. Result: The results showed a significant decrease in airborne bacteria, ranging from 92.057% to 99.999%, with an average ozone concentration of 0.04 ppm, below the reference value. Conclusion: The study suggests that plasma discharge can be used as a means of preventing the spread of airborne bacteria and viruses, while ensuring safety for human exposure.

Pharmacokinetics of Uridine Following Ocular, Oral and Intravenous Administration in Rabbits

  • Kim, Eunyoung;Kang, Wonku
    • Biomolecules & Therapeutics
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    • v.21 no.2
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    • pp.170-172
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    • 2013
  • The pyrimidine nucleoside uridine has recently been reported to have a protective effect on cultured human corneal epithelial cells, in an animal model of dry eye and in patients. In this study, we investigate the pharmacokinetic profile of uridine in rabbits, following topical ocular (8 mg/eye), oral (450 mg/kg) and intravenous (100 mg/kg) administration. Blood and urine samples were serially taken, and uridine was measured by high-performance liquid chromatography-tandem mass spectrometry. No symptoms were noted in the animals after uridine treatment. Uridine was not detected in either plasma or urine after topical ocular administration, indicating no systemic exposure to uridine with this treatment route. Following a single intravenous dose, the plasma concentration of uridine showed a bi-exponential decay, with a rapid decline over 10 min, followed by a slow decay with a terminal half-life of $0.36{\pm}0.05$ h. Clearance and volume of distribution were $1.8{\pm}0.6$ L/h/kg and $0.58{\pm}0.32$ L/kg, respectively. The area under the plasma concentration-time curves (AUC) was $59.7{\pm}18.2{\mu}g{\cdot}hr/ml$, and urinary excretion up to 12 hr was ~7.7% of the dose. Plasma uridine reached a peak of $25.8{\pm}4.1{\mu}g/ml$ at $2.3{\pm}0.8$ hr after oral administration. The AUC was $79.0{\pm}13.9{\mu}g{\cdot}hr/ml$, representing ~29.4% of absolute bioavailability. About 1% of the oral dose was excreted in the urine. These results should prove useful in the design of future clinical and nonclinical studies conducted with uridine.

Effects of Antihypertensive Diets Mainly Consisting of Buckwheat, Potato, and Perilla Seed on Blood Pressures and Plasma Lipids in Normotensive and Spontaneously Hypertensive Rats (메밀, 감자, 들깨를 이용한 항고혈압 기능성 식이가 정상혈압쥐 및 본태성고혈압쥐에서 혈압 및 혈장지질에 미치는 영향)

  • 한찬규
    • Journal of Nutrition and Health
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    • v.29 no.10
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    • pp.1087-1095
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    • 1996
  • The study was carried out to investigate a new type of functional foods with hypotensive effect which is critical in the prevention and treatment of hypertension and related circulatory diseases. The experimental diets(A, B, C) were prepared from plant based ingredients such as buckwheat, potato, perilla seed with different ratios formulated as an edible form appropriate for human consumption according to AIN-77 standard. Control group(D) was fed commercial rat chow. Twety-four 15-week-old SDR(Sprague Dawley rats) and twenty-four 20-week-old SHR(spontaneously hypertensive rats), weighing 200g respectively, were assigned to 4 treatments of 6 rats each in a completely randomized design. Blood pressure was measured at 7 day interval by tail-cuff sphygmomanometer using an IITC cuff pump and amplifier. The growth rates of both SHR and SDR were not statistically different in comparions with the control except those of diet B and C in SDR and SHR, respectively(p<0.05). When exprimental diets were fed, systolic blood pressure of SDR and SHR at day 28 had been lowered by 17-20 mmHg(ave. of 19.6mmHg for both strains) compared with the reference pressure at day 0. The levels of HDL-cholesterol were increased, while the levels of LDL-cholesterol consistently decreased in both strains when experimental diets were fed(p<.05). Plasma total cholesterol levels were not different among treatments. Plasma triglyceride levels were higher in control diet(commercial rat chow) due to two times higher fat content of control diet itself(p<.05). Atherogenic indices were lower compared to those of control when the experimental diets were fed in both strains (p,.05). The results suggest that the antihypertensive diets mainly consisting of plant-based ingredients may possibly effective in relieving hypertension as well as circulatory diseases.

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