• 제목/요약/키워드: serum separation

검색결과 92건 처리시간 0.028초

Alpha-hCG 측정을 위한 섬광 근접 측정법 (Scintillation Proximity Assay)에 관한 연구 (Studies on Scintillation Proximity Assay for the mesurement of alpha-hCG)

  • 최태현;임상무;최창운;정위섭;임수정;이수진;이태섭;오옥두
    • 대한핵의학회지
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    • 제36권2호
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    • pp.133-139
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    • 2002
  • 목적: 섬광 근접측정법은 항원 항체 반응 후 결합 분획과 유리분획을 분리하는 과정이 필요없다. 이러한 원리를 검체 내 hCG와 항 ${\alpha}$ hCG 항체간의 항원 항체 반응에 적용하고자 한다. 대상 및 방법: 항 ${\alpha}$ hCG 항체를 biotin과 결합시켜 SPA bead에 부착된 streptavidin과 부착 가능하게 만들었다. 이 측정법은 항 ${\alpha}$ hCG 항체가 부착된 SPA beads에 대해 혈청내 hCG와 표지항원인 $[^{125}I]hCG$간의 경쟁 반응을 기본 원리로 이용하였다. Biotin 표지 항 ${\alpha}$ hCG 항체를 $[^{125}I]hCG\;100{\mu}{\ell}$와 표준용액이나 환자 혈청 $200{\mu}{\ell}$이 들어있는 실온에서 20분 방치하였다. 그리고 streptavidin이 붙은 SPA beads $20{\mu}{\ell}$를 바이알에 넣고 10분 더 방치한다. 환자 혈청의 수치를 표준 응답곡선을 통해 계산하였다. 결과: SPA측정법에 사용되는 방사성 핵종의 방사능 양에 따라 반응용액 속에서 SPA bead와 자유 방사성 핵종에 의한 배후 방사능이 측정값에 영향이 없음을 확인하였다. SPA 방법을 응용한 측정에서 적합한 표준 응답곡선을 얻었고, 실제 환자혈청에서의 hCG 농도를 결정할 수 있었다. 결론: 이 실험을 통해 SPA 방법을 이용한 측정법이 임상진단에 유용하게 사용될 수 있을 것으로 사료된다.

The Effects of Flutamide and Diethylstilbestrol Exposure on the Reproductive Organs and Thyroid of Male Rats by the Rodent 20-day Thyroid/Pubertal Assay

  • Moon, Hyun-Ju;Shin, Jae-Ho;Kim, Hyung-Sik;Lee, Su-Jung;Kang, Il-Hyun;Kim, Tae-Sung;Seok, Ji-Hyun;Kim, In-Young;Han, Soon-Young
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 2002년도 추계국제학술대회
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    • pp.168-168
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    • 2002
  • To establish a test protocol for the rodent 20-day thyroid/pubertal assay, flutamide and diethylstilbestrol (DES) were administered to intact male Sprague-Dawley rats from postnatal day 33 for 20 days. Flutamide (1, 5, and 25 mg/kg/day) or DES (10, 20, and 40 ug/kg/day) was given once daily by oral gavage to immature male rats. Prepuce separation was significantly delayed in flutamide group and in DES group. One day after the last dose, the rats were killed and pituitary, thyroid, and reproductive organs were removed and weighed. Flutamide treatment resulted in a significant reduction in the weights of epididymides, ventral prostate, seminal vesicles plus coagulating glands and fluid (SVCGF), levator ani. bulbocarvenus muscles (LABC), Cowper's glands, and glans penis. The weight of adrenal glands decreased at % mg/kg/day, while testes and any other organ weights were unaffected. No microscopic changes were observed in the thyroid glands. Serum levels of testosterone wert significantly increased in the flutamide-treated groups and serum levels of estradiol were also increased. A significant reduction in the weights of testes, epididymides, ventral prostate, SVCGF, LABC, Cowpers glands, and glans penis of DES treated group. Serum testosterone and LH decreased significantly in DES group. Decrease of estradiol was observed, but not significant. These results indicate that flutamide and DES delay puberty in the male rat and its mode of action appears to be via altered secretion of steroids, which subsequently affect the development of the reproductive tract. (Supported by the grant from NITR/Korea FDA for Endocrine Disrupter Research.)

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여지전기영동법(濾紙電氣泳動法)에 의(依)한 한우(韓牛) 급(及) 돈(豚)의 정상혈청단백질분획(正常血淸蛋白質分劃)에 대(對)한 연구(硏究) (PAPER ELECTROPHORETIC SEPARATION OF SERUM PROTEINS IN CATTLE AND SWINE)

  • 임봉호
    • 대한수의학회지
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    • 제4권1호
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    • pp.1-6
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    • 1964
  • The ratios of cattle and swine serum proteins taken from the slaughter house were studied by Paper Electrophoresis. 1. Of 79 cattle and 53 swine, 49 cattle and 32 swine were observed in this studying as normal animals, the rest which was over 60% of albumin, globulin values and 1/2 of A/G (albumin/globulin) ratio was observed separately as abnormalities, because physiological examination was not made before slaughter. The ratios of the normal serum proteins were A (albumin) 58.8, ${\alpha}$(alpha-globulin) 13.7, ${\beta}$(beta-globulin) 11.9, ${\gamma}$(gamma-globulin) 28.6, G(total globulin) 49.2, A/G 1.03 in cattle and A 48.4, ${\alpha}$ 18.0, ${\beta}$ 13.6, ${\gamma}$ 20.0, G 51,6, A/G 0.93 in swine, the result including abnormalities showed A 45.5, ${\alpha}$ 14.8, ${\beta}$ 12.5, ${\gamma}$ 26.7, G 54.5, A/G 0.83 in cattle and A 44.5, ${\alpha}$ 19.8, ${\beta}$ 13.7, ${\gamma}$ 21.8, G 55.3, A/G 0.80 in Swine. 2. The A/G ratio of cattle and swine were 1.03 and 0.93 respectively, the A/G ratio of Korean cattle and swine are higher than the ration reported of others. Although A/G ratio of swine was below 1.00, and its value showed slightly higher than the others. The A/G ratio in this result including the abnormalities was relatively low but this ratio was higher than that values obtained by other reporters. 3. Twenty nine percent of cattles and 34 per cent of swines in this study, fluctuation of A/G ratio was great. The values of ${\alpha}$ and ${\gamma}$ globulins thought to be influenced by the amount of total globulin except ${\beta}$-globulin in swine. To obtain more occurate results, more sample size is required, in other hand some animals that is in subclinical condition might influence the values of this study. 4. The ratios of each fraction mobility which were regarded albumin as 100 were A 100, ${\alpha}$ 73, ${\beta}$ 47, ${\gamma}$ 30 in Cattle and A 100, ${\alpha}$ 71, ${\beta}$ 46, ${\gamma}$ 30 in Swine.

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염소의 피하농양 발생에 따른 혈액 내 염증지표 분석 (Analysis of inflammatory markers in blood related with the occurrence of subcutaneous abscesses in goats)

  • 구지영;박준환;김선호;조용일;김찬란;차승언;신기욱;박진호
    • 한국동물위생학회지
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    • 제45권1호
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    • pp.47-54
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    • 2022
  • Subcutaneous abscesses, which occur mainly in goats and sheep, are lymph node abscesses caused by Corynebacterium pseudotuberculosis infection, and are divided into internal, external, and mixed types depending on the type of occurrence. While diagnostic methods for subcutaneous abscesses have been continuously studied, research reports for effective treatment and management of subcutaneous abscesses are inadequate. Therefore, this study was conducted to determine the changes in biometric information related to the inflammatory markers of goats induced by subcutaneous abscesses by infection with C. pseudotuberculosis. For this, hematological tests, analysis of inflammatory indicators, and analysis of serum proteins through electrophoresis separation of goats with healthy goats and goats inoculated with C. pseudotuberculosis to induce subcutaneous abscesses were compared and analyzed by date, and the differences and characteristics were identified periodically. As a result, in goats induced with subcutaneous abscesses, anemia findings related to a rapid decrease in red blood cell (RBC), hematocrit (HCT), and hemoglobin (Hb) were observed, and a significant increase in inflammatory cells expressed in total white blood cell (WBC), neutrophil, and monocytes was observed. And the levels of acute phase protein (APP) such as fibrinogen, haptoglobin, and serum amyloid A (SAA) were observed to increase rapidly immediately after infection. In addition, in the results of electrophoretic analysis of serum proteins, it was observed that the levels of α-globulin and β-globulin were significantly increased in goats with subcutaneous abscesses. That is, when looking at these changes, it was found that the systemic inflammatory response of goats was rapidly induced immediately after infection with the C. pseudotuberculosis pathogen. Through this study, it was possible to identify changes in the biomarkers of goats with subcutaneous abscesses, which had not been reported. Furthermore, these analyzed data are thoughts to be of great help in identifying, treating, and managing the goats of subcutaneous abscesses.

Effect of Mori Cortex in the Cardiac Injury Induced by Skin Burn

  • Moon, Hye-Jung;Cho, Hyun-Gug;Park, Won-Hark
    • 대한의생명과학회지
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    • 제10권2호
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    • pp.107-113
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    • 2004
  • This study was conducted to investigate an effect of Mori Cortex in the cardiac injury following dermal scald burn in rats. Sprague-Dawley rats were induced scald bum (15% of total body surface area). Heart was removed at 5 h postburn and examined with biochemical assay, ultrastructural observations and stereological analysis. The activity of serum aspartate aminotransferase and creatinine was increased at 5 h postburn compared with them of control. Administration of heat extracts of Mori Cortex after scald burn inhibited the production of KC (neutrophil chemoattractant cytokine) and increased the activity of protein kinase C (PKC) in heart tissue. The activity of myeloperoxidase (MPO) in heart tissue was decreased both at 5 h postburn and in case of Mori Cortex administration after scald burn. Ultrastructurally, many contraction bands and separation of intercalated disk induced by scald burn were decreased by administration of heat extracts of Mori Cortex. In stereological analysis, administration of Mori Cortex after scald burn resulted the volume densities of myofibril and mitochondria were increased compared with them of burn control. These data suggest that Mori Cortex may be a useful stuff to the range of available treatments for cardiac injury induced by skin burn.

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Liquid Chromatography-Tandem Mass Spectrometry Analysis of Riboflavin in Beagle Dog Plasma for Pharmacokinetic Studies

  • Jeong, Hyeon Myeong;Shin, Beom Soo;Shin, Soyoung
    • Mass Spectrometry Letters
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    • 제11권1호
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    • pp.10-14
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    • 2020
  • Riboflavin is a water-soluble vitamin, which serves as a precursor to flavin mononucleotide and flavin adenine dinucleotide. This study aimed to develop a simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis for the quantification of riboflavin in the Beagle dog plasma. This method utilized simple protein precipitation with acetonitrile and 13C4, 15N2-riboflavin was used as an internal standard (IS). For chromatographic separation, a hydrophilic interaction liquid chromatography (HILIC) column was used with gradient elution. The mobile phase consisted of 0.1% (v/v) aqueous formic acid with 10 mM ammonium formate and acetonitrile with 0.1% (v/v) formic acid. Since riboflavin is an endogenous compound, 4% bovine serum albumin in phosphate buffered saline was used as a surrogate matrix to prepare the calibration curve. The quantification limit for riboflavin in the Beagle dog plasma was 5 ng/mL. The method was fully validated for its specificity, sensitivity, accuracy and precision, recovery, and stability according to the US FDA guidance. The developed LC-MS/MS method may be useful for the in vivo pharmacokinetic studies of riboflavin.

The Tonic Effect of the Extract from Male Silkworm (Bombyx mori L.) Pupae on Rats

  • Ryu, Kang-Sun;Ahn, Mi-Young;Lee, Heui-Sam;Kim, Iksoo;Kim, Jin-Won;Kim, Sang-Hyun;Park, Jin-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • 제5권1호
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    • pp.123-126
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    • 2002
  • More than three hundred million men in the world are suffering from erectile dysfunction. Korean ancient medical records say that the unmated male silkworm moth is effective in strengthening men's vitality, but no scientific examination has preyed its clinical efficacy. Currently, silkworm moths are not allowed for food-stuff. Thus, we selected silkworm of 14-days-after-metamorphosis, at which external morphology possesses that of pupae, but internal component is that of moths. To obtain the reliable source of unmated male silkworm moth, we used the sex-limited silkworm breed with larval marking. The body marker allows casual separation of sex during larval period, preventing mating right after emergence from cocoon. Using the extract prepared with the pupae of 14-days-after-metamorphosis, we investigated the tonic effect of the extract on the rat fed the extract for three weeks. The results showed that the testosterone levels in serum increased maximum by 19%, that of testicle increased maximum by 200%, and athletic endurance of the rats rose by 6%, suggesting positive tonic effect of the pupae prepared at the stage of 14 days after metamorphosis.

젤 전기영동 및 액체 크로마토그래피 분리 방법을 이용하여 지방 세포로부터 분비되는 단백질들에 대한 프로테오믹스 연구 방법 (Intensive Proteomic Approach to Identify Secreted Peptides/Proteins from 3T3-L1 Adipocytes using Gel Electrophoresis and Liquid Chromatograph Separation Methods)

  • 황현호;백문창
    • 약학회지
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    • 제55권3호
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    • pp.203-212
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    • 2011
  • Adipocytes have been known to secrete a number of important proteins called adipokines with roles in energy metabolism, reproduction, cardiovascular function and immunity. In this study we have attempted to identify intensively secretory proteins from 3T3-L1 adipocytes. 3T3-L1 preadipocytes were differentiated into mature adipocytes and then the cells were left in serum-free medium. The supernatant was filtrated and dialyzed. Lyophilized secretome was fractionated by two different methods, 1-D SDS PAGE and RP-FPLC. The tryptic peptides from the gel slices and the FPLC fractions were analyzed by nanoLC/ESI-MS/MS. We identified a total of 303 identical proteins from two methods, 251 proteins from 1-D gel and 184 proteins from RP-FPLC. 86 of them were listed as a secretory protein Finally, we identified many known or unknown secreted proteins existed in the low level including adiponectin, angiotensinogen, bone morphogenetic protein-1 (BMP-1), macrophage migration inhibitory factor (MIF), insulin like growth factor-II (IGF-II), interleukin-6 (IL-6), follistatin-related protein-1, minecan, and resistin. The existence of some of secreted proteins has been confirmed in RNA level. This proteomic experiment is useful for the intensive screening of secretory proteins in many kinds of other cells.

A Potent Anti-Complementary Acylated Sterol Glucoside from Orostachys japonicus

  • Yoon, Na-Young;Min, Byung-Sun;Lee, Hyeong-Kyu;Park, Jong-Cheol;Choi, Jae-Sue
    • Archives of Pharmacal Research
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    • 제28권8호
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    • pp.892-896
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    • 2005
  • In order to isolate substances that inhibit the hemolytic activity of human serum against eryth-rocytes, we have evaluated whole plants of the Orostachys japonicus species with regard to its anti-complement activity, and have identified its active principles following activity-guided isolation. A methanol extract of the O. japonicus, as well as its n-hexane soluble fraction, exhibited significant anti-complement activity on the complement system, which was expressed as total hemolytic activity. A bioassay-guided chromatographic separation of the constituents resulted in the isolation of three known compounds 1-3 from the active n-hexane fraction. The structure of these compounds were analyzed, and they were identified as hydroxyhopanone (1), $\beta-sitosteryl-3-O-\beta-D-glucopyranosyl-6'-O-palmitate$ (2), and $\beta-sitosteryl-3-O-\beta-D-glucopyranoside$ (3), respectively. Of these compounds, compound 2 exhibited potent anti-complement activity $(IC_{50}=1.0\pm0.1{\mu}M)$ on the classical pathway of the complement, as compared to tiliroside $(IC_{50}=76.5\pm1.1{\mu}M)$, which was used as a positive control. However, compounds 1 and 3 exhibited no activity in this system.

Metabolomics reveals potential plateau adaptability by regulating inflammatory response and oxidative stress-related metabolism and energy metabolism pathways in yak

  • Huang, Meizhou;Zhang, Xin;Yan, Wenjun;Liu, Jingjing;Wang, Hui
    • Journal of Animal Science and Technology
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    • 제64권1호
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    • pp.97-109
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    • 2022
  • Species are facing strong selection pressures to adapt to inhospitable high-altitude environments. Yaks are a valuable species and an iconic symbol of the Qinghai-Tibet Plateau. Extensive studies of high-altitude adaptation have been conducted, but few have focused on metabolism. In the present study, we determined the differences in the serum metabolomics between yaks and the closely related species of low-altitude yellow cattle and dairy cows. We generated high-quality metabolite profiling data for 36 samples derived from the three species, and a clear separation trend was obtained between yaks and the other animals from principal component analysis. In addition, we identified a total of 63 differentially expressed metabolites among the three species. Functional analysis revealed that differentially expressed metabolites were related to the innate immune activation, oxidative stress-related metabolism, and energy metabolism in yaks, which indicates the important roles of metabolites in high-altitude adaptation in yaks. The results provide new insights into the mechanism of adaptation or acclimatization to high-altitude environments in yaks and hypoxia-related diseases in humans.