• Title/Summary/Keyword: Serum samples

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Purification of IgG1 Type Mouse Monoclonal Antibodies with DEAE-Trisacryl Chromatography (DEAE-Trisacryl 크로마토그래피법에 의한 IgG1 Type 쥐 단일클론 항체의 분리정제)

  • 최태부;정용근
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.335-342
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    • 1988
  • An anion exchange chromatography was employed for the purification of mouse monoclonal antibodies from ascitic fluid and in vitro cultivation media. After cultivation of hybridomas, Alps 25-3, HCGK, A4W, and KW, producing IgG1, the culture supernatants were harvested by centrifugation, precipitated with 50-60% ammonium sulfate, and dialyzed against 0.025 M Tris-HCI buffer (pH 8.2). Then the dialyzed samples were loaded into a DEAE-Trisacryl M anion exchange column. Monoclonal antibodies bound to the DEAE-Trisacryl M were eluted with 0.025 M Tris-HCI buffer (pH 8.2) containing 30-40 mM NaCl. In ammonium sulfate precipitation, the recovery of the monoclonal antibody was shown to be 90% and 84% from in vitro culture media containing 10% and 2% fetal bovine serum, respectively. On the other hand, the pretreatment by ultrafiltration enhanced the yield up to 91% whereas the purity was lower than that by ammonium sulfate treatment. Subsequently, in the DEAE-Trisacryl M chromatographic separation, the purities and recoveries of all the monoclonal antibodies from both the in vitro culture supernatants and ascitic fluids were 70-80% and 65% respectively. The monoclonal antibody, Alps 25-3 could be further purified with a purity of 95% through an immunoadsorbent chromatography.

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An Evaluation Study on the Cardiovascular Risk Factors in a Rural Adult Population (농촌지역 주민의 심혈관 질환 위험요인 평가)

  • Na, Baek-Ju;Park, Kyung-Soo;Lim, Jung-Su;Sun, Byeong-Hwan;Nam, He-Sung;Sohn, Seok-Joon
    • Journal of agricultural medicine and community health
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    • v.23 no.2
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    • pp.193-204
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    • 1998
  • Cardiovascular diseases are the leading cause of death and disability in Korea. Their risk factors can be classified as either modifiable or nonmodifiable and among modifiable factors are high bood pressure, elevated blood cholesterol, obesity and cigarette smoking. The purpose of this study was to evaluate the risk factors for the cardiovascular diseases in a rural community and to get basic data for the development of a community-based rick reduction intervention program. Evaluation involved population-based, cross-sectional samples of adult residents in a rurual community. We measured blood pressure, body fat percent by bioelectric impedance fatness analyzer and serum cholesterol and interviewed adult residents over 20-year-old age. Blood pressure was checked twice and hypertension was classified by the sixth report of the Joint National Committee on Detection. Evaluation, and Treatment of High Blood Pressure. The Cutpoints for high blood cholesterol was used National Cholesterol Treatment Guidelines and those for obesity was 25% in male. 30% in female. The results were as follows: 1. Prevalence of definitive hypertension was 59.7% in males and 54.4% in female. 2. Prevalence of hypercholesterolemia was 14.3% in male and 18.2% in female. 3. Prevalence of obese was 10.7% in male and 41.1% in female. 4. Among definitive hypertension, hypercholesterolemia, and obesity 52.1% possessed one risk factor, 12.6% two risk factors and 2.5% three risk factors in males. In females 41.4% possessed one risk factor and 27.6%. 5.7% respectively. 5. The smoking rate was 65.8% in males and 5.2% in females. Our results are used effectively for the community-based intervention towards cardiovascukr diseases risk reduction. However, because of limitations in our study design, further datas are needed including other risk factors and in-person clinical datas.

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Bioequivalence Test of Rebamipide 100 mg Tablets (레바미피드 100 mg 정제의 새울학적동등성)

  • Kim, Se-Mi;Cho, Hea-Young;Kang, Hyun-Ah;Lee, Yong-Bok
    • Korean Journal of Clinical Pharmacy
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    • v.18 no.1
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    • pp.38-44
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    • 2008
  • Rebamipide, ($\pm$)-2-(4-chlorobenzoylamino)-3-[2(1H)-quinolinon-4-yl] propionic acid, is used for mucosal protection, healing of gastroduodenal ulcers, and treatment of gastritis. It works by enhancing mucosal defense, scavenging free radicals and temporarily activating genes encoding cyclooxygenase-2. The purpose of the present study was to evaluate the bioequivalence of two rebamipide tablets, $Mucosta^{(R)}$ (Korea Otsuca Pharmaceuticals Co., Ltd.) and Mustar (Korean Drug Co., Ltd.), according to the guidelines of the Korea Food and Drug Administration (KFDA). The release of rebamipide from the two rebamipide formulations in vitro was tested using KP VIII Apparatus II method with pH 6.8 dissolution medium. Twenty six healthy male subjects, $23.46{\pm}2.63$ years in age and $66.62{\pm}8.97\;kg$ in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After a single tablet containing 100 mg as rebamipide was orally administered, blood samples were taken at predetermined time intervals and the concentrations of rebamipide in serum were determined using HPLC with fluorescence detector. The dissolution profiles of two formulations were similar in the tested dissolution medium. The pharmacokinetic parameters such as $AUC_t$, $C_{max}$ and $T_{max}$ were calculated, and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_t$, $C_{max}$ and untransformed $T_{max}$. The results showed that the differences between two formulations based on the reference drug, $Mucosta^{(R)}$ were -5.08, 3.52 and -9.71 % for $AUC_t$, $C_{max}$ and $T_{max}$, respectively. There were no sequence effects between two formulations in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log 0.8 to log 1.25 (e.g., log 0.84$\sim$log 1.07 and log 0.90$\sim$log 1.17 for $AUC_t$ and $C_{max}$, respectively). Thus, the criteria of the KFDA bioequivalence guideline were satisfied, indicating Mustar tablet was bioequivalent to $Mucosta^{(R)}$ tablet.

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Effects of Insulin-like Growth Factor in Peritoneal Fluid of Patients with Endometriosis on the Proliferation of Endometrial Stromal Cells (자궁내막증 환자의 복강액내 IGF가 자궁내막 기질세포 증식에 미치는 영향)

  • Kim, Jung-Gu;Suh, Chang-Seok;Kim, Seok-Hyun;Choi, Young-Min;Moon, Shin-Yong;Lee, Jin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.3
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    • pp.331-338
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    • 1999
  • The purposes of this study were to evaluate the effects of insulin-like growth factor (IGF)s in peritoneal fluid (PF) from patients with and without endometriosis on the proliferation of endometrial stromal cells and to investigate the effects of type I IGF receptor antibody on the response of endometrial stromal cells to PF from patients with endometriosis. IGFs in PF from patients with endometriosis (n=14) and without endometriosis (n=10) were measured by immunoradiometric assay and PF samples were divided into low IGF-I PF group (less than 85 ng/ml) and high IGF-I PF group (more than 85 ng/ml). Endometrial stromal cells from patients without endometriosis were cultured in serum free media in the presence or absence of 1 % PF and thymidine incorporation test were used to evaluate the proliferation of endometrial stromal cells. Also cultures were incubated with type I IGF receptor monoclonal antibody (${\alpha}IR_3$) before adding PF. PF from patients with endometriosis and without endometriosis increased thymidine incorporation in endometrial stromal cells. In patients with endometriosis, high IGF-I PF group had high IGF-II levels and resulted in higher thymidine incorporation than low IGF-I PF group but no significant difference in increase in thymidine incorporation between high IGF-I and low IGF-I PF group was noted in patients without endometriosis. There was not a significant correlation between increase in thymidine incorporation and IGF-I levels in PF from patients without endometriosis but in PF from patients with endometriosis. Preincubation with ${\alpha}IR_3$ significantly inhibited the mitogenic response of endometrial stromal cells to PF. Our data indicate that IGF-I in PF may be involved in the growth of ectopic endometrium in patients with endometriosis.

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Effect of Sinapis alba L. on expression of interferon-gamma and interleukin-4 production in anti-CD3/anti-CD28-stimulated CD4(+) T cells (CD4+ T cells에서 백개자가 IFN-$\gamma$와 IL-4 생성에 미치는 영향)

  • Park, Dae-Jung;Lee, Jang-Cheon;Lee, Young-Cheol
    • The Korea Journal of Herbology
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    • v.25 no.2
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    • pp.129-136
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    • 2010
  • Objective : Sinapis alba L. (SA) is a korean traditional herbal medicine that is usually used to prevent or treat inflammatory diseases, such as respiratory infection and rheumatoid arthritis. However, the effects of SA supplementation in vitro on serum antibody levels, splenocyte and peritoneal macrophage immune responses have not yet been determined. In this study, we examined the effect of SA on the production of Th1/Th2 cytokines. Methods : Splenocytes were isolated from naive C57BL/6 mice. Cells were enriched for CD4+ cell populations by first staining the cells with anti-CD4 (BD PharMingen, Calif, USA). CD4+ T cells were selected on a (CS) column, and the flow-through was collected as CD4+ T cells. Isolated cells were activated by overnight incubation on 24-well plates coated with $1{\mu}g/mL$ anti-CD3, $1{\mu}g/mL$ anti-CD28 and with SA ($100{\mu}g/mL$). Primary macrophages were collected from the peritoneal cavities of mice (8-week-old female C57BL/6). The peritoneal macrophages were washed and plated with RPMI-1640 overnight for the experiments. After 48-hours cultures, samples were centrifuged at 2000 rpm for 10 minutes, and the supernatants were stored at $-80^{\circ}C$. Mouse IL-4, IFN-$\gamma$ and TNF-$\alpha$ were quantified using ELISA kits (BioSource International, Camarillo, Calif, USA) according to the manufacturer's protocols. Results : SA at 100ug/ml decreased the generation of Th1 cytokine (IFN-$\gamma$) by 0.5-fold. However, SA has no effect on Th2 (IL-4) production. Conclusions : These results suggest that SA may play an important role in the control of T-cell-mediated autoimmunity by down-regulation of Th1 cytokine (especially IFN-$\gamma$, TNF-$\alpha$). These data may contribute to the design of new immunomodulating treatments for a group of autoimmune diseases.

Disposable Glucose Sensor Based on Platinised Carbon Paste Electrode (백금 도금된 탄소반죽 전극을 이용한 일회용 글루코오스 센서)

  • Lee Dong Joo;Yoo Jae Hyun;Cui Gang;Choi Moon Hee;Kim Moon Hwan;Ryu Joon Oh;Han Sang-Hyun;Cha Geun Sig;Nam Hakhyun
    • Journal of the Korean Electrochemical Society
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    • v.2 no.4
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    • pp.227-231
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    • 1999
  • Disposable, amperometric glucose sensor was constructed using platinised carbon paste electrode. The sensor response was studied by amperometry and cyclic voltammetry applying sample solutions on the strip-type electrode. Platinization of screen-printed carbon paste electrode effectively improved the electrochemical reversibility of a mediator and the analytical characteristics of the sensor. The heterogeneous rate constant for $[Fe(CN)_6]^{4-/3-}$ was $1.45\times10^{-2}cm{\cdot}s^{-1}$. An applied potential of 0.3V vs. Ag/AgCl resulted in the best selectivity for glucose. The apparent Michaelis-Menten constant for glucose on the strip sensor, $K_m^{app}$, was 24.5 mM. To evaluate the analytical performance of the glucose sensor strip, a correlation study was performed with the NOVA S.P, Ultra M analyzer for 30 serum samples containing $80\~297mg/dL$ of glucose: the correlation coefficient value was 0.983. It can be seen that the strip sensor has satisfactory precision and accuracy.

Analysis of the Prognostic Factors for Abdominal Trauma (복부외상환자의 예후에 영향을 미치는 인자들에 대한 분석)

  • Kim, Hee Joon;Kim, Hyung Soo;Seo, Kyung Won;Ju, Jae Kyun;Ryu, Seong Yeop;Kim, Jeong Cheol;Kim, Hyung Rok;Park, Young Kyu;Kim, Dong Yi;Kim, Young Jin;Kim, Shin Kon
    • Journal of Trauma and Injury
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    • v.20 no.1
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    • pp.12-18
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    • 2007
  • Purpose: Recently, trauma is more frequent due to the increases in the population, the number of traffic accident, and the incidence of violence. Especially, abdominal trauma is a leading cause of morbidity and mortality. We analyzed the clinical features and the factors associated with morbidity and mortality. Methods: We analyzed 136 patients of abdominal trauma who were admitted at the Department of Surgery, Chonnam National University Hospital, from January 2003 to June 2005. We analyzed the cause of trauma, the injured organ, combined injuries, mental status, blood pressure, laboratory findings, morbidity, and mortality. The relationships between by variable were assesed by using the independent samples test and the Kruskal?Wallis test. Results: The causes of trauma were traffic accidents (98 cases, 72%), falling accidents (9 cases, 6.6%), violence (6 cases, 4.4%), and stab injuries (6 cases, 4.4%). The injured organs were the small intestines (47 cases, 34.6%), the liver (35 cases, 25.7%), the spleen (26 cases, 19.1%), the mesentery (17 cases, 12.5%), the large intestines (15 cases, 11.0%), the pancreas (14 cases, 10.3%), etc. The most common combined injury was chest injury (53 cases, 39%). Comatose or semicomatose mental status and shock on admission (<60 mmHg in systolic) were related to high mortality (85.7%). In laboratory findings, decreased hemoglobin (<8 g/dL), and platelet count (<$50,000/mm^3$), and increased creatinine level (>1.6 mg/dL) were significant prognostic factors. The incidence of postoperative complications was 40.4%, and frequent complications were wound infection (8.1%) and re-bleeding (8.1%). The overall mortality rate was 18.4%, and most common cause was hypovolemic shock (18 cases, 13.2%), however, there was no statistical difference according to injurd organ. Conclusion: In the multivariate analysis, mental status, hemoglobin, and serum creatinine level were the most significant prognostic factors. When an abdominal trauma patient arrives at the emergency room, a rapid and accurate evaluation of the patient's status and risk factors, and resuscitation, if necessary, have to be performed to lower the morbidity and mortality.

Attenuation of Renal Ischemia-Reperfusion Injury by Antioxidant Vitamins in Pigs (돼지의 신장에서 Antioxidant Vitamins에 의한 허혈 및 재관류 손상의 감소에 관한 연구)

  • Kim, Myung-Jin;Lee, Soo-Jin;Park, Chang-Sik;Son, Hwa-Young;Jun, Moo-Hyung;Jeong, Seong-Mok;Kim, Myung-Cheol
    • Journal of Veterinary Clinics
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    • v.24 no.2
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    • pp.94-98
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    • 2007
  • This study was to investigate the effects of ascorbic acid and alpha-tocopherol on the attenuation of renal ischemia-reperfusion (IR) injury in pigs. Ten pigs were subjected to 60 minutes of warm unilateral renal ischemia followed by removal of contralateral kidney and then divided into two groups. Treatment group was performed ascorbic acid and alpha-tocopherol pretreatment 2 days before operation and ascorbic acid with heparin-saline solution irrigation-aspiration. Otherwise, control group used only irrigation-aspiration of heparin-saline solution. Blood samples were collected from these pigs for measurement of serum blood urea nitrogen (BUN) and creatinine values, antioxidant superoxide dismutase (SOD) at pre, day 1, day 3, day 7 and day 14. The kidneys were taken for histopathologic evaluation after euthanasia on postoperative day 14. The levels of BUN were significantly increased in the control group on day 1, day 3 and day 7 (P<0.05). And the level of creatinine was significantly increased in the control group on day 3 (p<0.05). Activity of antioxidant enzymes in plasma revealed significant difference (p<0.05) between control and treatment group at day 14. In histopathologic findings, treatment group was showed less damage than that of control group on the basis of renal tubular damage. It was concluded that ascorbic acid and alpha-tocopherol attenuated renal I/R injury in the pigs.

An Outbreak of Ibaraki Disease in Korea (한국(韓國)에서의 이바라기 병(病)(Ibaraki disease)의 발생)

  • Bak, Ung-Bok;Cheong, Chang-Kook;Choi, Hee-In;Lee, Chang-Woo;Oh, Hyo-Sung;Rhee, Young-Ok;Cho, Myung-Rae;Lim, Young-Il
    • Korean Journal of Veterinary Research
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    • v.23 no.1
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    • pp.81-89
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    • 1983
  • Epizootics and sporadic outbreaks of the cattle disease characterized by fever, stomatitis and dysphagia were observed in Korea in September through November, 1982. The number of cattle showed typical symptoms of the disease was estimated to 73 and these animals were concentrated in Gyongido province, the central district of Korea. In other districts around Gyongido province the disease tended to be sporadic occurrence. The main lesions of the disease were defined as edema, hemorrhage, degeneration and necrosis of the oral mucosa and musculatures of the esophagus and laryngopharyngeal area, and abomasum. Of these lesions hyaline degeneration and hemorrhage of the striated muscle of the esophagus and laryngopharyngeal area were the most striking changes of the disease and rated to be the lesion of diagnostic significance. The serum samples of the cattle with typical symptoms showed significant positive titer of the antibodies against Ibaraki virus. On these clinical, epidemiological, pathological and serological findings of the disease it was diagnosed as Ibaraki disease presenting the only report on the epizootics in the countries other than Japan.

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Effect of Polysaccharides from Astragalus membranaceus on Exercise-Induced Fatigue and Oxidative Damage in Skeletal Muscle in Exhaustive Exercise Animal Models (과도 운동에 의해 유발되는 피로 및 골격근 산화적 손상에 대한 황기 다당체의 효과)

  • Go, Eun Ji;Lee, Hannah;Park, Hyun Su;Kim, Soo Jin;Park, Yeong Chul;Seong, Eun Soo;Yu, Chang Yeon;Lim, Jung Dae
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.4
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    • pp.271-280
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    • 2018
  • Background: Astragalus membranaceus is a well known oriental medicinal herb. The polysaccharides of the aboveground parts (AMA) and the radix (AMR) of A. membranaceus are the most important functional constituents. Methods and Results: The aim of this study was to determine the effects of AMA and AMR on the oxidative damage induced in the skeletal muscle of rats subjected to exhaustive exercise. Sprague-Dawley rats were randomly divided into exercise and non-exercise groups; in the groups receiving the test compounds, AMA and AMR were administered orally for 30 days. Skeletal muscle samples were collected from each rat after running to exhaustion on a treadmill to determine the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) and the concentation of malondialdehyde (MDA). The antioxidant enzyme activities of SOD and GSH-Px of skeletal muscle of AMA- and AMR-treated groups were significantly higher than those of the control and commercial sports drink (SPD)-treated groups in exhaustive exercise rats. In addition, MDA concentrations in the skeletal muscle of the AMA- and AMR-treated groups were significantly lower than those of the control and SPD-treated groups. In the present study, the effects of AMA and AMR on exercise endurance capacity were also evaluated in mice subjected to a swimming exercise test. AMA and AMR supplementation prolonged the swimming time of mice and enhanced exercise endurance capacity. AMA and AMR possess the ability to retard and lower the production of blood lactate, and prevent the decrease of serum blood glucose. Conclusions: These results showed that, AMR and AMA exerted beneficial effect in mice, increasing the activity of the antioxidant systems and inhibiting oxidative stress induced by exhaustive exercise. The compounds improved exercise performance and showed anti-fatigue effects against exhaustive exercise.