• Title/Summary/Keyword: ${gamma}-glutamyl$ transpeptidase

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Regulatory Mechanisms of Angiotensin II on the $Na^+/H^+$ Antiport System in Rabbit Renal Proximal Tubule Cells. I. Stimulatory Effects of ANG II on $Na^+$ Uptake

  • Han, Ho-Jae;Koh, Hyun-Ju;Park, Soo-Hyun
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.4
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    • pp.413-423
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    • 1997
  • The importance of the kidney in the development of hypertension was first demonstrated by Goldblatt and his colleagues more than fifty years ago. Many hormones and other regulatory factors have been proposed to play a major role in the development of hypertension. Among these factors angiotensia II (ANG II) is closely involved in renal hypertension development since it directly regulates $Na^+$ reabsorption in the renal proximal tubule. Thus the aim of the present study was to examine signaling pathways of low dose of ANC II on the $Na^+$ uptake of primary cultured rabbit renal proximal tubule cells (PTCs) in hormonally defined seum-free medium. The results were as follows: 1) $10^{-11}$ M ANG II has a significant stimulatory effect on growth as compared with control. Alkaline phosphatase exhibited significantly increased activity. However, leucine aminopeptidase and ${\gamma}-glutamyl$ transpeptidase activity were not significant as compared with control. In contrast to $10^{-11}$ M ANG II stimulated $Na^+$ uptake $(108.03{\pm}2.16% of that of control)$, $10^{-9}$ M ANG II inhibited ($92.42{\mu}2.23%$ of that of control). The stimulatory effect of ANG II on $Na^+$ uptake was amiloride-sensitive and inhibited by losartan (ANG II receptor subtype 1 antagonist) and not by PD123319 (ANG II receptor subtype 2 antagonist). 2) Pertussis toxin (PTX) alone inhibited $Na^+$ uptake by $85.52{\pm}3.52%$ of that of control. In addition, PTX pretreatment prevented the AMG II-induced stimulation of $Na^+$ uptake. 8-Bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP), forskolin, and isobutylmethylxanthine (IBMX) alone inhibited $Na^+$ uptake by $88.79{\pm}2.56,\;80.63{\pm}4.38,\;and\;84.47{\pm}4.74%$ of that of control, respectively, and prevented the ANG II-induced stimulation of $Na^+$ uptake. However, $10^{-11}$ M ANG II did not stimulate cAMP production. 3) The addition of 12-O-te-tradecanoylphorbol-13-acetate (TPA, 0.01 ng/ml) to the PTCs produced significant increase in $Na^+$ uptake ($114.43{\pm}4.05%$ of that of control). When ANG II and TPA were added together to the PTCs, there was no additive effect on $Na^+$ uptake. Staurosporine alone had no effect on $Na^+$ uptake, but led to a complete inhibition of ANG II- or TPA-induced stimulation of Na'uptake. ANG II treatment resulted in a $111.83{\mu}4.51%$ increase in total protein kinase C (PKC) activity. In conclusion, the PTX-sensitive PKC pathway is the main signaling cascade involved in the stimulatory effects of ANG II on $Na^+$ uptake in the PTCs.

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닭에서 고추씨박의 사료적 가치에 관한 연구

  • 임호중;강창원
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2001.11a
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    • pp.24-39
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    • 2001
  • These experiments were conducted to evaluate the nutritional and feeding value of red pepper seed oil meal (RPSOM) as local vegetable protein ingredients for poultry feeding. In the first experiment, nutritional values of RPSOM were evaluated by analyzing chemical compositions and determining true metabolizable energy (TME), nitrogen corrected TME (TMEn) and true available amino acid (TAAA) contents. According to the chemical analysis, RPSOM contained 22.50% of crude protein, 4.75% of ether extract, 27.70% of crude fiber, 0.34 mg/g of capsaicin and 49.97 ppm of xanthophylls. The values of TME and TMEn determined by force-feeding 16 roosters (ISA-Brown) were 1.73 kcal/g and 1.63 kcal/g on dry matter basis, respectively. The average TAAA value of 16 amino acids measured by the force-feeding technique was 81.70%. These values were used for formulating experimental diets containing various levels of RPSOM for broiler chicks and laying hens. Two feeding trials were made to investigate the effects of dietary incorporation of RPSOM into chicken feed performances of broiler chicks and laying hens. In the broiler feeding (Exp. 2), a total of two hundred twenty-five, 4 wk old male broiler chicks (Ross) were randomly divided into 9 groups of 25 birds each and assigned to three experimental diets containing 0, 5 and 10% RPSOM. The birds were fed ad libitum the diets for 3 wk and feed intake, weight gain and feed conversion rate were determined. At the end of the feeding, the blood levels of glutamate oxaloacetate transaminase (GOT), gamma glutamyl transpeptidase (GGT), blood urea nitrogen (BUN) and cholesterol, and the body and fatty acid compositions of leg muscle were measured. No significant differences were observed in weight gain, feed intake, feed conversion rate, body composition, serum levels of GOT, GGT and BUN among the treatments. However, blood cholesterol level was lower (P<0.05) in 10% RPSOM diet group than those in the other. The dietary RPSOM at 5 and 10% levels increased the content of linoleic acid (P<0.05) in leg muscle compared to that of control group. The results indicate that RPSOM can be used for broiler feed up to 10% without any significant negative effects on broiler performance. In the layer feeding (Exp. 3), the effects of dietary RPSOM on the performances of laying hen were investigated by feeding ninety 45 wk old laying hens (ISA-Brown) with experimental diets containing 0, 5 and 10% RPSOM for 4 wk (30 birds per treatment). Measurements were made on egg production rate, egg weight, feed intake, Haugh unit, egg shell strength which was higher (P<0.05) in layers fed 10% RPSOM diet compared to those fed 0 and 5% RPSOM diets. Thus, it can be concluded that RPSOM can be included into laying hen feed up to 10% without any harmful effects.

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Functional characterization of primary culture cells grown in hormonally defined, serum-free medium and serum-supplemented medium (호르몬 한정배지를 이용한 세포 초대배양계의 확립)

  • Han, Ho-jae;Kang, Ju-won;Park, Kwon-moo;Lee, Jang-hern;Yang, Il-suk
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.551-563
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    • 1996
  • This study investigated the properties of primary cultured proximal tubule cells in hormonally defined(insulin, transferrin, and hydrocortisone), serum-free medium or 10% serum-supplemented medium. The growth rate of the primary cultured proximal tubule cells was lower in the hormonally defined, serum-free medium than in the 10% serum- supplemented medium(p < 0.05), while the activities of brush border marker enzymes, alkaline phosphatase(AP), leucine aminopeptidase(LAP), and y-glutamyl transpeptidase(${\gamma}$-GTP) were increased(p < 0.05). The activities of these enzymes, however, decreased with the lapse of incubation time to 50-70% after 6 days culture compared to those of the freshly-prepared proximal tubules. The enzymatic activities of the primary cultured proximal tubul cells on 6, 9, 12, and 15 days of culture were significantly increased in the hormonally defined, serum-free medium compared to the 10% serum-supplemented medium(p < 0.05). The functional differentiation of the primary culture was examined by observing multicellular domes of the confluent monolayer, which is indicative of transepithelial solute transport. The dome formation by the proximal tubule cultures occurred at a higher frequency in the hormonally defined, serum-free medium than in the 10% serum-supplemented medium(p < 0.05). Upon electron microscopic examination, an increased density of the brush border was observed in the hormonally defined, serum-free medium compared to the cells grown in 10% serum-supplemented medium. The activities of $Na^+$glucose cotransporter($^{14}C$-a-MG uptake), $Na^+$phosphate cotransportere($^{32}P$ uptake) and $Na^+$ transporter($^{22}Na^+$ uptake) in the brush border membrane, and of $Na^+/K^+$-ATPase($^{86}Rb$ uptake) in the basolateral membrane were significantly stimulated in the hormonally defined, serum-free medium than in 10% serum-supplemented medium(p < 0.05). In conclusion, the primary cultured proximal tubule cells grown in the hormonally defined, serum-free medium demonstrated a slower growth rate, but the functions of cell were enhanced.

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An Empirical Study of the Clinically Reportable Range in Clinical Chemistry (임상보고 가능범위의 실증적 연구)

  • Chang, Sang-Wu;Lee, Sang-Gon;Choi, Ho-Seong;Song, Eun-Young;Park, Yong-Won;Lee, In-Ae
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.1
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    • pp.31-36
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    • 2007
  • The purpose of the clinically reportable range (CRR) in clinical chemistry is to estimate linearity in working range. The reportable range includes all results that may be reliably reported, and embraces two types of ranges: the analytical measurement range (AMR) is the range of analyte values that a method can directly measure on the specimen without any dilution, concentration, or other pretreatment not part of the usual assay process. CAP and JCAHO require linearity on analyzers every six months. The clinically reportable range is the range of analyte values that a method can measure, allowing for specimen dilution, concentration, or other pretreatment used to extend the direct analytical measurement range. The AMR cannot exceed the manufacturer's limits. Establishing AMR is easily accomplished with Calibration Verification Assessment and experimental Linearity. For example: The manufacturer states that the limits of the AST on their instrument are 0-1100. The lowest level that could be verified is 2. The upper level is 1241. The verified AMR of the instrument is 2-1241. The lower limit of the range is 2, because that is the lowest level that could be verified by the laboratory. The laboratory could not use the manufacturer's lower limit of 2 because they have not proven that the instrument values below 2 are valid. The upper limit of the range is 1241, because although the lab has shown that the instrument is linear to 1241, the manufacturer does not make that claim. The laboratory needs to demonstrate the accuracy and precision of the analyzer, as well the validation of the patient AMR. Linearity requirements have been eliminated from the CLIA regulations and from the CAP inspection criteria, however, many inspectors continue to feel that linearity studies are a part of good lab practice and should be encouraged. If a lab chooses to continue linearity studies, these studies must fully comply with the calibration/calibration verification requirements of CLIA and/or CAP. The results of lower limit and upper limit of clinically reportable range were total protein (2.1 - 79.9), albumin (1.3 - 39), total bilirubin (0.2 - 106.2), alkaline phosphatase (13 - 6928.2), aspartate aminotransferase (24 - 7446), alanine aminotransferase (13 - 6724.2), gamma glutamyl transpeptidase (16.64 - 9904.2), creatine kinase (15.26 - 4723.8), lactate dehydrogenase (127.66 - 13231.8), creatinine (0.4 - 129.6), blood urea nitrogen (8.67 - 925.8), uric acid (1.6 - 151.2), total cholesterol (48.52 - 3162), triglycerides (36.91 - 3367.8), glucose (31 - 4218), amylase (21 - 6694.2), calcium (3.1 - 118.2), inorganic phosphorus (1.11 - 108), HDL (11.74 - 666), NA (58.3 - 1800), K (1.0 - 69.6), CL (38 - 1230).

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Hematologic and biochemical reference intervals for natural monument Korean goral (Naemorhedus caudatus) (천연기념물 한국 산양의 혈액 및 혈액 화학적 정상표준범위)

  • Ahn, Sangjin;Choi, Youngjin;Kim, Jong-Taek
    • Korean Journal of Veterinary Service
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    • v.40 no.2
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    • pp.155-159
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    • 2017
  • Korean goral (Neamorhedus caudatus) is registered as a natural monument number 217 by South Korea Cultural Heritage Administration. It is also recognized as the endangered species I by Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). In this study, blood samples of Korean goral were collected to make reference intervals. Blood sampling was conducted on 19 numbers of Korean gorals (ten females, nine males), which were reared in Korean Goral Restoration Center. For total samples, the reference intervals of hematological parameters were: white blood cell $7.69{\sim}10.87K/{\mu}{\Lambda}$; hematocrit 36.73~46.18%; red blood cell $10.72{\sim}12.86K/{\mu}{\Lambda}$; hemoglobin 12.79~15.14 g/dL; mean corpuscular volume 33.15~36.75 fL; mean corpuscular hemoglobin 11.53~12.23 pg; mean corpuscular hemoglobin concentration 32.64-35.91 g/dL and red blood cell distribution width 39.2~43.46%. For total samples of each parameters, the following results were obtained for serum biochemistry: glucose 111.81~153.77 mg/dL; blood urea nitrogen 22.35~28.91 mg/dL; creatine 1.22~1.84 mg/dL; phosphate 4.57~6 mg/dL; calcium 8.7~9.1 mg/dL; total protein 6.53~6.92 g/dL; albumin 3.1~3.48 g/dL; globulin 3.26~3.62 g/dL; alanine aminotransferase 56.7~158.56 U/L; aspartate aminotransferase 230.35~473.06 U/L; alkaline phosphatase 178.06~332.47 U/L; gamma-glutamyl transpeptidase 131.6-~181.24 U/L; total bilirubin 1.47~2.12 mg/dL; cholesterol 46.48~71.52 mg/dL; amylase 16.3~26.03 U/L; sodium 150.43~153.88 mmol/L; potassium 3.98~4.6 mmol/L and chlorine 109.48~113.26 mmol/L. The ranges of values were similar campared to previous studies except in the case of RDW value, which showed higher range than the RDW value of a previous study. The reference intervals from this study will be useful data for treatment and management of gorals.

An Empirical Study of the Recovery Experiment in Clinical Chemistry (임상화학검사실에서 회수율 실험의 실증적 연구)

  • Chang, Sang-Wu;Lee, Sang-Gon;Song, Eun-Young;Park, Yong-Won;Park, Byong-Ok
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.184-188
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    • 2006
  • The purpose of the recovery experiment in clinical chemistry is performed to estimate proportional systematic error. We must know all measurements have some error margin in measuring analytical performance. Proportional systematic error is the type of error whose magnitude increases as the concentration of analyte increases. This error is often caused by a substance in the sample matrix that reacts with the sought for analyte and therefore competes with the analytical reagent. Recovery experiments, therefore, are used rather selectively and do not have a high priority when another analytical method is available for comparison purposes. They may still be useful to help understand the nature of any bias revealed in the comparison of kit experiments. Recovery should be expressed as a percentage because the experimental objective is to estimate proportional systematic error, which is a percentage type of error. Good recovery is 100.0%. The difference between 100 and the observed recovery(in percent) is the proportional systematic error. We calculated the amount of analyte added by multiplying the concentration of the analyte added solution by the dilution factor(mL standard)/(mL standard + mL specimen) and took the difference between the sample with addition and the sample with dilution. When making judgments on method performance, the observed that the errors should be compared to the defined allowable error. The average recovery needs to be converted to proportional error(100%/Recovery) and then compared to an analytical quality requirement expressed in percent. The results of recovery experiments were total protein(101.4%), albumin(97.4%), total bilirubin(104%), alkaline phosphatase(89.1%), aspartate aminotransferase(102.8), alanine aminotransferase(103.2), gamma glutamyl transpeptidase(97.6%), creatine kinase(105.4%), lactate dehydrogenase(95.9%), creatinine(103.1%), blood urea nitrogen(102.9%), uric acid(106.4%), total cholesterol(108.5), triglycerides(89.6%), glucose(93%), amylase(109.8), calcium(102.8), inorganic phosphorus(106.3%). We then compared the observed error to the amount of error allowable for the test. There were no items beyond the CLIA criterion for acceptable performance.

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An Empirical Study of the Analytical Measurement Range in Clinical Chemistry (분석측정범위의 실증적 평가)

  • Chang, Sang-Wu;Lee, Sang-Gon;Kim, Young-Hwan;Song, Eun-Young;Park, Yong-Won;Park, Byong-Ok;Lyu, Jae-Gi
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.2
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    • pp.117-124
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    • 2006
  • The analytical measurement range (AMR) is the range of analyte values that a method can directly measure on a specimen without any dilution, concentration, or other pretreatment not part of the usual assay process. The linearity of the AMR is its ability to obtain test results which are directly proportional to the concentration of analyte in the sample from the upper and lower limit of the AMR. The AMR validation is the process of confirming that the assay system will correctly recover the concentration or activity of the analyte over the AMR. The test specimen must have analyte values which, at a minimum, are near the low, midpoint, and high values of the AMR. The AMR must be revalidated at least every six months, at changes in major system components, and when a complete change in reagents for a procesure is introduced; unless the laboratory can demonstrate that changing the reagent lot number does not affect the range used to report patient test results. The AMR linearity was total protein (0-16.6), albumin (0-8.1), total bilirubin (0-18.1), alkaline phosphatase (0-1244.3), aspartate aminotransferase (0-1527.9), alanine aminotransferase (0-1107.9), gamma glutamyl transpeptidase (0-1527.7), creatine kinase (0-1666.6), lactate dehydrogenase (0-1342), high density lipoprotein cholesterol (0.3-154.3), sodium (35.4-309), creatinine (0-19.2), blood urea nitrogen (0.5-206.2), uric acid (0-23.9), total cholesterol (-0.3-510), triglycerides (0.7-539.6), glucose (0-672.7), amylase (0-1595.3), calcium (0-23.9), inorganic phosphorus (0.03-17.0), potassium (0.1-116.5), chloride (3.3-278.7). We are sure that materials for the AMR affect the evaluation of the upper limit of the AMR in the process system.

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Comparison of hematologic and biochemical values in htPA transgenic pigs (사람 조직 플라스미노겐 활성인자 생산용 형질전환 돼지에서의 혈액학적 성상 비교)

  • Park, Mi-Ryung;Hwang, In-Sul;Lee, Seunghoon;Lee, Hwi-Cheul
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.12
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    • pp.395-400
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    • 2020
  • Pigs have been used widely in biomedical research owing to their physiologic and anatomic similarities to humans. Analysis of the hematologic and biochemical values in pigs is an important basis for biomedical research and veterinary clinical diagnosis, but research on transgenic pigs has been sparse. This study was conducted to obtain basic data on transgenic pigs and to describe and compare the reference values for hematologic and biochemical parameters in human tissue plasminogen activator (htPA) transgenic pigs vs normal pigs. Blood samples were obtained from 7 normal LY (Landrace-Yorkshire crossbred) pigs and 8 transgenic pigs and 16 hematologic and 15 serum biochemical parameters were tested. Among the hematologic parameters tested, significant differences were observed in the red blood cells (RBC), mean red blood cell hemoglobin (MCH), and lymphocytes (LYM), between the non-transgenic and transgenic pigs. Among the biochemical parameters tested, the blood urea nitrogen (BUN), total protein (TP), cholesterol (CHOL), alanine aminotransferase (ALT), creatinine (CREA), gamma glutamyl transpeptidase (GGT), globin (GOB), and amylase (AMYL) showed significant differences between the two groups. Thus, the values determined in this study can be used as basic reference values for transgenic pigs and will contribute to their use in biomedical research.

Effect of fermented Hovenia dulcis Thunb fruit water extract on biomarker for liver injury and body weight changes in rats given oral administration of ethanol (헛개열매추출액발효물이 흰쥐의 에탄올 경구투여에 의한 간손상 지표와 체중 감량 완화에 미치는 영향)

  • Choi, Ji-Young;Kim, Jun-Han;Kim, Gho;Kim, Choon-Kyung;Choi, Myung-Sook
    • Food Science and Preservation
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    • v.21 no.3
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    • pp.412-420
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    • 2014
  • This study was performed to investigate the effect of fermented Hovenia dulcis Thunb fruit water extract on biomarkers for acute (a) ethanol-induced hangover and chronic (c) ethanol-induced liver injury in rats. For acute ethanol-induced hangover, the rats were administered distilled water (D.W., 10 mL/kg body wt.), Hovenia dulcis Thunb fruit water extract (HWE, 400 mg/10 mL/kg body wt.) and fermented HWE (FHWE, 400 mg/10 mL/kg body wt.), respectively, before 40% ethanol (5 g/kg body wt.) was administered. For chronic ethanol-induced liver injury, the rats were randomly divided into the normal control (cNC), ethanol (cET), cET-HWE and cET-FHWE groups. The cNC and cET groups were administered D.W. (10 mL/kg body wt.) before 40% alcohol (5 g/kg body wt.) was administrered for 21 days. The cET-HWE and cET-FHWE groups were administered HWE (400 mg/10 mL/kg body wt.) and FHWE (400 mg/10 mL/kg body wt.), respectively before 40% ethanol (5 g/kg body wt.) administration for 21 days. For acute ethanol-induced hangover, the serum alcohol and acetaldehyde concentrations were more significantly reduced in the aHWE and aFHWE groups than in the aET group. Moreover, the effect of FHWE was greater than that of HWE. For chronic ethanol-induced liver injury, the serum ethanol, acetaldehyde, ${\gamma}$-glutamyl transpeptidase (${\gamma}$-GTP) levels and the hepatic lipids concentration more significantly dropped in the cET-HWE and cET-FHWE groups than in the cET group. The FHWE administration showed faster recovery of the serum glucose concentration than in the cET and cET-HWE groups. The body weight reduction tended to normalize in the cET-HWE and cET-FHWE groups, which is ideal for chronic ethanol administration. These results suggest that FHWE has a protective effect against ethanol-induced liver damage, as evidenced by its ability to lower the serum ethanol and acetaldehyde concentrations after alcohol administration, and by its ability to decrease the level of ${\gamma}$-GTP and hepatic lipids. FHWE also elevated serum glucose concentration. Therefore, FHWE is effective in reducing ethanol-induced hangover and can play a beneficial role in the treatment of ethanol-induced liver damage as well as body weight reduction.

Lymphocyte Subpopulations and Proliferation of T cells, Phagocytic Activity of Leukocytes on Alcoholics (알코올중독자의 백혈구탐식능, 림프구아형 및 증식능)

  • 김용호;서병배;이정녀;김영훈
    • Biomedical Science Letters
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    • v.2 no.2
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    • pp.167-174
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    • 1996
  • Alcoholics increased susceptibility to microbial infection that is associated with decreased immunity. but there has been little experimental evidence to support alcoholics-induced increase of microbial infection directly in non-specific immunity. Therefore, we were used the method of phagocytic-plaque including all the stimulating factors for the phagocytosis, subtypes of lymphocytes and T-lymphocyte proliferation. The experimental groups were divided into 3 groups: (1) alcoholics who were hospitalized less than 1 week (newly hospitalized alcoholics), (2) alcoholics who were hospitalized more than 2 weeks (old hospitalized alcoholics), (3) healthy blood donors. We have studied 98 alcoholics and 35 healthy blood donors and control groups. A physician has checked the biological markers and diagnosed the body-condition alcoholics. The immunity and non-specific immunity on the alcoholics were analyzed by using the simultest kit and flow cytometry. Proliferation of the lymphocytes was analyzed by the phytohemmagglutinine mitogen. Phagocytosis and migration properties of leukocytes were identified on the layer formed by Staphylococcus aureus Cowan I strain. Biological markers of alcoholics and control groups, by such as blood glucose, ${\gamma}$-glutamyl transpeptidase and mean corpuscular volumes of red blood cells, were determined by biochemical and hematological methods. Compared with control groups, cluster of differentiation (CD)3+, CD8+ and CD19+ in alcoholic were more decreased except CD4+/CD8+ ratio. Proliferation of the T-lymphocytes, phagocytosis and migration properties of the leukocytes in alcoholics were decreased compared with those of control groups. According to the results observed in our experiment, they can be summerized as follows: 1, Cellular, humoral and non-specific immunities, are markedly decreased in alcoholics than those in control groups. 2. It is inferred that Phagocytic plaque formation is a very useful method to evaluate phagocytosis and migration properties of the alcoholic leukocytes 3. It is thought that the subtypes of lymphocytes, especially CD4+/CD8+ ratio, are essential methods to analyzed the alcoholic immunity. 4. Specific and non-specific immunity on the old hospitalized alcoholics was slightly increased, which depends upon the alcoholic medication.

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