• Title/Summary/Keyword: Human red blood cell

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Comparative Analysis of Screening Results from Various ELISA Formats Used for Detection of Anti-Erythropoietin Antibodies in Korean Patients

  • Ha, Sung-Kyu;Yang, Seung-Ju;Shin, Sug-Kyun;Jo, Young-Il;Baek, Kyung-Min;Hong, Seung-Hwa;Pack, Seung-Pil;Kim, Sung-Jo;Heo, Tae-Hwe
    • Biomolecules & Therapeutics
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    • v.18 no.2
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    • pp.184-190
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    • 2010
  • Clinical cases of pure red cell aplasia (PRCA) have been reported during the recombinant human erythropoietin (EPO) therapy for the anemia patients. PRCA is a rare hematological disorder leading to a severe anemia due to an almost complete stop of red blood cell production. Antibody (Ab)-associated PRCA is caused by the EPO-neutralizing Abs that eliminate the biological activity of EPO. In order to detect anti-EPO Abs in human sera, we performed conventional ELISA, directly coated bridging ELISA, and streptavidin coated bridging ELISA, and compared their sensitivity and specificity. Some false positive results were obtained in the conventional ELISA. One positive sample was detected successfully by streptavidin coated bridging ELISA, which was not appeared in the directly coated bridging ELISA. In conclusion, streptavidin coated bridging ELISA was substantially sensitive and specific format and one out of sixty-eight serum samples was proved to be anti-EPO positive.

Inhibitory Effect of Injinchunggantang(Yinchenqinggan-tang) on Hepatic Sclerosis (인진청간탕(茵蔯淸肝湯)이 간보호(肝保護) 및 섬유화(纖維化) 억제(抑制)에 미치는 영향(影響))

  • Seung Hyun-Suk;Lee Jang-Hoon;Woo Hong-Jung;Kim Young-Chul
    • The Journal of Internal Korean Medicine
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    • v.24 no.1
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    • pp.21-32
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    • 2003
  • Objective : The aim of this study is to investigate the inhibitory effect of Injinchunggantang on hepatic sclerosis induced by $CCl_4$. Methods : Weight, liver function test and complete blood cell count, gross findings, and findings on liver tissue of the past(Hematoxylin & Eosin stain, Masson-Trichrome stain) were studied. Results : When it comes to the change of rats' body weight, The $CCl_{4^+}$Injinchunggantang group lost far less weight than The $CCl_{4^-}$only group. In the liver function test, which is focused on various areas such as total cholesterol, alkaline phosphotase, albumin, aspartate transaminase, alanine transaminase, The $CCl_{4^+}$Injinchunggantang group was much more closer to normal limit than the $CCl_4$ only group. In the complete blood cell count, including white blood cell, red blood cell, hemoglobin, hematocrite, platelet, The $CCl_{4^+}$Injinchunggantang group significantly closer to normal limit than $CCl_{4^-}$only group. In the gross findings of hepatic fibrosis models, Injinchunggantang showed inhibitory effect on hepatic fibrosis in the order. In the past findings of hepatic fibrosis models in Hematoxylin & Eosin, Masson-Trichrome staining, the liver in $CCl_{4^-}$only group showed atrophy and necrotic change with white nodules, whereas that of $CCl_{4^+}$Injinchunggantang group showed lesser significant change with the well_preserved tone of the tissue. In the extent of the inhibition of the hepatic fibrosis, the Injinchunggantang group showed statistically significant inhibitory effect(p<0.05) in the sclerosis model. Conclusions : These results show that Injinchunggantang have inhibitory effect on hepatic sclerosis induced by $CCl_4$ and further ultimately prevent liver cirrhosis. To obtain more credible results in this experiment, the invention of a new experimental model more similar to human hepatic sclerosis is still needed.

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Anti-apoptotic Effects of Red Ginseng on Oxidative Stress Induced by Hydrogen Peroxide in SK-N-SH Cells

  • Kim, Eun-Hye;Lee, Mi-Jeong;Kim, In-Hye;Pyo, Suhk-Neung;Choi, Kwang-Tae;Rhee, Dong-Kwon
    • Journal of Ginseng Research
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    • v.34 no.2
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    • pp.138-144
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    • 2010
  • Ginseng (Panax ginseng C.A. Meyer) has been shown to have anti-stress effects in animal studies. However, most studies have only managed to detect altered levels of biomarkers or enzymes in blood or tissue, and the actual molecular mechanisms by which ginseng exerts these effects remain unknown. In this study, the anti-oxidative effect of Korean red ginseng (KRG) was examined in human SK-N-SH neuroblastoma cells. Incubation of SK-N-SH cells with the oxidative stressor hydrogen peroxide resulted in significant induction of cell death. In contrast, pre-treatment of cells with KRG decreased cell death significantly. To elucidate underlying mechanisms by which KRG inhibited cell death, the expression of apoptosis-related proteins was examined by Western blot analysis. KRG pre-treatment decreased the expression of the pro-apoptotic gene caspase-3, whereas it increased expression of the anti-apoptotic gene Bcl-2. Consistent with this, immunoblot analysis showed that pre-treatment of the SK-N-SH cells with KRG inhibited expression of the pro-inflammatory gene cyclooxygenase 2 (COX-2). RT-PCR analysis revealed that the repression of COX-2 expression by KRG pre-treatment occurred at the mRNA level. Taken together, our data indicate that KRG can protect against oxidative stress-induced neuronal cell death by repressing genes that mediate apoptosis and inflammation.

A Characteristics of Phagocytic Plaque on Staphylococcus aureus Layer Formed by Leukocytes of the Alcoholics (Staphylococcus aureus를 이용한 주정중독환자 백혈구의 Phagocytic Plaque 형성력)

  • 성희경;서병배;김용호
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.205-212
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    • 2002
  • This study was conducted to develop a method for direct determination of phagocytic activities in human circulatic systems and to measure the phagocytic activities in human leukocytes from the alcoholics, since phagocytic activity was considered to be very important in human immune mechanism at early stage for the health care of the alcoholics. The subjects for this study were 130 among which 95 males and 3 females were diagnosed as alcoholism and 32 was healthy blood donors. A thin layer of heat-killed Staphylococcus aureus Cowan I was placed on a plastic dish and reacted with whole blood to measure the phagocytic plaque formation by human leukocytes. In order to determine the health conditions of the subjects, some clinical laboratory tests, such as white blood cell counts, hemoglobin contents (Hgb), mean corpuscular volume of red blood cells(MCV), serum electrophoresis, B and T-lymphocytes, T-lymphocyte subtypes and phytohemagglutination test were also implemented. Compared to the non-alcoholism, new and old alcoholic inpatients showed statistically significant differences on levels of Hgb and MCV (p<0.05), but showed that T and B-lymphocyte numbers decreased and Helper T cell/Suppressor T cell ratio ($1.6{\pm}0.8$%) increased. Compared to non-alcoholism, phagocytic plaque activities of leukocytes from alcoholic patients decreased significantly and an unusual pattern in phagocytic plaque was observed, showing a strange body and chain shaped phagocytosis. Based upon these results, it is concluded that a phagocytic-plaques of Staphylococcus aureus Cowan I by leukocytes was very simple and useful method for the early immunological determination of phagocytic activities in alcoholic patients without requiring any special equipments.

Investigations into the immunomodulatory activity of Ulmus davidiana Planch extracts

  • Lee, Eon-Do;Yoon, Jong-Hwa;Lee, Seung-Deok;Kim, Kap-Sung;Kim, Kyung-Ho
    • Journal of Acupuncture Research
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    • v.22 no.2
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    • pp.83-92
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    • 2005
  • Objective: Although the effect of Ulmus davidiana Planch (UD) extracts on collagen-induced-arthritis (CIA) and bone metabolism has been studies, research on its effect on human immunomodulatory activity is further a due. The objective of the present study was to investigate the immunomodulatory activity of UD on cellular and humoral immunity. Methods : Oral administration of the ethanolic and water extracts of UD, at doses of 20, 100 and 200 mg/kg in mice, dose dependently potentiated the delayed type hypersensitivity reaction induced both by sheep red blood cells (SRBC) and oxazolone. Results : It significantly enhanced the production of circulating antibody titre in response to SRBC in mice. Extracts of UD failed to show any effect on macrophage phagocytosis. Chronic administration of UD extracts significantly ameliorated the total white blood cell count and also restored the myelosuppressive effects induced by cyclophosphamide. Conclusion : The present investigation reveals that UD extracts possesses immunomodulatory activity.

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Cell Selectivity of an Antimicrobial Peptide Melittin Diastereomer with D-amino Acid in the Leucine Zipper Sequence

  • Zhu, Wan Long;Nan, Yong Hai;Hahm, Kyung-Soo;Shin, Song-Yub
    • BMB Reports
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    • v.40 no.6
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    • pp.1090-1094
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    • 2007
  • Melittin (ME), a linear 26-residue non-cell-selective antimicrobial peptide, displays strong lytic activity against bacterial and human red blood cells. To design ME analogue with improved cell selectivity, we synthesized a melittin diastereomer (ME-D) with D-amino acid in the leucine zipper sequence (Leu-6, Lue-13 and Ile-20). Compared to ME, ME-D exhibited the same or 2-fold higher antibacterial activity but 8-fold less hemolytic activity. Circular dichroism analysis revealed that ME-D has much less $\alpha$-helical content in $\alpha$-helical content in the presence of zwitterionic EYPC/cholesterol (10 : 1, w/w) liposomes compared to negatively charged EYPE/EYPG (7 : 3, w/w) liposomes. The blue shift of the fluorescence emission maximum of ME-D in zwitterionic EYPC/cholesterol (10 : 1, w/w) liposomes was much smaller than in negatively charged EYPE/EYPG (7 : 3, w/w) liposomes. These results suggested that the improvement in therapeutic index/cell selectivity of ME-D is correlated with its less permeability to zwitterionic membranes.

Development and Immunochemical Properties of Two Monoclonal Antibodies Specific to Human Chorionic Gonadotropin

  • Kim, You-Hee;Koh, Kwan-Sam
    • BMB Reports
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    • v.32 no.5
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    • pp.474-479
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    • 1999
  • Using a hybridoma technique, spleen cells of Balb/c mice immunized with human chorionic gonadotropin (hCG) were fused with NS-1 mouse myeloma cells. Two hybrid cell lines, clones KS-8 and KS-19, secreting monoclonal antibodies to hCG, were isolated. KS-8 and KS-19 belong to the immunoglobulin $G_1$ subclass. With the aid of a double-antibody radioimmunoassay, it was established that the KS-8 monoclonal antibody recognizes an immunodeterminant of the $\beta$-subunit of hCG, whereas the KS-19 monoclonal antibody recognizes an epitope present on the $\alpha$-subunit of hCG. The KS-8 monoclonal antibody specifically reacts with human chorionic gonadotropin and shows cross-reactivity of less than 0.3% to other related human glycoprotein hormones. On the other hand, using a hemagglutination test based on antibody-induced agglutination of sheep red blood cells coated with hCG, It was shown that only the KS-19 monoclonal antibody was capable of inducing a positive reaction, although both monoclonal antibodies had similar binding capacity to the coated cells. The results from the dual screening procedures demonstrate that KS-8 and KS-19 monoclonal antibodies show high sensitivity in two different assays, and are hence useful for the qualitative and quantitative determination of hCG by both radioimmunoassay and hemagglutination inhibition tests.

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Comparison of Dangguibohyel-tang and Erythropoietin on Cyclophosphamide-induced Anemia in Rats (당귀보혈탕(當歸補血湯)과 eryhropoietin이 cyclophosphamide로 유도된 흰쥐의 빈혈에 미치는 영향 비교 연구)

  • Kang Soon-Ah;Chang Mun-Seog;Oh Myung-Sook;Kim Do-Rim;Kim Ji-Sook;Park Seong-Kyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.1
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    • pp.31-36
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    • 2006
  • The aim of this study was to elucidate the mechanism of anemia associated with Dangguibohyel-tang (DBT) in rats. Using cyclophosphamide-induced (30mg/kg BW) anemic rats, changes in weight gain, the levels of red blood cell (RBC), hematocrit (Hct), platelet and hemoglobin (Hgb), serum vitamin B12, ALT(GPT) levels and erythropoietin (EPO) gene expression were monitored, and compared with DBT (1,000mg/kg BW, 14d)-treated and EPO (1,000IU/kg BW, 14d, s.c.)-treated rats. Food efficiency ratio (FER) were 31.6%, in normal group, 28.1% in cyclophosphamide-induced control group, 31.7% in DBT-treated group and 25.1%, in EPO-treated group after 14 days. The levels of red blood cell (RBC), hematocrit (Hct), platelet and hemoglobin (Hgb) of DBT-treated group were significantly higher than those of control. And DBT extract administered group showed dominant effects on the recovery of Hgb level. Serum vitamin B12 and ALT(GPT) levels were significantly increased at DBT-treated groups. EPO gene expression was decreased 91,9% in control group, 79.6% in DBT-treated group and 53.9% in EPO-treated group, respectively. These results suggest that administration of DBT could prevent human patient from chemotherapy derived anemia by improving hematological value and EPO status.

Erythrocyte Sedimentation Rate: Its Determinants and Relationship with Risk Factors Involved in Ischemic Stroke

  • Kaur, Kirandeep;Kaur, Amandeep;Kaur, Anupam
    • Korean Journal of Clinical Laboratory Science
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    • v.54 no.1
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    • pp.1-8
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    • 2022
  • Erythrocyte sedimentation rate (ESR) evaluation is a useful tool for monitoring disease activity in various inflammatory and non-inflammatory conditions. ESR is known to be influenced by a multitude of confounding factors. The present study aimed to assess the possible determinants of the ESR and its relationship with various risk factors involved in ischemic stroke. ESR and other hematological and biochemical parameters were investigated in 163 ischemic stroke patients (107 males and 56 females) selected based on imaging techniques including magnetic resonance imaging (MRI) or computed tomography (CT) scans. Statistical analysis was performed using the SPSS 16.0 software. Linear regression analysis showed a significant inverse relationship of hemoglobin (Hb) and hematocrit or packed cell volume (PCV) (P<0.001 for females; P<0.01 for males) with the ESR. It was observed that the red blood cell (RBC) count was not strongly correlated with the ESR (P<0.05 for both males and females). It was also observed that sex significantly affected the variables determining the ESR levels, whereas age had no effect. Gender differences were also observed with respect to Hb, RBC, PCV, mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and ESR. The possible determinants of higher ESR levels in ischemic stroke may be sex, Hb, hematocrit, and RBC count, but the role of other clinical and laboratory parameters cannot be underestimated.

Ice-Binding Protein Derived from Glaciozyma Can Improve the Viability of Cryopreserved Mammalian Cells

  • Kim, Hak Jun;Shim, Hye Eun;Lee, Jun Hyuck;Kang, Yong-Cheol;Hur, Young Baek
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.1989-1996
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    • 2015
  • Ice-binding proteins (IBPs) can inhibit ice recrystallization (IR), a major cause of cell death during cryopreservation. IBPs are hypothesized to improve cell viability after cryopreservation by alleviating the cryoinjury caused by IR. In our previous studies, we showed that supplementation of the freezing medium with the recombinant IBP of the Arctic yeast Glaciozyma sp. (designated as LeIBP) could reduce post-thaw hemolysis of human red blood cells and increase the survival of cryopreserved diatoms. Here, we showed that LeIBP could improve the viability of cryopreserved mammalian cells. Human cervical cancer cells (HeLa), mouse fibroblasts (NIH/3T3), human preosteoblasts (MC3T3-E1), Chinese hamster ovary cells (CHO-K1), and human keratinocytes (HaCaT) were evaluated. These mammalian cells were frozen in dimethyl sulfoxide (DMSO)/fetal bovine serum (FBS) solution with or without 0.1 mg/ml LeIBP at a cooling rate of -1℃/min in a -80℃ freezer overnight. The minimum effective concentration (0.1 mg/ml) of LeIBP was determined, based on the viability of HeLa cells after treatment with LeIBP during cryopreservation and the IR inhibition assay results. The post-thaw viability of mammalian cells was examined. In all cases, cell viability was significantly enhanced by more than 10% by LeIBP supplementation in 5% DMSO/5% FBS: viability increased by 20% for HeLa cells, 28% for NIH/3T3 cells, 21% for MC3T3-E1, 10% for CHO-K1, and 20% for HaCaT. Furthermore, addition of LeIBP reduced the concentrations of toxic DMSO and FBS down to 5%. Therefore, we demonstrated that LeIBP can increase the viability of cryopreserved mammalian cells by inhibiting IR.