• Title/Summary/Keyword: immunoglobulin G (IgG)

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Engineering Human-like Sialylation in CHO Cells Producing hCTLA4-Ig by Overexpressing α2,6-Sialyltransferase (α2,6-Sialyltransferase 과발현을 통한 인간형 시알산 부가 hCTLA4-Ig 생산 CHO 세포주 제작)

  • Lim, Jin-Hyuk;Cha, Hyun-Myoung;Park, Heajin;Kim, Ha Hyung;Kim, Dong-Il
    • KSBB Journal
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    • v.32 no.3
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    • pp.193-198
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    • 2017
  • Sialylation is important in producing therapeutic proteins such as antibody, cytokine and fusion protein. Thus, enhancement of sialylation is usually performed in CHO cell cultures. ${\alpha}2,6$-Sialyltransferase (ST), which plays a key role in the attachment of ${\alpha}2,6-sialic$ acid, is present in human cells but not in Chinese hamster ovary (CHO) cells. Overexpression of ${\alpha}2,6-ST$ can be used for enhancing the degree of sialylation and achieving human-like glycosylation. In this study, we constructed CHO cells producing human cytotoxic T-lymphocyte antigen4-immunoglobulin (hCTLA4-Ig) as well as ${\alpha}2,6-ST$. Transfected CHO cells were selected using G418 and stable cell line was established. Profiles of viable cell density and hCTLA4-Ig titer in an overexpressed cell line were similar to those of a wild-type cell line. It was confirmed that the total amount of sialic acid was increased and ${\alpha}2,6-sialic$ acid was attached to the terminal residues of N-glycan of hCTLA4-Ig by ESI-LC-MS. Compared to 100% of ${\alpha}2,3-sialic$ acid in wild type cells, 70.9% of total sialylated N-glycans were composed of ${\alpha}2,6-sialic$ acid in transfected cells. In conclusion, overexpression of ${\alpha}2,6-ST$ in CHO cells led to the increase of both the amount of total sialylated N-glycan and the content of ${\alpha}2,6-sialic$ acid, which is more resemble to human-like structure of glycosylation.

A Study on the Effect of Erycibae Caulis and Corydalis Tuber Pharmacopuncture on a Mouse Model with Collagen Induced Rheumatoid Arthritis (정공등현호색(丁公藤玄胡索) 약침(藥鍼)이 류마티스 관절염 생쥐 모델에 미치는 영향)

  • Kim, Hyun Ji;Jeon, Ju Hyun;Kim, Young Il
    • Journal of Acupuncture Research
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    • v.33 no.2
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    • pp.21-34
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    • 2016
  • Objectives : The aim of the present study is to examine the effect and mechanism of Erycibae Caulis and Corydalis Tuber Pharmacopuncture (ECP) on a mouse model with collagen induced rheumatoid arthritis (CIA). Methods : We evaluated the Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), Creatinine, and the Blood urea nitrogen (BUN) of serum to examine the safety of this study. In vivo, we compared the results of the non-treated group, the normal saline pharmacopuncture treated control group, the indomethacin treated group and the ECP group. We evaluated rheumatoid arthritis manifestation and the Rheumatoid Arthritis Index (AI). Also, immune cells in blood affected by ECP were evaluated by calculating the level of white blood cells (WBC), neutrophil, lympocytes and monocytes. Next, the level of Immunoglobulin M (IgM), Immunoglobulin G (IgG), Interleukin (IL)-$1{\beta}$, IL-6, IL-17, Tumor Necrosis Factor (TNF)-${\alpha}$ and Granulocyte-macrophage Stimulating Factor (GM-CSF)in serum were measured. We examined the imaging of cartilage degeneration using micro CT-arthrography of the hind paw. Additionally, we examined the effects of reducing bone volume (BV) ratio and bone surface/bone volume (BS/BV) ratio with 3D Micro-CT. Finally, we did a histopathologic examination analysis. Results : The absence of liver and kidney toxicity was evident. In vivo, edema of the joints of the ECP group decreased greatly in macroscopic observation. AI measurement of the ECP group also decreased significantly compared to the control group. The level of WBC, neutrophil, lympocytes, and monocytes in the blood decreased but there was no statistical significance of this data. IgM of the ECP group decreased significantly compared to the control group. IL-$1{\beta}$, IL-6, TNF-${\alpha}$, and GM-CSF production of the ECP group decreased significantly compared to the control group. As a result of examining joint condition with 3D micro CT, deformation and destruction of the joint was shown to have decreased. Bone density of ECP group increased at a statistically significant level compared to the control group. Degree of joint inflammation of ECP group decreased significantly compared to the control group. After H&E and M-T staining, infiltration of immune cells, subsidence of the cartilage, damage to the synovial cells and joint erosion decreased. Conclusion : This study showed that ECP hindered the process of rheumatoid arthritis and protected joints and cartilage.

Effects of Diaphragm Breathing and Garlic Powder Intake on Body Composition, Heart Rate, Blood Pressure and Immunoglobulin in Middle-aged Male smokers. (횡격막 호흡과 마늘 분말 섭취가 중년 남성 흡연자의 신체조성, 심박수, 혈압 및 면역글로불린에 미치는 영향)

  • Choi, Seung-Uk;Baek, Yeong-Ho;Kwak, Yi-Sub
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1266-1271
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    • 2007
  • The purpose of this study was to investigate combined effects of diaphragm breathing and garlic powder intake on body composition, heart rate, blood press and immunoglobulin levels in middle-aged male smokers from the age 40-49. Diaphragm breath training was 2-5 grade intensity on dyspnea scale for 20 minutes four times a week for 4 weeks and subjects were given garlic at 3 g of powder after breakfast and dinner two times a day during the 4 weeks. The conclusions of this study are as follows; Garlic intake group decreased in percentage of body fat, in the comparison between groups, garlic intake group had a lower percentage of body fat than control group. Heart rate was decreased in Diaphragm breathing group at rest. SBP was decreased in Diaphragm breathing+garlic intake group. Garlic intake group and diaphragm breathing+garlic intake group increased in IgG.

Oocyst production and immunogenicity of Cryptosporidium muris (strain MCR) in mice (마우스에 있어서 쥐와포자충(MCR주)의 오오시스트 배설상황과 면역원성)

  • Lee, Jae-Gu;Yok, Sim-Yong;Park, Bae-Geun
    • Parasites, Hosts and Diseases
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    • v.33 no.4
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    • pp.377-382
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    • 1995
  • Three-week-old ICR SPF mice were orally inoculated with one of 5 doses ranging from $2{\;}\times{\;}10^2{\;}to{\;}2{\;}\times{\;}10^6$ oocysts of Crwptosporidium tsuris (strain MCR) per mouse. Oocyst inoculation was directly proportional to the amount of oocysts shed and was inversely proportional to the period required for peals oocyst production and to the prepatent period. Peak oocyst production occurred between fifteen and thirty-one days with a patent period from 61 to 64 days. Three days after all mice stopped shedding oocysts, they were orally challenged with a single dose of $2{\;}\times{\;}10^6$ oocysts or the same species. Marked seroconversion for IgG antibody accompanied recovery from mice inoculated with $5{\;}\times{\;}10^5$ oocysts. Mice administered with carrageenan excreted a small number of oocysts for 49.0 days on the average after challenge inoculation (ACI) and control mice for 14.2 days in a dose-independent fashion. Just before challenge infection, phagocytic activity of peritoneal macrophages ($M{\phi}$) and the number of peripheral $M{\phi}$ were dramatically decreased. Mild challenge infection implies that the immunogenicity of C. nuris (strain MCR) is very strong, despite $M{\phi}$ blocker carrageenan administration.

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Effect of Partial Substitution of Dietary Spray-dried Porcine Plasma or Fishmeal with Soybean and Shrimp Protein Hydrolysate on Growth Performance, Nutrient Digestibility and Serum Biochemical Parameters of Weanling Piglets

  • Sun, Zhantian;Ma, Qiugang;Li, Zhongrong;Ji, Cheng
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.7
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    • pp.1032-1037
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    • 2009
  • The present experiment was conducted to study the effects of partial replacement of spray-dried porcine protein (SDPP) or fish meal with soybean and shrimp protein hydrolysate (SSPH) on growth performance, nutrient digestibility and serum biochemical parameters in weaned pigs. Two hundred and forty 21${\pm}$2 d old pigs ((Pietrain${\times}$Duroc)${\times}$(Landrace${\times}$Large Yorkshire)) with initial weight of 6.9${\pm}$0.5 kg were randomly allocated to five dietary treatments with six replicates per treatment and eight piglets per replicate. The control diet (T1) contained 2% SDPP and 6% fishmeal, and SDPP for experimental diets T2 and T3 was replaced with 1% and 2% SSPH, respectively, on an iso-nitrogenous basis. The fishmeal for experimental diets T4 and T5 was replaced with 1% and 2% SSPH, respectively, also on an iso-nitrogenous basis. The experimental period was 21 days. The results showed that weaned piglets fed the diets containing 1% and 2% SSPH as a replacement for SDPP had similar average daily gain (ADG), average daily feed intake (ADFI), feed/gain (F/G), diarrhea rate and serum biochemical indices e.g. blood urea nitrogen (BUN), total serum protein (TP), albumin to globulin ratio (A/G), globulin (GLO), serum glucose (GLU), and immunoglobulin G (IgG) to those fed the control diet during 0-10 d and 0-21 d of the experiment. The substitution of 1% SSPH on an iso-nitrogenous basis for fish meal appeared to be beneficial for ADG (p = 0.59) and ADFI (p = 0.23) of piglets during the overall period. The digestibility of calcium was higher (p<0.01) in pigs fed diets containing SSPH than on the control diet. Addition of 1% SSPH on an iso-nitrogenous basis for fish meal could increase the digestibilities of dry matter and energy of the diet. Dietary replacement of fish meal with 1% and 2% SSPH had no effect on the concentrations of BUN, TP, A/G, GLO, GLU, and IgG. In conclusion, dietary SDPP or fish meal could partially replace SSPH without any adverse effect on growth performance, nutrient digestibility and serum biochemical parameters in weaned piglets.

Effects of Feeding Lactic Acid Bacteria-Based Direct-Fed Microbial Complex on Growth Performance and Blood Characteristics in Growing Pigs (Lactic acid bacteria 계열의 복합 생균제가 육성돈의 성장 능력과 혈액 특성에 미치는 영향)

  • Yoo, Jong-Sang;Kim, In-Ho
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.7 no.6
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    • pp.1416-1420
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    • 2006
  • This study was conducted to investigate the effects of direct-fed microbial(DFM) complex on the growth performance and blood characteristics in growing pigs. A total of 72 growing pigs with an average initial BW of 24.64$\pm$2.46 kg were used in 28 days experiment trial. There were six pens per treatment with three pigs per pen. Dietary treatments included 1)NC(basal diet; antibiotics free diet), 2) PC(NC diet with 0.1% antibiotics; chlortetracycline 0.05% +neomycin 0.05%), 3)DFM0.1 UC diet + 0.1% DFM)and 4)DFM0.3 (NC diet + 0.3% DFM). During the entire experimental period, hnG(Average daily gain) was increased NC treatment compared with NDFM0.1 and NDFM0.3 treatments. However, there was not significantly effect by treatments(P>0.05). Also, there was not significantly effect in ADFI(average daily feed intake) among the treatments. Blood characteristics(RBC, WBC and IgG) tended to improved, however, no significant differences were observed(P>0.05). In our experiment, DFM had not effects on growth performance and blood characteristics in growing pigs.

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Development of Chicken Immunoglobulin Y for Rapid Detection of Cronobacter muytjensii in Infant Formula Powder

  • Kim, Yesol;Shukla, Shruti;Ahmed, Maruf;Son, Seokmin;Kim, Myunghee;Oh, Sejong
    • Food Science of Animal Resources
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    • v.32 no.6
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    • pp.706-712
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    • 2012
  • The present study was aimed to produce a chicken polyclonal antibody against Cronobacter muytjensii and to develop an immunoassay for its detection. Purification of anti-C. muytjensii IgY from egg yolk was accomplished using various methods such as water dilution and salt precipitation. As a result, sodium dodecyl sulfate-polyacrylamide gel electrophoresis produced two bands around 30 and 66 kDa, corresponding to a light and a heavy chain, respectively. Indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was performed to determine the effectiveness of the chicken IgY against C. muytjensii. The optimum conditions for detecting C. muytjensii by indirect ELISA and checkerboard titration of the antigen revealed an optimum average absorbance at the concentration of 18 ${\mu}g/mL$, having ca. $10^8$ coated cells per well. The anti-C. muytjensii IgY antibody had high specificity for C. muytjensii and low cross-reactivity with other tested pathogens. In this assay, no cross-reactivity was observed with the other genera of pathogenic bacteria including Escherichia coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus, Enterobacter aerogenes, Salmonella Enteritidis and Listeria monocytogenes. In addition, detection of C. muytjensii in infant formula powder showed a low matrix effect on the detection curve of IC-ELISA for C. muytjensii, with similar detection limit of $10^5$ CFU/mL as shown in standard curve. These findings demonstrate that the developed method is able to detect C. muytjensii in infant formula powder. Due to the stable antibody supply without sacrificing animals, this IgY can have wide applications for the rapid and accurate detection of C. muytjensii in dairy foods samples.

Effect of negative therapy at back meridian points on blood gas components and immune functions in male college students (배부(背部) 경혈(經穴)에 부항요법(附缸療法) 시술(施術)이 남자대학생(男子大學生)의 면역기능(免疫機能)에 미치는 영향(影響))

  • Oh, Jae-Keun;Kim, Sung-Soo
    • The Journal of Korean Medicine
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    • v.20 no.1 s.37
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    • pp.75-83
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    • 1999
  • To investigate the effects of negative therapy at back meridian points on blood gas components and immune functions in male college students, this study was conducted on treatment types(abdomen group and back group) at three sampling times (before, post-2 wks and post-4 wks) by using $2{\times}3$ factoral design. Blood gas $components(pH,\;PCO_2,\;PO_2,\;HCO_3^-,\;O_2SAT,\;BE)$, red blood cell, hematocrit, hemoglobin, white blood cell and subsets(neutrophil, basophil, eosinophil. lymphocyte, monocyte), total T cells, helper T cells, suppressor T cells, Th/Ts ratio, total B cells, serum immunoglobulin levels (IgG, IgA, IgM, IgD, IgE), Cytokines(Interlukin$-1{\beta}$, -2, -4, 2 receptor, -6 and ${\gamma}$-interferon), NK cells were measured. Collected with data were analyzed statistically by repealed measured ANOVA. The pattern of change between two groups for hematocrit, hemoglobin, suppressor T cells, interleukin-6, ${\gamma}-interferon$, NK cells at post-2 weeks and BE, lymphocyte, basophil at post-4 weeks was significantly different(p<0.05) And also the pattern of change over time for ${HCO_3}^-$(2 wks vs 4 wks), WBC, neutrophil, lymphocyte(0 wks vs 2 wks and 2 wks vs 4 wks) was significantly different(p<0.05). In summary, these data suggest that negative therapy at back meridian points had an effect on blood gas components and immune functions in male college students because practicing negative therapy at back meridian points was not associated with changes of all blood gas components and immune factors but associated with changes of BE, hematocrit, hemoglobin, WBC. neutrophil, lymphocyte, interleukin-6. ${\gamma}-interferon$, NK cells.

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A Study on the Growth Inhibition of Salmonella by Secretory Immunoglobulin A in Human and Bovine Milk (인유(人乳)와 우유(牛乳) 중에 함유(含有)되어 있는 Secretory Immunoglobulin A의 Salmonella속(屬) 성장억제(成長抑制)에 관(關)한 연구(硏究))

  • Bae, Heoung Churl;Kim, Jong Woo
    • Korean Journal of Agricultural Science
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    • v.21 no.2
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    • pp.122-132
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    • 1994
  • Bacteriostatic activity of secretory immumoglobulin A (SIgA) in human and bovine colostrums on enterotoxigenic type Salmonella was tested in the tissue culture medium. SIgA was used in $0.1{\sim}5.0mg/m{\ell}$ concentration with or without the addition of egg lysozyme tested for theirs bacteriostatic activites. 1. Bovine SIgA rich fraction with a large amount of $IgG_1$-dimer could be prepared from bovine colostrum of Holstein cows by anion exchange chromatography using DEAE-Sephadex A-50 and gel filtration on Sephadex G-200 and Sepharose 6B. 2. Human SIgA appeared to be the most bacteriostatic effect for all varieties of Salmonella in a range of $0.5{\sim}1.0mg/m{\ell}$ Bovine SIgA showed a marked bacteriostatic effect increased by increasing the concentration. Bovine IgG had not show bacteriostatic effect against both enterotoxigenic type Salmonella. Egg lysozyme as well as bovine SIgA also showed a marked bacteriostatic effect increased by increasing the concentration. 3. When the growth inhibition of human SIgA was tested by adding egg lysozyme with time interval, egg lysozyme showed bacteriostatic effect as compared with control. But human SIgA and adding with lysozyme showed slight the bacteriostatic effect. 4. When the growth inhibition of bovine SIgA was tested by adding egg lysozyme with time interval, all treatment against S. enteritidis showed bacteriostatic effect as compared with control In the case of S. typhinwrium, egg lysozyme showed a marked bacteriostatic effect as compared with control.

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International Scoring System in Symptomatic Multiple Myeloma: Experience from a Tertiary Care Center

  • Sultan, Sadia;Irfan, Syed Mohammed;Parveen, Saira;Taufiq, Ufaq
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.2031-2033
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    • 2016
  • Background: Symptomatic multiple myeloma (MM) is an acquired B-cell malignant proliferation of antibody secreting plasma cells, characterized by end organ damage due to monoclonal immunoglobulin secretion. The aim of this study wa to determine the stage stratification according to an international scoring system in adult Pakistani MM patients at presentation. Materials and Methods: This single centre retrospective study extendedfrom January 2012 to December 2015. Data were retrieved from the departmental maintained records. Results: A total of 39 patients were diagnosed at our center with MM during the period of the study, 25 males and 14 females. Age ranged between 36 and 81 with a mean of $54.5{\pm}14.8$ and a median of 57 years. Common presenting complaints included fatigue (80.9%), backache (79.3%) and bone pain (66.2%). Overall, 9 patients were in ISS stage I (23%), 12 were in stage II (30.7%) and 18 were in stage III (46.1%). Out of the total, 29 (74.3%) had kappa immunoglobulin andthe remaining 10 (25.6%) had lambda type myelomas. IgG myeloma was commonest, seen in 26 (66.6%) followed by IgA in 11 (28.2%) with non secretory myeloma in one (2.5%) and light chain disease also in one patient (2.5%). Conclusions: MM in Pakistani patients is seen in a relatively young population with male predominance. Primarily patients are symptomatic and risk stratification revealed a predominance of advanced stage III disease in our setting.