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The Lung Expression of Proinflammatory Cytokines, TNF-$\alpha$ and Interleukin 6, in Early Periods of Endotoxemia (내독소혈증 유발 급성폐손상에서 폐장내 Proinflammatory Cytokines 발현에 관한 고찰)

  • Moon, Seung-Hyug;Kim, Yong-Hoon;Park, Choon-Sik;Lee, Shin-Je
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.3
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    • pp.553-564
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    • 1998
  • Background: The immediate hoot response to LPS is the production of proinflammatory cytokines that act as intercellular mediators in inflammatory reactions, including acute lung injury. These "early response" cytokines transmit signals from recognition cells to target or effector cells. This host response is further amplified by the expression of leukocyte chemoattractants, growth factors, and adhesion molecules, resulting in an array of proinflammatory events. This experiment was performed to define the lung origin of proinflammatory cytokines, such as TNF-$\alpha$, IL 6 in early periods of endotoxin induced acute lung injury (ALI). Method: The healthy male Sprague-Dawley, weighted 150 - 250g, were divided into saline control (NC) and endotoxemia-induced ALI (ETX-), and leukopenic endotoxemia-induced ALI (CPA-ETX-Group) which was induced by cyclophosphamide, 70 mg/kg i.p. injection. Acute lung injury was evoked by LPS, 5 mg/kg, intravenously administered. Bronchoalveolar lavage was performed at 0, 3, 6 h after LPS-treated to estimate the influx of phagocytes and concentration of total protein, and cytokines as TNF-$\alpha$ and IL 6 by a bioassy using MIT method. We also examined the localization of TNF-$\alpha$ and IL 6 protein in endotoxemia-challenged lung tissue by immunohistochemical stain (IH). Results: The total cell, macrophage and PMN count in BALF were elavated in ETX group compared to NC(p<0.05). In CPA-ETX group, total cell and macrophage count in BALF were not changed compared to NC. but PMN count was markedly reduced and it took part in less than 0.1 % of total BAL cells (p<0.01). The protein concentration in BALF were significantly increased in ETX and CPA-ETX group Compared to NC (p<0.05), but there was significant difference between ETX- and CPA-ETX group only at 6 h (p<0.05). This observation suggested that even if PMNs are involved in the pathogenesis of acute lung injury, their role cannot be viewed as essential The concentration of TNF-$\alpha$ and IL 6 in BALF was significantly increased in the ETX- and CPA-ETX group compared to NC. There was no difference between ETX- and CPA-ETX group. In IH, anti-TNF-$\alpha$- and anti-IL 6 antibody was strongly localized at interstitial monocytes and alveolar macrophages in endotoxemia-challenged lung tissue. From above point of view, activated alveolar macrophage/monocyte considered as a prominent source of proinflammatory cytokines in endotoxemia-challenged lung injury. Conclusion: The prominent source of proinflammatory cytokines in early periods of endotoxemia-induced lung injury will be the activated resident macrophages like an alveolar macrophage and interstitial monocytes. The pulmonary macrophage/monocyte will impact the initiation and continuance of lung injury without PMNs's certain inflammatory role, particularly in endotoxemia-induced acute lung injury.

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The Ability of Anti-tumor Necrosis Factor Alpha(TNF-${\alpha}$) Antibodies Produced in Sheep Colostrums

  • Yun, Sung-Seob
    • 한국유가공학회:학술대회논문집
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    • 2007.09a
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    • pp.49-58
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    • 2007
  • Inflammatory process leads to the well-known mucosal damage and therefore a further disturbance of the epithelial barrier function, resulting abnormal intestinal wall function, even further accelerating the inflammatory process[1]. Despite of the records, etiology and pathogenesis of IBD remain rather unclear. There are many studies over the past couple of years have led to great advanced in understanding the inflammatory bowel disease(IBD) and their underlying pathophysiologic mechanisms. From the current understanding, it is likely that chronic inflammation in IBD is due to aggressive cellular immune responses including increased serum concentrations of different cytokines. Therefore, targeted molecules can be specifically eliminated in their expression directly on the transcriptional level. Interesting therapeutic trials are expected against adhesion molecules and pro-inflammatory cytokines such as TNF-${\alpha}$. The future development of immune therapies in IBD therefore holds great promises for better treatment modalities of IBD but will also open important new insights into a further understanding of inflammation pathophysiology. Treatment of cytokine inhibitors such as Immunex(Enbrel) and J&J/Centocor(Remicade) which are mouse-derived monoclonal antibodies have been shown in several studies to modulate the symptoms of patients, however, theses TNF inhibitors also have an adverse effect immune-related problems and also are costly and must be administered by injection. Because of the eventual development of unwanted side effects, these two products are used in only a select patient population. The present study was performed to elucidate the ability of TNF-${\alpha}$ antibodies produced in sheep colostrums to neutralize TNF-${\alpha}$ action in a cell-based bioassay and in a small animal model of intestinal inflammation. In vitro study, inhibitory effect of anti-TNF-${\alpha}$ antibody from the sheep was determined by cell bioassay. The antibody from the sheep at 1 in 10,000 dilution was able to completely inhibit TNF-${\alpha}$ activity in the cell bioassay. The antibodies from the same sheep, but different milkings, exhibited some variability in inhibition of TNF-${\alpha}$ activity, but were all greater than the control sample. In vivo study, the degree of inflammation was severe to experiment, despite of the initial pilot trial, main trial 1 was unable to figure out of any effect of antibody to reduce the impact of PAF and LPS. Main rat trial 2 resulted no significant symptoms like characteristic acute diarrhea and weight loss of colitis. This study suggested that colostrums from sheep immunized against TNF-${\alpha}$ significantly inhibited TNF-${\alpha}$ bioactivity in the cell based assay. And the higher than anticipated variability in the two animal models precluded assessment of the ability of antibody to prevent TNF-${\alpha}$ induced intestinal damage in the intact animal. Further study will require to find out an alternative animal model, which is more acceptable to test anti-TNF-${\alpha}$ IgA therapy for reducing the impact of inflammation on gut dysfunction. And subsequent pre-clinical and clinical testing also need generation of more antibody as current supplies are low.

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Effects of Ojeoksangamibang on the Lipid Metabolism, Anti-oxidation and Concentration of Proinflammatory Cytokines in Rat Fed High Fat Diet (오적산가미방(五積散加味方)이 고지방식이 유도 비만쥐의 지질대사, 항산화계 및 전염증성 cytokine 생산에 미치는 영향)

  • Kong, In-Pyo;Park, Won-Hyung;Cha, Yun-Yeop
    • Journal of Korean Medicine Rehabilitation
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    • v.21 no.4
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    • pp.23-40
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    • 2011
  • Objectives: This study was designed to examine the effects of extracts of Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) on the lipid lowering, anti-oxidation and concentration of proinflammatory cytokines and was investigated on hyperlipidemic rats. Methods: Male rats weighing $182.39{\pm}4.71g$ were fed high fat diet for 8 weeks and 36 rats(above 400 g) were divided into 4 groups. Each of 9 rats was divided a control group and experimental groups. We fed a control group of rats a basal diet and administered normal saline(100 mg/kg, 1 time/1 day) for 4 weeks. And we fed each experimental group of rats basal diet and administered an extract of Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) extracts(100 mg/kg, 200mg/kg, 300 mg/kg, 300 mg/kg, 1 time/1 day) for 4 weeks. At the end of the experiment, the rats were sacrificed to determine their chemical composition. We measured lipid of plasma and liver, concentration of proinflmmatory cytokines, anti-oxidative activity and $TNF-{\alpha}$, Apo-B, Apo-E and leptin gene expression. Results: 1. Concentration of plasma free fatty(FFA) showed no significant difference in all the treatment groups. Concentration of plasma triglyceride(TG) showed a significant decrement in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. 2. Concentration of plasma total cholesterol showed a significant decrement in the 200 and 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. Concentration of plasma low density lipoprotein(LDL)-cholesterol showed a Significant decrement in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. Concentration of plasma high density lipoprotein(HDL)-cholesterol showed a significant increment in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) group. 3. Concentration of liver total cholesterol showed a tendence to decrease in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups. Concentration of liver TG showed a significant decrement in all Ojeoksangamibang groups than that of control group. 4. Concentration of plasma and liver thiobarbituric acid reactive substance(TBARS) showed a tendence to decrease in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups. 5. The values of glutathione peroxidase(GSH-Px), superoxide dismutase(SOD) and catalase(CAT) activity showed a significant increment in all Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. 6. The values of plasma aspartate aminotransferase(AST) and alanine aminotransferase(ALT) activity showed no significant different in all treatment group. 7. Concentration of plasma $interleukin(IL)-1{beta}$ showed no significant difference in all the treatment groups. Concentration of plasma IL-6 showed a significant decrement in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) group than that of control group. Concentration of plasma tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ a siginifant decrement in the 200 and 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) group than that of control group. However the concentration of plasma IL-10 in the 300 mg/kg Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups showed a significant increment than that of control group. 9. In the analysis of reverse transcription-polymerase chain reaction(RT-PCR), gene expression of $TNF-{\alpha}$, Apo-B and Apo-E in the Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups showed a lower expression than that of control group. However the gene expression of leptin showed no difference in the treatment groups. 10. The ratio of $TNF-{\alpha}$, Apo-B, and Apo-E per ${\beta}-actin$ expression in the Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups showed a significant decrement than that of control group. However The ratio of leptin expression per ${\beta}-actin$ expression showed no significant difference among all the treatment groups. Conclusions: According to above results, in lowering lipid effect, anti-oxidation and control of pro-inflammatory cytokines production, Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) gives effect.

CELL CULTURE STUDIES OF MAREK'S DISEASE ETIOLOGICAL AGENT (조직배양(組織培養)에 의한 Marek 병(病) 병원체(病原體)의 연구(硏究))

  • Kim, Uh-Ho
    • Korean Journal of Veterinary Research
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    • v.9 no.1
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    • pp.23-62
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    • 1969
  • Throughout the studies the following experimental results were obtained and are summarized: 1. Multiplication of agents in primary cell cultures of both GF classical and CR-64 acute strain of Marek's disease infected chicken kidneys was accompanied by the formation of distinct transformed cell foci. This characteristic nature of cell transformation was passaged regularly by addition of dispersed cell from infected cultures to normal chicken kidney cell cultures, and also transferred was the nature of cell transformation to normal chick-embryo liver and neuroglial cell cultures. No cytopathic changes were noticed in inoculated chick-embryo fibroblast cultures. 2. The same cytopathic effects were noticed in normal kidney cell monolayers after the inoculation of whole blood and huffy coat cells derived from both forms of Marek's disease infected chickens. In these cases, however, the number of transformed cell foci appearing was far less than that of uninoculated monolayers prepared directly from the kidneys of Marek's disease infected chickens. 3. The change in cell culture IS regarded as a specific cell transformation focus induced by an oncogenic virus rather than it plaque in slowly progressing cytopathic effect by non-oncogenic viruses, and it is quite similar to RSV focus in chick-embryo fibroblasts in many respects. 4. The infective agent (cell transformable) were extremely cell-associated and could not be separated in an infective state from cells under the experimental conditions. 5. The focus assay of these agents was valid as shown by the high degree of linear correlation (r=0.97 and 0.99) between the relative infected cell concentration (in inoculum) and the transformed cell foci counted. 6. No differences were observed between the GF classical strain and the CR-64 acute strain of Marek's disease as far as cell culture behavior. 7. Characterization of the isolates by physical and chemical treatments, development of internuclear inclusions in Infected cells, and nucleic acid typing by differential stainings and cytochemical treatments indicated that the natures of these cell transformation agents closely resemble to those described fer the group B herpes viruses. 8. Susceptible chicks inoculated with infected kidney tissue culture cells developed specific lesions of Marek's disease, and in a case of prolonged observation after inoculation (5 weeks) the birds developed clinical symptoms and gross lesions of Marek's disease. Kidney cell cultures prepared from those inoculated birds and sacrificed showed a superior recovery of cell transformation property by formation of distinct foci. 9. Electron microscopic study of infected kidney culture cells (GF agent) by negative staining technique revealed virus particles furnishing the properties of herpes viruses. The particle was measured about $100m{\mu}$ and, so far, no herpes virus envelop has been seen from these preparations. 10. No relationship of both isolates to avian leukosis/sarcoma group viruses and PPLO was observed.

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Studies on the Meat Production and Woolskin Processing of Sheep and Korean Native Goats for Increasing Farm Income as a Family Subsidiary Work (농가부업(農家副業)의 소득향상(所得向上)을 위한 양육생산(羊肉生産) 및 모피가공(毛皮加工)에 관(關)한 연구(硏究))

  • Kwon, Soon-Ki;Kim, Jong-Woo;Han, Sung-Wook;Lee, Kyu Seung
    • Korean Journal of Agricultural Science
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    • v.5 no.2
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    • pp.93-114
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    • 1978
  • The purpose of the study was to find out possible ways for increasing farm income through the sheep and Korean native goats farming, and to investigate meat productivity, wool productivity; woolskin utility, physiological characteristics and correlation between economical college animal farm of the Chungnam National University and sample farms in the suburbs of Dae jeon City were selected for feeding 20 heads of Corriedale wethers and another 20 heads Korean native kids as research materials for the periods of 5th May-26th November, 1977. The data such as growth rate, carcass, viscera weight, blood picture and plamsa components, hebage intake and economic traits were obtained and analysed. The result of the study are summarized as follows: 1. Meat production and quality 1) After 196days of feeding, the body weight of sheep and Korean native goats was increased by two times of those at the beginning of the trial, i.e. 20kg and 8kg respectively. 2) There was no significance of growth rates of sheep in housing and grazing. 3) The growth rate of Korean native goats were excellent at the mountainous areas of Gong ju-Gun where infectious diseases were not found 4) Accroding to the body measurements of 18-month-old sheep, percentages of hip height, body length, rump length, chest depth, chest width, hip width, chest girth and forearm circumference to the withers height were 103,%, 104%, 33%, 44%, 31%, 23%, 135% and 15% respectively, and those of hip height, body length, chest depth and chest girth of 8-month-old native goats to the withers height were 106%, 109%, 46% and 122,% respecitively. As a result, it was found that the percentage of hip height, body length and chest depth of Korean native goats were higher than those of sheep while that of the chest girth of goats was lower. 5) In the carcass data, 47, $52{\pm}2.27%$ of carcass percentage, $34.61{\pm}1.62%$ of lean meat, $26.07{\pm}2.51%$ of viscera, $9.75{\pm}1.4%$ of bone, and $20.95%{\pm}2.14%$ of woolskin for sheep, and $45.58{\pm}5.63%$ of carcass percentage, $27.62{\p}3.81%$ of meat, $34.86{\pm}4.16%$ of viscera, $11.66{\pm}1.83%$ of bone, $3.63{\pm}1.61%$ of skull and $9.26{\pm}2.41%$ of woolskin for native goats were obtained. 6) The contents of moisture, crude protein, crude fat and crude ash in native goat meat were much similar in both plots of housing and grazing. It was, however, known that the contents of moisture and protein were higher in grazinrg than in housing, while fat content was lower in grazing plots. 7) The weights of visceral organs shown similar tendency for both of sheep and native goats. For the weights of liver, heart, kidney and spleen, significance was not reconized among the treatments. Those of rumen, reticulum, small and large intestine were heavier in grazing than in housing, while the amount of visceral fat was heavier in housing. 2. Wool productivity and woolskin 1) The wool production of sheep for 7 months was $3.88{\pm}1.02kg$, and wool percentage, staple length, straighten length, wool growth per day and number of crimps were $9.27{\pm}1.48%$, 8. $47{\pm}1.00cm$, $10.63{\pm}0.99cm$, $0.40{\pm}0.04cm$ and $2.78{\pm}0.40$ respecitively. 2) The tensile strength and tear strength of woolskin treated by alum tanning were highest on the skin obtained from rump, i.e. $1,351kg/mm^2$ and $2,252kg/mm^2$ respectively, and they are in order of loin and shoulder. 3. Utilization and improvement of pasture. 1) The difference of herbage intake of native goats was not recognized between grazing and tethering, but the intake in the afternoon was s lightly higher than that in the morning. However the hervage intake of sheep was superior in grazing and in the afternoon. 2) The cultivation effect was lower in the native goat plots due to their cultivation abilities, in other words, the establishment rates of pasture by hoof cultivation were 60.25% in the goat plots and 77.35% in the sheep plots. 4. Correlation among economical traits. 1) The correlation between live weight of sheep and daily gain was higher. On the other hand, the correlation between other traits was not significant except that live weight, daily gain and lean meat percentage to the length of thoracic vertebrae. The live weight of native goats and meat production were highly correlated, and high correlation was also found between weights of carcass and meat. However, negative correlation was shown between viscera weight and live weight as well as daily gain. 2) The correlatoin between fleece weight of sheep and other traits such as live weight, daily gain and fleece percentage is very high at the 1% siginficant level, and this means that rapid-growth individuals can produce much fleece. 3) The correlation between the factors such as weights of live body, lean meat and viscera of sheep and body measurements, i. e. chest girth and body length was highest, and weights, of carcass and lean meat was highly correlated to chest width and depth. It will be therefore reasonable that the meat productivity estimates will have to be made on the basis of chest girth and body length. The meat production traits of native goats were highly correlated to the most of body measurement data, and the correlation coefficient between chest girth and weights of live body, carcass, lean meat and bone percentage was very high, i. e. 0.992-0.974 in particular. The correlations of meat production traits to chest depth, forearm circumference, body length were 0.759-0.911, 0.759-0.909 and 0.708-0.872 respectively. Therefore, the meat production of native goats will have to be estimated on the basis of chest data. 5. Blood picture and plasma components. 1) The number of erythrocyte and MCHC of native goats were $12.93{\times}10^6/mm^3$ and 36.14%, and those of sheep were $10.68{\times}10^6/mm^3$ and 36.26 respectively. The values of native goats were significantly higher than those of sheep. 2) The hemoglobin concentration, PVC, MCV and MCR of native goats were 10.92 g/100ml, $23.40{\mu}^3$ and 10.94 pg, and those of sheep were 11.73 g/100ml, 36.25 ml/100ml, $33.97{\mu}^3$ and 30.2 ml/100ml 8.43 pg respectively. The values of native goats were significantly lower those of sheep. 3) The number of leukocytes of native goats was significantly higher than that of sheep, that is, $11.64{\times}10^3/mm^3$ in native goats and $9.32{\times}10^3/mm^3$ in sheep. 4) In differential count of leukocyte, neutrophil was significantly high in native goats while lympocyte in sheep. On the other hand, the basophil, eosinophil and monocyte were not significant between native goats and sheep. 5) The amounts of total protein and glucose in the plasma of native goats were 6.2g/100ml and 53.6mg/100ml, and those of sheep were 5.6g/100ml and 45.7mg/100ml, which means that the values of native goats were significantly higher that those of sheep. The amount of total-lipid of native goats(127.6mg/100ml) was significantly than that of sheep(149.6mg/100ml). 6) The amount of non-protein nitrogen, cholesterol, Ca, P, K, Na and Cl were not different between native goats and sheep. 6. Economic analysis. 1) The gross revenue of a farm which fed native goats and sheep was 4,000won per head and the optimum size for feeding them in a farm as a subsidiary work is 5-10 heads. 2) Since there was no difference between housing and grazing, they can be fed in group for farm's subsidiary work. 3) They can be also fed by youths and house wives in the suburbs of cities, because labour requirement is estimated as only two hours per days for feeding 5 heads of native goats and sheep.

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