• Applied Microscopy
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• 제24권2호
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• pp.78-92
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• 1994

Lysozyme has been reported to be present in the secretory granules of the Paneth cell, and lysozyme immunoreactivity has been detected by immunogold method in Paneth cells of the intestine of human, mouse and rat. The present study was aimed at clarifying the intracellular distribution and changes of the lysozyme immunoreactivity in rabbit Paneth cell after common bile duct ligation of rabbit, using the electron microscope immunogold technique. Healthy adult rabbits weighing about 2kg body weight were divided into normal and bile duct ligated groups. Common bile duct ligation was performed aseptically under ether anesthesia. Experimental animals were sacrificed on the 1st, the 3rd, the 5th, the 7th and the 14th day after the operation. Mucosal specimens from the intestinal gland of ileum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome. Ultrathin sections were placed on parlodion coated nickel grids (200mesh). The section-bearing grids were floated upside down on the added substance in a moist chamber at room temperature except for the primary antibody step, which was at $4^{\circ}C$. Sections were etched with a saturated solution of sodium m-periodate for 60min. After etching, sections were pretreated with 0.02M tris buffered saline (TBS), pH 8.4, with 1% bovine serum albumin (BSA, Sigma) for 60min, then treated polyclonal rabbit anti-human lysozyme (Dakipatts) diluted 1 : 50 in TBS with 0.1% BSA for 20hr. Subsequently, grids were incubated 60min in biotinylated goat anti rabbit IgG (Amersham) diluted 1 : 100 in TBS with 0.1% BSA. After this, sections were incubated 60min on streptavidin gold G10 (Amersham) diluted 1 : 50 in TBS with 0.1% BSA. After each step, the grids were briefly rinsed with TBS with 0.1% BSA. After the strepavidin gold step, the sections were jet washed with distilled water. Counterstain of the sections performed by uranyl acetate and lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow; 1. Secretory granules of mouse Paneth cells have a lysozyme immunoreactivity and also eosinophil leucocyte of rabbit applied for the positive-control stain, are well labeld with gold particles. 2. Normal rabbit Paneth cells have a lysozyme immunoreactivity restricted on the secretory granules. 3. Amount lysosomes containing myelin figures in the Paneth cells were significantly increased from 5th day after the common bile duct ligation. 4. Immunoreactivity of Paneth cell secretory granules were more activated on the 3rd day after the common bile duct ligation as compared with those of the normal animal. But the lysozyme immunoreactivity were decreased from the 5th day after the common bile duct ligation. 5. Considering the above finding, lysozyme contained Paneth cell are affected following of common bile duct ligation, whereas lysosomes containing myelin-figure do not exhibit any immunoreactive relationship with those of secretory granules.

• 대한수의학회지
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• 제14권1호
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• pp.115-133
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• 1974

Although considerable research has been done on the changes associated with age in the blood picture of domestic and laboratory animals, little work has been made of the changes occurring at different age in the blood picture of goats. And a comprehensive survey of the bood picture of Korean native goats has not been made. The object of the present investigation was to suggest standards for the blood picture of Korean native goats at frequent intervals from birth to maturity. The goats were kept under average farming conditions in this country. Observations were made at the following ages: at birth; 2,4 and 7 days; 2, 3, 4, 5, 6, 7, 8 and 9 weeks; 2.5, 3,6 and 12 months. Blood samples were drawn from the jugular vein. It was taken between 8 and 9 a.m. except those for the at-birth period. Erythrocyte and leukocyte enumerations and, determinations of hemoglobin in blood and hematocrit value were made in the usual manner. Reticulocytes were enumerated per 1,000 erythrocytes in blood smears stained with briIliant cresyl blue and counterstained with Wright's stain. Erythrocytes counts declined from $8.7{\times}10^8/mm^3$ at birth to a low of 7.0 at 4 days of age. These values increased to 11.5 at 5 weeks and reached a maximum of 14.0 at 3 months of age; it then fell to 11.5 at 12 months of age. Concentrations of hemoglobin in blood and hematocrit values were not related to the changes of erythrocyte counts. The values at birth were higher than at any other period during the first year of life. These fell from highs of 12.3 g/100 ml and 38.0 ml/100 ml to lows of 9.2 and 29 at 4 weeks for concentration of hemoglobin in blood and hematocrit value, respectively. There was a common pattern for the hematocrit value and hemoglobin in blood which showed three phases-a fall during the first month, a rise to the third month, and a fall to the mature level at 12 months of age. Mean corpuscular volume and mean corpuscular hemoglobin showed a common pattern. The values were $44.2{\mu}m^3$ and 14.2 pg at birth and fell, at first slowly and then rapidly, to reach adult levels of 24.1 and 7.9 at 6 weeks of age for mean corpuscular volume and mean corpuscular hemoglobin, respectively. Mean corpuscular hemoglobin concentration was little affected by age. Reticulocyte was observed from birth to 4 weeks of age. Percentage of reticulocyte decreased from 0.85% at birth to 0.06% at 4 weeks of age. Total leucocyte counts increased from $5.64{\times}10^3/mm^3$ at birth to a maximum of 13.4 at 3 months; it then fell to 11.5 at 12 months of age. In differential counts myelocyte, juvenile and band form decreased with advancing age. No myelocyte and juvenile were seen after the age of 7 and 9 weeks, respectively, and band forms were rare after the age of 3 months. Percentage of mature neutrophil showed a quick decline from 52.5% at birth to reach a minimum level (34.5%) at 3 months of age; it then rose to 38% at 12 months of age. Percentage of lymphocyte increased from 39.2% at birth to maximum of 59% at 3 month of age; it then fell to 54.9% at 12 months of age. Percentage of monocyte was not affected with advance of age. Percentage of eosinophil and basophil were increased with advance of age to reach a maximum at 2 to 3 month of age. It then fell to adult level at 12 month of age.

• 대한미생물학회지
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• 제21권1호
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• pp.133-144
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• 1986

This study was undertaken to assess the effect of ginseng administration on T lymphocyte induced local xenogenic graft-versus-host(GVM) reactions which were induced with thymocyte, spleen cell and lymph node cell of ICR mice. Mice received daily 10mg of 70% alcohol ginseng extract oral1y for 100days and control mice remained untreated for the same period of time. The cells from donor mice were injected intradermally into the closely shaven abdominal skin of Sprague-Dawley rats for GVH tests. The thymocyte from control(ginseng-untreated) mice showed a negative local GVH reaction, whereas thymocyte from experimental(ginseng-treated) mice showed a positive reaction with the rate of 17.4%. When spleen cells were injected, the incidence of positive local GVH reaction was 66.7% among ginseng-treated mice, as opposed to incidence of 45.5% of positive local GVH reaction among control mice. The incidence of positive local GVH reaction of the lymph node cells when injected into a recipient was 71.4% among ginseng-treated mice as compared with that of 18.9% among control mice. The relationship between spleen cell inoculum and intensity of the local GVH reaction was assessed in ginseng-untreated mice. The intensity of GVH reaction clearly appears to be dose related. In ginseng-treated mice, a minimum of $1{\times}10^7$ spleen cell was required for production of positive local GVH reaction with almost linear relationship up to an inoculum of $5{\times}10^8$ cells. In control mice, however, a minimum of $1{\times}10^8$ spleen cells was required for positive GVH reaction. These results strongly suggest that the ginseng administration augments significantly the local xenogenic GVH reaction which was used to assess T lymphocyte function and immunocompetence of mice and in addition to this, these results appear to support previous suggestions that the local GVH reaction consitutes a qualitative test of the functional activity of T lymphocytes. These results may be the first to induce local GVH reaction, employing rats as recipient and mice as donor. This study was also desingned to investigate some of the effects of ginseng extract on lymphocyte-macrophage interactions. This was accomplished by in vitro quantification of 1) migratory inhibitory factor(MIF) synthetic capacity of splenic lymphocytes in mice previously primed with ginseng 2) MIF responsiveness of mouse peritoneal macrophages or chicken peripheral leucocytes under the presence of ginseng extract 3) migration ability of chicken peripheral leucocytes by direct stimulation of ginseng extract or ginseng saponin and 4) immunosuppressive effects of immunosuppressants such as cyclophosphamide, cyclosporin A or dexamethasone. Mice divided equally into the ginseng and the saline groups, which received intraperitoneally daily 0.2ml of ginseng absolute alcohol-extract(5mg/ml) and same amount of saline for 15 days, respectively. The cellular immune responsiveness of these mice was assayed 15 days after ginseng pretreatment. Splenic lymphocytes of mice treated with ginseng, when stimulated with sensitized specific-antigen such as sheep red blood cells or toxoplasmin, or with polyclonal activator concanavalin A, produced significantly more MIF than those of control saline group. MIF responsiveness of normal mouse macrophages was significantly augmented when assayed under the presence of ginseng extract (1mg/ml). The migratory ability of normal chicken leucocytes in the absence of MIF was significantly decreased by the stimulation of ginseng extract alone. MIF response was significantly decreased by immunosuppressants and this impaired response was not restored by ginseng pretreatment. This study was additionally performed to evaluate the effect of ginseng on the expulsion of adult Trichinella spiralis in mice. ICR mice were infected experimentally by esophageal incubation of 300 T. spiralis infective muscle larvae prepared by acid-pepsin digestion of infected mice. and received oral administration of 70% alcohol ginseng extract(10mg/mouse/day) for the indicated days plus 4 days before infection. At various times after infection, the number of adult T. spiralis worms in small intestines was determined. Interestingly, ginseng-treatment was accompanied by accelerated expulson of T. spiralis. These results led to the conclusion that Panax ginseng caused some enhancing effect on GVH reaction, macrophage migration inhibition reaction and expulsion of T. spiralis. In addition these results suggested that the mechanisms responsible for this enhancement of ginseng may be chiefly or partially due to nonspecific stimulation of cell-mediated immune response.