• Korean Journal of Veterinary Service
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• v.45 no.1
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• pp.47-54
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• 2022

Subcutaneous abscesses, which occur mainly in goats and sheep, are lymph node abscesses caused by Corynebacterium pseudotuberculosis infection, and are divided into internal, external, and mixed types depending on the type of occurrence. While diagnostic methods for subcutaneous abscesses have been continuously studied, research reports for effective treatment and management of subcutaneous abscesses are inadequate. Therefore, this study was conducted to determine the changes in biometric information related to the inflammatory markers of goats induced by subcutaneous abscesses by infection with C. pseudotuberculosis. For this, hematological tests, analysis of inflammatory indicators, and analysis of serum proteins through electrophoresis separation of goats with healthy goats and goats inoculated with C. pseudotuberculosis to induce subcutaneous abscesses were compared and analyzed by date, and the differences and characteristics were identified periodically. As a result, in goats induced with subcutaneous abscesses, anemia findings related to a rapid decrease in red blood cell (RBC), hematocrit (HCT), and hemoglobin (Hb) were observed, and a significant increase in inflammatory cells expressed in total white blood cell (WBC), neutrophil, and monocytes was observed. And the levels of acute phase protein (APP) such as fibrinogen, haptoglobin, and serum amyloid A (SAA) were observed to increase rapidly immediately after infection. In addition, in the results of electrophoretic analysis of serum proteins, it was observed that the levels of α-globulin and β-globulin were significantly increased in goats with subcutaneous abscesses. That is, when looking at these changes, it was found that the systemic inflammatory response of goats was rapidly induced immediately after infection with the C. pseudotuberculosis pathogen. Through this study, it was possible to identify changes in the biomarkers of goats with subcutaneous abscesses, which had not been reported. Furthermore, these analyzed data are thoughts to be of great help in identifying, treating, and managing the goats of subcutaneous abscesses.

• Parasites, Hosts and Diseases
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• v.23 no.1
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• pp.156-164
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• 1985

To elucidate the effect of splenectomy on the development of experimental primary amoebic meningoencephalitis in mice, the death rate and survival time of mice infected intranasally with Naegleria fowleri trophozoites $5{\times}10^4$ cultivated in CGVS medium were compared according to the mouse age when splenectomy was done, and post-operation until experimental infection. Immunodigusion was undergone to detect the presence of serum antibod). due to N, fowleri infecttion in mice. Polyacrylamide gel electrophoresis was done to compare the protein fractions of mouse serum in each experimental groups. In experiment I, splenectomy was done 3 weeks and infection 4 weeks after birth, the death rate of control, sham operated and splenectomized group were 100%, 85% and 95%, and the mean survival time after infection 7.3 days, 7.5 days and 7.8 days, respectively. In experiment II, splenectomy was undergone 3 weeks and infection 6 weeks after birth, the death rate of of control, sham operated and splenectomized group were 95%, 95% and 95%, and the mean survival time after infection 12.1 days, 11.5 days and 11.5 days, respectively. In experiment III, splenectomy was done 5 weeks and infection 6 weeks after birth, the death rate of control, sham operated and splenectomized group were 95%, 90% and 95%, and the mean survival time after infection 8.1 days, 8.3 days and 8.5 days, respectively. By Ouchterlony immunodigusion, anti-JV. fowleri antibody in the serum of mouse with primary amoebic meningoencephalitis was detected against a N. fowleri antigen, which was prepared by ultrasonication of N, fowleri trophozoites, each reacting two lines of precipitation. The patterns of serum fractions by polyacrylamide gel electrophoresis were different between control and sham operated groups from splenectomized group in fraction II, III and V, the sera of which were collected after N. fowleri infection. This results may be summarized as that splenectomy has no effect on the development of primary amoebic meningoencephalitis in mice.

• Journal of Pharmacopuncture
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• v.18 no.4
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• pp.7-11
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• 2015

Objectives: Bee venom (BV) is a complex mixture of proteins and contains proteins such as phospholipase and melittin, which have an effect on blood clotting and blood clots. The mechanism of action of honey bee venom (HBV, Apis mellifera) on human plasma proteins and its anti-thrombotic effect were studied. The purpose of this study was to investigate the anti-coagulation effect of BV and its effects on blood coagulation and purification. Methods: Crude venom obtained from Apis mellifera was selected. The anti-coagulation factor of the crude venom from this species was purified by using gel filtration chromatography (sephadex G-50), and the molecular weights of the anti-coagulants in this venom estimated by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Blood samples were obtained from 10 rabbits, and the prothrombin time (PT) and the partial thromboplastin time (PTT) tests were conducted. The approximate lethal dose (LD) values of BV were determined. Results: Crude BV increased the blood clotting time. For BV concentrations from 1 to 4 mg/mL, clotting was not observed even at more than 300 seconds, standard deviations $(SDs)={\pm}0.71$; however, clotting was observed in the control group 13.8 s, $SDs={\pm}0.52$. Thus, BV can be considered as containing anti-coagulation factors. Crude BV is composed 4 protein bands with molecular weights of 3, 15, 20 and 41 kilodalton (kDa), respectively. The $LD_{50}$ of the crude BV was found to be $177.8{\mu}g/mouse$. Conclusion: BV contains anti-coagulation factors. The fraction extracted from the Iranian bees contains proteins that are similar to anti-coagulation proteins, such as phospholipase $A_2(PLA_2)$ and melittin, and that can increase the blood clotting times in vitro.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.5
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• pp.1108-1117
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• 2007

To investigate the possible mechanism of ${\alpha}-spinasterol$ as a candidate of anti-cancer drug, I examined the effects of ${\alpha}-spinasterol$ on the apoptosis of U937 cells MTT assay, flow cytometric analysis, SDS-polyacrylamide gel electrophoresis, Western blot analysis, and RT-PCR were performed. ${\alpha}-spinasterol$ treatment reduced the cell viablilty of U937 cells in a dose-dependent manner, which was associated with the induction of apoptotic cell death. ${\alpha}-spinasterol$ treatment also reduced the levels of Bcl-xL anti-apoptotic protein expression and increased the levels of caspase-3, p53, pro-apoptotic protein, in U937 cells. After treatment the level of Bcl-xL, anti-apoptotic gene expression was decreased and the level of ICE pro-apoptotic gene expression was increased. These findings suggest that ${\alpha}-spinasterol$ induced the apoptotic cell death via regulation of several growth regulatory gene products. The abbreviations used are: FBS, fetal bovine serum; PBS, phosphate buffered saline; PI, propidium iodide; OD, optical density; DiOC6, 3,3-dihexyloxa carbcyanine iodide; MTT, 3 [4-5-dimethylthiazol-2-yl] -2-diphenyltetrazolium bromide.

• Journal of Life Science
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• v.20 no.2
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• pp.153-160
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• 2010

This study was conducted to examine the utilization of immunohistochemistry using the bovine anti-brucella immunoglobulin G (IgG) antibody in the diagnosis of brucellosis and to develop a functional biomarker relation for the progress of the disease. Anti-brucella IgG antibody was purified from the affected bovine serum using an affinity chromatography. We performed our investigation on 17 cases of brucellosis and 19 control cases with negative Rose-Bengal test results. Our purified anti-brucella IgG antibody showed a positive immunoreactivity in cytoplasmic hepatocytes of the centrilobular region, and glomeruli and tubular epithelium of the kidney. The protein pattern of the affected liver versus control was analyzed by two-dimensional electrophoresis, showing a different expression pattern of proteins between the two. Five protein spots were up-regulated and another were five down-regulated in the brucellosis liver. Significant upregulaton of catalase and 3-hydroxyacyl-CoA dehydrogenase might be due to a compensatory reaction in response to the endotoxic shock of brucella. In conclusion, the anti-brucella IgG antibody may be a good tool for discriminative diagnosis of the affected tissues and proteomics data suggest new target proteins underlying a possible pathogenic mechanism of brucellosis.

• Parasites, Hosts and Diseases
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• v.27 no.1
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• pp.9-14
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• 1989

Enzyme-linked immunoelectrotransfer blot (EITB) using crude worm antigen of adult Paragonimus westermani was performed for human patients sera to identify the species-specific components. Crude antigen was obtained by homogenizing and centrifuging 24-week old adult worms at 10,000 rpm for 60 minutes in phosphate buffered saline (PBS, PH 7.2) containing: Phenyl methyl sulfonyl auoride (PMSF). Gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) was performed and blotted electrophoretically onto a sheet of nitrocellulose paper. The sheet was cut into strips and exposed to sera diluted 1 : 200 with PBS. SDS-PAGE showed 26 protein bands ranging 229 to 10 kDa. Of them 229, 91, 60, 50, 35∼31, 27, 25, 21, 17, 11 and 10 kDa components showed positive reaction with serum antibody of patients with p. westermani. Sera of patients infected with Clcnorchis sinensis reacted with 35∼31, 19, and 11 kDa bands. Human sera from cysticercosis and diphyllobothriasis cases showed non-specific cross reactions with 229, 35∼31, 27, 25 and 17kDa bands. Protein bands of 91, 60, 21 and 10kDa showed strong positive reaction without cross reactions with sera from other helminthic infections.

• Parasites, Hosts and Diseases
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• v.32 no.4
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• pp.249-258
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• 1994

Recently, the importance of toxoplasmosis is raised as a complication in immunosuppressed or AIDS patients. Our study focused on the identification of a variety of Toxoplasma antigens by immunoblotting. Rabbits and BALB/c mice were immunized with Toxoplosmo Iysate (RH strain) , frozen tachyzoites (RH strain) or cysts (Beverly and Fukaya strain) . Blood were collected from ear vein, heart or orbital plexus for detecting the serum antibody levels. For excretory-secretory (E.S) antigens, T gondii (RH) tachyzoite were cultured in CHL (Chinese hamster lung) cells with MEM containing of 5% FCS. After 72hrs, culture supernatant was collected. BALB/c mice were inoculated with RH tachyzoite intraperitoneally and peritoneal fluids were extracted three days later. E.S antigens were detected in culture supernatant and infected mouse peritoneal fluid by EITB. Serum IgG levels in rabbit were 1 :512 of 10 days after primary immunization, 1 : 2,048 of 10 days after secondary immunization, 1: 1,024 of 20 days after secondary immunization by IFAT, respectively. Serum IgG levels of immunized mice were 1:128 after 7 weeks. Tachyzoite antigens of the RH strain were detected 25 protein bands ranging 10 kDa-220 kDa of molecular weights with Coomassie blue stain. Toxoplcsma major antigens corresponding to n of 24 kDa, 27 kDa,30 kDa, 35 kDa, 38 kDa were recognized by IgG and IgM antibodies. Excretory-secretory antigens present in culture supernatant with M. W. of 20, 30 kDa and in infected mouse peritoneal fluid with M.W. of 33 (P30), 45 kDa. When RH tachyzoite antigen was probed with different mice sera immunized with 2 strains of T gondii, the IgG antibody bud of Fukaya and Beverly strain (8 week-serum) is identical to those of RH strain. It is considered that the 30 kDa polypeptide detected in excretory- secretory materials and Iysate was important major antigen of T gondii (RH).

• Tuberculosis and Respiratory Diseases
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• v.51 no.2
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• pp.155-160
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• 2001

Cryoglobulinemia is the presence of globulins in the serum that precipitate on exposure to cold temperatures(cryoglobulins). Pulmonary complications of cryoglobulinemia include interstial infiltration, impaired gas exchange, small airway disease and pleurisy. Only one other acute respiratory distress syndrome(ARDS) case has been described in patients with cryoglobulinemia. A 55-years old man was admitted with dyspnea. He had been diagnosed as being a hepatitis B virus antigen carrier 15 years ago. On the first admission, chest radiography showed a bilateral pleural effusion and a patchy infiltration on both lungs. On protein- and immuno-electrophoresis, cryoglobulinemia was confirmed. The patient was treated with corticosteroid and plasmapheresis. Forty-five days after the diagnosis, the patient complained of progressive dyspnea and showed a diffuse bilateral pulmonary infiltration on chest radiography. Despite intensive care with mechanical ventilation, the patient died as consequence of hypoxemia and multiple systemic organ failure. On a pathologic examination of the postmortem lung biopsy, multiple necrotizing vasculitis and increased infiltration of the lymphocytes and monocytes were observed. In conclusion, ARDS developed as a result of pulmonary hemorrhage due to cryoglobulinemia-associated vasculitis.

• Journal of Dental Rehabilitation and Applied Science
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• v.19 no.2
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• pp.87-96
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• 2003

The purposes of this study were to find out the optimum intensity of magnetic field where magnetism could promote the activity of osteoblast, and to discover the possibility of clinical application in the areas of dental implants and bone grafts by confirming the effect of clinically increasing bone formation. In this experiment, we used the Neodymium magnet, which had magnetic power six times as strong as the current ones and enabled the resistances against the demagnetization up to 20 to 50 times to be minimized with the size of 1mm in sight. In order to culture cells, a specially designed device was used. It was made to adjust the distance and accordingly to control the intensity of the magnetic field, by placing the cell culture plate in the center with a magnet of 1mm long and thick installed on the both ends. Using MC3T3-E1 cell, a kind of osteoblast-like cell, we cultured, for 24 hours, not only the test group which had been cultured under the magnetic fields with different intensity of 5, 10, 50, 100, 500, and 1000 Gauss, but also the control group excluding the influences of the magnetic field. After observing the cell's form and the density of the culture medium through an inverted microscope, we made a series of proceedings needed for the immunofluoroscence staining, such as fixation, normal serum reaction, primary antibody reaction, and secondary antibody reaction. And with a fluorescence microscope, we observed those-above and compared the frequency of expression of IFG-1 receptor. To make a Western immunoblotting analysis, the cells cultured under the same condition as the above had the procedure of the lysis buffer and the acrylamide gel electrophoresis was carried out. Protein transferred into the nitrocellulose membrane and tested on the primary and the secondary antibody reactions was observed and compared. The results were as follows: When observed through an inverted microscope, the nuclear divisions of the cells under the magnetic field of 10 Gauss were the most active, and the density of the cells could be observed the most enormously. As the result of an immunofluoroscence staining of IGF-1 receptor, the expression of IFG-1 was the most frequently observed under the magnetic field of 10 Gauss. On the other hand, few differences of consideration were made between the test group cultured under the magnetic fields of 5, 500, and 1000 Gauss and the control group. In respect of the expression of IFG-1 receptor, the test group cultured under the magnetic fields of 50 and 100 Gauss were higher than the control group, and lower than that cultured under the magnetic field of 10 Gauss.(p<0.05) According to the Western immunoblotting analysis, the band of IFG-1 receptor which had 85KDa of molecular weight was the darkest. Judging from the above-mentioned results, the growth factor receptor of an osteoblast cell which was an important criterion for the bone formation was increased in maximum under the magnetic field of 10 Gauss. Moreover it was observed that the optimum intensity of magnetic field in which magnetism made the activity of the osteoblast cell increase was about 10 Gauss.

• Journal of radiological science and technology
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• v.28 no.4
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• pp.333-340
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• 2005

In this study, the radioprotective effects of Grifola umbellata hot water extracts (Gu-extract) on mice were investigated. Single pre-administration of Gu-extract increased the 40-day survival ratio of irradiated mice from 65.5％ to 78.6％. The growth of 3 week old male mice in the irradiated group was slightly retarded as compared to those of the control and Gu-extract treated mice. The average spleen and thymus weights of the irradiated mice were lower than those of the control and Gu-extract treated mice. The weight reduction of testis in the irradiated mice was significant. While it was relatively slight in the Gu-extract treated mice as compared to that of control mice. No significant difference in the weight was observed in heart, kidney or liver among three groups. The leukocytes of the Gu-extract treated mice did not decrease dramatically as in the irradiated group, but recovery patterns were similar in both groups. Reduction of erythrocytes were similar in both groups but its recovery occurred more rapidly in the extract treated group. The glucose level of the Gu-extract treated group did not change during the period examined, while it was still higher in the irradiated group than the level in the control group in two weeks. The cholesterol levels in the irradiated and the Gu-extract treated groups were higher than that of control group on day 7, but decreased to the level of the control group on day 14. No difference was observed in total protein amount of the serum among the three groups. SDS-polyacrylamide gel electrophoresis of the soluble proteins extracted from various organs did not reveal differences to any extent in all groups except in the livers of the irradiated and extract treated groups, in which some proteins were missing or less present.