• Korean Journal of Poultry Science
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• v.17 no.3
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• pp.225-232
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• 1990

The immune responses of commercial layer chickens against Newcastle disease (ND) were compared among different administration methods and times of vaccination during 4 weeks of age. A total of 372 day-old chickens were devided into 4 groups of 93 birds each. Each of 3 groups was received a commercially available B$_1$ live vaccine via drinking water, eye instillation or spray method at one, 14 and 28 days of age. One group was used as an unvaccinated control. At two and 4 weeks after each time of vaccination, 15 birds from each group were challenged with virulent ND virus at the dose of 10$^{5}$ EID$_{50}$ per bird to examine the pretection rate. Ten to 15 birds from each group were bled at two weeks intervals from day old to 8 weeks of age to determine hemagglutination inhibition antibody titer. The protection rate was generally low regardless of the times of vaccination although two or more times vaccination gave higher protection than once vaccination. The low protection was considered due to low titer of the vaccine used since the vaccine titer was less than 10$^{2.5}$ EID$_{50}$ per bird. Spray method gave better protection compared to eye instillation of drinking water method which resulted in lowest response. When birds were challenged majority showed clinical signs on ND between 3 and 6 days after challenge. Death occured one or two days after onset of symptoms. Major clinical signs observed were depression (96%), drowsy(90%), anorexia (84%), diarrhoea (29%), difficult breath (15%) and torticollis (10%). Hemorrhagic lesions on post mortem were seen in duodenum (51%), trachea(36%), illeum (13%), ceacal tonsil (11%), proventriculus (10%) and some other organs. When birds were challenged majority showed clinical signs on ND between 3 and 6 days after challenge. Death occured one or two days after onset of symptoms. Major clinical signs observed were depression (96%), drowsy(90%), anorexia (84%), diarrhoea (29%), difficult breath (15%) and torticollis (10%). Hemorrhagic lesions on post mortem were seen in duodenum (51%), trachea(35%), illeum (13%), ceacal tonsil (11%), proventriculus (10%) and some other organs.

• Korean Journal of Poultry Science
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• v.40 no.4
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• pp.379-388
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• 2013

Avian paramyxovirus (APMV) type 4 and 6 were isolated during an avian influenza (AI) surveillance program of wild birds. This study also conducted experimental infection of wild-bird-origin APMV type 4 and 6 in specific pathogen free (SPF) chickens to study pathogenicity and transmission within domestic flocks. In addition, serological prevalence data of APMV type 4 and 6 in domestic fowls was conducted with chicken sera collected from 2007 to 2009 in order to understand infection status. The results of the animal experiment showed that APMV type 4 and 6 had the ability to infect chickens with sero-conversion and to transmit the virus from infected birds to contacted birds, but showed low pathogenicity. Serological tests revealed that APMV type 4 was widespread in the poultry industry, especially in layer flocks, but the positive rate for APMV type 6 was very low. This study concluded that wild bird-origin APMV type 4 and 6 could infect the chickens by inter-species transmission and the seroprevalence of APMV type 4 was quite high in Korean poultry. However, since almost all the chicken flocks had a high level of antibody titer against APMV type 1, there was possibility of cross reaction between APMV type 1 and 4, which made the interpretations more complicated. In order to understand infection status in the natural environment, additional study is necessary regarding the seroprevalence of APMV type 4 and 6 in the wild bird population.

• Korean Journal of Veterinary Research
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• v.43 no.4
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• pp.555-561
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• 2003

Incidence of estrum or abortions in pregnant cows may be affected by large follicles developed together with corpus luteum in pair ovaries of pregnant cows. But the follicles of pregnant phase were not assessed about histological findings. Determination of the healthy and atretic follicles by presence of proliferative cells or apoptotic cells and histological compositions of follicles would be used as important data on measurements of ovarian functions. This study was focussed mainly to investigate macroscopical, histological and immunohistochemical findings of ovarian follicles of pregnant Korean native cows and dairy cows (Holstein). In immunohistochemical methods, assessments of proliferative cells using PCNA antibody and apoptotic cells using TUNEL methods were performed. The follicles were observed on all 24 pregnant cows (17 Korean native cows and 7 Holstein cows). Follicles of greater than 10 mm in daimeter were developed in 37.5% (9/24 heads) of these pregnant cows. largest follicles from in these cows were $16.0{\times}15.0mm$ in diameter in a Korean native cow(l20 days of gestation), $13.4{\times}10.1mm$ in a Korean native cow(50 days of gestation), $12.9{\times}11.5mm$ in a Holstein cow (120 days of gestation). 40.5% among all follicles having diameter of greater than 1.0 mm in pregnant cows were assessed as atretic follicles and in addition, healthy follicles also showed less in number and smaller in size and thinner in wall layer compared with those of cyclic phase ovaries. In immunohistochemical findings, also proliferative positive cells and apoptotic positive cells on the granulosa cell layers in the healthy follicles of pregnant cows appeared less than on those of cyclic follicles. So these follicles were assessed as weakly active follicles. In large follicles, above positive cells were not nearly appeared but granulosa cell debris were more appeared among the granulosa cells. So these large follicles were assessed as inactvie or atretic follicles. The above findings suggest that small follicles of pregnant phase were weakly active or atretic and large follicles were inactive or atretic.

• IMMUNE NETWORK
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• v.6 no.1
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• pp.33-42
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• 2006

Background: Calcineurin plays a crucial role in T cell activation, cell growth, apoptosis, and angiogenesis, and its over-expression has been implicated in the pathogenesis of cardiomyopathy and stroke. However, the expression and function of calcineurin in the pathologic lesion of chronic inflammatory diseases, like rheumatoid synovium, remain to be defined. This study was aimed to determine the role of calcineurin in inflammatory arthritis and investigate the expression and function of calcineurin in the rheumatoid synovium and synoviocytes, the actual site of chronic inflammation. Methods: Immuno-histochemical staining using specific antibody to calcineurin was perfomed in the synovium of rheumatoid arthritis (RA). Fibroblast-like synoviocytes (FLS) from RA and osteoarthritis (OA) patients were isolated from RA and OA patients, and cultured with IL-1${\beta}$ and TNF-${\alpha}$ in the presence or absence of cyclosporin A, a calcineurin inhibitor. The calcineurin expression was assessed by phosphatase assay and Western blotting analysis. IL-6, -10, -17, matrix metalloproteinase (MMP)-1, -2, -3, and -9 released into the culture supernatants were measured by ELISA. After transfection with GFP-Cabin 1 gene into synoviocytes, the levels of IL-6 and MMPs were measured by ELISA. Results: Calcineurin was highly expressed in the lining layer of synovium and cultured synoviocytes of RA patients. The elevated calcineurin activity in the rheumatoid synoviocytes was triggered by proin flammatory cytokines such as IL-1${\beta}$ and TNF-${\alpha}$. In contrast, IL-10, an anti-inflammatory cytokine, failed to increase the calcineurin activity. The targeted inhibition of calcineurin by the over-expression of Cabin 1, a natural calcineurin antagonist, inhibited the production of IL-6 and MMP-2 by rheumatoid synoviocytes in a similar manner to the calcineurin inhibitor, cyclosporin A. Conclusion: These data suggest that abnormal activation of calcineurin in the synoviocytes may contribute to the pathogenesis of chronic arthritis, and thus provide a potential target for controlling inflammatory arthritis.

• The Korean Journal of Pharmacology
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• v.31 no.2
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• pp.241-250
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• 1995

Protein B23 is one of the major nucleolar phosphoproteins associated with pre-ribosomal particles, and is localized in the granular region of the nucleolus. Recent studies suggest that protein B23 shuttles between nucleus and cytoplasm and also interacts with HIV Rev. These findings indicate that protein B23 is important in nucleocytoplasmic relationship and viral replication. However, the exact function of protein B23 is not clear yet. In acute nucleolar hypertrophy of rat liver, treated with thioacetamide, there was observed an increase of not only protein B23 but also B23-like protein p45 when anti-B23 monoclonal antibody (MAb) was used for identification. On the basis of the large B23 specific epitope structure composed of 68 amino acids, a hypothesis was formulated to examine that p45 is the pre-B23 resulting from excessive production of B23. In an attempt to investigate the precursor of B23, we analyzed the subcellular fractions and microsomal subfractions. Subsequently, we analyzed the finger printings of B23-like proteins using the tryptic peptide mapping. The results are summarized: 1) Using B23 MAb, we observed the presence of B23-like proteins in nucleolar fraction, nucleoplasmic fraction and microsomal fraction. 2) In the further microsomal subfractionation, we could partially purify B23-like protein in 2M layer of sucrose gradient. 3) When ion exchange chromatography was employed, there were protein species 80kDa(p80), 65kDa(p65) and 60kDa(p60). 4) Based on the tryptic map analysis of $^{125}I$ labeled proteins, the similarity between B23 and p80 was found only in 9 out of 14(B23) and 21(p80) peptides, and difference was found in the remaining peptides. p80 and p60 had 18 common peptides, and all the peptides of p60 were similar to those of p80. From these results, it is proposed that p45 is an abnormal metabolite resulting from carcinogenesis by thioacetamide, and it is not the precursor of B23. In addition, we suggest that p80 may be a precursor of p45.

• Journal of Animal Science and Technology
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• v.46 no.4
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• pp.571-584
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• 2004

It is known widely that granulosa cell apoptosis leads follicular atresia and macrophages exert their effects directly and/or indirectly from the initiation to the completion of follicular atresia by phagocytosis of apoptotic bodies and secretion of various cytokines. However, the site of initiation, propagation routes and the elimination methods of apoptotic bodies, and the time and methods of penetration of macrophages into the follicles are not known completely. Using pig(Yorkshire-breed) ovary, immunohistochemical studies with TUNEL for apoptotic bodies and pig macrophage monoclonal antibody 4E9 for macrophages, and light and transmission electron microscopic observations were performed. In the pig, follicular atresia began with the granulosa cell apoptosis, and the apoptosis of theca intema cells occured at the same time. The apoptosis of granulosa cells initiated randomly within the granulosa cell layer and propagated rapidly into the whole layer. Ultrastructura1ly, apoptotic granulosa cells showed characteristic pyknotic and deformed nucleus and intracytoplasmic vesicles. Apoptotic bodies were eliminated by intact granulosa cells and macrophages. Intact granulosa cells ingested apoptotic bodies transiently, soon after they fell into the apoptosis. Finally, apoptotic bodies and degenerated oocyte were phagocytosed by macrophages. Macrophages entered the ovarian follicle at the time of initiation of granulosa cell apoptosis, and migrated with the progression of apoptosis. By elimination of theca cells, macrophages contributed the completion of follicular atresia These results will provide valuable informations on the study of the interrelation between macrophage and ovarian follicular atresia.

• Tuberculosis and Respiratory Diseases
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• v.42 no.2
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• pp.149-155
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• 1995

Background: Cytokeratin 19 is 40KD acidic molecule whose distribution is restricted to simple or pseudo-stratified epithelia, such as the epithelial layer of the bronchial tree. Immunohistochemical study have shown that cytokeratin 19 is overexpressed in lung carcinoma tissue. An immunoradiometric assay, CYFRA 21-1 has been developed using two monoclonal antibody, BM 19-21 and KS 19-1, reactive to different epitopes on cytokeratin 19. We studied the diagnostic value of CYFRA 21-1 in lung cancer. Method: The serum CYFRA 21-1 level using immunoradiometric kit(ELSA-CYFRA 21-1) was measured in 54 patients who admit to Yeungnam University Hospital from April, 1993 to August, 1994. Lung cancer group was 39 primary lung cancer patients(19 patients with squamous cell carcinoma, 11 patients with adenocarcinoma and 9 patients with small cell carcinoma). Control group was 15 patients with non malignant lung diseases(8 patients with pulmonary tuberculosis, 3 patients with chronic obstructive pulmonary disease, 2 patients with pneumonia and 2 patients with chronic obstructive pulmonary disease combined with pulmonary tuberculosis). Results: The mean serum value of CYFRA 21-1 was $20.2{\pm}4.7ng/ml$ in squamous cell carcinoma, $7.2{\pm}1.6ng/ml$ in adenocarcinoma and $15.5{\pm}4.7ng/ml$ in non-small cell lung cancer. The serum value of CYFRA 21-1 in control group was $1.7{\pm}0.5ng/ml$. All of the serum values of 3 histologic types were significantly higher than that of control group(p<0.01). The serum value of CYFRA 21-1 of squamous cell carcinoma was significantly higher than that of adenocarcinoma(p <0.05). Serum value of CYFRA 21-1 in small cell lung cancer was $2.9{\pm}0.9ng/ml$ and not significantly different compared with control group. Using cut off value of 3.3ng/ml, sensitivity and specificity was 11.1%, 65.2% in small cell lung cancer, 70.0%, 62.5% in non-small cell lung cancer, 73.7%, 75% in squamous cell carcinoma and 63.6%, 78.9% in adenocarcinoma, respectively. Conclusion: The serum levels of CYFRA 21-1 may be useful in diagnosis of non-small cell lung carcinoma, especially in squamous cell carcinoma with its high specificity.

• Applied Biological Chemistry
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• v.47 no.3
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• pp.315-320
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• 2004

The aim of this study was to develop an activator, 3-10 kDa fraction from radish water extracts, that will improve the intestinal function and bowel movement in the colons. Radish water extracts were investigated for their intestinal function effects according to the charcoal meal transit method, employing Balb/c mice: also, their anti-constipation activities were compared utilizing the loperamide-induced constipation method, employing SD rats. The result suggested that the effects of the charcoal meal transit increased remarkably in radish water extract administrated rats in comparison to loperamide administrated rats. Futhermore, the effects of various solvent extracts of radish on charcoal meal transit in Balb/c mice increased remarkably in radish water fraction administrated rats than in different solvent fraction administrated rats. Radish extraction was tested and isolated into 4 groups: below 3 kDa, 3-10 kDa, 10-300 kDa, and over 300 kDa. 3-10 kDa was the most effective on the intestinal function and bowel movement in the colons; also, 3-10 kDa fraction of radish water extraction was found to be the most effective charcoal meal transit. The dry weight and moisture content of feces remarkedly increased in the 3-10 kDa administrated rats group than in the loperamide only group. Experimental results revealed that 3-10 kDa fraction of radish water extract was the most effective on the intestinal function and bowel movement was the crypt epithelial cells that contained more MUC2 in the 3-10 kDa administrated group than the loperamide only group: in addition, the thickness of mucus layer stained with alcian blue was significantly thicker in 3-10 kDa administrated rats than in loperamide administrated rats. Crypt epithelial cells secreted more MUC2 in the 3-10 kDa administrated group than the loperamide only group and the stained cells clearly showed the MUC2 with antibody Biogenex AM358.

• Journal of the Korean Society of Food Science and Nutrition
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• v.24 no.3
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• pp.470-486
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• 1995

Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.