• Korean Journal of Veterinary Research
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• v.40 no.3
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• pp.563-573
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• 2000

Periodic outbreaks of Newcastle disease (ND) caused by velogenic viscerotropic ND virus (vvNDV) has become a major concern in Korea nowadays. Throughout last epidemic, the winter season in 2000, most chicken flocks infected early, under 2-4 weeks of age, showed high mortality up to 50-100%. Serum samples collected from 201 breeder, 284 layer and 112 broiler chicken flocks were examined to evaluate the efficacy of various vaccination methods and programs routinely used for mass vaccination in the field poultry farms. Despite repeated live vaccination, most poultry flocks vaccinated by drinking water route using nipple water supply system failed to produce solid active immune response to NDV during the growing time. In the present study, we applied the spray vaccination technique using Ulvavac or Desvac sprayer to the experimental poultry flocks and examined the efficacy of live vaccination effects induced by it under field condition. Measurable antibody to NDV as well as early protection against vvNDV challenge were found in poultry flocks vaccinated by spray route. Further, we did not found significant post vaccination reactions caused by spray vaccination if properly administered. These data indicate that the spray vaccination will be safe and reliable mass vaccination method for the prevention of ND.

• Journal of Microbiology and Biotechnology
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• v.16 no.1
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• pp.126-135
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• 2006

In a previous study, a biological response modifier (BRM) specifically enhancing the function of B-cells was isolated from Korean fermented soybean paste (Kfsp), but not from non-fermented soybeans. In this study, we attempted to isolate the bacteria producing the BRM from Kfsp (KfspBRM) by ELISA using anti-KfspBRM and by B-cell proliferation. Five bacteria whose culture supernatants showed the BRM activities were isolated, and one of them was identified as Bacillus licheniformis E1. The bacterial BRM (bBRM) originated from a slime layer of B. licheniformis El had a molecular weight of 1,594 kDa, and contained $33\%\;(w/w)$ of reduced sugar and $4.6\%\;(w/w)$ of protein content. The bBRM appeared to be a glycoprotein that is physically, structurally, and functionally similar to the KfspBRM, suggesting that the isolates including B. licheniformis El may produce the KfspBRM in the fermentation process of soybean paste. The mass production of the BRM by the bacterium may help to study B-cells in immunology, and the enrichment of the BRM in Kfsp may help patients in future who are medically in need of potentiation of B-cell proliferation and antibody production.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.6
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• pp.948-953
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• 2007

The relative performance and immune response was evaluated in White Leghorn layers fed liquid DL-methionine hydroxyl analogue-free acid (MHA-FA) relative to dry DL-methionine (DLM) in maize-soybean-sunflower based diets. Three graded levels of methionine (Met) from DLM or MHA-FA were added to the basal diet containing 0.27% Met on an equimolar basis to achieve 0.30, 0.36 and 0.42% Met in the diet. Each diet was fed ad libitum to 25 replicates of one bird (individual feeding) each, from 24 to 40 weeks of age. A regime of 16 h light was provided and all the layers were kept under uniform management throughout the experimental period. None of the parameters studied were influenced by the interaction between source and level of Met in diets. Similarly, the majority of parameters, except for daily feed consumption and immune response (influenced by level) and egg specific gravity and shell thickness (influenced by source), were not affected by either source or level of Met in the diets. Feed consumption was significantly lower in the birds fed a diet containing 0.42% Met compared to those fed lower levels of Met. The cutaneous basophilic hypersensitivity response to PHA-P and antibody titre (32 and 40 wk) to inoculation of sheep red blood cells increased significantly by increasing the concentration of Met in the diet from 0.30 to 0.36%. Thus, the Met requirement for immune competence was higher than for optimum production. The source of Met significantly influenced the egg specific gravity and shell thickness. The specific gravity and shell thickness of eggs increased significantly when MHA-FA was used as the source of Met in the diet compared to DLM. From the study it is concluded that Met requirement for immune competence (360 mg/b/d) is higher than for optimum production (300 mg/b/d). MHA-FA was comparable with DLM as a source of Met for production performance and immunity, when the bioavailability of MHA-FA was considered as 88% of DLM. Further, MHA-FA improved egg shell quality compared to DLM.

• Journal of Microbiology and Biotechnology
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• v.19 no.1
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• pp.55-64
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• 2009

A B cell-specific growth substance (BGS) was isolated from the slime layer of Bacillus licheniformis E1. Unlike LPS, the BGS was not affected by polymixin B, an inhibitor of LPS, or by TLR4, and resulted in the growth of B cells. When BALB/c mice were treated with the BGS, the B cell population was found to increase in both the bone marrow and the spleen, with a marked increase after 24 h in the bone marrow and after 48 h in the spleen. When using antibodies to B cell lineage-restricted surface molecules to analyze the B cell population changes resulting from treatment with the BGS, an increase in immature B cells ($IgM^+$ and $AA4.1^+$) and mature B cells ($IgM^+$ and $IgD^+$) was found in the bone marrow 24 h after treatment with the BGS, whereas a decrease in mature B cells and increase in $IgG^+$ B cells were found in the spleen. When the BGS and OVA antigen were injected into the peritoneal cavity of BALB/c mice, this resulted in a high OVA-specific antibody titer in the sera, similar to that induced by aluminum hydroxide. Therefore, it is anticipated that the mass production of the BGS by B. licheniformis E1 could be used for studies of B cells in immunology, and contribute to the development of a new adjuvant for vaccine manufacture.

• Korean Journal of Veterinary Research
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• v.36 no.1
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• pp.101-108
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• 1996

This study was desinged to investigate the effect of estrogen(Est) on the proliferating of progesterone(Prog) target cells. The spayed 13 rats(Wistar, approximately 300gm) were randomly alloted into 3 groups. One group was the control group and another Prog-treated group was injected with 1mg of Prog/rat/day for 2 consecutive days, and Estand Prog-treated group was injected intramuscularly with $17{\beta}$-estradiol $20{\mu}g/rat/day$ for 3 consecutive days and then with Prog for 2 days as above from 4th day. Rats were administrated intraperitoneally with bromodeoxyuridinc(Brdur,0.2mg/BW once) befero 2 hours of exanguination. In gross finding, the groups with more level of dimension and weight on the uterus were ordered as Est- and Prog-treated group, Prog-treated group and control group. The investigation by immunohistochemical methods using paraffin sections of the uteri was performed by using anti-Brdur antibody for labeling proliferating cells of Prog target cells. The groups with higher labeling index(LI) were ordered as Prog-treated grop, Est- and Prog- treated group and control group. The number of proliferating cells from Prog target cells in the rats were rather deceased by Prog injection following Est injection than prog injection only. The cell types with higher LI in the wall layers of all 3 groups were ordered as endometrial stromal cells, glandular epithelial cells, luminal epithelial cells, myometrial muscle cells and serosa methodelial cells, and the region with highest LI was functional zone of the endometrium and the region with lower LI was muscular layer and then those with lowest LI was serosa and also the considerable different LI from individual rat were observed.

• Journal of Sensor Science and Technology
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• v.22 no.2
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• pp.144-149
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• 2013

In this research, we developed a biosensor to detect lung cancer-specific biomarker using Anodic Aluminum Oxide (AAO) chip based on interference and nano surface plasmon resonance (nanoSPR). The nano-porous AAO chip was fabricated $2{\mu}m$ of pore-depth by two-step anodizing method for surface uniformity. NanoSPR has sensitivity to the refractive index (RI) of the surrounding medium and also provides simple and label-free detection when specific antibodies are immobilized to the Au-deposited surface of nano-porous AAO chip. To detect the lung cancer-specific biomarker, antibodies were immobilized on the surface of the chip by Self Assembled Monolayer (SAM) method. Since then lung cancer-specific biomarker was applied atop the antibodies immobilized layer. The specific reaction of the antigen-antibody contributed to the change in the refractive index that cause shift of resonance spectrum in the interference pattern. The Limit of Detection (LOD) was 1 fg/ml by using our nano-porous AAO biosensor chip.

• Applied Microscopy
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• v.30 no.4
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• pp.377-387
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• 2000

There are a few tanycytes between the general ependymal cells lining the ependymal layer of the brain ventricle. These cells are considered as modified ependymal cells which possess a long basal process. Tanycytes are known to have an ability to communicate by absorbing substances from cerebrospinal fluid and transporting them to the blood vessels and/or to the neurons in the CNS. The third and fourth ventricular tanycytes were mainly studied as subjects but it's rare to find reports about the tanycytes of the area postrema. Glial fibrillary acidic protein is an intermediate filament protein that is expressed especially in astrocytes of the CNS. But GFAP is also found in filament of the tanycytes and its process. Therefore this study was carried out for the examination of the GFAP immunoreactive tanycytes lining the area postrema of the bat, and we also examined the ultrastructure of tanycytes using electron microscope. GFAP immunoreactive tanycytes were located in the caudal portion of the fourth ventricle, and especially mainly in the transitional zone between the floor of the caudal fourth ventricle and ependymal layer lining the area postrema. A few GFAP immunoreactive tanycytes were also found in the ependymal layer lining the area postrema, and some groups of tanycytes were found in the ependymal layer of the area postrema near the floor of the caudal fourth ventricle , The processes of tanycytes were stained deeply with anti-GFAP antibody. Especially the GFAP immunoreactive tanycytes lining the area postrema had very long processes that cross the whole width of the area postrema. In the electron microscope, the cell body of ependymal tanycyte was located on the ependymal layer and had a long basal process. Intermediate filaments were observed around the nucleus and well developed in the process of tanycrte. Longitudinal oriented long mitochondria and a few lipid droplets were also found in this process. After immunocytocheical staining, the gold particles were found only in the intermediate filaments. We could not determine the function of the tanycytes in the area postrema. Thus, further investigation is required to determine the functional relationship between the tanycytes and the area postrema in hibernating animal, the bat.

• Korean Journal of Veterinary Research
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• v.35 no.2
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• pp.217-228
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• 1995

The study was designed to investigate the effects of progesterone on the reproductive system. This investigation was performed by immunohistochemical methods using anti-bromodeoxyuridine-antibody following bromodeoxyuridine(Brdur) injection for labeling proliferating cells in the uterus and ovary of rats. Sixteen female rats(Wistar), weighing initially 300g, were randomly allotted into ovariectomized and unovariectomized large groups. These two large groups were subdivided into three subgroups of control, 3-day and 6-day groups, respectively. 3-days and 6-days group were injected with 1mg of progesterone/rat/day for 3 or 6 days, respectively. In gross findings, the uterus of ovariectomized groups markedly atrophied, and were not hypertrophied by progesterone injection for 3 days or 6 days and the uterus of unovariectomized groups also were not hypertrophied. Labeling index(LI, %) was measured by counting the number of Brdur-positive cells from 300 to 3,000 cells per layer in the uterus tissue. The average LI of the uterus in unovariectomized groups was higher than that of ovariectomized groups. The subgroups with higher LI in unovariectomized groups were ordered as 6-day group, 3-day group. So progesterone considerably effected to the proliferating of the cells in the uterus of unovariectomized groups. The layers with higher LI in the uterus wall were ordered as the functional zone of endometrium, epithelial layer of endometrium, basal zone of endometrium, myometrium and perimetrium. The cell types with higher LI in the uterus of unovariectomized groups were ordered as the surface epithelial cells, stromal cells, glandular epithelial cells and muscle cells. Growing follicles with proliferating cells from secondary and tertiary follicles in the ovary of unovariectomized groups appeared to be 37.66% in control group, 39.23% in 3-day groups, 39.47% in 6-day groups. Mature follicles in the ovary were more number in control group than those in 3-day groups but not appeared in 6-day groups. So progesterone not nearly effects to the number of the growing follicles but appeared to be related to suppression of the development and protrusion of the mid-tertiary and mature follicles on the ovary surface. The cell types with higher LI in the ovary of unovariectomized groups were respectively ordered as granulosa cells, theca interns cells in secondary follicles; theca interna cells, granulosa cells, theca externa cells in tertiary follicles; fibroblasts, theca in terns cells in atretic follicles; fibroblasts, luteal cells in corpus luteum.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.3
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• pp.191-198
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• 2007

The p53 which is well known as tumor suppressor gene is located at 17p13. p53 is a sequence-specific DNA binding transcription factor that responds to certain cytotoxic stresses, such as DNA damage, by enhancing the transcription of genes that regulate cell-cycle progression as well as programmed cell death. The p63 gene that is located at 3q27-29, is recognized members of the p53 family, and responsible for the transcription of 6 isoforms. Three isoforms ($TAp63{\alpha}$, $TAp63{\beta}$, $TAp63{\gamma}$) contain an N-terminal transactivation (TA) domain and can induce apoptosis. The other 3 isoforms (${\Delta}Np63{\alpha}$, ${\Delta}Np63{\beta}$, ${\Delta}Np63{\gamma}$) lack the TA domain and may function in a dominant-negative fashion by inhibiting the transactivation functions of p53 and TAp63 proteins, and thus act as oncoproteins. A number of studies have investigated the role of p63 in human squamous cell carcinomas from different organs. Only a few studies have examined ${\Delta}Np63$ isoform in oral squamous cell carcinoma including normal epithelium. This study aimed to evaluate expression of ${\Delta}Np63$ isoform in human oral squamous cell carcinoma tissue and normal mucosa. The 3 cases of well differenciated oral squamous cell carcinoma specimen including adjacent normal mucosa were examined, and immunohistochemical study with monoclonal antibody(4A4) and tumor cell apoptosis analysis with Transmission Electon Microscopy were studied. And, RT-PCR analysis was done for expression of ${\Delta}Np63$ isoform. The results were as followed. 1. Normal gingiva showed the restricted p63 expression in basal cell layer. 2. Well differentiated squamous cell carcinoma showed mainly p63 expression in overall area of malignancy, especially in basal cell layer to adjacent stromal tissue. 3. Tumor cells around keratinized area with no p63 expression disclosed less micro-organelle in decreased size cytoplasm and severe chromatin margination with nuclear destruction that means apoptosis. 4. Comparison of mRNA expression of ${\Delta}Np63$ isoform by RT-PCR showed variable expression of ${\Delta}Np63$ isoform, but ${\Delta}Np63{\alpha}$ was most highly expressed in all 3 tumor specimen. From theses results, it should be suggested that ${\Delta}Np63$ isoform expression in well differentiated squamous cell carcinoma was closely related to tumor oncogenesis, expecially overexpression of ${\Delta}Np63{\alpha}$ is a most important factor in tumor genesis of oral squamous cell carcinoma.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.129-142
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• 2002

Epithelial-mesenchymal interaction plays a important role in cell growth and differentiation. This interaction is already well known to have an importance during the organ development as well as cell growth and differentiation. However, in vitro experimental model is not well developed to reproduce in vivo cellular microenvironment which provide a epithelial-mesenchymal interaction. Because conventional monolayer culture lacks epithelial-mensenchymal interaction, cultivated cells have an morphologic, biochemical, and functional characteristics differ from in vivo tissue. Moreover, it's condition is not able to induce cellular differention due to submerged culture condition. Therefore, the aims of this study were to develop and evaualte the in vitro experimental model that maintains epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally reconstituted oral keratinocytes by histological and immunohistochemical analysis. The results were as follow; 1. Gingival keratinocytes reconstituted by three-dimensional organotypic culture revealed similar morphologic characteristics to biopsied patient specimen showing stratification, hyperkeratinosis, matutation of epithelial architecture. 2. Connective tissue structure was matured, and there is no difference during stratification period of epithelial 3-dimensional culture. 3. The longer of air-exposure culture on three-dimensionally reconstituted cells, the more epithelial maturation, increased epithelial thickness and surface keratinization 4. In reconstitued mucosa, the whole epidermis was positively stained by anti-involucrin antibody, and there is no difference according to air-exposured culture period. 5. The Hsp was expressed in the epithelial layer of three-dimensionally cultured cells, especially basal layer of epidermis. The change of Hsp expression was not significant by culture stratification. 6. Connexin 43, marker of cell-cell communication was revealed mild immunodeposition in reconstitued epithelium, and there is no significant expression change during stratification. These results suggest that three-dimensional oragnotypic co-culture of normal gingival keratinocytes with dermal equivalent consisting type I collagen and gingival fibroblasts results in similar morphologic and immunohistochemical characteristics to in vivo patient specimens. And this culture system seems to provide adequate micro-environment for in vitro tissue reconstitution. Therefore, further study will be focused to study of in vitro gingivitis model, development of novel perioodntal disease therapeutics and epithelial-mensenchymal interaction.