• Journal of Embryo Transfer
• /
• v.21 no.2
• /
• pp.137-146
• /
• 2006

The objective of this study was to examine the effect of donor cell types, the source of recipient oocytes and estrous synchronization on pregnancy and delivery rates of somatic cell nuclear transfer (SCNT) embryos in Korean native goats. Recipient oocytes were surgically collected after superovulation. Ear cells and fetal fibroblasts were collected and cultured in serum-starvation condition (TCM-199 + 0.5% FBS) for cell confluence. The zonae pellucidae of in vivo- and in vitro-matured oocytes were partially drilled using a laser system. Single somatic cell was transferred into the enucleated oocyte. The reconstructed oocytes were electrically fused with 0.3 M mannitol. After the fusion, embryos were activated by Ionomycin+6-DMAP. NT embryos were cultured in mSOF medium supplemented with 0.8% BSA at $39^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $O_2$, 90% $N_2$ for 12 to 20 hr. One hundred and two SCNT embryos were transferred into 20 recipients and pregnancy rate at days 30 was 20.0%. Of them, one developed to term and delivered 1 kid. Ear cells showed significantly higher fusion (63.8 vs. 26.5%) and pregnancy rates (20.0 vs. 0.0%) than those of fetal fibroblast (p<0.05). The recipients synchronized by CIDR showed significantly lower pregnancy rates compared to that of recipient in natural estrus ($0.0{\sim}25.0%$ vs. 100%) (p<0.05). Cloned kid was born from the recipient in natural estrus. For the synchronization of estrus between recipient and donor, there was no difference between treatments (${\pm}0$ vs. +12 hr) in pregnancy rate. The first healthy cloned kid (Jinsoonny) was produced by transfer of SCNT embryos derived from in vivo oocytes and ear cells into a recipient goat whose estrus was synchronized with the donor. These results imply that donor cells for nuclear transfer may affect the success rate, and the estrus synchronization between donor and recipient animals can also be important.

• Tuberculosis and Respiratory Diseases
• /
• v.40 no.2
• /
• pp.98-103
• /
• 1993

Background: Inactivation of retinoblastoma gene (Rb) has been observed in a variety of human cancers. Loss of heterozygosity (LOH) of Rb which is a common mode of allelic inactivation of Rb, has been known as a frequent genetic event in small cell lung cancer but it has been detected less frequently in non-small cell lung cancer. To define the role of Rb deletion in lung cancer, we investigated the genomic DNAs of 43 non-small cell lung cancers and 1 small cell lung cancer paired with normal lung tissues obtained by thoracotomy. Methods: The genomic DNAs were obtained by the digestion with proteinase K followed by phenol-chloroform extraction method. The genomic DNAs were digested by restriction endonuclease (EcoRI), separated by agarose gel electrophoresis, transferred to nylon membrane by Southern blot transfer and then hybridized with labelled Rb 1 probe which contains. 1.4 kb sized DNA sequence containing N-terminal portion of Rb. Results: In 26 squamous cell lung cancers, 16 cases were informative after EcoRI digestion and LOH of Rb was found in 10 cases (62.5%). In 17 adenocarcinomas of lung, 11 cases were informative and LOH of Rb was found in five cases (45.4%). The analysis of clinical parameters revealed no significant differences between the two groups with or without LOH of Rb in the aspects of age, sex, degree of differentiation, stage and smoking amount. Conclusions: These results suggest that Rb inactivation is also significantly involved in the molecular pathogenesis of non-small cell lung cancer.

• Journal of fish pathology
• /
• v.19 no.1
• /
• pp.7-15
• /
• 2006

Amyloodinium sp. was found on the gills of mullet (Mugil cephalus) cultured on land. No external symptoms in the diseased fish were found except decoloration of the gills. In fresh preparations of the gills the parasites were opaque round or oval shape with a bright nucleus and 43.5 ㎛ (18.2～72.7, n=20) in size. In preparations added a drop of Lugol solution, they were black with the same shapes in fresh preparations and 43.5 ㎛ (n=20) in size. The parasites were stained black and blue in a droplet of Lugol solution and Diff-Quick III solution, respectively and their sizes were a little larger than in wet preparations. After stained with May-Grunwald Giemsa, the parasites appeared granular eosinophlic in the peripheral cytoplasm and granular strong basophilic in the center. In silver impregnated specimens the peripheral granules were negative and the central ones positive. The granules appeared brown in purplish cytoplasm after staining with Lugol solution. The parasites developed by binary division when they were cultivated in filtered seawater at 20℃. Histopathologically severe epithelial hyperplasia and fusion in the gill filaments resulted in clubbing, especially the proximal region of the filament. Epithelial hyperplasia was also found in the basal regions of the gill filaments and some epithelial cells were occasionally detached from the filaments. Some pear-shaped trophonts of the parasites with rhizoid attached on the gill filaments showing hyperplasia of the epithelial cells and mucous cells.

• Korean Journal of Medicinal Crop Science
• /
• v.4 no.1
• /
• pp.78-85
• /
• 1996

The effects of growth regulators, sucrose and gelling agents were investigated to increase the efficiency of the callus growth and plant regenerarion in tissue culture of Angelica koreana Max. The fresh weight and dry weight of subcultured callus was highest in MS medium supplemented with 1 mg/l 2,4-D. Callus growth was excellent in 2% sucrose, but it was inhibited in propotion to sucrose content. Effect of gelling agents on callus growth was highest on 1.2% agar and 0.4% Gelrite medium, respectively. The browning of callus was protected on the media supplemented with 10 mg/l ABA and 5 or 10 mg/l $AgNO_3$. In the callus induction and growth from the peduncle of immature inflorescence, 2,4-D was more effective than NAA, and the frequency of callus induction was highest as 81.7% in 2 mg/l 2,4-D. Plant was not regenerated from the callus derived from young leaf. Somatic embryos were developed from the surface of callus drived from the peduncle of immature inflorescence in the medium containing 0.5 mg/l 2,4-D, 1 mg/l kinetin, 5 mg/l ABA and 5 mg/l $AgNO_3$. Plants were developed from the matured somatic embryos in the medium supplemented with 0.2 mg/l 2,4-D and 1 mg/l kinetin.

• Korean Journal of Food Science and Technology
• /
• v.34 no.4
• /
• pp.625-630
• /
• 2002

Raw pine-mushrooms (Tricholoma matsutake Sing.) of four grades and those frozen were analyzed for proximate composition, smell pattern, volatile flavor compounds, and sensory evaluation. Proximate compositions of raw pine-mushrooms (A-C, regular grade) were $89.48{\sim}90.77%$ moisture, 6.81% ash excluding D (below regular grade) sample, $2.24{\sim}2.52%$ crude lipid, and $16.19{\sim}20.01%$ crude protein. Proximate compositions of frozen pine-mushrooms preserved for 6 months at -20 and $-70^{\circ}C$ showed no difference compared with raw pine-mushrooms. Results of smell pattern and multidimensional analysis revealed raw pine-mushrooms showed no differences among samples, but frozen pine-mushrooms differed significantly depending on the grade. Volatile flavor compounds of pine-mushrooms were analyzed using a purge and trap method with GC/MSD. Twenty-nine volatile compounds were identified, among which alcohols such as 1-octen-3-ol, 2-octen-1-ol, 3-methyl-butanol, and n-octanol were commonly found in all pine-mushroom samples. In sensory attributes, raw pine-mushrooms were not significantly different at 5% level, and sample D of frozen pine-mushrooms scored lower than samples $A{\sim}C$.

• KSBB Journal
• /
• v.14 no.1
• /
• pp.82-90
• /
• 1999

In order to eventually fabricate an analytical system for infectious microorganisms, we synthesized major immunochemical components, utilized them for the construction of model system, and investigated an assay concept for bacterial whole cells. For the preparation of system components, a polyclonal antibody, against Salmonella thompson as model analyte, purified by immuno-affinity chromatography was used to chemically link to streptavidin or an enzyme, horseradish peroxidase(HRP). The antibody and streptavidin was modified with sulfosuccinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxylate and N-succinimidyl-3-[2-pyridyldithio]propionate(subsequently activated by dithiotheritol), respectively. The modified components were reacted to synthesize antibody-streptavidin conjugates which were then purified on a two-layer chromatography column of diaminobiotin gel and Sephadex G-100. For antibody-HRP conjugates, HRP molecules were activated by $NalO_4$ oxidation and then coupled to immunoglobulin. After stabilizing with ($NaCNBH_3$, the conjugates were purified by size exclusion chromatography on Biogel A5M column. To devise a model system, such produced components were combined with a dot-blotter in which a nitrocellulose membrane($12{\mu}m$ pre size) with immobilized biotin was already located. The analyte (S. thompson cells) was reacted with the both antibody conjugates in a liquid phase, and the complexes formed were captured on the membrane surfaces by applying vacuum in the bottom compartment of the blotter to invoke biotin-streptavidin reaction. Under optimal conditions, the system enabled to identify the analytical concept for bacterial whole cells, and the lower limit of detection was approximately $1{\mu}g/m{\ell}$($10^5-10^6$ cells/m$m{\ell}$). The controlling factors were the concentrations of each antibody conjugate that caused agglutination in the presence of analyte as they increased.

• Applied Biological Chemistry
• /
• v.41 no.1
• /
• pp.60-66
• /
• 1998

During the screening for anti-complementary activity from 10 kinds of edible mushrooms, an alkali extract of Agaricus bisporus showed the highest activity through the complement fixation test. The crude anti-complementary material(AB-0) from Agaricus bisporus was obtained by the alkali extraction using 1 N NaOH containing 5% urea$(65^{\circ}C)$, followed by methanol reflux, dialysis and lyophilization. The fraction AB-O showed potent anit-complementary and anti-tumor activity against sarcoma-180 injected mice. The fraction AB-0 was divided into 5 fractions(AB-20, AB-40, AB-60, AB-80, AB-A) by gradual acetone precipitation. Among them fraction AB-20 having the highest activity and yield was found to contain 39% carbohydrate and 46% protein. The anti-complementary protein-bound polysaccharide AB-20 consisted of glucose, arabinose, xylose, galactose and mannose in a molar ratio of 6.49 : 1.98 : 1.24 : 1.00 : 0.71, respectively and its main component amino acids were alanine(20.59%), isoleucine(16.85%), glutamine+glutamic acid(14.12%) and leucine (13.83%). The anti-complementary activity of AB-20 was decreased greatly by periodate oxidation, but decreased slightly by pronase digestion. This indicates that polysaccharide moiety is corelated with the anti-complementary activity and that protein is also involved in the activity.

• Journal of Ecology and Environment
• /
• v.29 no.6
• /
• pp.521-529
• /
• 2006

This study was carried out to classify all forest vegetation types in Ulleung Island, Korea using the methodology of the Z.-M. school's phytosociology, and map out the spatial distribution patterns of those vegetation types. The forest vegetation was classified into the mountain forest type (Acer okamotoanum community group) and maritime forest type (Artemisia scoparia community group). Vegetation units at the community level were divided into three categories; six communities, sixteen groups, and seven subgroups, giving a total of 22 communities. Total area for mapping was 5,544.9 ha, of which Fagus engleriana community accounted for 1,952 ha (35.2%), Hedera rhambea community ror 1,196 ha (21.6%), Camellia japonica community for 1,104 ha (19.9%), Sambucus sieboldiana var. pendula community for 612 ha (11.0%), Aster spathulifolius community for 506 ha (9.1%), and Tsuga sieboldii - Pinus parviflora community for 174 ha (3.1%). According to canonical correspondence analysis (CCA), variables such as $Mg^{2+}$, pH, $Ca^{2+}$ were positively correlated in maritime vegetation types, and variables such as total nitrogen, carbon content, C/N ratio and ration exchange capacity (CEC) were highly correlated in mountain vegetation types, respectively. The sea level and the slope direction were not showing regular trend as a factor to decide on species diversity, evenness and richness in this research area. But it seems to be affected by topography, slope degree and dominance vegetation.

• Korean Journal of Animal Reproduction
• /
• v.26 no.2
• /
• pp.95-104
• /
• 2002

This study was performed during the four seasons for the production of transgenic pigs containing the human erythropoietin(hEPO) transgene. Purebred Landrace gilts and sows approximately 8∼15 months of age (n=42) were used fur the collection of 1-cell zygotes for DNA microinjection and transfer. Retrospectively, estrus synchronization and superovulation schemes were evaluated to assess practicality for zygote collection. Synchronization and superovulation procedures were used that cyclic gilts were synchronized with 20mg altrenogest (ALT) per day for 9days after PG600 administration followed by superovulation with 1500IU pregnant mares serum gonadotropin (PMSG) and 500IU human chorionic gonadotrophin (hCG). Preparation of recombinant gene for microinjection is mice whey acidic protein promoter (mWAP) linked to human erythropoietin (hEPO) gene. After hormone treatment, 650 embryos were collected from 23 donors and 83.1% (540/650) embryos were in 1-cell stage which can be visualized the pronuclei for DNA microinjection. A total of 543 DNA microinjected embryos fiom donors were transferred to 19 synchronized recipients, seven of them maintained pregnancy and delivered 47 piglets. One of the 47 offsprings were determined to have transgene by PCR analysis. The overall rate of transgenic production was 2.13% (tansgenic/offspring). This study provides the success and useful information regarding production of transgenic pig for bioreactor research.

• Korean journal of applied entomology
• /
• v.31 no.1
• /
• pp.50-68
• /
• 1992

The results of survey on the nematodes associated with sapling from 8 Forestry Experiment Stations(F.E.S.) in Korea are reported herein. A total of 35 species under 20 genera, 11 families, and 2 orders were identified from the soils around the roots of 119 tree species and amongst them, Xiphinema insigne is unrecorded species for Korea and Geocenamus sp. is a new species. Tylenchorhynchus claytoni, which showed the highest detection rate (D.R.) from all F.E.S., except Nambu F.E.S, was the dominant species among the nematodes associated with saplings. The D.R. of T. claytoni was the highest in Ch’ungnam F.E.S. by 100% from 15 saplings sampled, followed by 87.5% in Kangwon, 66.6% in Kyongbuk, 58.3% in Cheonbuk, 48.1% in Ch’ungbuk, 41.3% in Kyongki, and 40% in Cheonnam F.E.S. Other important nematode species which showed very high densities by stations are; Criconemella morgensis (D.R.: 37.5%), Helicotylenchus digonicus (D.R. : 35%), Pararotylenchus pini (D.R.: 37%), Tylenchorhynchus nudus (D.R.: 37.9%), Criconemella informis (D.R.: 27.5%), and Meloidogyne sp. (D.R.: 12.5), from Chunbuk, Chunnam, Ch’ungbuk, Kyongki, and Kangwon F.E.S., respectively.