• Journal of Microbiology and Biotechnology
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• v.8 no.6
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• pp.650-655
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• 1998

The chromosomal DNA fragment from Phaffia rhodozyma CBS 6938 which is able to autonomously replicate in the yeast Saccharomyces cerevisiae was cloned on an integrative URA3 plasmid. Its minimal fragment exhibiting autonomously replicating activiy in the S. cerevisiae gave a higher frequency transformation efficiency than that found for centromere-based plasmid, and enabled extrachromosoma1ly stable transmission of the plasmids in one copy per yeast cell under non-selective culture condition. The 836-bp DNA element lacked an ORF and did not contain any acceptable match to an ARS core consensus. Sequence analysis, however, displayed a cluster of three hairpin-Ioop-sequences with individual $\triangle {G_{25}}^{\circ}C$ free energy value of -10.0, -17.5, and -17.0 kcal. $mor^{-l}$as well as a 9-bp sequence with two base pair mismatches to the S. cerevisiae/E. coli gyrase-binding site. This 836-bp sequence also included one 7-bp sequence analogous to the core consensus of centromeric DNA element III (CDEIII) of S. cerevisiae, but CDEIII-like 7 bp sequence alone did not give a replicative function in this yeast.

• Applied Biological Chemistry
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• v.36 no.6
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• pp.562-566
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• 1993

The conditions required for plant transformation through the electroporation system and for the electrofusion of the prtoplasts were investigated for geranium (Pelargonium zonale hybrids). The optimum condition for electroporation was 1.77 kV/cm for $40\;{\mu}sec$ under which 70% of the protoplasts were viable and 58% of the viable protoplasts were stained with methylene blue. The pBin19 DNA plasmid used as a carrier vector was isolated from E.coli $DH5{\alpha}$ strain, purified, identified by the electrophoresis on agarose gel and electroporated into the protoplasts. The KM8 liquid medium gave better cell division than any other media. One MHz of AC frequency with 40 V/cm of amplitude for 15 sec followed by 0.5 kV/cm of DC amplitude for $60\;{\mu}sec$ was most efficient for the electrofusion of protoplasts.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.12
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• pp.7523-7527
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• 2013

Thymidylate synthase (TS) catalyzes the transfer of a methyl group from methylenetetrahydrofolate to dUMP to form dTMP. It is a primary target in the chemotherapy of colorectal cancers and some other neoplasms. In order to obtain pure protein for analysis of structure and biological function, an expression vector TS-pET28b (+) was constructed by inserting wild-type human thymidylate synthase (hTS) cDNA into pET28b (+). Then an expression strain was selected after transformation of the recombined plasmid into Rosetta (DE3). Fusion protein with His-tag was efficiently expressed in the form of inclusion bodies after IPTG induction and the content was approximately 40.0% of total bacteria proteins after optimizing expression conditions. When inclusion bodies were washed, dissolved and purified by Ni-NTA under denatured conditions, the purity was up to 90%. On SDS-PAGE and West-blotting, the protein band was found to match well with the predicted relative molecular mass-36kDa. Bioactivity was 0.1 U/mg. The results indicated that high-level expression of wild-type hTS cDNA can be achieved in prokaryotes with our novel method, facilitating research into related chemotherapy.

• The Journal of Korean Oriental Chronic Disease
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• v.10 no.1
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• pp.21-31
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• 2005

We have analyzed 247 patients who were admitted to the 3rd ICU for intensive care between January 2003 and December 2003 to investigate characteristics of ICU patients and obtain better guidance and management in the ICU. The results are as follows: 1) The total number of patients was 247. The proportion of males was 59.1 %, of females was 40.9%, and the ratio of male to female was about 1.45:1. 2) The most prevalent age group is those in their seventies, the proportion is 33.6%. 3) The total admission days was 3,438days, and mean patient stay being 13.9days. 4) The total number of expired patients was 26, the mortality rate was 10.5%. The mortality in distribution by disease was higest in Cerebral infarction, rate was 53.8%. the proportion of raised intracranial pressure was 42.3%, of hemorrhagic transformation was 11.5%. 5) In distribution by admission channel, Oriental hospital E.R. was 60.7%, Oriental hospital ward was 38.5%, and etc. was 0.8%. 6) The number of patients of Cerebral infarction was 146(59.1%), being the highest in distribution by chief disease and the proportion of MCA territory infarction is highest in the Cerebral infarction(61.6%). 7) In distribution by Oriental Medicine therapy, the proportion of Acupuncture treatment was 15.8%, of Acupuncture treatment combination Herb medication was 78.5%, and of Acupuncture, Herb medication plus subcutaneous Acupuncture treatment was 5.7%.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.7
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• pp.2883-2887
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• 2015

Background: The calcium-binding S100A4 protein is involved in epithelial to mesenchymal transition, oncogenic transformation, angiogenesis, cytoskeletal integrity, mobility and metastasis of cancer cells. This study aimed to clarify the roles of S100A4 in genesis and progression of glioma. Materials and Methods: S100A4 expression was examined by real-time RT-CPR and Western blot in glioma and paired normal brain tissue (n=69), and compared with clinicopathological parameters of tumors. In addition, glioma U251 cells transfected with an S100A4-expressing plasmid were examined for proliferation by MTT, apoptosis by Annexin V-FITC, and migration and invasion with Transwell chambers. Results: Increased S100A4 mRNA expression was found in gliomas, compared with paired non-tumor tissue (p<0.001). Gradual elevation of overexpression of S100A4 was observed with increasing glioma grade (p<0.001). Astrocytoma showed lower S100A4 mRNA expression than oligodendrogliomas, with glioblastomas having highest values (p<0.001). Similar results were obtained for S100A4 protein, a positive link being found between mRNA and protein expression in gliomas (p<0.001). There was higher growth, lower apoptosis, stronger migration and invasion of S100A4 transfectants than control and mock transfected cells (p<0.001). Conclusions: These findings indicate that up-regulated S100A4 expression is positively linked to pathogenesis, progression and histogenesis of glioma by modulating proliferation, apoptosis, migration and invasion.

• Journal of the Korean Ceramic Society
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• v.40 no.6
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• pp.566-571
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• 2003

Hydroxyapatite (HAp)/Polyacrylic Acid(PAA) homogeneous composites of four different composition ratio were preparation by co-precipitation process with synthetic HAp and PAA as a binder. HAP/PAA composites were molding by cold isostatic pressing and were sintering by various condition in air. Crystallinity and structure of sintered HAp/PAA composites were investigated by XRD and FT-IR. Also, the compressive strength and the fracture surface of sintered specimens were measured by UTM and SEM. HAp/PAA composites were showed phase transformation of partially ${\alpha}$, ${\beta}$-tricalcium phosphate at sintering condition of 1200$^{\circ}C$ and 3 h. The pore size and porosity of sintered body were showed the range of 0.2∼3.0 $\mu\textrm{m}$ and 0.49∼13.43%, respectively. The compressive strength of sintered specimens were appeared the range of 36.6∼58.2 MPa. From these results, the sintered HAp/PAA comosites can be accounted for the microporous HAp having a good compressive strength due to homogeneous pore morphology.

• Journal of the Korean Ceramic Society
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• v.25 no.3
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• pp.225-230
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• 1988

The strength and thermal shock resistance of $Al_2O_3-ZrO_2$ composites have been studied. The tetragonal $ZrO_2$ powder containing 1 mol.% $Y_2O_3$ and monoclinic $ZrO_2$ powder were prepared by coprecipitation method and subsequently mixed with $Al_2O_3$ powder and granulated by sieving. Duplex composites were prepared by dry mixing matrix agglomerate with 15 to 30 vol.% of dispersion agglomerate, followed by pressing and sintering at 1$600^{\circ}C$ for1 hr. These $Al_2O_3-ZrO_2$ 2 composites having heterogeneous structure showed improved thermal shock behaviors because of the microcracking and pores in dispersed granules, and compressive stresses around dispersed granules resulting from $ZrO_2$ transformation.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1491-1499
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• 2010

Rhizosphere microorganisms possessing phytase activity are considered important for rendering phytate-phosphorus (P) available to plants. In the present study, the Citrobacter braakii phytase gene (appA) was overexpressed in rhizobacteria possessing plant growth promoting (PGP) traits, for increasing their potential as bioinoculants. AppA was cloned under the lac promoter in the broadhost-range expression vector pBBR1MCS-2. Transformation of the recombinant construct pCBappA resulted in high constitutive phytase activity in all of the eight rhizobacterial strains belonging to genera Pantoea, Citrobacter, Enterobacter, Pseudomonas (two strains), Rhizobium (two strains), and Ensifer that were studied. Transgenic rhizobacterial strains were found to display varying levels of phytase activity, ranging from 10-folds to 538-folds higher than the corresponding control strains. The transgenic derivative of Pseudomonas fluorescens CHA0, a well-characterized plant growth promoting rhizobacterium, showed the highest expression of phytase (~8 U/mg) activity in crude extracts. Although all transformants showed high phytase activity, rhizobacteria having the ability to secrete organic acid showed significantly higher release of P from Ca-phytate in buffered minimal media. AppA overexpressing rhizobacteria showed increased P content, and dry weight (shoot) or shoot/ root ratio of mung bean (Vigna radiata) plants, to different extents, when grown in semisolid agar (SSA) medium containing Na-phytate or Ca-phytate as the P sources. This is the first report of the overexpression of phytase in rhizobacterial strains and its exploitation for plant growth enhancement.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.1
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• pp.43-51
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• 2002

The cloning and expression of $\beta-glucosidase$ II, encoded by the gene ${\beta}glu2$, from thermotolerant yeast Pichia etchellsii into Escherichia coli is described. Cloning of the 7.3 kb BamHI/SalI yeast insert containing ${\beta}glu2$ in pUC18, which allowed for reverse orientation of the insert, resulted in better enzyme expression. Transformation of this plasmid into E. coli JM109 resulted in accumulation of the enzyme in periplasmic space. At $50^{\circ}C$, the highest hydrolytic activity of 1686 IU/g protein was obtained on sophorose. Batch and fed-batch techniques were employed for enzyme production in a 14 L bioreactor. Exponential feeding rates were determined from mass balance equations and these were employed to control specific growth rate and in turn maximize cell growth and enzyme production. Media optimization coupled with this strategy resulted in increased enzyme units of 1.2 kU/L at a stabilized growth rate of $0.14\;h^{-l}$. Increased enzyme production in bioreactor was accompanied by formation of inclusion bodies.

• The Korean Journal of Zoology
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• v.34 no.2
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• pp.217-227
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• 1991

Ultrastmctura changes of multivesicular bodies and mitochondria of oocytes of PseudopotamU- Ia occelata Moore were examined with transmission electron microscope in order to follow the process of yolk formation. Yolk granules begin to form at the previtellogenic stages of 50 $\mu$m diameter from multivesicular bodies. Small vesicles and membranous structures within the multivesicular bodies are fused to form the precursors of core bodies of yolk granules. Some vesicles from cytoplasm are also coalesced into the multivesicular bodies. Mature yolk granules are composed of electron-dense core bodies which are seperated from each other by electronopaque small vesicle-like structures. Structural changes of cristae into vesicular shapes and increase in electron density of matrix in mitochondria strongly suggested that mitochondria are in the process of transformation. The transformed mitochondria appear to be basic structures which later become multivesicular bodies.