• Asian-Australasian Journal of Animal Sciences
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• v.8 no.6
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• pp.641-645
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• 1995

This study was conducted to develop a method for production of nuclear transplant bovine embryos using in vitro-matured (IVM) oocytes and to examine the effect of different conditions of electrofusion on fusion rate and developmental capacity of donor nucleus transplanted to enucleated oocytes. Eight- to sixteen-cell embryos derived from oocytes matured and fertilized in vitro used as donor blastomeres and IVM oocytes were used as recipient oocytes. Oocytes were enucleated immediately after 23-24 h IVM and then reconstituted with a donor blastomere in two different micromanipulation media. Fusion rate and subsequent development of the reconstituted oocytes was compared under the different electric stimuli and recipient oocyte ages. Success rate of enucleation was significantly higher in TCM-199 medium containing FCS than in DPBS. The high fusion rate(75-94%) and development (6.4-14.8%) to morulae and blastocyst (M + B) were obtained from 0.6-0.75 kV/cm DC voltage, although total cleavage was not different among the electric pulses. Most optimal condition of electric stimulation for fusion and development was 1 DC voltage of 0.75 kV/cm, in which 80.5% of oocytes were fused, 80.0% and 31.7% of which was cleaved and developed to M + B, respectively. No M + B was obtained from 1.2 kV/cm DC voltage regardless of pulse frequency. Recipint oocyte age at electrofusion greatly affected the cleavage and subsequent development to M + B, showing high rate at 40-41 h oocyte maturation. These results suggest that a suitable condition of electrofusion for donor nuclei derived from IVF may be 1-2 DC pulses of 0.7 kV/cm for $70{\mu}sec$ and that processing of a transplanted nucleus in IVM oocytes may be affected by maturation age of recipient oocytes.

• Polymer(Korea)
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• v.32 no.5
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• pp.415-420
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• 2008

The porcine small intestinal submucosa (SIS) without immunorejection responses and hyalunonic acid (HA) can be used as biomaterials. In this study, we tried to design and characterize novel sponge. SIS- HA sponge was prepared by freeze-drying after addition 1wt% HA solution into fabricated SIS sponge. Sponge was crosslinked with 1-ethyl-(3-3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC) solution with 100mM concentration for 24 hrs and lyophilized. SIS-HA sponge was characterized by scanning electron microscopy and fourier transform infrared spectrometer. And water absorption ability of sponge was evaluated. We seeded NIH/3T3 cells in SIS-HA sponge and cellular attachment was assayed by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltertazolium-bromide (MTT) test. We demonstrated presence of HA in SIS-HA sponge from C-O functional group observed by the FT-IR analysis. Moreover, we confirmed low cytotoxicity and high cell viability of the SIS-HA sponges. Therefore, we could expect that SIS- HA scaffolds are applicable for the tissue regeneration.

• Journal of Korean Neurosurgical Society
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• v.65 no.4
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• pp.549-557
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• 2022

Objective : This study analyzed the risk factors in patients who developed distal junctional kyphosis (DJK) after posterior cervical fusion. Methods : We retrospectively analyzed the clinical and radiographic outcomes of 64 patients, aged ≥18 years (51 and 13 male and female patients, respectively), who underwent single-staged multilevel (3-6 levels) posterior cervical fusion surgery due to multiple cervical spondylotic myelopathy. The surgeries were performed by a single spinal surgeon between January 2012 and December 2017. Demographic data, clinical outcomes, and radiological results were collected. We divided the patients into a DJK group and a non-DJK group according to the presence of DJK and investigated the risk factors by comparing the differences between the two groups. Results : Of the 64 patients, 13 developed DJK. No significant differences in clinical results were observed between the two groups before and immediately after the surgery. At the final follow-up, a higher visual analog score for neck pain was observed in the DJK group compared to the non-DJK group (p<0.01). The DJK group had a significantly lower T1 slope and a significantly higher C2-7 sagittal vertical axis (SVA) before surgery compared to the non-DJK group (p=0.03 and p<0.01, respectively). Immediately after surgery, the difference between the two groups decreased and no significant difference was observed. However, at the last follow-up, a significantly higher C2-7 SVA was observed in the DJK group (p<0.01). At the last follow up, there is no discrepancy in T1S-CL. In multiple logistic regression analysis, preoperative higher C2-7 SVA and preoperative lower T1 slope were identified as independent risk factors (p=0.03 and p<0.01, respectively). As a result, it was confirmed that DJK occurred along the process of returning to preoperative values. Conclusion : DJK can be considered to be caused by cervical misalignment due to excessive change in the surgical site in patients with low T1 slope and high C2-7 SVA before surgery. This also affects the clinical outcome after surgery. It is recommended to refrain from excessive segmental lordosis changes during multilevel cervical post fusion surgery, especially in patients with a small preoperative T1 slope and a large SVA value.

• Applied Biological Chemistry
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• v.32 no.4
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• pp.435-440
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• 1989

To improve alkaline protease producing strain by protoplast fusion, a strain of Bacillus sp. was treated with $100{\mu}g/ml$ NTG (N-methyl-N'-itro-N-nitrosoguanidine) for 45 minutes and mutants of Bacillus subtilts $Arg^-,\;Try^-,\;His^-$ and $Ade^-$ were isolated. The frequency of protoplast formation was about 99%, when teas of exponential phase were treated with $200{\mu}g/ml$ lysozyme at $42^{\circ}C$ for $10{\sim}30$ minutes. In a regeneration medium containing 0.3M sodium succinate, 2.0% polyvinylpyrrolidone, 0.5% casamino acid, 10mM $MgCI_2$ and 20mM $CaCI_2$, regeneration frequency cf the isolated Bacillus subtilis strains was 25.2%. The fusion frequency between mutant was from $2.1{\times}10^{-5}$ to $8.1{\times}10^{-5}$ under optimum condition.

• Journal of Welding and Joining
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• v.30 no.1
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• pp.64-71
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• 2012

The characteristics of high heat input (342kJ/cm) EG (Electro Gas Arc) welded joint of EH36-TM steel has been investigated. The weld metal microstructure consisted of fine acicular ferrite (AF), a little volume of polygonal ferrite (PF) and grain boundary ferrite (GBF). Charpy impact test results of the weld metal and heat affected zone (HAZ) met the requirement of classification rule (Min. 34J at $-20^{\circ}C$). In order to evaluate the relationship between the impact toughness property and the grain size of HAZ, the austenite grain size of HAZ was measured. The prior austenite grain size in Fusion line (F.L+0.1 mm) was about $350{\mu}m$. The grain size in F.L+1.5 mm was measured to be less than $30{\mu}m$ and this region was identified as being included in FGHAZ(Fine Grain HAZ). It is seen that as the austenite grain size decreases, the size of GBF, FSP (Ferrite Side Plate) become smaller and the impact toughness of HAZ increases. Therefore, the CGHAZ was considered to be area up to 1.3mm away from the fusion line. Results of TEM replica analysis for a welded joint implied that very small size ($0.8\sim1.2{\mu}m$) oxygen inclusions played a role of forming fine acicular ferrite in the weld metal. A large amount of (Ti, Mn, Al)xOy oxygen inclusions dispersed, and oxides density was measured to be 4,600-5,300 (ea/mm2). During the welding thermal cycle, the area near a fusion line was reheated to temperature exceeding $1400^{\circ}C$. However, the nitrides and carbides were not completely dissolved near the fusion line because of rapid heating and cooling rate. Instead, they might grow during the cooling process. TiC precipitates of about 50 ~ 100nm size dispersed near the fusion line.

• Microbiology and Biotechnology Letters
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• v.28 no.2
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• pp.80-86
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• 2000

To develop thermotolerant ethanol producing yeast strains, the protoplasts of Saccharomyces carlsbergensis having good fermentability at $30^{\circ}C$ and Kluyveromyces marxianus able to grow at $42^{\circ}C$ were fused. Under the optimal conditions for protoplast formation, the frequency of protoplast formation of S. carlsbergensis was 92 - 94% and that of K. marxianus was 98%. Fusion frequency between S. carlsbergensis and K. marxianus was $1.4\times10^{-6}-4.8$\times10^{-7}$. Among the 27 fusants obtained, 6 fusants were able to grow at $42^{\circ}C$. While the parental strains produced 3.2-3.4%(w/v) ethanol after 3 days from the fermentation medium containing glucose, fusants SK41-4 and SK53-22 produced 5.2%(w/v) ethanol in the same condition. The thermotolerance of SK53-22 was not high, but that of SK41-4 was quite high.

• Journal of Life Science
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• v.29 no.3
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• pp.376-381
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• 2019

To construct useful yeast strain for bioethanol production, we improved yeast harboring various phenotypes by using yeast protoplast fusion method. In this study, S. cerevisiae BYK-F11 strain which have ethanol tolerance, thermotolerance and ${\beta}-glucanase$ activity and P. $stipitis{\Delta}ura$ strain which has xylose metabolism pathway were fused by genome shuffling. P. $stipitis{\Delta}ura$ strain was constructed for protoplast fusion by URA3 gene disruption, resulting in uracil auxotroph. By protoplast fusion, several fused cells were selected and BYKPS-F8 strain (fused cell) showing both karyotypes from two parent strains (S. cerevisiae BYK-F11 and P. $stipitis{\Delta}ura$ strain) among 22 fused cells was finally selected. Sequentially, various phenotypes such as ${\beta}-glucanase$ activity, xylose utility, ethanol tolerance, thermotolerance and ethanol productivity were analyzed. The BYKPS-F8 strain obtained ${\beta}-glucanase$ activity from BYK-F11 strain and 1.2 fold increased xylose utility from P. $stipitis{\Delta}ura$ strain. Also, the BYKPS-F8 strain showed thermotolerance at $40^{\circ}C$ and increased ethanol tolerance in medium containing 8% ethanol. In this fused cell, 7.5 g/l ethanol from 20 g/l xylose was produced and the multiple phenotypes were stably remained during long term cultivation (260 hr). It was proved that novel biological system (yeast strains) is easily and efficiently bred by protoplast fusion among yeasts having different genus.

• Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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• v.13 no.2
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• pp.113-122
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• 2015

⁵⁵Fe and ⁶³Ni are key factors in deciding the proper handling of the decommissioning of radioactive waste from nuclear facilities. For determining beta emitting radionuclides, the dismantled waste samples should be completely decomposed and separated from the sample matrix. This study reports the comparison results of the recovering efficiencies of Iron and Nickel with wet digestion methods that use various acids and alkali-fusion methods. Various matrices of NIST SRMs (1646a, 1944, 8704, 2709a, and 1633c), the recovering efficiencies of using alkali-fusion methods ranged from 95.3 to 98.3% for Iron, and from 86.6 to 88.1% for Nickel within about 2% of relative standard deviation. On the other hand, those using one of the three wet digestion methods ranged from 77.9 to 105.3% for Iron and from 40.1 to 78.5% for Nickel with over 10% of relative standard deviation. Therefore, one may draw the conclusion that the analytical results derived from Iron and Nickel using alkali-fusion methods are fairly reliable due to the recovering efficiencies observed.

• Microbiology and Biotechnology Letters
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• v.14 no.4
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• pp.311-318
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• 1986

Glucoamylase and ethanol productivities of HSDD-170 and HSDM-119 formed by S. cerevisiae and S. diastaticus protoplast fusion were investigated. For the production of the glucoamylase, soluble starch as carbon source, yeast extract and C. S. L as nitrogen source added into the basal medium were favorable. The production of the enzyme reached at maximum after cultivation of the fusant for 4 days at 3$0^{\circ}C$, aerobically. The properties of glucoamylase produced by fusants were very similar to those produced by S. diastaticus as based on optimum temperature, pH stability. In alcohol fermentation from starch, strain HSDD-170 fermented starch faster than either of its parental strains. The maximum of alcohol yield in 15% of liquefied potato starch was 7.5% (v/v).

• Korean Journal of Microbiology
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• v.31 no.4
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• pp.279-285
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• 1993

Alterations of the p53 gene arc among the most frequent genetic changes in human cancer and often result in increased levels of p53 protein within the malignant cells. Detection of accumulated p53 protein can be a useful prognostic tool in human cancer. In order to make polyclonal antibodies for immunohistochemical screening. human p53 gene was expressed in E. coli in the form of GST (glutathione S-transfi.:rase) fusion proteins. Two p53 gene fragments. which were N('()I small fragment encoding amino acid residues of 1-151-: and Ncol large fragment of 159-393. were subeloned into the unique BamHI site present within the pGEX-2T vector using BamHI linker and recombinant plasmids pGTNS and pGTNL were constructed. respectively. The p53 cDNA fragment (from pC53-$SN_3$,) encoding amino acid 38-145 (proline at residue 72) was amplified by polymerase chain reaction(PCR). The amplified DNA was digested with BamHI and Prull and inserted into the BamHI-Smal sites of pG EX-2T and recombinant plasmid pGTBP was constructed. After IPTG induction of these plasmids for 4 hours. fusion proteins were purified from E. coli extracts with glutathione Sepharose beads. The bound proteins were resolved by 10% SDS-polyacrylamide gel electrophoresis and the molecular weights were 54 kDa. 53 kDa and 40 kDa. respectively. Approximately one milligram of fusion proteins were purified from 1 -liter cultures.