• Journal of Chest Surgery
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• v.28 no.2
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• pp.125-130
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• 1995

Autotransfusion system is a common method of reducing the need of intraoperative and postoperative homologous blood transfusion in cardiac operation. Between August 1991 and August 1993, a series of 51 adults undergoing open heart surgery was selected. Autotransfusion using Cell Saver [COBE Baylor Rapid Autologous Transfusion System was done with homologous blood transfusion in 15 cases [Group II or without homologous blood transfusion in 17 cases [Group III . The other 19 cases were taken without Cell Saver for control [Group I . The shed blood in the operative field, remained blood in the oxygenator after cardiopulmonary bypass, and blood drained from chest tubes in postoperative care were aspirated by means of a locally heparinized collection system. After the salvaged blood was washed and centrifuged, the processed blood subsequently reinfused. Composition of processed blood by Cell Saver was hemoglobin 16.9gm%, hematocrit 49%, RBC 5,140,000/ml, WBC 670/ml, and platelet 30,000/ml. In three group, hemoglobin, hematocrit, and platelet counts were decreased postoperatively, but no significant differences between three group. Postoperatively, the amounts of drainage from chest tubes was 543$\pm$121ml in Group I, 809$\pm$201ml in Group II, and 631$\pm$147ml in Group III. In Group II, there was large amount of drainage compared with Group I [p<0.05 . The amount of homologous blood transfused was 1116$\pm$219 ml in Group I, 791$\pm$183 ml in Group II [p<0.05 . The homologous blood was not transfused in 17 cases [53% with Cell Saver.Preoperative and postoperative, coagulation parameters showed no significant differences between three group. And there was no complication related to Cell Saver. We conclude that the autotransfusion using Cell Saver is effective for reducing the homologous blood transfusion in cardiac surgery.

• Journal of Crop Science and Biotechnology
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• v.10 no.4
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• pp.231-236
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• 2007

Differentially expressed genes(DEG) were identified in a rice variety, Sathi, an indica type showing high allelopathic potential against barnyardgrass(Echinochloa crus-galli(L.) Beauv. var. frumentaceae). Rice plants were grown with and without barnyardgrass and total RNA was extracted from rice leaves at 45 days after seeding. DEG full-screening was performed by $GeneFishing^{TM}$ method. The differentially expressed bands were re-amplified and sequenced, then analyzed by Basic Local Alignment Search Tool(BLAST) searching for homology sequence identification. Gel electrophoresis showed nine possible genes associated with allelopathic potential in Sathi, six genes(namely DEG-1, 4, 5, 7, 8, and 9) showed higher expression, and three genes(DEG-2, 3 and 6) showed lower expression as compared to the control. cDNA sequence analysis showed that DEG-7 and DEG-9 had the same sequence. From RT PCR results, DEG-6 and DEG-7 were considered as true DEG, whereas DEG-1, 2, 3, 4, 5, and 8 were considered as putative DEG. Results from blast-n and blast-x search suggested that DEG-1 is homologous to a gene for S-adenosylmethionine synthetase, DEG-2 is homologous to a chloroplast gene for ribulose 1,5-bisphosphate carboxylase large subunit, DEG-8 is homologous to oxysterol-binding protein with an 85.7% sequence similarity, DEG-5 is homologous to histone 2B protein with a 47.9% sequence similarity, DEG-6 is homologous to nicotineamine aminotransferase with a 33.1% sequence similarity, DEG-3 has 98.8% similarity with nucleotides sequence that has 33.1% similarity with oxygen evolving complex protein in photosystem II, DEG-7 is homologous to nucleotides sequence that may relate with putative serin/threonine protein kinase and putative transposable element, and DEG-4 has 98.8% similarity with nucleotides sequence for an unknown protein.

• Korean Journal of Microbiology
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• v.41 no.4
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• pp.239-245
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• 2005

Transformation-Associated Recombination (TAR) cloning technique allows selective isolation of chromosomal regions and genes from complex genomes. The procedure requires knowledge of relatively small genomic sequences that reside adjacent to the chromosomal region of interest. This technique involves homologous recombination during yeast spheroplast transformation between genomic DNA and a TAR vector that has 5' and 3' gene targeting sequences. In this study, we examined the effect of non-homologous spacing sequence in target hooks on homologous recombination using a plasmid model system. The efficiency of homologous recombination between the modified his3-TRP1-his3 fragments and HlS3 gene on plasmid were analyzed by the characterization of $Ura^+$ transformants. The numbers of $Ura^+$ transformant showed same level when seven different modified his3-TRP1-his3 fragments were used. But the percentage of positive recombinants. $Trp^+His^-$, dramatically decreased when used the modified his3-TRP1-his3 fragments contained incorrect spacing of nonhomologous region. As a result, we suggest that incorrect spacing inhibits the homologous recombination between target hook and substrate DNA. Therefore, we should consider the correct spacing in target hook when the target hook are used for cloning of orthologue gene.

• Korean Journal of Veterinary Research
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• v.29 no.3
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• pp.283-289
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• 1989

In order to measure in vitro cell mediated immunity in the guinea pigs sensitized with the killed bacilli of Mycobacterium bovis ($AN_5$), M avium (serotype 2), M tuberculosis and M intracellulare (serotype 8), leukocyte adherence inhibition (LAI) test was established using the antigens of purified protein derivatives (PPD) tuberculin. By using LAI test, specificity of cell-mediated immune responses of the guinea pigs inoculated with various Mycobacterium spp was investigated, and comparison between values of LAI and skin test was also made to evaluate the specificity of the newly designed test. The results obtained throughout the experiments were summarized as follows; 1. The optimal concentration of PPD antigens for LAI test was 1 to 2mg per ml of medium. 2. When the leukocytes of guinea pigs sensitized with both M bovis($AN_5$) and M avium (serotype 2) for 2 to 8 weeks were incubated with homologous or heterologous PPD antigens, mean values of LAI test were $61.2{\pm}11.2$ and $65.6{\pm}5.1%$ in homologous PPD antigens respectively, while $30.0{\pm}3.7$ and $32.8{\pm}5.7%$ in heteNlogous PPD antigens, showing the prominently high value of LAI in the homologous syst,em (p<0.01). 3. When the leukocytes of guinea pigs sensitized with both M tuberculosis and M intracellulare (serotype 8) for 2 to 8 weeks were incubated with homologous and heterologous PPD antigens, mean values of LAI test were $67.9{\pm}2.9$ and $66.9{\pm}5.0%$ in homologous PPD antigens, while $27.4{\pm}7.4$ and $24.4{\pm}7.1%$ in heterologous PPD antigens, showing the prominently high value of LAI in the homologous system (p<0.01). 4. Comparing with the specificity of LAI and skin tests on the basis of the value obtained from the homologous system, deviation of reaction was revealed to be 49.5 to 100.2 in LAI test, and -15.9 to 52.0 in skin test.

• Reproductive and Developmental Biology
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• v.41 no.1
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• pp.7-16
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• 2017

The knock-in efficiency in the fibroblast is very important to produce transgenic domestic animal using nuclear transfer. In this research, we constructed three kinds of different knock-in vectors to study the efficiency of knock-in depending on structure of knock-in vector with different size of homologous arm on the ${\beta}-casein$ gene locus in the somatic cells; DT-A_cEndo Knock-in vector, DT-A_tEndo Knock-in vector I, and DT-A_tEndo Knock-in vector II. The knock-in vector consists of 4.8 kb or 1.06 kb of 5' arm region and 1.8 kb or 0.64 kb of 3' arm region, and neomycin resistance gene(neor) as a positive selection marker gene. The cEndo Knock-in vector had 4.8 kb and 1.8 kb homologous arm. The tEndo Knock-in vector I had 1.06 kb and 0.64 kb homologous arm and tEndo Knock-in vector II had 1.06 kb and 1.8 kb homologous arm. To express endostatin gene as transgene, the F2A sequence was fused to the 5' terminal of endostatin gene and inserted into exon 7 of the ${\beta}-casein$ gene. The knock-in vector and TALEN were introduced into the bovine fibroblast by electroporation. The knock-in efficiencies of cEndo, tEndo I, and tEndo II vector were 4.6%, 2.2% and 4.8%, respectively. These results indicated that size of 3' arm in the knock-in vector is important for TALEN-mediated homologous recombination in the fibroblast. In conclusion, our knock-in system may help to create transgenic dairy cattle expressing human endostatin protein via the endogenous expression system of the bovine ${\beta}-casein$ gene in the mammary gland.

• 한국균학회소식:학술대회논문집
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• 2006.10a
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• pp.32-44
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• 2006

To investigate non-aflatoxin-production of A. oryzae at the molecular level, an aflatoxin biosynthesis gene homolog cluster of RIB 40 was analyzed. Although most genes in the corresponding cluster exhibited from 97 to 99 % similarity to those of Aspergillus flavus, three genes shared 93 % similarity or less. In addition, although slight expression of aflR, positive transcriptional regulator gene, was detected in some A. oryzae strains having seven aflatoxin biosynthesis homologous genes, other genes related to aflatoxin production were not detected. RIB strains were mainly divided into group 1, having seven aflatoxin biosynthesis homologous genes (aflT, nor-i, aflR, norA, avnA, verB, and vbs), and group 2, having three homologous (avnA, verB, and vbs). Partial aflatoxin homologous gene cluster of RIB62 from group 2 was sequenced and compared with that of RIB40 from group 1. RIB62 showed a large deletion upstream of ver-1 with more than half of the aflatoxin homologous gene cluster missing including aflR, a positive transcriptional regulatory gene. Adjacent to the deletion of the aflatoxin homologous gene cluster, RIB62 has a unique sequence of about 8kb and a telomere. Southern analysis of A. oryzae RIB strains with four kinds of probe derived from the unique sequence of RIB62 showed that all group 2 strains have identical hybridizing signals. Polymerase chain reaction with specific primer set designed to amplify the junction between ver-1 and the unique sequence of RIB62 resulted in the same size of DNA fragment only from group 2 strains. Based on these results, we developed a useful genetic tool that distinguishes A. oryzae group 2 strains from the other groups' strains and propose that it might have differentiated from the ancestral strains due to chromosomal breakage.

• Genomics & Informatics
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• v.12 no.3
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• pp.80-86
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• 2014

Foldback intercoil (FBI) DNA is formed by the folding back at one point of a non-helical parallel track of double-stranded DNA at as sharp as $180^{\circ}$ and the intertwining of two double helixes within each other's major groove to form an intercoil with a diameter of 2.2 nm. FBI DNA has been suggested to mediate intra-molecular homologous recombination of a deletion and inversion. Inter-molecular homologous recombination, known as site-specific insertion, on the other hand, is mediated by the direct perpendicular approach of the FBI DNA tip, as the attP site, onto the target DNA, as the attB site. Transposition of DNA transposons involves the pairing of terminal inverted repeats and 5-7-bp tandem target duplication. FBI DNA configuration effectively explains simple as well as replicative transposition, along with the involvement of an enhancer element. The majority of diverse retrotransposable elements that employ a target site duplication mechanism is also suggested to follow the FBI DNA-mediated perpendicular insertion of the paired intercoil ends by non-homologous end-joining, together with gap filling. A genome-wide perspective of transposable elements in light of FBI DNA is discussed.

• Environmental Mutagens and Carcinogens
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• v.17 no.1
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• pp.1-6
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• 1997

The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. The insert DNA of the RAD4 homolog was contained 3.2 kb. Here, we report the partial cloning and characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the sequence homologous DNA to RAD4 gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 1.2 kb PvuII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. The level of the transcript did not increase upon UV-irradiation, suggesting that the RAD4 homologous gene in C. cinereus is not UV-inducible.

• Journal of Chest Surgery
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• v.26 no.2
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• pp.108-114
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• 1993

The risks of homologous blood transfusion are well documented and recently increased with the emergence of acquired immunodeficiency syndrome. This is a report concerning 29 consecutive adult with no blood conservation patients (Group I) who had an elective cardiac operations. A similar group of 29 patients operated on 1992 but with blood conservation (Group II). In I of Group I patients and 15 of Group II patients, no homologous blood products were required. Group II patients used significantly less fresh frozen plasma (2.05${\pm}$0.68 unit versus 6.52${\pm}$0.72 unit, p<0.05) and the homologous blood transfusion (0.42${\pm}$0.9 unit versus 3.64${\pm}$0.17 unit, p<0.05) than Group I patients. Group II patients had also significantly less postoperative bleeding (338${\pm}$39.9 ml versus 585${\pm}$93.0 ml, p<0.05) than Group I patients. Group II patients recieved 460${\pm}$62.6 ml of mediastinal shed blood in acquired group. In conclusion, a simple and inexpensive blood conservation program, mainly combining autologous blood removal before bypass, retransfusion of the volume remaining in the oxygenator, and consistent autotransfusion of mediastinal shed blood has enabled us to avoid infusion of homologous blood in 15/29 patients of Group II patients. No side effects or complicatinos could be related to the blood conservation program.

• Bulletin of the Korean Chemical Society
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• v.18 no.10
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• pp.1085-1089
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• 1997

A new HPLC chiral stationary phase (CSP 3) has been prepared by connecting N-phenyl N-propyl amide of (S)-naproxen to silica gel through the 6-methoxy-2-naphthyl group of (S)-naproxen. The new CSP has been applied in resolving a homologous series of N-(3,5-dinitrobenzoyl)-α-amino acid esters and a homologous series of N-(3,5-dinitrobenzoyl)-α-(4-alkylphenyl)alkylamines. The separation factors, α, for resolving a homologous series of N-(3,5-dinitrobenzoyl)-α-amino esters and a homologous series of N-(3,5-dinitrobenzoyl)-α-(4-alkylphenyl)alkylamines on the new CSP have been found to remain almost constant throughout the wide range of the length of the alkyl substituent of the analytes while those on the previously reported CSPs (CSP 1 and 2) which were prepared by connecting N-phenyl N-propyl amide of (S)-naproxen to silica gel through the N-propyl group increase or decrease continuously. These results are concluded to support the chiral recognition models which utilize the intercalation of the alkyl substituent of the racemic analytes between the adjacent strands of CSP 1 or 2 to rationalize the increasing or decreasing trends of separation factors.