• Korean Journal of Clinical Laboratory Science
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• v.39 no.2
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• pp.113-121
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• 2007

Hearing loss is a common congenital disorder that is frequently associated with mutations in the Cx26 gene (GJB2). Recently, the mutation analysis of GJB2 has been used in a newborn screening test for the detection of hearing impairment. Population-based studies should be performed before the application of genetic testing for the identification of deaf newborns. In this study, 8 positions of GJB2 mutations-including 35delG, 167delT, 235delC, V27I, V37I, M34T, E114G, and I203T-were analyzed using PCR-direct sequencing in a total of 437 healthy Korean neonates. DNAs from dried blood spots were extracted using a commercial DNA extraction kit. The PCR-amplified products (783 bps) of the GJB2 gene were detected using 2% agarose gel electrophoresis and subjected to direct sequencing. The sequences were compared with those in the GenBank database by using the BLAST program. In this study, 5 GJB2 mutations -including V27I (79G>A), V37I (109G>A), E114G (341A>G), I203T (608T>C), and 235delC- were found. Of the 437 neonate samples, 301 subjects showed GJB2 mutations (68.9%, 301/437). The V27I mutation was found in 271 subjects and was the most frequent (62.0%, 271/437). The E114G, I203T and V37I mutations were shown in 146, 17 and 14 subjects, respectively. The 235delC mutation was found in 1 subject. The E114G mutation was frequently accompanied by the V27I mutation. V27I/E114G (97.2%, 143/147) was the most common double mutation and 3 subjects had the double mutation V27I/I203T. A triple mutation, V27I/E114G/I203T, was found in 1 subject. In conclusion, PCR-direct sequencing is a convenient tool for the rapid detection of GJB2 mutations and this data might provide information for the genetic counseling of the GJB2 gene.

• Radiation Oncology Journal
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• v.21 no.2
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• pp.149-157
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• 2003

Purpose: The purpose of this study was to determine if there were prognostic differences between BRCA related and BRCA non-related Korean patients with early-onset breast carcinomas. Materials and Methods: Sixty women who had developed breast cancers before the age of 40, and who were treated at the Soonchunhyang University Hospital, were studied independently of their family histories. The age range was 18 to 40 with a median of 34.5 years. Lymphocyte specimens from peripheral blood were studied for the heterozygous mutations of BRCA1 and BRCA2 using direct sequencing methods. Immunohistochemistry was peformed on the paraffin-embedded tissue blocks that were available. Results: Eleven deleterious mutations (18.3%, 6 in BRCA1 and 5 in BRCA2) and 7 missense mutations of unknown significance (11.7%), were found among the 60 patients. More than half of the mutation were novel, and were not reported in the database. Most of the BRCA-associated patients had no history of breast cancer. No treatment related failures were observed in the BRCA carriers, with the exception of one patient that had experienced a new primary tumor of the contralateral breast. The seven year relapse free survival rate were 50 and 79% In the BRCA carrier and BRCA negative patients, respectively. Although the expression of estrogen and progesterone receptors were less common, and histological features more aggressive, in the BRCA associated tumors, the outcome of the patients with BRCA mutations was not poorer than that on the patients without deleterious mutations. Conclusion.: Despite the BRCA mutation carriers having adverse prognostic features, the recurrence rate was relatively lower than that in the BRCA non-carrying Korean patients wi4h early-onset breast carcinomas. In addition, although the prevalence of the BRCA mutation in Korean patients was higher than that in white patients, the penetrance of the cancer seemed to be relatively low in Korean women carrying BRCA mutations. A large population based study of the BRCA mutation, with a long-term follow-up of the study patients will be required to confirm these results.

• Nuclear Medicine and Molecular Imaging
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• v.43 no.5
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• pp.487-494
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• 2009

Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FIT( conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt's lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt's lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.

• Journal of Mushroom
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• v.12 no.1
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• pp.41-51
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• 2014

Cauliflower mushroom widely known high concent of ${\beta}$-glucan for farm cultivation invigoration verified characteristics of mycelia growth, genetic diversity, resistance to Trichoderma by replacement culture with Trichoderma and growth characteristics of new variety crossbleeding strain. The result of replacement culture with Trichoderma for verification resistance about Trichoderma, 6951 (T. viride) strain did not show special change after formation of confrontation line and 6952 (T. spp.) strain was showed more formation of spore after formation of confrontation line. But 6426 (T. harzianum) strain found to encroach part of growth area of cauliflower mushroom mycelia. Among 10 kinds cauliflower mushroom strain, JF02-06 strain collected by Gurye, found did not spore of Trichoderma and thought to be resistant to Trichoderma. The result of crossbleeding after selected that mother strain good growth and formation of fruit body, verified good mycelia growth at JF02-47, 49 and 50 strain in Korean pine of wood-chip media. The result of gene sequence about ITS1, 5.8S and ITS4 for analysis of genetic diversity at crossbleeding strain, found high significance to other cauliflower mushroom in registered Genebank. The result of growth characteristic of spore and mycelia of cauliflower mushroom by observation microscope, size of spore showed water drop shape to major axis $6{\mu}m$ and minor axis $5{\mu}m$ and clamp showed 3 types in mycelia. The wide of mycelia was $3{\mu}m$. The characteristic of mycelia of cauliflower mushroom found to grow mycelia in clamp at approximately 50%. The growth speed of mycelia was $0.507{\mu}m/min$ and 2nd mycelia grown similar speed to mother mycelia at parallel with mother mycelia after growth speed at $0.082{\mu}m/min$. The formation of clamp made small clamp for 5 hours after shown transfer of electrolyte in mycelia inside. The septum formation started after 3 hours and then finally completed after 2 hours. In this study, strain of cauliflower mushroom verified resistance of Trichoderma, genetic diversity and characteristic of mycelia growth. Therefore, basic knowledge of cauliflower mushroom will improve and further contribute to development of mushroom industry.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.295-301
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• 2004

Recently, the rapid increase and global spread of extended-spectrum $\beta-lactamase$ producing clinical isolates has become a serious problem. The incidence of extended-spectrum $\beta-lactamase$ producing clinical isolates of Escherichia coli in Korea and susceptibility to antimicrobial agents were investigated. Total 233 isolates of E. coli were obtained from urine from hospitalized patients in Guro hospital, Korea University in 2001. One hun­dred and eighty four isolates $(78.9\%)$ were resistant to ampicillin, 80 isolates $(34.3\%)$ were resistant to ceph­alothin, 93 isolates $(39.9\%)$ were resistant to gentamicin, and 64 isolates $(27.5\%)$ were resistant to norfloxacin. Among 233 isolates, 17 isolates $(7.3\%)$ were positive as determined by the double disk synergy test. When min­imal inhibitory concentrations were assayed with additional 6 antimicrobial agents, 13 isolates $(76.5\%)$ were multi-drug resistant to at least four different class antimicrobial agents. Extended-spectrum $\beta-lactamase$ were characterized with isoelectric focusing gel electrophoresis and DNA sequencing. They were TEM-1 in 5 iso­lates, TEM-15 in 1 isolate, TEM-20 in 1 isolate, TEM-52 in 4 isolates, TEM-1 and AmpC in 2 isolates, TEM-1 and OXA-30 in 1 isolate, TEM-1 and OXA-33 in 1 isolate, TEM-1, CTX-M-3, and AmpC in 1 isolate, but SHV was not detected. Antimicrobial resistance genes were transferred to animal isolate of E. coli (CCARM No. 1203) by the filter mating method. Extended spectrum $\beta-lactamase$ producers studied in the current study have low correlation to each other as determined by random amplified polymorphic DNA and pulsed field gel elec­trophoresis. This is a contradictory result from the general hypothesis that extended-spectrum $\beta-lactamase$ pro­ducers in one hospital is a result from a clonal spread.

• Korean Journal of Microbiology
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• v.39 no.4
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• pp.260-266
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• 2003

In this study bacterial community was analyzed to evaluate the impacts of long-term use of agricultural chemicals on a soil ecosystem as well as to obtain fundamental data on the relationship. Sequences of 16S rRNA clones from a non-agricultural site and a tangerine orchard soil which has a history of long-term use of agricultural chemicals over 30 years were analyzed. This revealed that bacterial community containing 5 divisions and 18 genera was distributed in a tangerine orchard soil, while bacterial community containing 9 divisions and 44 genera was distributed. In a tangerine orchard soil site, the most abundant bacteria in subdivision level were placed into Proteobacteria γ group which occupied 56% of total clones. The other bacterial clones from the ocrhcard soil exposed to agricultural chemicals over 30 years were Acidobacteria group (25%), Fimicutes group (5%), Planctomycetes group (2%), Proteobacteria α (1%), δ group (1%), and Cyanobacteria group (1%). Whereas, the clones were from the non-agricultural site were distributed among the division or subdivision Acidobacteria group (14%), Planctomycetes group (13%), Proteobacteria α (10%), β (9%), δ (9%), Fimicutes group (8%), Verrucomicrobia group (8%), Actinobacteria group (6%), Proteobacteria γ group (3%), Bacteroidetes group (3%), Gemmatimonadetes group (3%), and Cyanobacteria group (1%). This finding suggests the possibility that long-term application of agricultural chemicals or fertilizers on a tangerine orchard might result in drastic reduction or alteration in the composition of the bacterial community in the contaminated soil site.

• Korean Journal of Microbiology
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• v.43 no.3
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• pp.173-178
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• 2007

The occurrence of acute respiratory infections caused by the influenza virus are particularly high during the winter season in Busan, Korea. In 2004 and 2005, a study of the rate of occurrences of the influenza virus was conducted. The results reveal that in 2004, of the 1,869 people with an acute respiratory infection that 154 (8.2%) people were infected by the influenza virus. In 2005, of the 1,579 people infected with an acute respiratory infection that 19 people (1.2%) were infected with the influenza virus. The study shows a decrease in the numbers of an influenza virus infection from 2004 to 2005. Data was collected by inspecting throat swabs and nasal discharge from those with an acute respiratory infection. Further inspection of the throat swab and nasal discharge from the infected individuals during 2004 and 2005 study show the occurrence of the different types of influenza virus in the population: 6 cases (3.5%) of influenza type A/H1N1, 129 cases (74.5%) of A/H3N2, and 38 cases (22.0%) of type B. The study conducted in 2004 and 2005 reveal that children between the ages of two and five were more likely to be infected than any other age group. In the study, about 62.2% of the infected individuals were between two and five years old. The detection rates between males and females are similar. However, it is notable that females are slightly more likely to develop an acute respiratory infection caused by the influence virus compared to their male counterparts.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.178-185
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• 2008

Avian influenza virus (AIV) is recognized as key to the emergence of pandemic influenza for humans; there are growing concerns that AIV H9N2 may become more efficient to transmit to humans in the near future, since the infection of poultry with AIV H9N2 has been common in recent years. In this study, we aimed to produce antisera recognizing the HA and NA proteins of AIV H9N2. Initially, coding sequences corresponding to the N-terminal regions of the HA and NA proteins of the Korean AIV H9N2 (A/Ck/Kr/MS96/96) isolated from a domestic chicken were amplified from the genomic RNA. Following cloning of the amplified cDNA fragments into pGEX4T-1 vector, two GST-fusion proteins (GST-HAln and GST-NAn) were expressed in E. coli BL21 and purified with glutathione sepharose columns; the recombinant GST-HAln and GST-NAn proteins were both used as immunogens in rabbits. The antigenicity of the rabbit antisera was analyzed by immunoblotting of the cell lysates prepared from AIV H9N2-infected MDCK cells. Overall, the recombinant HAln and NAn proteins fused to the C-terminus of GST and the rabbit antisera raised against the corresponding recombinant proteins would provide a valuable reagent for AIV diagnosis and basic research.

• Development and Reproduction
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• v.11 no.3
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• pp.263-272
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• 2007

Contrast to mouse where its in vitro maturation rates are high without specific supplements or presence of the cumulus cells, there are some species, such as porcine, where its in vitro oocyte maturation rates are still very low. This comparative study was conducted to investigate the role of malate dehydrogenase(Mor2) during oocyte maturation by RNAi in the mouse and porcine. The Mor2 double-stranded RNA(dsRNA) was prepared speciesspecifically and microinjected into the cytoplasm of denuded germinal vesicle(GV) oocytes. Oocytes were cultured for 48 h(porcine) and 16 h(mouse) in M199 with 10% porcine follicular fluid, pyruvate, p-FSH, EGF, cystein, and estradiol-$17{\beta}$. We measured changes in oocyte morphology, maturation rates and mRNA levels after Mor2 RNAi. We confirmed gene sequence-specific knock down of Mor2 mRNA in both species after Mor2 RNAi. In contrast to our previous finding that mMor2 RNAi resulted in GV arrest in the mouse, we found that pMor2 RNAi resulted in MI arrest in denuded porcine oocytes(58%), but developed to MII(84.4%) in COCs. To determine whether this difference between mouse and porcine RNAi is due to differences in culture media, we cultured mouse oocytes in the M199 media for 16 h after mMor2 RNAi. Mouse oocytes were developed to MII stage(62%) and there was no statistical difference compared to that of non-injected(76.8%) and buffer-injected(73.3%) control groups. Therefore, we concluded that the mouse and porcine oocytes are having different metabolic systems in relation to malate dehydrogenase for oocyte maturation. This could be a basis for differences in maturation rates in vitro in two species. Further scrutinized studies on the metabolic pathways would led us in finding better culture system to improve oocyte maturation rates in vitro, especially in more challenging species like the porcine.

• Korean Journal of Ecology and Environment
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• v.50 no.1
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• pp.137-147
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• 2017

The identity of toxin producers remains only hypothesis unless there were identified by strain isolation and analytical confirmation of both the cyanotoxin production and the genetic identity of the monoculture. The purposes of this study were to identify a morphologic and phylogenetic classification in Aphanizomenon flos-aquae strains isolated from the Nakdong River and to investigate the potential ability of the strains to produce toxins such as saxitoxin and cylindrospermopsin using target genes. The 16S rRNA and sxtA, sxtI, cyrA, cyrJ genes were analyzed on two strains (DGUC001, DGUC003) isolated from the Nakdong River. Morphological features of the strains were observed a shape of aggregated trichomes in parallel fascicles which can reach up to macroscopic size and a hyaline terminal cell without aerotope. In addition, the 16S rRNA phylogenetic analyses showed that the strains were identified as the same species with high genetic similarity of 98.4% and grouped within a monospecific andsupported cluster I of Aphanizomenon flos-aquae selected from GenBank of the NCBI. The cyrA and cyrJ genes encoding for the cylindrospermopsin-biosynthesis were not detected in the present study. The sxtA gene was in detected both the two strains, whereas the sxtI gene which had been suggested as a suitable molecular marker to detect saxitoxin-producing cyanobacteria was not found both the strains. Thus, the two strains isolated from Nakdong River were identified as the same species of Aphanizomenon flos-aquae Ralfs ex Bornet et Flahault 1888, the two strains were confirmed as potential non-producing strains of the saxitoxin and cylindrospermopsin.