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Morphological and Genetic Stock Identification of Todarodes pacificus in Korean Waters (한국 주변해역에 서식하는 살오징어(Todarodes pacificus)의 형태 및 유전학적 계군분석)

  • Kim, Jeong-Yun;Yoon, Moon-Geun;Moon, Chang-Ho;Kang, Chang-Keun;Choi, Kwang Ho;Lee, Chung Il
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.18 no.3
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    • pp.131-141
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    • 2013
  • Stock identification of Todarodes pacificus collected in the East Sea, Yellow Sea and East China Sea during the period from September to December in 2011 was analyzed by morphometric characters and mitochondrial DNA (mtDNA) cytochrome oxidase subunit I (COI) gene nucleotide variations. Frequency distributions of mantle length was analyzed by morphological method with measuring size of T. pacificus. Then each stock was estimated to confirm their maturation for mean mantle length comparing with mean mature mantle length 20-22 cm. According to morphologic stock identification, it is estimated that the northern part of East Sea is categorized as summer stock and the rest parts, including mid /southern part of the East Sea, northern part of the East China Sea and northern part of the West Sea were autumn stock. For genetic analysis, a total 49 haplotypes were defined by 33 variable nucleotide sites. From the extensive haplotype diversity, limited nucleotide diversity and star-like shape of haplotype network, T. pacificus appears to have undergone rapid population expansion from an ancestral population with a small effective population size. Although pair-wise Fst estimates which represent genetic difference among groups were low, there are relatively remarkable difference of Fst between middle and southern part of the East Sea. Although middle part of the East Sea and southern part of the East Sea were situated at the East Sea, genetically separated groups were appeared.

Clinical Features, Response to Treatment, Prognosis, and Molecular Characterization in Korean Patients with Inherited Urea Cycle Defects

  • Yoo, Han-Wook;Kim, Gu-Hwan;Seo, Eul-Ju
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.2 no.1
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    • pp.77-79
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    • 2002
  • The urea cycle, consisting of a series of six enzymatic reactions, plays key roles to prevent the accumulation of toxic nitrogenous compound and synthesize arginine de novo. Five well characterized diseases have been described, resulting from an enzymatic defect in the biosynthesis of one of the normally expressed enzyme. This presentation will focus on two representative diseases; ornithine transcarbamylase(OTC) deficiency and citrullinemia(argininosuccinate synthetase deficiency). OTC deficiency is one of the most common inborn error of urea cycle, which is inherited in X-linked manner. We identified 17 different mutations in 20 unrelated Korean patients with OTC deficiency; L9X, R26P, R26X, T44I, R92X, G100R, R141Q, G195R, M205T, H214Y, D249G, R277W, F281S, 853 del C, R320X, V323M and 10 bp del at nt. 796-805. These mutations occur at well conserved nucleotide sequences across species or CpG hot spot. The L9X and R26X lead to the disruption of leader sequences, required for directing mitochondrial localization of the OTC precursor. Their phenotypes are severe, and neonatal onset. The G100R, R277W and V323M mutations were uniquely identified in patients with late onset OTC deficiency. The other genotypes are associated with neonatal onset. Out of 20 patients with OTC deficiency, only 6 patients are alive; two were liver transplanted, and normal in growth and development at 2, 4 years after transplantation respectively. Citrullinemia is an autosomal recessive disease, caused by the mutations in the argininosuccinate synthetase(ASS) gene. We identified in 3 major mutations in 11 unrelated Korean patients with citrullinemia; G324S, $IVS6^{-2}$ A to G, and 67 bp ins at nt 1125-1126. Among these, the 67 base pair insertion mutation is novel. The allele frequency of each mutation is; G324S(45%), IVS6-2 A to G(32%), and 67 base pair insertion(14%). All patients are diagnosed at neonatal or infantile age. Interestingly, two patients presented with stroke like episode. Out of 11 patients, 5 patients died. Among 6 patients alive, one patient was successfully liver transplanted.

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Expression of MicroRNA-221 in Korean Patients with Multiple Myeloma (한국인의 다발성골수종 환자에서 MicroRNA-221의 발현)

  • Choi, Woo-Soon
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.2
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    • pp.197-204
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    • 2018
  • Multiple myeloma (MM) is the leading cause of death among hematologic neoplasms. Recently, microRNA has been reported to be useful in the diagnosis of multiple myeloma. This study examined whether miR-221 could be used as a diagnostic marker for multiple myeloma. The study was performed on 20 patients with multiple myeloma without any other hematological diseases. MicroRNA extraction was performed using formalin-fixed paraffin-embedded (FFPE) tissues obtained from the bone marrow of patients with multiple myeloma. miR-15a, miR-16, miR-21, miR-181a, and miR-221 were selected as the microRNA target genes for multiple myeloma. The significance of microRNA was based on a fold change of <1.5. To quantify the fold changes, data normalized to the human gene, SNORD43, were used as the values of the patient group. Fold change values greater than 1.5 were defined as "overexpression", whereas values less than -1.5 were defined as "underexpression". Of note, 65.0% (13/20) of samples showed significant "overexpression" in the levels of miR-221 expression and plasma cells with a group of more and less than 30% in MM patients did not show any significance of plasma cell (P<0.05). The results of other studies showing a correlation between the expression of miR-221 and MM in Caucasians were confirmed. These results suggest that miR-221 may be a useful indicator for diagnosing patients with MM. In conclusion, miR-221 is useful in the diagnosis and determining the prognosis of multiple myeloma in Koreans.

Identification and Chararterization of Stationary-phase Specific Cytosolic Protein in Salmonella typhimurium (정지기 Salmonella typhimurium 세포에서 특이적으로 발현되는 세포질 단백질의 동정 및 발현조절에 대한 연구)

  • Yoo, Ah-Young;Kim, Young-Hee;Yu, Jong-Earn;Kim, Sam-Woong;Baik, Hyung-Suk;Kang, Ho-Young
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.298-304
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    • 2007
  • Salmonella is facultative intracellular pathogen that can survive and replicate in macrophages even though these cells are equipped with a plethora of anti-microbial mechanisms. To survive in this hostile intracellular environment, Salmonella has evolved numerous defense mechartisms. An approximately 20 kDa protein was detected as a stationary-phase specific protein band in cytosolic fraction. It was identified as a DNA binding protein in stationary phase (Dps) by analysis of MALDI-TOF assay. It has been known that Dps, the protein produced in the stationary phase of bacteria, allows DNA to form chromatin by binding to DNA nonspecifically and protects DNA from reactive oxidative species (ROS). For further study, Dps specific polyclonal antibodies were generated by injection of purified Dps protein into rabbit. To examine the Finfluence of several regulatory proteins in the expression dps gene, Dps protein level in various S. typhimurium mutants defecting regulatory proteins were investigated by Westernblot using Dps specific polyclonal antibodies.

Expression and Production of Human Granulocyte Colony Stimulating Factor (G-CSF) in Silkworm Cell Line (누에세포를 이용한 인간 G-CSF의 발현 및 생산)

  • Park, Jeong-Hae;Jang, Ho-Jung;Kang, Seok-Woo;Goo, Tae-Won;Chung, Kyung-Tae
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1577-1581
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    • 2010
  • Granulocyte colony stimulating factor (G-CSF) is a hematopoietic cytokine that stimulates bone marrow cells to proliferate and differentiate into granulocytes. G-CSF is approved and used for therapeutic purposes. The endoplasmic reticulum (ER) signal peptide of hG-CSF was replaced with silkworm-specific signal peptides to express and efficiently secrete recombinant hG-CSF by silkworm cells. Plasmids that contain cDNAs for hG-CSF and hG-CSF fused with silkworm- specific signal peptides of prophenoloxidase activating enzyme (PPAE), protein disulfide isomerase (PDI), and bombyxin (BX) were constructed. The G-CSF protein was expressed in insect cell line BM5 and was detected by western blot analysis. The cells transfected with plasmids containing rhG-CSF genes with silkworm-specific signal sequences released mature rhG-CSF protein more efficiently than the cells transfected with pG-CSF, the plasmid containing human G-CSF gene, including its own signal sequence. The production of hG-CSF reached maximal level at four days post-transfection and remained at a high level until 7 days post-transfection. These data demonstrate that the modification of the human G-CSF mimic to insect proteins synthesized in ER greatly improves the production of the protein.

Identification of Methicillin-Resistant Staphylococcus aureus by Polymerase Chain Reaction (중합효소 연쇄반응을 이용한 메치실린 내성균주의 동정)

  • Park, In-Cheol;Kim, Gwang-Su;Park, Myeong-Jin;Lee, Seung-Hun;Hong, Seok-Il;Choe, Tae-Bu
    • KSBB Journal
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    • v.14 no.4
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    • pp.460-464
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    • 1999
  • Methicillin-resistant Staphyloccus aureus (MRSA) has been known to be resistant to many kinds of antibiotics and causes a problem of nosnocomial infection since the third generation of cephalosporines has been introduced in the 1980s. As antibiotic sensitivity tests which have been routinely used to detect MRSA in the laboratory depend on the culture conditions such as, pH, temperature, and time, etc., it is difficult to decide in the case of borderline- or low-level of MRSA. Therefore it would be necessary to develope a new method based on the molecular biological technique to overcome these problems. In this study, we extracted DNA from S. aureus and performed polymerase chain reaction (PCR) to amplify mec A gene, encoding penicillin-binding protein 2' (PBP-2'), which is known to confer bacteria resistance to the bacteriostatic action of methicillin. The results were compares with those of minimal inhibitory concentration (MIC) test. When MIC test with oxacillin was performed on the 120 isolates of S. aureus from each patient's specimens, 64 of them were MRSA and 56 of them were methicillin-sensitive Staphylococcus aureus (MSSA). In pus specimen, more precisely, 61.9% (26/42) of MRSA was detected, and 44.2% (19/43), 60% (9/15) and 50% (10/20) of MRSA were detected in sputum, body fluid, and other specimen respectively. When 40 isolates of MRSA and MSSA were tested by PCR method and compares with the results of MIC method, different results were obtained from 1 isolate of MRSA (2.5%) and in 2 isolates of MSSA (5%) suggesting that PCR method should be performed at the same time for more accurate clinical test of MRSA.

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Biological Characteristics and Tissue Structure of a Crustose Coralline Lithophyllum Alga (해조류 무절산호조 혹돌잎의 생물학적 특성 및 조직구조)

  • Kang, Ji-Young;Benliro, Ianthe Marie P.;Lee, Ik-Joon;Choi, Ji-Young;Joo, Jin;Choi, Yoo Seong;Hwang, Dong Soo;Hong, Yong-Ki
    • Journal of Life Science
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    • v.23 no.3
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    • pp.341-346
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    • 2013
  • The disappearance of seaweed flora in some rocky areas, which is known as algal whitening, barren ground, coralline flats, or deforested areas, is associated with some species of coralline algae. To determine the biological characteristics of a representative species of crustose coralline alga, the 18S rDNA gene was sequenced to identify the genus Lithophyllum. According to its morphological and distributional characteristics, it was deduced to be L. yessoense. Viability was measured using triphenyl tetrazolium chloride and showed high viability from December to February. Culture conditions of $16^{\circ}C$, a 16 hr light, 8 hr dark cycle, and 30 ${\mu}E/m^2/s$ light intensity were optimal for maintaining the viability of the alga for up to five days. Included in the fatty acids was 9.7% ${\omega}$-3 eicosapentaenoic acid. An electron microscopy scan of the surface structure revealed round craters about 3.6 ${\mu}m$ in diameter, which were covered with rough, irregular, and angular polygon-shaped structures about 1.0 to 3.7 ${\mu}m$ in size. Based on the composition and structure found in our study, biomimetic coralline alga might become an environmentally friendly antifouling material against the attachment of soft foulants.

Isolation and Characterization of Plant-Derived Lactic Acid Bacteria as Potential Probiotic (잠재적 생균제로서 식물 젖산균의 분리 및 특성)

  • Kim, Jeong-Do;Park, Sung-Bo;Lee, Na-Ri;Jeong, Jin-Ha;Lee, Hee-Seob;Hwang, Dae-Youn;Lee, Jong-Sup;Jeong, Seong-Yun;Son, Hong-Joo
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.308-312
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    • 2011
  • Plant lactic acid bacteria were isolated from plant-associated fermentative foods and crops, and their probiotic properties were investigated. Isolates K27 and O2 were isolated from Kimchi and onion, and identified as Lactobacillus plantarum on the basis of 16S rRNA gene analysis. The two strains were highly resistant to acid (an MRS broth at pH 2.5), where the survival rates of L. plantarum K27 and L. plantarum O2 were 90.2% and 97.3%, respectively. L. plantarum K27 and L. plantarum O2 also showed high bile resistance to 0.5% oxgall, with a more than 70% survival rate. They showed an inhibitory effect against pathogenic strains of Escherichia coli KCCM 40880 and Pseudomonas aeruginosa ATCC 10145. The antibacterial effect of the two strains was probably due to the presence of lactic acid. ACE inhibitory activities of the two strains ranged from 72.8% to 80.6% in MRS broth. Notably, the two strains showed high ACE inhibitory activity (89.2~98.2%) in MRS broth containing 10% skim milk. Antioxidant activity was tested by DPPH radical scavenging activity, with antioxidant activities of the strains being in the range of 56.8~61.5%. The results obtained in this study suggest that L. plantarum K27 and L. plantarum O2 may be potential probiotic starter cultures with applications with fermentative products.

Gut Bacterial Diversity of Insecticide-Susceptible and -Resistant Nymphs of the Brown Planthopper Nilaparvata lugens Stål (Hemiptera: Delphacidae) and Elucidation of Their Putative Functional Roles

  • Malathi, Vijayakumar M.;More, Ravi P.;Anandham, Rangasamy;Gracy, Gandhi R.;Mohan, Muthugounder;Venkatesan, Thiruvengadam;Samaddar, Sandipan;Jalali, Sushil Kumar;Sa, Tongmin
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.976-986
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    • 2018
  • Knowledge about the gut bacterial communities associated with insects is essential to understand their roles in the physiology of the host. In the present study, the gut bacterial communities of a laboratory-reared insecticide-susceptible (IS), and a field-collected insecticide-resistant (IR) population of a major rice pest, the brown planthopper Nilaparvata lugens, were evaluated. The deep-sequencing analysis of the V3 hypervariable region of the 16S rRNA gene was performed using Illumina and the sequence data were processed using QIIME. The toxicological bioassays showed that compared with the IS population, IR population exhibited 7.9-, 6.7-, 14.8-, and 18.7-fold resistance to acephate, imidacloprid, thiamethoxam, and buprofezin, respectively. The analysis of the alpha diversity indicated a higher bacterial diversity and richness associated with the IR population. The dominant phylum in the IS population was Proteobacteria (99.86%), whereas the IR population consisted of Firmicutes (46.06%), followed by Bacteroidetes (30.8%) and Proteobacteria (15.49%). Morganella, Weissella, and Enterococcus were among the genera shared between the two populations and might form the core bacteria associated with N. lugens. The taxonomic-to-phenotypic mapping revealed the presence of ammonia oxidizers, nitrogen fixers, sulfur oxidizers and reducers, xylan degraders, and aromatic hydrocarbon degraders in the metagenome of N. lugens. Interestingly, the IR population was found to be enriched with bacteria involved in detoxification functions. The results obtained in this study provide a basis for future studies elucidating the roles of the gut bacteria in the insecticide resistance-associated symbiotic relationship and on the design of novel strategies for the management of N. lugens.

Effects of Yuldahansotang after kainate administration in the mouse hippocampus area (열다한소탕(熱多寒少湯)이 kainic acid에 의해 유발된 mouse의 해마체 손상에 미치는 영향)

  • Kim, Il-hwan;Kim, Kyung-yo
    • Journal of Sasang Constitutional Medicine
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    • v.11 no.2
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    • pp.283-299
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    • 1999
  • 1. Purpose : Systemic injection of kainic acid in experimental animals induces the limbic seizure and structural damages in hippocampus and amygdala which resembles the changes in human temporal lobe epilepsy. The author performed this study to investigate the neuroprotective effects of Yuldahansotang, on the neurotoxicity induced by kainic acid in the hippocampus in rats. 2. Method : Kainic acid was administered intraperitoneally. And feeding with Yuldahansotang for 3 weeks after kainic acid administration. Seizure were induced in male mice (kainate 10-40 mg/kg i.p) and animals were sacrified at various time-points after injection. The experimental animals were sacrificed at 1, 2, 3day and 1, 3weeks while Yuldahansotang administrations. Seizure were graded using a behavioral scale developed in our laboratory. c-fos belong to immediate early genes(IEGs) known to have rapid and brief responses. And neuronal injury was assayed by examining DNA fragmentation using in situ nick translation histochemistry. 3. Results & Conclusion : Seizure severity paralled kainate dosage. At higher doses DNA fragmentation is seen mainly in hippocampus in area CA3, and variable in CA1, thalamus, amygdala within 24 h, is maximal within 72 h, and is large gene by 7 days after administration of kainate. And we can't see the expression of DNA fragmentation and c-fos in the mice what feeded by Yuldahansotang after 7 days from kainic acid administration. It is consequently suggested that Yuldahansotang may attenuate the kainic acid-induced neuronal degeneration and increase the immunoreactivity of hippocampus in mouse.

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