• Journal of Life Science
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• v.21 no.12
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• pp.1666-1677
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• 2011

The regulation of gene expression plays an important role in cell cycle controls. In this study, a novel gene, the $mas1^+$($\underline{mi}$tosis $\underline{as}$sociated protein) gene, a homolog of human CIP29/Hcc1, was isolated and characterized from fission yeast Schizosaccharomyces pombe (S. pombe) using a gene-specific polymerase chain reaction. The isolated gene contained a complete open reading frame capable of encoding 245 amino acid residues with a typical promoter, as judged by nucleotide sequence analysis. It was also found that a PCB ($\underline{p}$ombe cell $\underline{c}$ycle $\underline{b}$ox) is located in the promoter region, which controls M-$G_1$ specific transcription in S. pombe. The quantitative analysis of the $mas1^+$ transcript against $adh1^+$ showed that the pattern of expression is similar to that of the septation index. Cytokinesis of mas1 mutant was greatly delayed at $25^{\circ}C$ and $36^{\circ}C$, and a large number of multi-septate cells were produced. The mas1 mutant had 2C, 4C and 6C DNA contents, as determined by FACS analysis. In addition, the number of multi-septate cells significantly increased. When cells were cultured in nitrogen starvation medium to increase proliferation, the abnormal phenotypes of mas1 mutant dramatically increased. These phenotypes could be rescued by an overexpression of the $mas1^+$ gene. The mas1 protein localized in the nuclei of S. pombe and human HeLa cells, as evidenced by Mas1-EGFP signals. The abnormal growth pattern and the morphology of mas1 mutant were complemented by a plasmid carrying human CIP29/Hcc-1cDNA. In addition, CIP29 /Hcc-1 transcript level increased in active cell proliferation stages in the developing mouse embryos. These results indicate that the $mas1^+$ ishomologous to the human CIP29/Hcc1 gene and is involved in cytokinesis and cell shape control.

• Journal of Life Science
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• v.16 no.1
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• pp.101-107
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• 2006

We have recently discovered that mycelium of Phellinus linteus, popular medical mushrooms in Korea, possess alcohol dehydrogenase and produce alcohol. In the present study, it was examined that the effect of fermented rice wine made by using mycelium of P. linteus (FLMP) on the expression of in-flammation-related proteins in both $HepG_2$ cells and rats. To examine the effect of FLMP on the morphology and expression of inflammatory proteins in $HepG_2$ cells, the cells were incubated with ethanol, and FLMP for 24 hours, and then analyzed by microscopic observation and Western blot and reverse transcription polymerase chain reaction (RT-PCR). While ethanol induced the morphological change accompanied with cell debris formation and scattering on $HepG_2$ cells, FLMP had no effect. The results of Western blot and RT-PCR analyses showed that the level of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-1 and COX-2 was induced by ethanol, however, FLMP inhibited the expression of these proteins and its mRNAs. In the animal model, the value of flutamate oxaloacetate transaminase and glutamate pyruvate transaminase was significantly increased by administration with ethanol. But the group administrated with FLMP showed lower levels on the changes of these markers compared with ethanol-administrated group. Besides, the results of Western blot and RT-PCR analyses showed that the expression of inflammatory proteins such as iNOS, COX-1 and COX-2 was not affected by FLMP administration in rat liver. About histopathological and immunohistochemical observations, inflammatory loci were markedly decreased in the FLMP-administrated rat compared to ethanol-administrated rats and showed weaker COX-2 and iNOS jmmunoreactions. These results suggested that FLMP showed slight changes on the inflammatory proteins expression compared to ethanol and FLMP may be used as a functional alcoholic beverage.

• Tuberculosis and Respiratory Diseases
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• v.60 no.6
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• pp.638-644
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• 2006

Backgrounds: This study investigated whether or not a polymorphism in the gene encoding the surfactant protein A(SP-A) has any bearing on the individual susceptibility to the development of chronic obstructive pulmonary disease(COPD) in a genetically homogenous Korean population. Methods: The genotypes of 19 COPD patients and 20 healthy neonates as controls were tested using a polymerase chain reaction followed by restriction fragment length polymorphism analysis for the SP-A gene. Results: The specific frequencies of the 6A2 and 6A18 alleles of SP-A1 and the 1A2 allele of SP-A2 were much higher in the COPD group than control group (p<0.05). However, the frequencies of the 6A3 and 6A4 alleles of SP-A1 and the 1A0 allele of SP-A2 in the COPD group were significantly lower than the control group. In the COPD group, the frequencies of the +50 locus genotypes GG of SP-A1 and the +9 locus genotypes CC of SP-A2 were 85.0% and 60.6%, respectively, and 19.7% and 24.8% in the control group, respectively. The frequencies of the polymorphic genotypes or alleles showed a statistically significant difference between the COPD group and the control group (P<0.05). Conclusion: A genetic polymorphism in SP-A is associated with the development of COPD in the Korean population.

• Applied Microscopy
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• v.37 no.2
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• pp.93-102
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• 2007

Secretory leukocyte protease inhibitor (SLPI) was known as one of bacterial lipopolysaccharide (LPS)-induced products of macrophage. Macrophages play an important role in the development of inflammatory responses by secreting an array of cytokines and chemokines in a tissue microenvironment. To identify the function and relationship between potent growth factors and SLPI after LPS stimulation, we conducted reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of SLPI and growth factors such as VEGF, PDGF, bFGF after 100 ng LPS stimulation on the RAW264.7 cells. The result of RT-PCR was showed SLPI mRNA expression was increased from 60 min to 48h in RAW 264.7 cells after incubation with LPS. VEGF and PDGF mRNA was expressed highly at initial stage by LPS stimulation. The mRNA of bFGF and type I collagen was very weakly expressed after LPS stimulation. SLPI protein level was increased likely the mRNA levels in RAW 267.7 cells. Additionally, phase contrast and scanning electron microscopic observation demonstrated that the LPS induce the change of morphology of the RAW264.7 cells. From these results, it suggest that expression of SLPI by LPS treatment may associate with VEGF and PDGF expression in RAW264.7 cells.

• Development and Reproduction
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• v.10 no.4
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• pp.255-261
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• 2006

Although some phytoes rogens might have beneficiary rather than adverse effects, most endocrine disrupting compounds(EDCs) are considered to be harmful to human and wildlife health through interfering the endocrine system. Previously we found that prepubertal exposure to genistein(GS), a well-known isoflavone phytoestrogen, could activate the reproductive system of immature female rats resulting precocious puberty. Interestingly, di(2-ethyl hexyl) phthalate(DEHP) exposure brought inverse result, a delayed puberty, in the same experimental regimen. In this study, we examined whether prepubertal exposure to GS or DEHP affect the gene expressions of estrogen receptors($ER\;{\alpha}$ and $ER\;{\beta}$) and LH receptor(LHR) which represent the maturational status of ovary and uterus in immature rats. GS (100 mg/kg/day) was administered daily from postnatal day 25 to the day when the first vaginal opening(VO) was observed, and the animals were sacrificed on the next day(day 32). Similarly, DEHP(l00 mg/kg/day) was administered daily from postnatal day 25 through the day when the first V.O. in control group was observed, and the animals were sacrificed on the next day(day 36). To determine the transcriptional changes in the hormone receptors, total RNAs were extracted from ovary and uterus and were applied to semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). In the GS group, the transcriptional activities of $ER\;{\alpha}$, $ER\;{\beta}$ and LHR in uterus and LHR in ovary were significantly increased when compared to those of control group. In the DEHP group, the transcriptional activities of all the hormone receptors measured were significantly lowered when compared to those of control group. These alteration of the reproductive hormone receptor expressions in ovary and uterus might be represent the phenotypic aspects(secondary sexual characteristics) such as tissue weights and reproductive hormone levels during perinatal period in immature female rats.

• Korean Journal of Poultry Science
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• v.43 no.2
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• pp.77-88
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• 2016

To establish a new synthetic Korean meat chicken breed, we tested $5{\times}5$ diallel cross mating experiment with domestic chicken breeds. Comparing stress responses among diallel crossed chicken breeds, we analyzed telomere length, DNA damage and expressions of heat shock protein genes (HSPs) as the markers of the stress response. The telomere length was measured by quantitative fluorescence in situ hybridization on the nuclei of lymphocytes. The expression levels of HSP-70, $HSP-90{\alpha}$ and $HSP-90{\beta}$ genes were analyzed by quantitative real-time polymerase chain reaction in lymphocytes. The DNA damage rate of lymphocytes was quantified by the comet assay known as the single cell gel electrophoresis. In results, there were significant differences in the values of the stress markers such as telomere length, HSPs and DNA damage rate, and also were significant differences in viabilities and body weights among the $5{\times}5$ diallel crossed chicken breeds. The telomere shortening rate, expression values of HSPs and DNA damage rate were significant low in W and Y crossed chickens compare to the others, but GG pure breed showed the highest values in the 25 crossed chickens. Estimating correlation coefficient, the survival rate positively correlated to telomere length, but negatively correlated to the expression levels of HSP-70, $HSP-90{\alpha}$, $HSP-90{\beta}$ genes and to the value of % DNA in tail as DNA damage rate. The expression levels of HSP-70, $HSP-90{\alpha}$ and $HSP-90{\beta}$ genes of dead chickens had significantly higher than those of survival chickens. According to the results on the stress marker analysis, it would be considered that the crossed breeds had more stress resistant than the pure breeds, and the crossed chickens with a light strain such as W or Y were relatively resistant to stress, but the crossed chickens with a heavy strain such as G, H, F were susceptible to stress.

• Neonatal Medicine
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• v.15 no.1
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• pp.44-53
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• 2008

Purpose : Epidemic keratoconjunctivitis (EKC) caused by adenovirus is a highly contagious disease, which has been reported as outbreaks involving adults in the community. However, there has been no report on EKC outbreak by adenovirus in a neonatal intensive care unit (NICU) in Korea. Aims of this study were to investigate the EKC outbreak by adenovirus type 8 in NICU and to confirm an effectiveness of polymerase chain reaction (PCR) for diagnosis. Methods : Conjunctival swab or nasopharyngeal aspirate specimens were taken from all patients and tested by viral culture and PCR. Adenovirus serotype was determined by sequencing of PCR product of selected region of hexon gene using the virus isolates or specimens. Results : An outbreak of EKC occurred which was involving 12 preterm infants in the NICU of the Seoul National University Children's Hospital between July 12th and August 1st, 2005. Three hospital staffs and one family member of the neonate were also affected. Adenovirus was detected in 12/12 (100%), 6/11 (54.5%) by PCR and virus culture, respectively. Eleven PCR-positive neonates were identified as serotype 8 by sequencing. The first affected 4 babies have had routine ROP (retinopathy of prematurity) examinations one week ago. While previous outbreaks were sustained for a few months, the event in our unit was controlled without complications in 3 weeks. Conclusion : We analyzed the EKC outbreak by adenovirus type 8 in NICU. Adenovirus serotype was identified by PCR and sequencing with high sensitivity for the first time in Korea, so we suggest this method can be very useful for rapid diagnosis and infection control.

• Research in Plant Disease
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• v.12 no.2
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• pp.139-143
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• 2006

Virion captured reverse transcription polymerase chain reaction (VC/RT-PCR) could detect plant virus quickly and accurately. In the VC/RT-PCR, no antibody is needed unlike immuno-captured RT-PCR (IC/RT-PCR) which had been improved method of RT-PCR for plant viruses, and virus nucleic acids can be obtained easily within 30minutes by property of polypropylene PCR tube which is hold and immobilized viral particles on its surface. For the virion capture of Tomato spotted wilt virus (TSWV), the extraction buffer was tested. The optimum macerating buffer for TSWV was 0.01M potassium phosphate buffer, pH 7.0, containing 0.5% sodium sulfite. The viral crude sap was incubated for 30 min at $4^{\circ}C$. The virions in the PCR tubes were washed two times with 0.01M PBS containing 0.05% Tween-20. The washed virions were treated at $95^{\circ}C$ immediately for 1 min containing RNase free water and chilled quickly in the ice. Disclosed virions' RNAs by heat treatment were used for RT-PCR. Dilution end point of $10^{-5}$ from plant's crude sap infected with TSWV showed relatively higher detection sensitivity for VC/RT-PCR. During multiple detection using two or more primers, interference was arisen by interactions between primer-primer and plant species. The result of multiplex RT-PCR was influenced by combinations of primers and the kind of plant, and the optimum extraction buffer for the multiplex detection by VC/RT-PCR should be developed.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.9 no.1
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• pp.1-13
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• 2006

Purpose: Acute gastroenteritis in children is one of the frequently encountered diseases with relatively high admission rate. The aim of this study is to determine the isolation trends of common and emerging pathogens in acute gastroenteritis in children over a 12-month period in a community hospital. Methods: The study group included the children who were hospitalized to Seoul National University Boramae Hospital from April, 2003 to March, 2004 or visited outpatient clinic from April, 2003 to July, 2003 with presenting features of acute gastroenteritis. Stool specimens were obtained within 2 days after the visit and examined for the following pathogens: rotavirus, adenovirus, Salmonella, Shigella, Vibrio, pathogenic Escherichia coli (E.coli), Campylobacter and Yersinia species. Viral study was done with commercial kits for antigen detection. Identification of the bacterial pathogens was done by culture using selective media. For pathogenic E.coli, polymerase chain reaction (PCR) was done with the target genes related to the pathogenecity of enterotoxigenic E.coli (ETEC), enteropathogenic E.coli (EPEC) and enterohemorrhagic E.coli (EHEC). Results: The 130 hospitalized children and 28 outpatients were included in this study. The majority of children (>93%) were less than 6 years. Pathogens were isolated in 47% of inpatients and 43% of outpatients, respectively. Rotavirus was the most frequently identified pathogen, accounting for 42.3% of inpatients and 29.6% of outpatients. Nontyphoidal salmonella is the most commonly isolated bacterial pathogen (3.9%) in hospitalized children. Pathogenic E.coli (EPEC, ETEC) was detected in 2.1% (2/97) of inpatients and 25% (3/12) of outpatients. EHEC, adenovirus, Campylobacter, Yersinia and Shigella species were not detected in this study. Conclusion: Rotavirus is the most common enteropathogen in children with acute gastroenteritis. Nontyphoidal salmonella and pathogenic E.coli are important bacterial pathogens. Campylobacter species may not be commonly detected organism in hospitalized children with acute diarrhea.

• Pediatric Infection and Vaccine
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• v.18 no.2
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• pp.154-162
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• 2011

Purpose : The aim of this study was to identify the clinical characteristics of lower respiratory tract infection due to respiratory syncytial virus (RSV) in young children and to provide information for an effective guideline for palivizumab administration in Korea. Methods : We reviewed medical charts of 167 patients under 3 years of age who were hospitalized in Dongguk University Ilsan Hospital for lower respiratory tract infection between January 2007 and February 2011. Diagnosis of the virus was made based on the multiplex real time polymerase chain reaction. Results : There were 113 patients who were infected by respiratory syncytial virus. 90 patients were term infants and 23 patients were preterm infants. No difference was shown between term and preterm infants except the days of admission which was 9.0${\pm}$6.0 days and 12.6${\pm}$21.0 days respectively. In the preterm group their mean age at the time of admission was 5.21${\pm}$4.9 months and the mean gestational age was 33.1${\pm}$4.3 weeks, and the mean birth weight was 2,152${\pm}$950 g. Only 4 patients were born under 28 weeks gestational age and were candidates for palivizumab administration. Conclusion : Most of the patients with severe RSV lower respiratory tract infection were term or near term infants who were not candidates for palivizumab prophylaxis. A nationwide study is needed to make a new risk stratified guideline for RSV prophylaxis for our country.