• Tuberculosis and Respiratory Diseases
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• v.57 no.5
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• pp.449-460
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• 2004

Background : PS-341 is a novel, highly selective and potent proteasome inhibitor, which showed cytotoxicity against some tumor cells. Its anti-tumor activity has been suggested to be associated with modulation of the expression of apoptosis-associated proteins, such as p53, $p21^{WAF/CIP1}$, $p27^{KIP1}$, NF-${\kappa}B$, Bax and Bcl-2. c-Jun N-terminal kinase (JNK) and glycogen synthase kinase-$3{\beta}$ (GSK-$3{\beta}$) are important modulators of apoptosis. However, their role in PS-341-induced apoptosis is unclear. This study was undertaken to elucidate the role of JNK and GSK-$3{\beta}$ in the PS-341-induced apoptosis in lung cancer cells. Method : NCI-H157 and A549 cells were used in the experiments. The cell viability was assayed using the MTT assay and apoptosis was evaluated by proteolysis of PARP. The JNK activity was measured by an in vitro immuno complex kinase assay and by phosphorylation of endogenous c-Jun. The protein expression was evaluated by Western blot analysis. Dominant negative JNK1 (DN-JNK1) and GSK-$3{\beta}$ were overexpressed using plasmid and adenovirus vectors, respectively. Result : PS-341 reduced the cell viability via apoptosis, activated JNK and increased the c-Jun expression. Blocking of the JNK activation by overexpression of DN-JNK1, or pretreatment with SP600125, suppressed the apoptosis induced by PS-341. The activation of caspase 3 was mediated by JNK activation. Blocking of the caspase 3 activation suppressed PS-341-induced apoptosis. PS-341 activated the phosphatidylinositol 3-kinase (PI3K)/Akt pathway, but its blockade enhanced the PS-341-induced cell death via apoptosis. GSK-$3{\beta}$ was inactivated by PS-341 via the PI3K/Akt pathway. Overexpression of constitutively active GSK-$3{\beta}$ enhanced PS-341-induced apoptosis; in contrast, this was suppressed by dominant negative GSK-$3{\beta}$ (DN-GSK-$3{\beta}$). Inactivation of GSK-$3{\beta}$ by pretreatment with lithium chloride or the overexpression of DN-GSK-$3{\beta}$ suppressed both the JNK activation and c-Jun up-regulation induced by PS-341. Conclusion : The JNK/caspase pathway is involved in PS-341-induced apoptosis, which is negatively regulated by the PI3K/Akt-mediated inactivation of GSK-$3{\beta}$ in lung cancer cells.

• Development and Reproduction
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• v.17 no.4
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• pp.299-309
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• 2013

Transforming growth factor (TGF) family is well known to induce the chondrogenic differentiation of mesenchymal stem cells (MSC). However, the precise signal transduction pathways and underlying factors are not well known. Thus the present study aims to evaluate the possible role of C2 domain in the chondrogenic differentiation of human mesenchymal stem cells. To this end, 145 C2 domains in the adenovirus were individually transfected to hMSC, and morphological changes were examined. Among 145 C2 domains, C2 domain of protein kinase C eta ($PKC{\eta}$) was selected as a possible chondrogenic differentiation factor for hMSC. To confirm this possibility, we treated $TGF{\beta}3$, a well known chondrogenic differentiation factor of hMSC, and examined the increased-expression of glycosaminoglycan (GAG), collagen type II (COL II) as well as $PKC{\eta}$ using PT-PCR, immunocytochemistry and Western blot analysis. To further evaluation of C2 domain of $PKC{\eta}$, we examined morphological changes, expressions of GAG and COL II after transfection of $PKC{\eta}$-C2 domain in hMSC. Overexpression of $PKC{\eta}$-C2 domain induced morphological change and increased GAG and COL II expressions. The present results demonstrate that $PKC{\eta}$ involves in the TGF-${\beta}3$-induced chondrogenic differentiation of hMSC, and C2 domain of $PKC{\eta}$ has important role in this process.

• BMB Reports
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• v.45 no.9
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• pp.509-514
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• 2012

We have found that the previously uncharacterized bone morphogenetic protein-9 (BMP9) is one of the most osteogenic factors. However, it is unclear if BMP9 cross-talks with $TGF{\beta}1$ during osteogenic differentiation. Using the recombinant BMP9 adenovirus, we find that low concentration of rh$TGF{\beta}1$ synergistically induces alkaline phosphatase activity in BMP9-transduced C3H10T1/2 cells and produces more pronounced matrix mineralization. However, higher concentrations of $TGF{\beta}1$ inhibit BMP9-induced osteogenic activity. Real-time PCR and Western blotting indicate that BMP9 in combination with low dose of $TGF{\beta}1$ potentiates the expression of later osteogenic markers osteopontin, osteocalcin and collagen type 1 (COL1a2), while higher concentrations of $TGF{\beta}1$ decrease the expression of osteopontin and osteocalcin but not COL1a2. Cell cycle analysis reveals that $TGF{\beta}1$ inhibits C3H10T1/2 proliferation in BMP9-induced osteogenesis and restricts the cells in $G_0/G_1$ phase. Our findings strongly suggest that $TGF{\beta}1$ may exert a biphasic effect on BMP9-induced osteogenic differentiation of mesenchymal stem cells.

• Journal of Life Science
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• v.18 no.10
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• pp.1395-1399
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• 2008

We have recently reported the PRC1 promoter as a promoter candidate to control expression of transcriptionally targeted genes for breast cancer gene therapy. We tested whether the PRC1 promoter could be also applied for the lung cancer gene therapy. In the transient transfection assay with naked plasmids containing the luciferase fused to the PRC1 promoter, the promoter showed little activity in the normal lung cell line, MRC5. However, in the lung cancer A549 cells, PRC1 showed approximately 30-fold activation which was similar to the survivin promoter, the gene whose promoter has been already reportedas a candidate for the gene therapy of lung cancer. In viral systems, the PRC1 promoter showed approximately 75% and 66% of transcriptional activity compared to the CMV promoter in the adeno-associated virus (AAV) and the adenovirus (AV) systems, respectively. However, the PRC1 promoter in either AAV or AV showed approximately 20% activity compared to the CMV promoter in the normal lung cells. In addition, human lung tumor xenograft mice showed that the PRC1 promoter activity was as strong as the CMV activity in vivo. Taken together, these results suggested that PRC1 might be a potential promoter candidate for transcriptionally targeted lung cancer gene therapy.

• Pediatric Infection and Vaccine
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• v.23 no.1
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• pp.40-45
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• 2016

Purpose: The purpose of this study was to investigate the etiology of acute pharygotonsillitis in pediatric patients. Methods: Pharyngeal swabs from patients with acute pharyngotonsillitis were evaluated for viruses and bacterial organisms from March 2010 through March 2011. Results: Of 615 patients, potentially pathogenic bacteria were isolated in 40 (6.5%), viruses were isolated in 310 (50.4%), and no pathogens were isolated in 267 patients (43.4%). Both viral and bacterial pathogens were found in 2 (0.3%). Of 40 patients with bacterial pathogens, group A streptococci were found in 31 (77.5%). Among 310 patients with virus infection, adenovirus was the most frequently recovered (203 patients; 65.5%), followed by rhinovirus (65 patients; 21.0%), enterovirus (43 patients; 13.9%) and coronavirus (18 patients; 5.8%). There were 25 patients who had been coinfected with 2 viruses. In viral pharyngotonsillitis, cough, rhinorrhea, conjunctivitis and diarrhea were prominent. On the other hand, pharyngeal injection and pharyngeal petechiae were prominent in bacterial pharyngotonsillitis. Conclusions: Virus infection was a big part of acute pharyngotonsillitis and there were differences in clinical manifestations among viral and bacterial infections. Therefore, we need to distinguish between virus infection and bacterial infection using clinical signs for preventing the abuse of antibiotics.

• Korean Journal of Poultry Science
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• v.38 no.3
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• pp.181-189
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• 2011

A 5-month (May to November in 2009) monitoring program for five viral pathogens in litter, such as avian influenza virus A (AIV), infectious bronchitis virus (IBV), infectious bursal disease virus (IBDV), fowl adenovirus (FAdV), and chicken infectious anemia virus (CIAV) was conducted in 62 flocks at 31 broiler farms (two flocks in each farm) in Korea in 2009. Viral pathogens were examined twice (before and at the end of the rearing period) at 31 broiler farms, and included fresh litter (n = 16) and recycled litter (n = 15) farms. Thirty-seven viruses (14 IBVs, 2 IBDVs, 9 FAdVs, and 12 CIAVs) were isolated from 75% (12/16) and 73% (11/15) of fresh litter and reused litter farms during the period, respectively, indicating no difference in viral contamination rate between farms using new and reused litter. Of these isolates, three (two CIAVs and one IBDV) were isolated from recycled litter samples collected before the rearing period at three broiler farms, whereas the others (n=34) were isolated from fresh and recycled litter samples collected at the end of the rearing period. When the performances, involving viability, body weight, and feed conversion ratio, were compared, no significant differences were found between farms using fresh and recycled litter during the period.

• Nuclear Medicine and Molecular Imaging
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• v.41 no.1
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• pp.54-58
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• 2007

Purpose: Luciferase is one of the most commonly used reporter enzymes in the field of in vivo optical imaging. D-luciferin, the substrate for firefly luciferase has very high cost that allows this kind of experiment limited to small animals such as mice and rats. In this current study, we validated local injection of D-luciferin in the articular capsule for bioluminescence imaging in rabbits. Materials and Methods: Chondrocytes were cultured and infected by replication-defective adenoviral vector encoding firefly luciferase (Fluc). Chondrocytes expressing Fluc were injected or implanted in the left knee joint. The rabbits underwent optical imaging studies after local injection of D-luciferin at 1, 5, 7, 9 days after cellular administration. We sought whether optimal imaging signals was could be by a cooled CCD camera after local injection of D-luciferin. Results: Imaging signal was not observed from the left knee joint after intraperitoneal injection of D-luciferin (15 mg/kg), whereas it was observed after intraarticular injection. Photon intensity from the left knee joint of rabbits was compared between cell injected and implanted groups after intraarticular injection of D-luciferin. During the period of imaging studies, photon intensity of the cell implanted group was 5-10 times higher than that of the cell injected group. Conclusion: We successfully imaged chondrocytes expressing Fluc after intraarticular injection of D-luciferin. This technique may be further applied to develop new drugs for knee joint disease.

• Pediatric Infection and Vaccine
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• v.18 no.1
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• pp.61-67
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• 2011

Purpose : This study was performed to investigate the epidemiologic characteristics of human bocavirus (HBoV)-associated lower respiratory tract infections (LRTIs) in children. Methods : Nasopharyngeal aspirate samples were obtained from 658 children who had been hospitalized for LRTIs in Seoul National University (SNU) Children's Hospital and SNU Bundang Hospital from March 2000 to September 2005. Multiplex RT-PCR was performed to detect 11 respiratory viruses including respiratory syncytial virus, adenovirus, rhinovirus, parainfluenza viruses 1 and 3, influenza viruses A and B, human metapneumovirus, HBoV, human coronavirus (HCoV) OC43/ 229E, and HCoV-NL63. Clinical data were reviewed retrospectively. Results : Overall, respiratory viruses were detected in 325 (49.4%) among 658 patients. HBoV was detected in 62 cases (9.4%) and was responsible for 19.1% of virus-positive cases. HBoV was prevalent among infants and young children aged from 3 months to 5 years with the mean age of 25.3 months. Co-detection of HBoV and other respiratory viruses was observed in 37.1% which is significantly higher than average co-detection rate (12.3%) among overall virus-positive cases (P=0.000). HBoV was identified mainly in late spring and early summer from May to July. Conclusion : This study describes epidemiologic features of HBoV in Korean children compared with those associated with other respiratory viruses. HBoV was prevalent among LRTIs in childhood, especially in late spring and early summer season in Korea.

• The Korean Journal of Physiology and Pharmacology
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• v.13 no.6
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• pp.449-454
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• 2009

5'-AMP-activated protein kinase (AMPK) is a heterotrimeric complex consisting of a catalytic ($\alpha$) and two regulatory ($\beta$ and $\gamma$) subunits. Two isoforms are known for catalytic subunit (${\alpha}1$, ${\alpha}2$) and are encoded by different genes. To assess the metabolic effects of $AMPK{\alpha}1$, we examined the effects of overexpression of adenoviral-mediated $AMPK{\alpha}1$ in hyperlipidemic type 2 diabetic rats. The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is an established animal model of type 2 diabetes that exhibits chronic and slowly progressive hyperglycemia and hyperlipidemia. Thirty five-week-old overt type 2 diabetic rats (n=10) were administered intravenously with Ad.$AMPK{\alpha}1$. AMPK activity was measured by phosphorylation of acetyl CoA carboxlyase (ACC). To investigate the changes of gene expression related glucose and lipid metabolism, quantitative real-time PCR was performed with liver tissues. Overexpression of $AMPK{\alpha}1$ showed that blood glucose concentration was decreased but that glucose tolerance was not completely recovered on 7th day after treatment. Plasma triglyceride concentration was decreased slightly, and hepatic triglyceride content was markedly reduced by decreasing expression of hepatic lipogenic genes. Overexpression of $AMPK{\alpha}1$ markedly improved hepatic steatosis and it may have effective role for improving hepatic lipid metabolism in hyperlipidemic state.

• Korean Journal of Microbiology
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• v.39 no.2
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• pp.89-94
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• 2003

Respiratory viruses were isolated from patients with acute respiratory infections in Busan during 2000-2001 and characterized for their antigenic properties. In 2000, 39 out of 43 isolated viruses were identified as influenza viruses and the others were adenoviruses. Among the isolated influenza viruses,23 were type A influenza viruses and 16 were type B influenza viruses. As a result of antigenic characterization, the influenza viruses were determined to A/Sydney/05/97(H3N2)-like, A/Beijing/262/95(H1N1)-like, and B/Harbin07/94-like viruses and serotypes of the isolated adenoviruses were type 1, 2, and 5. In 2001, 56 viruses were isolated and all of the viruses were identified as influenza viruses. They were A/panama/253/99(H3N2)-like and A/Newcaledonia/2007/99(H1Nl)-like viruses when determined by their antigenic properties. The sex distribution of the patients is as follows, 14 males (32.56%),23 females (67.44％) in 2000, and 23 males (41.07%), 33 females (58.93%) in 2001. Occurrence rate was found to be higher in female patients in both years. Age distribution of patients, in 2000, 48.84％ of infection occurred in 0 to 1 year old while in 2002, 33.93% occurred among 11-20 year olds. In 2000, occurrence rate was found to be high in January and again in April and various types of viruses were isolated. These results may be useful for vaccine development and establishment of reliable epidemic data.