HtrA2/Omi, a mitochondrial trypsin-like serine protease, is pivotal in regulating apoptotic cell death. Several lines of recent evidence suggest that HtrA2 is associated with the pathogenesis of neurodegenerative disorders; however, the physiological function of HtrA2 still remains elusive. For studying physiological function of HtrA2 in depth, it is necessary to develop a suitable expression system in the model animal. We therefore utilized the zebrafish as a model animal to establish expression of human HtrA2 (hHtrA2) in vivo. For expression of mature HtrA2 as GFP fusion in zebrafish embryos, the HtrA2 (WT) or (S306A) cDNAs with the C-terminal GFP tag were inserted into the pCS2+ plasmid. Expression patterns of HtrA2 in HEK293 cells were first monitored by immunofluorescence staining and immunoblot assays, showing approximately 64 kDa of the HtrA2-GFP fusion proteins. Subsequently, the hHtrA2 plasmid DNA or in vitro transcribed mRNA was microinjected into zebrafish embryos. The expression patterns of HtrA2 in Zebrafish embryos were monitored by GFP fluorescence in 24 hours-post-fertilization (hpf). Although expression patterns of HtrA2-GFP in developing embryos were different between the injected DNA and mRNA, both nucleic acids revealed good expression levels to further study the physiological role of HtrA2 in vivo. This study provides a suitable condition for expressing hHtrA2 in the zebrafish embryos as well as a method for generating useful system to investigate physiological properties of the specific human genes.
The Journal of Korean Institute of Electromagnetic Engineering and Science
/
v.18
no.7
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pp.774-783
/
2007
This paper presents an integrated packaging antenna-diplexer module for wireless communication systems in the Cellular and SDMB band. The design and the realization of the proposed one are experimentally analyzed and discussed. It consists of a dual-resonance antenna and a diplexer with a multi-layer LTCC(${\varepsilon}_r=7.8,\;tan\;{\delta}=0.0043$) technology with integration capability and low loss. The dual-resonance antenna of the proposed module has the meander line structure for size reduction and has the shorting structure of an inverted F antenna to achieve good impedance matching. The diplexer of the proposed module was designed with the combination of low pass filter(LPF) and high pass filter(HPF). Decreasing the mutual interference between them provides a high isolation characteristic. The proposed antenna-diplexer module with dimensions of $27.5{\times}12.0{\times}2.2mm$ operates within a range from 813 MHz to 902 MHz for the cellular band and from 2,586 MHz to 2,655 MHz for the SDMB band. And the measured gain of the fabricated module is -1.96 dBi for Cellular band and -5.43 dBi for SDMB band. The parameters for the antenna-diplexer module are investigated and the several performances are discussed.
An, Jae-Min;Kim, Yu-Seon;Pyo, Hyun-Seong;Lee, Hye-Sun;Lim, Yeong-Seog
The Journal of Korean Institute of Electromagnetic Engineering and Science
/
v.21
no.1
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pp.28-35
/
2010
In this paper, we designed and fabricated a ultra-wideband(UWB) bandpass filter with a good performance of a stopband, and a notched band in passband. The transformed equivalent circuit of the highpass filter was realized by distributed element. A wide-passband with 3-dB fractional bandwidth of more than 100 % was achieved by using optimum response of the HPF. For improving lower and upper stopband characteristic, a cross coupling between feed lines was employed, which was analyzed by desegmentation technique. In order to reject interference of Wireless LAN and Hyper LAN(5.15~5.825 GHz), the narrow notched(rejection) band was realized by a spurline. The fabricated BPF indicated the passband from 3.1 to 10.55 GHz and the flat group delay of less than 0.94 ns over the entire passband except the rejection band. The filter shown sharp attenuation both inside and outside the band and notched band from 5.2 to 6.12 GHz.
Infection of Epstein-Barr virus(EBV) gives rise to a broad spectrum of clinical manifestations in children. Although renal involvement is rare, diverse renal manifestations are known from hematuria to acute renal failure. Secondary membranous nephropathy(MN) associated with systemic EBV infection is an uncommon renal pathology and only two cases have been reported. We are adding another case of MN associated with EBV infection in a child. An 8-year-old girl was admitted for renal biopsy. She had been followed up for microscopic hematuria and intermittent proteinuria for 5 months. There had been no specific findings in serology and radiology. Tonsil biopsy had been done due to exudative tonsillar hypertrophy and enlarged multiple cervical lymph nodes. And it showed EBV-associated lymphoproliferative findings. Serologic tests for EBV showed positive evidence of recent infection; viral capsid antigen(VCA) IgM was borderline positive, VCA IgG and early antigen IgG were positive, and EB nuclear antigen IgG was negative. In Situ Hybridization of tonsil for EBV mRNA was positive. Because her proteinuria and hematuria were aggravated at that time(protein 3 +, RBC >60/HPF), renal biopsy was done. Renal biopsy showed the findings of MN, characterized by thickened capillary walls with epimembranous spikes on light microscopy and subepithelial, mesangial and subendothelial electron dense deposits on electron microscopy. On immunofluorescence microscopy, IgG, C1q, kappa and lambda chains were positive. After steroid administration, proteinuria and hematuria resolved gradually within 6 months.
Urinalysis is considered to be easier and simpler than other tests. It has been known to cause no burden to patients, while offering important information on diagnosing, treating, and determining the prognoses of kidney and urinary tract diseases. Urinary sediments are usually performed by microscopic examination of centrifuged urine by technologist. The guidelines proposed by the Korean Association of External Quality Assessment Service are actually different from those actually practiced by medical institutions and taught to biomedical students in textbooks. Therefore, we verified whether different sediment preparation methods lead different test results. Specimens that tested positive from the occult blood and leukocyte esterase in the urine dipstick test were randomly selected for a microscopic examination. The differences in the urine sediment preparation affected the sediment concentrations, which influenced the cell grade and cell number per HPF. The first factor in determining the sediment concentration is the centrifugal force. Many medical institutions use 1,500 rpm as the centrifugal speed without considering the radius of the centrifuge; such a value may not be accurate for 400 G. Consequently, there were differences in urine concentrations, which influenced the results. The second factor is the amount of sediment in urine. Different amounts of the remaining supernatant led to different sediment concentration factors, again, causing different results. Furthermore, not only by using a pipette to obtain an accurate amount as stipulated, but also by roughly obtaining a drop, the microscopic examination using such a volume of sediment examined affected the results. Therefore, this study highlights the importance of standardization of urine sediment preparation procedures to promote consistency and accuracy across institutions.
Kim, In-Sun;Park, Sang-Chan;Han, Sung-Sik;Kim, Eun-Soo
Applied Microscopy
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v.36
no.3
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pp.217-226
/
2006
Image processing by ultra high voltage electron microscopy (UHVEM) and tomography has offered major contributions to research in the field of cellular ultrastructure. Furthermore, such advancements also have enabled the improved analysis of three-dimensional cellular structures in botany. In the present study. using UHVEM and tomography, we attempted to reconstruct the three-dimensional images of plastid inclusions that probably differentiate during photosynthesis. The foliar tissues were studied Primarily with the TEM and further examined with UHVEM. The spatial relationship between tubular elements and the thylakoidal membrane and/or starch grains within plastids mainly have been investigated in CAM-performing Sedum as well as in $C_4$ Salsola species. The inclusion bodies were found to occur only in early development in the former, while they were found only in mesophyll cells in the latter. The specimens were tilted every two degrees to obtain two-dimensional images with UHVEM and subsequently comparison has been made between the two types. Digital image processing was performed on the elements of the inclusion body using tilting, tomography, and IMOD program to generate and reconstruct three-dimensional images on the cellular level. In Sedum plastids, the inclusion bodies consisted of tubular elements exhibiting about 20 nm distance between elements. However, in Salsola, plastid inclusion bodies demonstrated quite different element structure, displaying pattern, and origin relative to those of the Sedum. The inclusion bodies had an integrative relationship with the starch grains in both species.
Urine sediments are performed by a microscopic examination of centrifuged urine by medical technologists. This study examined different urine sediment preparation procedures. The 107 fresh urine specimens that tested positive from white blood cells (WBCs) and red blood cells (RBCs) in the urine dipstick test and the cobas u 701 analyzer, respectively, were selected for manual microscopy. This study evaluated an automated urine sediment analyzer and three manual microscopy methods for WBCs and RBCs. The methods were performed according to the test guidelines. The coefficients of determination between the cobas u 701 analyzer and the Korean Association of Quality Assurance for Clinical Laboratory (KAQACL) for WBCs and RBCs were r2=0.977 and r2=0.970, respectively. The concordance rates between the cobas u 701 analyzer and KAQACL for WBCs and RBCs were 74.8% and 77.6%, respectively. A good correlation and concordance with the automatic analyzer were shown when the specimens were prepared and examined using the KAQACL method. Consequently, the differences in the urine sediment preparation procedures affected the sediment concentrations, influencing the cell number per high power field (HPF).
Ha, Chang Woo;Kim, Ji Young;Lee, Jeong Nyeo;Lee, Jeong Hwa;Chung, Woo Yeong
Clinical and Experimental Pediatrics
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v.45
no.7
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pp.884-890
/
2002
Purpose : Henoch-Schonlein purpura(HSP) nephritis has been reported to vary from 25 to 50% among HSP patients and is a common cause of chronic glomerulonephritis in children. In our study, we evaluated the distribution and the association of the Insertion/Deletion(I/D) polymorphism of angiotensin converting enzyme(ACE) gene with clinical manifestations, particularly proteinuria in children with HSP nephritis, compared with that in HSP. Methods : ACE gene polymorphism was determined in children with HSP nephritis(n=33) and HSP(n=28) who were diagnosed in Busan Paik hospital from January 1996 to June 2001. The I/D polymorphism of ACE gene was determined by PCR amplication of genomic DNA. Results : The ACE I/D genotype frequency was DD : 25%, ID : 50%, II : 25% in HSP and DD : 24 %, ID : 46%, II : 30% in HSP nephritis, there was no significant difference in the genotype and allele frequencies between two groups. When statistical analysis was done according to the presence of D allele, the amount of 24-hour urinary protein excretion and the incidence of moderate to heavy proteinuria(>$500mg/m^2/day$) at onset and last follow-up were higher in DD/ID genotype than in those in II genotype, but these differences were not statistically significant. Conclusion : We suggest a lack of association between I/D polymorphism of ACE gene and clinical manifestations in children with HSP nephritis. However, further follow-up studies based on a sufficient number of patients and long term follow up periods are necessary to confirm the role of I/D polymorphism of ACE gene in children with HSP nephritis.
Jo, Yun Ju;Lee, Eun Jeong;Choi, Kyong Min;Eun, Young Min;Yoo, Hwang Jae;Kim, Cheol Hong;Lee, Hyun Hee;Kim, Pyung Kil
Pediatric Infection and Vaccine
/
v.17
no.1
/
pp.30-35
/
2010
Purpose : We investigated the causative organism and its antibiotic susceptibility of community acquired urinary tract infection (UTI) in children at a secondary hospital to test the adequacy of the current guidelines. Methods : Children diagnosed with UTI at the Department of Pediatrics, Kwandong University MyMyongji Hospital by pyuria and bacterial growth of greater than $1.0{\times}10^5CFU/mL$ on clean catch midstream urine from January 2005 to December 2008 were studied retrospectively. The epidemiologic data, causative organism, and the antibiotic susceptibility were analyzed. Results : Sixty two children were diagnosed with sixty four cases of UTI's. Two bacteria were isolated in one case and thus data on 65 urine cultures were analyzed. The male:female ratio was 1.6:1 and 78.1% were less than 12 months of age. Escherichia coli was the predominant cause consisting of 53 cases (82.8%) of the cases. K. pneumoniae (5), Enterobacter (4), Enterococcus (1), $\beta$-streptococcus (1), Diphtheroides (1) were isolated. The antibiotic resistance of E. coli were as follows; ampicillin 69.8%, cefotaxime 1.9%, gentamicin 15.1%, amikacin 0.0%, levofloxacin 1.9%, and trimethoprim/sulfamethoxazole 26.4 %. Only one case of the E. coli was extended spectrum $\beta$-lactamase (ESBL) positive. Conclusion : Compared to prior reports from other tertiary hospitals in Korea, E. coli was the predominant cause in childhood UTI and the rate of ESBL positivity was low. The antibiotic resistance was also different compared to prior reports. We conclude that a difference in the cause and antibiotic resistance of childhood UTI exists between centers and this should be taken into consideration when prescribing antibiotics for childhood UTIs.
Bak, Sang Myeon;Park, Soo Yeon;Hur, Gyu Young;Lee, Seung Heon;Kim, Je Hyeong;Lee, Sang Yeub;Shin, Chol;Shim, Jae Jeong;In, Kwang Ho;Kang, Kyung Ho;Yoo, Se Hwa
Tuberculosis and Respiratory Diseases
/
v.54
no.1
/
pp.80-90
/
2003
Background : Goblet cell hyperplasia is a critical pathological feature in hypersecretory diseases of the airways. A bacterial infection of the lung is also known to induce inflammatory responses, which can lead to the overproduction of mucus. Recently, mucin synthesis in the airways has been reported to be regulated by neutrophilic inflammation-induced epidermal growth factor receptor (EGFR) expression and activation. In addition, it was reported that migration of the activated neutrophils is dependent on the matrix metalloproteinases (MMPs), especially MMP-9. In this study, bacterial lipopolysaccharide (LPS)-induced goblet cell hyperplasia and mucus hypersecretion by EGFR cascade, resulting from the MMPs-dependent neutrophilic inflammation were investigated in the rat airways. Methods : Pathogen-free Sprague-Dawley rats were studied in vivo. Various concentrations of LPS were instilled into the trachea in $300{\mu}{\ell}$ PBS (LPS group). Sterile PBS ($300{\mu}{\ell}$) was instilled into the trachea of the control animals (control group). The airways were examined on different days after instilling LPS. For an examination of the relationship between the LPS-induced goblet cell hyperplasia and MMPs, the animals were pretreated 3 days prior to the LPS instillation and daily thereafter with the matrix metalloproteinase inhibitor (MMPI; 20 mg/Kg/day of CMT-3; Collagenex Pharmaceuticals, USA). The neutrophilic infiltration was quantified as a number in five high power fields (HPF). The alcian blue/periodic acid-Schiff (AB/PAS) stain were performed for the mucus glycoconjugates and the immunohistochemical stains were performed for MUC5AC, EGFR and MMP-9. Their expressions were quantified by an image analysis program and were expressed by the percentage of the total bronchial epithelial area. Results : The instillation of LPS induced AB/PAS and MUC5AC staining in the airway epithelium in a time- and dose-dependent manner. Treatment with the MMPI prevented the LPS-induced goblet cell hyperplasia significantly. The instillation of LPS into the trachea induced also EGFR expression in the airway epithelium. The control airway epithelium contained few leukocytes, but the intratracheal instillation of LPS resulted in a neutrophilic recruitment. A pretreatment with MMPI prevented neutrophilic recruitment, EGFR expression, and goblet cell hyperplasia in the LPS-instilled airway epithelium. Conclusion : Matrix metalloproteinase is involved in LPS-induced mucus hypersecretion, resulting from a neutrophilic inflammation and EGFR cascade. These results suggest a potential therapeutic role of MMPI in the treatment of mucus hypersecretion that were associated with a bacterial infection of the airways.
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