• Title/Summary/Keyword: Indicator assay

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Identification of Daphne Mottle Virus Isolated from Daphne odora, a New Member of the Genus Potyvirus (서향에서 분리한 신종 포티바이러스(Daphne Mottle Virus)의 동정)

  • Park, Chung Youl;Park, Jungan;Lee, Boo-Ja;Bak, Sangmin;Lee, Hong-Kyu;Kim, Jeong-Sun;Yoon, Youngnam;Suh, Sang Jae;Lee, Su-Heon
    • Research in Plant Disease
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    • v.22 no.1
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    • pp.59-63
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    • 2016
  • A new poty-like virus was isolated from plants of winter daphne (Daphne odora) that showed virus-like symptoms on leaves, from four regions of Korea during 2014. Filamentous-shaped particles were observed by transmission electron microscopy of preparations extracted from symptomatic leaves and examined by the direct negative stain method. RT-PCR assay showed that three samples were positive for both Cucumber mosaic virus and potyvirus, and only one sample was positive for potyvirus only. A BLAST comparison to partial sequences from helper-component proteinase, cylindrical inclusion and coat protein genes detected the highest nucleotide identity of 76%, 72%, and 72% with Daphne mosaic virus, respectively, levels below the potyvirus species discrimination threshold. The new potyvirus was isolated using indicator plants (Chenopodium amaranticolor), in which local lesions were produced. In this study, we identified a novel potyvirus from winter daphne, which we have named Daphne mottle virus (DapMoV).

Cytokeratin 15 is an Effective Indicator for Progression and Malignancy of Esophageal Squamous Cell Carcinomas

  • Shen, Yu-Hong;Xu, Cui-Ping;Shi, Zhi-Meng;Zhang, Yan-Jiao;Qiao, Ya-Guang;Zhao, He-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.9
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    • pp.4217-4222
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    • 2016
  • Purpose: To compare the expression level of CK 15 in normal esophageal and esophageal squamous-cell carcinoma (ESCC) tissues and analyse possible functions of CK15 in occurrence and development. Materials and Methods: Immunohistochemistry was used to compare CK14, CK15 and proliferating cell nuclear antigen (PCNA) expression levels in ESCCs. Expression level of CK15 was also assessed by Western blotting. In addition, levels of CK15, cytokeratin 19 fragment antigen 21-1 (CYFRA21-1) and PCNA were detected in serum by enzymelinked immunosorbent assay (ELISA) and chemiluminescence methods. Relationships between clinicopathological parameters and CK14 and CK15 expression were then analyzed. Results: According to immunohistochemistry, in esophageal and intraepithelial neoplasia (SIN) tissues, the expression of CK14, CK15 and PCNA localized to basal layer of the epithelium. CK14 and CK15 levels were higher in normal esophageal squamous epithelial tissue than in SIN and ESCC, and greater in highly differentiated than poorly differentiated carcinoma tissue. By Western blotting, we found more pronounced expression of CK15 in normal esophageal tissue, compared with carcinoma tissue. The specificity of changed CK15 and CYFRA21-1 expression was respectively 90.0% and 96.7% in serum of ESCC patients. Joint detection could improve the sensitivity of esophageal carcinoma diagnosis. Relationships between CK14, CK15 expression and clinical parameters were not statistically significant (P>0.05). Postoperative survival in patients of CK14, CK15 positive expression was longer than with negative expression ($x^2=4.35$, P=0.037; $x^2=9.852$, P=0.002). Conclusions: CK15 expression decreased in esophageal squamous cell carcinoma tissue and serum of esophageal squamous carcinoma patients. We infer that CK15 may play an important role for the occurrence and development of esophageal squamous-cell carcinoma. In the future, CK15 may be used for the diagnosis, treatment and prognostic evaluation of esophageal squamous-cell carcinoma.

Screening of the total phenol content and analysis of phenolic compound in rice (Oryza saiva L.) genetic resources

  • Lee, Ji-Hee;An, Min-Jeong;Kim, Seung-Hyun;Chung, Ill-Min
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.290-290
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    • 2017
  • Rice (Oryza sativa L.) is one of the most consumed staple food crop which is energy source as carbohydrate and also is considered as the important antioxidant sources including various phenolic compounds. According to the increasing demand of healthy life, the concern to antioxidant also is increasing because of its health-promoting effect. Phenolic compounds are one of the plant secondary metabolites class, which shows various benefits to preventing or treating chronic diseases. In this study, we have measured the total phenol content from total 647 rice samples using the Floin-Ciocalteau method, and then were selected 30 rice genetic resources classified with high, middle, and low group on the basis of total phenol content. The average of the total phenol content of each group was high-group ($6892.9{\pm}488.5{\mu}g\;GAE/g$) > middle-group ($1428.1{\pm}76.0{\mu}g\;GAE/g$) > low-group ($97.6{\pm}11.4{\mu}g\;GAE/g$). The selected rice samples were analyzed with LC-MS/MS to find the composition and concentration of individual phenolic in rice grain. High-group and middle-group contained large amounts of protocatechuic acid and (+)-catechin whereas low-group showed limited amount. Among high-group samples, rice samples with black pericarp color (IT 174089, IT 220079, and IT 259958) had high content of peonidin-3-O-glucoside. Further, these black rice samples were special since polydatin, rarely found stilbenoid in rice grain, was detected. Overall, both the sum of phenolic acid and the sum of flavonoid were high-group > middle-group > low-group. Also, each group exhibited different phenolic compositions; high-group consisted of flavonoid more than phenolic acid, middle-group and low-group was comprised of phenolic acid rather than flavonoid, and non-pigmented rice was composed by fully phenolic acid. The total phenol content had positive relationships with the sum of phenolic compound (r = 0.64), the sum of flavonoid (r = 0.74) at the significance level of p < 0.0001. In addition, protocatechuic acid and quercetin showed positive correlation with above phenolic composition parameters; in order, r = 0.98, 0.65 for protocatechuic acid and r = 0.73, 0.78 for quercetin (p < 0.0001). In conclusion, the total phenol content assay showed the possibility of utilization as a phenolic composition indicator in rice grain. Also, this result was suggested study pigment on other material.

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Diurnal Variations in Milk and Blood Urea Nitrogen and Whole Blood Ammonia Nitrogen in Dairy Cows

  • Hwang, Sen-Yuan;Lee, Mei-Ju;Peh, Huo-Cheng
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.12
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    • pp.1683-1689
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    • 2001
  • The levels of urea nitrogen both in blood (BUN) and milk (MUN), and milk protein (MP) reflect protein and energy intake in dairy herd feeding. Blood and milk constituents may be changes rhythmically and influence by different sampling time within a day and after feeding. Trials were conducted using five dietary treatments in both lactating and dry cows to study the effects of sampling time on concentrations of BUN, MUN and whole blood ammonia nitrogen (BAN) in practical dairy cow feeding in Taiwan. The conventional feed ingredients and forages including corn silage, alfalfa hay, timothy or pangola hay and corn grain were used as major source of the diet to follow practical dairy cow feeding. Five different diets were varying in amounts (low=L; standard=S; high=H) of crude protein (P) and energy (E) according to the NRC (1989). The energy to protein ratios in kcal/kg for the PSES, PLES, PHES, PSEH and PSEL were 10.82, 12.54, 9.41, 12.53 and 9.13 in lactating cows, and 11.38, 13.33, 9.78, 13.28 and 9.74 in dry cows, respectively. Results showed that after feeding at 9:30, BUN reached peak at 13:30 and was significantly higher than those to that sampled at 14:30 to 18:30 (p<0.05) in dry cows. Therefore the best blood sampling time for urea nitrogen assay in dry cows is 4 hours after morning feeding. In lactating cows, BUN of 13:30 was significantly higher than those of 8:30 to 11:30 (p<0.05), but there were no significant difference between the BUN values of other sampling time. Hence the suitable blood sampling time for BUN value in lactating cows was located on 3 to 8 hours after morning feeding, but the best time was 4 hours after morning feeding. MUN content is significantly higher in the afternoon collected bulk milk than the fore-strip morning milk (p<0.05), therefore the best sampling time for MUN is from afternoon collected bulk milk. Diurnal BAN changed without traceable rhythmic pattern and was negatively correlated to the BUN (r = -0.78). It is suggested that BAN may not be a good indicator for monitoring dairy cow feeding.

Characterization of Prunus necrotic ringspot virus Isolate from Peach in Korea (국내 복숭아에서 분리한 Prunus necrotic ringspot virus의 특성)

  • Kim, Hyun-Ran;Lee, Sin-Ho;Shin, Il-Sheob;Kim, Jeong-Hee;Cho, Kang-Hee;Heo, Seong;Kim, Jeong-Soo;Choi, Yong-Mun
    • Research in Plant Disease
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    • v.15 no.3
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    • pp.170-174
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    • 2009
  • In this paper, we report a characterization of Prunus necrotic ringspot virus (PNRSV) isolate. The virus was identified from 'Yumyeong' peach showing mild mosaic on leaves in commercial orchard of 'Umsung', Chungbuk province in Korea. The virus isolate produced ringspot symptom on the inoculated cotyledons and systemic mosaic and malformation on the upper leaves of Cucumis sativus. Systemic mottles were appeared in Chenopodium quinoa. When the buds of the virus infected stem were grafted on the healthy young Prunus persica GF305 seedlings, line pattern with mosaic appeared within 3 months. Isometric virus-like particles were found in parenchyma cells and plasmodesmata of C. sativus leaves inoculated mechanically with the virus. The cDNA fragments of PNRSV coat protein (CP) region, approximately 675bp, were synthesized from genomic RNA extracted from virus-infected leaves by RT-PCR using specific primer pairs. Partial nucleotide sequences of the CP regions were determined and analyzed with the known PNRSV. The CP gene of PNRSVKorea isolates showed 93.9~94.7% similarity to the 4 known PNRSV isolates.

Adverse Reproductive Effects on Plasma Vitellogenin and Sex Steroid Levels, and Gonadosomatic Index in Juvenile Common Carps (Cyprinus carpio) Exposed to 17$\beta$-Estradiol and D-2-Ethylhexyl Phthalate

  • Seo, Jinwon;Park, Kyung-Seo;Moon, Woon-Gi;Lee, Sung-Kyu
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2002.11a
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    • pp.141-144
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    • 2002
  • Environmental estrogens are natural or synthetic substances present in the aquatic environment, especially in effluent from sewage treatment. However, the adverse effects of these estrogenic substances on fish reproduction are unknown. Di-2-ethylhexyl phthalate (DEHP) is the most common phthalate, which Ps used as a plasticizer in polyvinylchloride (PVC), and it is widespread in the environment and has been found in aquatic organisms and sediments. Therefore, juvenile common carps (Cyprinus carpio) were exposed to nominal concentrations of 17$\beta$-estradiol (E2) (0.5, 5, 50 $\mu\textrm{g}$/L) and DEHP (10, 100, 500 $\mu\textrm{g}$/L) for 21 days, to determine the adverse reproductive effects of these compounds on plasma vitellogenin (VTG) induction, sex steroid level, and gonad weight. Electrophoresis (SDS-PAGE) revealed that much of VTG was induced in fish exposed to 5 and 50 E$_2$ $\mu\textrm{g}$/L, but none of DEHP exposure showed induction. Enzyme-linked immunosorbent assay (ELISA) revealed that VTG was significantly induced in fish exposed to 5 and 50 E$_2$ $\mu\textrm{g}$/L, and combination of 50 E$_2$ $\mu\textrm{g}$/L with 10 and 500 DEHP $\mu\textrm{g}$/L, but none of DEHP exposure showed induction. Analysis of sex steroid levels in some fish revealed that testosterone (T) was detected in both male and female fish of the control and DEHP exposures, but none of fish exposed to 22 concentrations had detectable testosterone level. On the other hand, E$_2$ exposure induced 17$\beta$-estradiol in plasma of male fish, but there was no induction of 17$\beta$-estradiol in plasma of male fish exposed to DEHP. Comparison of gonadosomatic index (GSI) revealed that maximal E$_2$ exposure inhibited ovarian growth, but maximal DEHP exposure stimulated testicular growth. The results indicated that those comparisons can be a useful bio-indicator for determining adverse reproductive effect of endocrine disrupting chemicals (EDCs).

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Antimicrobial Activities of Root Surfaces Treated with Tetracycline-containing gel and a Mixture of tetracycline and citric acid-containing gel;in vivo study (테트라싸이클린 및 테트라싸이클린-구연산 혼합젤로 처리한 치근면의 항 미생물 활성 변화에 관한 연구;In Vivo Study)

  • Cheong, Hee-Sun;Han, Soo-Boo;Nam, Seok-Woo;Shim, Chang-Koo;Kye, Seung-Beom
    • Journal of Periodontal and Implant Science
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    • v.27 no.1
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    • pp.79-90
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    • 1997
  • The purpose of this study was to evaluate the substantivity of experimentally developed gel type tetracycline HCl and a mixture of tetracycline-citric acid gel, and compare to those of solution type tetracycline HCl. 11 extracted anterior teeth were subjected to this study. After scaling and root planing teeth were randomly divided into 3 treatments groups : group 1; 3 teeth were irrigated with tetracycline HCl(50mg/ml) solution , group 2; tetracycline gel (5%) was inserted in the periodontal pockets of 3 teeth, group 3; a mixture of tetracycline and citric acid gel was inserted in the pockets of 3 teeth. And 2 teeth treated in 0.9 % sterile saline served as controls. After 5-minute exposure, each tooth immediately extracted and incubated at room temperature for 22 days in tris-buffered saline as a desorption media. The total volume of TBS was removed and replaced with fresh TBS, at 24-h intervals. Removed desorption media transferred to a sterile vial and stored at -70 oC. This procedure was repeated every 24 h throughout the 22-day desorption period. Using Porphyromonas gingivalis as an indicator organism, a microtiter assay was used to evaluate antimicrobial activity desorbed from the teeth. 1. 50mg/ml tetracycline HCl solution exhibited the longest antimicrobial activity. Compared to saline treated group, it showed significant difference on the day 1 and day 2 desorption period. 2. The ODs of 5% tetracycline gel and a mixture of tetracycline-citric acid gel were significantly different during the first 24 hour only. 3. There was no statistically significant difference after the day 3 between the groups.(p<0.05). Despite our expectation a mixture of tetracycline-citric acid gel did not show longer antimicrobial activities than those of tetracycline gel, and the solution type exhibited the longest activities. Because the gel type agents may stay in the subgingival environment longer than the solution, if the teeth were not extracted immediately after the delivery of the agent, the result could be different. hus this result suggests the possibilities of practical use of these kind of gel type agents.

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A Study on the Development of Urine Analysis System using Strip and Evaluation of Experimental Result by means of Fuzzy Inference (스트립을 이용한 요분석시스템의 개발과 퍼지추론에 의한 검사결과 평가에 관한 연구)

  • Jun, K. R.;Lee, S. J.;Choi, B. C.;An, S. H.;Ha, K.;Kim, J. Y.;Kim, J. H.
    • Journal of Biomedical Engineering Research
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    • v.19 no.5
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    • pp.477-486
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    • 1998
  • In this paper, we implemented the urine analysis system capable of measuring a qualitative and semi-quantitative and assay using strip. The analysis algorithm of urine analysis was adopted a fuzzy logic-based classifiers that was robust to external error factors such as temperature and electric power noises. The spectroscopic properties of 9 pads In a strip were studied to developing the urine analysis system was designed for robustnesss and stability. The urine analysis system was consisted of hardware and software. The hardware of the urine analysis system was based on one-chip microprocessor, and Its peripherals which composed of optic modulo, tray control, preamplifier, communication with PC, thermal printer and operating status indicator. The software of the urine analysis system was composed of system program and classification program. The system program did duty fort system control, data acquisition and data analysis. The classification program was composed of fuzzy inference engine and membership function generator. The membership function generator made triangular membership functions by statical method for quality control. Resulted data was transferred through serial cable to PC. The transferred data was arranged and saved be data acquisition program coded by C+ + language. The precision of urine analysis system and the stability of fuzzy classifier were evaluated by testing the standard urine samples. Experimental results showed a good stability states and a exact classification.

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Etiological Properties and Coat Protein Gen Analysis of Potato Virus Y Occuring in Potatoes of Korea (우리나라 감자에 발생하는 PVY의 병원학적 특성 및 외피단백질 유전자 분석)

  • ;Richard M. Bostock
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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    • pp.77-96
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    • 1995
  • To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

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Clinical Characteristics, Prognostic Factors and Influence of Prophylaxis in Children with Pneumocystis jirovecii Pneumonia (소아 주폐포자충 폐렴의 임상양상, 위험인자 및 예방요법의 효과 연구)

  • Kim, Seohee;Yoo, Reenar;Sung, Hungseop;Lee, Jina
    • Pediatric Infection and Vaccine
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    • v.23 no.1
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    • pp.31-39
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    • 2016
  • Purpose: The aim of this study was to investigate the prognostic factors for Pneumocystis jirovecii pneumonia (PCP) and to evaluate the influence of PCP prophylaxis in pediatric patients. Methods: From January 2002 to April 2015, patients aged <18 years with a diagnosis of confirmed PCP at our institute were reviewed retrospectively. Clinical characteristics and outcomes were compared according to the groups with or without PCP prophylaxis. Risk factors associated with PCP-related death were analyzed by logistic regression analysis. Results: During study period, a total of 24 patients were diagnosed with PCP by immunofluorescence assay and/or PCR. The median age of the patients was 5 years (range, 3 months-18 years) and 23 (96%) had immunocompromised conditions including hematologic disorders with or without hematopoietic stem cell transplantation (n=15), solid organ transplantation (n=4), and primary immune deficiency (n=4). Most common presenting symptoms were tachypnea and cough (92%, each). At the time of diagnosis, 79% (19/24) and 25% (6/24) suffered from respiratory failure and multi-organ dysfunction syndrome (MODS), respectively. Mechanical ventilation was required in 8 (33%) patients and 5 (21%) patients died of PCP. Multivariate analysis showed that MODS at initial presentation was an indicator of poor prognosis (OR, 17.1 [95% CI 1.13-257.67]; P=0.04). Compared to the patients without PCP prophylaxis, the frequency of MODS at diagnosis, need for mechanical ventilation and length of hospital days were significantly less common in the children who received PCP prophylaxis. Conclusions: MODS at presentation was a significant predictor for poor outcome and PCP prophylaxis could alleviate the clinical courses of pediatric PCP. Prospective study will be mandatory to determine the risk factors for development and deterioration of PCP in children.