• Title/Summary/Keyword: cationic protein

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The Effect of Root Exudate and Chemotaxis on Host Recognition in Soybean-Bradyrhizobium Symbiosis (대두(大豆)-근류균(根瘤菌) 공생(共生)에서 뿌리분비물(分泌物)과 화학주성(化學走性)이 숙주인식(宿主認識)에 미치는 영향(影響))

  • Kang, Sang Jai;Park, Woo Churl
    • Current Research on Agriculture and Life Sciences
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    • v.11
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    • pp.121-132
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    • 1993
  • This study was carried out to research the effect of the chemotaxis of Bradyrhizobium japonicum KCTC 2422 and its mutant toward soybean root exudate and to elucidate the effect of the lectin of host specificity (Host Recognition) in soybean-Bradyrhizobium symbiosis. The results obtained were as follows: The homogeneities of the purified lectins from soybean and pea seed was ascertained chromatographically and electrophoretically. Gel electrophoresis of soybean lectin in the presence of sodium dodecyl sulfate appeared a single protein band, whereas pea lectin appeared two protein bands. Soybean lectin from 2 cultivars formed immunoprecipitin arcs at same position with anti-soybean lectin rabbit IgG, but pea lectin did not form immunoprecipitin lines with anti-soybean lectin rabbit IgG. Chemotactic responses of KCTC 2422, LPN-100 and LCR-101 toward proline in capillary assays were 3.1, 1.3 and 1.0-fold above background, respectively. The chemotactic responses of KCTC 2422, LPN-100, and LCR-101 toward Paldal crude root exudate in capillary assays were 3.5, 1.4 and 1.4-fold above background, respectively. The present work shows that B. japonicum and its mutants are capable of very different responses toward root exudate fraction. The chemotactic responses of KCTC 2422 was most with neutral fraction, least with anionic fraction and intermediate with cationic fraction. The nitrogenase activity of soybean nodule was shown in 15days after inoculation with LCR-101. However, we couldn't find out the nodules when soybean was inoculated with LPN-100. From these result we can suppose that the chemotaxis of Bradyrhizobium plays inportant the role of forming the nodule (host recognition) in the soybean-B. japonicum symbiosis.

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Biochemical Analysis of Interaction between Kringle Domains of Plasminogen and Prion Proteins with Q167R Mutation

  • Lee, Jeongmin;Lee, Byoung Woo;Kang, Hae-Eun;Choe, Kevine K.;Kwon, Moosik;Ryou, Chongsuk
    • Journal of Microbiology and Biotechnology
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    • v.27 no.5
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    • pp.1023-1031
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    • 2017
  • The conformational change of cellular prion protein ($PrP^C$) to its misfolded counterpart, termed $PrP^{Sc}$, is mediated by a hypothesized cellular cofactor. This cofactor is believed to interact directly with certain amino acid residues of $PrP^C$. When these are mutated into cationic amino acid residues, $PrP^{Sc}$ formation and prion replication halt in a dominant negative (DN) manner, presumably due to strong binding of the cofactor to mutated $PrP^C$, designated as DN PrP mutants. Previous studies demonstrated that plasminogen and its kringle domains bind to PrP and accelerate $PrP^{Sc}$ generation. In this study, in vitro binding analysis of kringle domains of plasminogen to Q167R DN mutant PrP (PrPQ167R) was performed in parallel with the wild type (WT) and Q218K DN mutant PrP (PrPQ218K). The binding affinity of PrPQ167R was higher than that of WT PrP, but lower than that of PrPQ218K. Scatchard analysis further indicated that, like PrPQ218K and WT PrP, PrPQ167R interaction with plasminogen occurred at multiple sites, suggesting cooperativity in this interaction. Competitive binding analysis using $\small{L}$-lysine or $\small{L}$-arginine confirmed the increase of the specificity and binding affinity of the interaction as PrP acquired DN mutations. Circular dichroism spectroscopy demonstrated that the recombinant PrPs used in this study retained the ${\alpha}$-helix-rich structure. The ${\alpha}$-helix unfolding study revealed similar conformational stability for WT and DN-mutated PrPs. This study provides an additional piece of biochemical evidence concerning the interaction of plasminogen with DN mutant PrPs.

The Effect of Glucocorticoid on the Change of Nitric Oxide and Cytokine Levels in Induced Sputum from Patients with Bronchial Asthma (기관지 천식 환자에서 부신피질 스테로이드 투여 전후 유도객담내 Nitric Oxide 및 Cytokine의 변화)

  • Kim, Tae-Yon;Yoon, Hyeong-Kyu;Choi, Young-Mee;Lee, Sook-Young;Kwon, Soon-Seog;Kim, Young-Kyoon;Kim, Kwan-Hyoung;Moon, Hwa-Sik;Park, Sung-Hak;Song, Jeong-Sup
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.6
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    • pp.922-931
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    • 2000
  • Background : It has been well known that bronchia1 asthma is a chronic airway inflammatory disorder. Recently, sputum induced with hypertonic saline was introduced as a simple and useful nonivasive medium to investigate airway inflammation and symptom severity in patients with asthma. We examined the eosinophil, eosinophil cationic protein (ECP), interleukin(IL)-3, IL-5, granulocyte-macrophage colony-stimulating facta (GM-CSF), and nitric oxide (NO) derivatives in induced sputum from patients with bronchia1 asthma in order to determine the role of NO and various inflammatory cytokines as a useful markers of airway inflammation or changes in pulmonary function tests and symptoms. Methods : A total 30 patients with bronchia1 asthma received oral prednisolone 30 mg daily for 2 weeks. Forced expiratory volume in one second ($FEV_1$), total blood eosinophil count and induced sputum eosinophil count, ECP, IL-3, IL-5, GM-CSF, and NO derivatives were determined before and after the administration of prednisolone. Results : Of the 30 patients, 13 (43.3%) were male and 17 (56.7%) were female. The mean age of patients was 41.8 years (range 19-64 years). Two patients could not produce sputum at the second study and 3 could not be followed up after their first visit. Two weeks after the prednisolone administration, there was a significant increase in $FEV_1$ (% of predicted value) from 78.1$\pm$20.6 % to 90.3$\pm$ 18.3 % (P<0.001). The eosinophil percentages in induced sputum were significantly decreased after treatment with prednisolone, with values of 56.1$\pm$27.2 % versus 29.6$\pm$21.3 % (P<0.001), and ECP were $134.5\pm68.1\;{\mu}g/L$ versus $41.5\pm42.4\;{\mu}g/L$ (P<0.001) respectively. After the prednisolone treatments, the eotaxin concentration also showed a decreasing tendency from 26.7$\pm$12.8 pg/ml to 21.7$\pm$8.7 pg/ml. There was a decreasing tendency but no significant differences in total blood eosinophil count (425.7$\pm$265.9 vs 287.7$\pm$294.7) and in the concentration of NO derivatives ($70.4\pm44.6{\mu}mol/L$ vs $91.5\pm48.3\;{\mu}mol/L$) after the prednisolone treatments. IL-3, IL-5, GM-CSF were undetectable in the sputum of most subjects either before the prednisolone treatments or after the treatments. Before the prednisolone treatments, a significant inverse correlation was observed between FEV1 and sputum ECP (r=-D.364, P<0.05) and there was a significant correlation between sputum eosinophils and eotaxin (r=0.369, P<0.05) Conclusion : The eotaxin and ECP concentration in induced sputum may be used as markers of airway inflammation after treatments in bronchia1 asthma. In addition, the measurement of sputum eosinophil percent ages is believed to be a simple method displaying the degree of airway inflammation and airway obstruction before and after the prednisolone treatment in bronchia1 asthma. However, unlike exhaled NO, the examination of NO derivatives with Griess reaction in induced sputum is considered an ineffective marker of changing airway inflammation and obstructing symptoms.

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Therapeutic Effects of Probiotics in Patients with Atopic Dermatitis

  • Yim, Jun-Hee;Kim, Duk-Han;Ku, Ja-Kyung;Kang, Yoon-Sung;Kim, Mi-Yeon;Kim, Hyung-Ok;Chung, Myung-Jun;Park, Young-Min
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1699-1705
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    • 2006
  • Recent studies have suggested that oral bacteriotherapy with probiotics might be useful for preventing and managing childhood atopic dermatitis (AD). The purpose of this investigation was to evaluate the efficacy and safety of oral treatment with probiotics for adolescent and adult AD patients as well as for childhood AD patients. Sixty-four patients with mild to moderate AD were recruited for treatment with a mixture of four probiotic strains (Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus casei, and Biftdobacterium lactis) twice daily for 8 weeks. The degree of pruritus was determined by a 10-point visual analog scale every other week, and the patients' global assessments of their clinical responses (i.e., better, unchanged, or worse) was done at the end of intervention. The clinical severity of the eczema was evaluated by eczema area and severity index (EASI) score every other week. As laboratory markers, total immunoglobulin E (IgE), eosinophil cationic protein (ECP) in the serum, and cytokine production [interleukin-4 (IL-4), interleukin-10 (IL-10), and $interferon-{\gamma}\;(IFN-{\gamma})$ by the peripheral blood mononuclear cells (PBMCs) were measured at the beginning and at the end of intervention. Of the 64 enrolled AD patients, only 50 patients finally completed the 8-week study. After 8-week treatment with probiotics, the EASI score was significantly improved (p<0.0001), 50% of the patients experienced improvement of their eczema, and significant improvement of the pruritus was also observed (p=0.0002). The effect was more pronounced for the patients with very high IgE levels (>1,000 ku/l) or for the patients with moderate disease severity. There was no significant difference in the therapeutic effects between the childhood AD and adolescent and adult AD patients. There were no significant changes of cytokines, as well as the total IgE and ECP levels, in the patients' serum. Treatment with the mixture of four probiotic strains was generally well tolerated. Our results suggest that the treatment with the mixture of four probiotic strains is beneficial for the management of the adolescent and adult AD patients, as well as for the childhood AD patients.

Biodistribution of [S-35] Labeled Antisense Oligodeoxynucleotides Increased Tumor Targeting With Microsphere Coinjection

  • Choe, Jae-Gol;Park, Gil-Hong;Claudio Nastruzzi;Yoon S. Cho-Chung;Kim, Meyoung-Kon
    • Environmental Mutagens and Carcinogens
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    • v.22 no.2
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    • pp.65-69
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    • 2002
  • To elucidate the effect of microsphere coinjection on the administration of oligodeoxynucleotides (ODN), we have investigated biodistribution of [S-35]-labeled antisense ODN targeted to cAMP-dependent protein kinase (PKA) RI-$\alpha$ subunit in nude mice xenografted with WiDr (human colon cancer, ATCC CCL218). The strategy of using microsphere has been proposed for cancer treatment as a carrier of therapeutic ODN so that it could offer an advantage with respect to maintaining constant ODN levels in blood and obtaining higher therapeutic ODN concentration at tumor sites. Comparative biodistribution studies were performed in nude mice (female, 20 g of body weight, n = 4-6) xenografted with WiDr cancer cells, when 0.1 $\mu$Ci (specific activity, 2.94 mCi/$\mu$mole) of [S-35]-labeled RI-$\alpha$ antisense ODN was injected alone or with microsphere (PLG-18, polylactic copolymer with cationic surfactant DDAB18). Peak tumor uptake of [S-35]-labeled ODN was significantly increased from 17.7% (at 6 h) of injected dose per gram of tissue (ID/g) to 42.5% (at 24 h) ID/g when microsphere was coinjected with ODN. The different biodistribution in the kidney accumulation (e.g., 100.2% ID/g for ODN alone and 54.9%/ID/g for microshpere coinjection) may contribute to higher blood concentration (e.g., 21.5%ID/$m\ell$ for ODN alone and 37.5%ID/$m\ell$ for microsphere coinjection) of radiolabeled ODN. Of importance is the fact that the whole body retention of radioactivity increased with microsphere coinjection from 50.8%ID/g to 68.0%ID/g after 24-h of injection. This decreased kidney accumulation and increased whole body retention of [S-35]-labeled ODN resulted in a significant improvement of ODN targeting to the tumor site. In conclusion, the coinjection of microsphere appears to be an important carrier system in vehiculation of antisense oligonucleotide to the tumor tissue in vivo.

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Comparison of Eosinophil Markers between Acute and Recovery Stages in Children with Mycoplasma pneumoniae Pneumonia (Mycoplasma pneumoniae 폐렴 환아에서 급성기 및 회복기의 호산구 지표의 비교)

  • Nah, Kyu Min;Kang, Eun Kyeong;Kang, Hee;Park, Yang;Koh, Young Yull
    • Clinical and Experimental Pediatrics
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    • v.45 no.10
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    • pp.1227-1233
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    • 2002
  • Purpose : Several studies have shown that increases of eosinophil markers are common findings of asthma and Mycoplasma pneumoniae infection, and eosinophil markers reflect the clinical stage of asthma. The purpose of this study was to investigate the change of eosinophil markers according to the clinical stage of Mycoplasma pneumonia. Methods : The patient group consisted of 33 outpatient children with Mycoplasma pneumonia. Peripheral blood total eosinophil count(TEC) and serum eosinophilic cationic protein(ECP) level were measured at both acute and recovery stages and were compared between both stages. The patient group was subdivided into the wheezing(n=16) and the nonwheezing group(n=17), and the TECs and the ECPs of one group were compared with those of the other group. The correlation between Mycoplasma antibody titer and the eosinophil markers of acute stage were analyzed. Results : In the whole patient group, the TECs and the ECPs of the acute stage were significantly higher than those of the recovery stage(P=0.018, P=0.005), but there were no differences in the TEC and the ECP between the wheezing and the nonwheezing group. In the wheezing group, there were no significant differences in the TEC and the ECP between acute and recovery stages. There were no correlations between acute stage Mycoplasma antibody titer and the eosinophil markers. Conclusion : Eosinophil markers reflect the clinical stage of Mycoplasma pneumonia and eosinophilic inflammations may continue even after the acute stage in wheezing patients with Mycoplasma pneumonia.

A Low Irritant Liquid Cleanser Composition Developed by Multi-Screening Methods (다탐색(多探索)법을 통한 저자극성 액체 세정제 조성물 개발)

  • Kim Peter;Hyeon Ki-An;Chung Ji-Youn;Yoon Sam-Sook;Kang Han Chyul;Park Sun Hee;Ko King Il;Kim Ki Ho
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.51-58
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    • 2005
  • Alkyl ethoxy sulfate type surfactants, widely used in commercial cleansers, are easily adsorbed to skin to often cause skin irritation and inflammation if not thoroughly rinsed nut. In order to replace or complement existing surfactants, we screened the existing surfactants through protein denaturation method, cell cytotoxicity assay and human IL-1$\alpha$ assay, etc. Fourteen surfactants have been chosen from among too irritant anionic, cationic and/or zwitter-ionic ones and investigated for cell cytotoxicity in human fibroblast cell lines using monolayer culture with the thirteen commercially available cleansers for sensitive skin. From these results, we selected 5 surfactants and 2 commercial cleansers (names not shown), such as sodium laureth sulfate (anionic), sodium cocoyl isethionate (anionic), sodium lauroamphoacetate (zwitter-ionic), and cocamidopropyl betaine (zwitter-ionic), alkyl polyglycoside (non-ionic). 20 formulations were made out of 5 surfactants and five of them were chosen through a protein denaturation method (lower than 3 M sodium dodecyl sulfate solution ($13.2\%$)), cell cytotoxicity and human patch test. These five selected formulations containing preservatives were compared to two selected commercial cleansers by cell cytotoxicity and human IL-1$\alpha$ ELISA assay using dermal equivalent. Finally, we selected the best formulation. To this formulation, fructan ($3\%$ or $5\%$) or/and portulaca extract ($3\%$ or $5\%$) well known for its anti-inflammatory and moisturizing effects were added and investigated for cell cytotoxicity using dermal equivalent. In cytotoxicity assay using dermal equivalent, two formulations containing $5\%$ fructan and $3\%$ or $5\%$ portulaca extract were less toxic than the others. In cytotoxicity assay and human IL-1$\alpha$ ELISA using 3D culture, the selected formulation containing $5\%$ fructan and $5\%$ portulaca extract showed better efficiency than those of the others and 2 commercial cleansers. As a result, we could develop a low irritant and safe liquid cleanser.

Utility of a forced expiratory flow of 25 to 75 percent as a predictor in children with asthma (소아 천식에서 최대호기중간유량의 기관지 과민성 예측인자로서의 의의)

  • Kang, Jung Wan;Kim, Kyung Won;Kim, Eun Soo;Park, Jun Young;Sohn, Myung Hyun;Kim, Kyu-Earn
    • Clinical and Experimental Pediatrics
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    • v.51 no.3
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    • pp.323-328
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    • 2008
  • Purpose : Asthma is defined as chronic inflammation of the lower small airways, and bronchial hyperreactivity (BHR) is a pathophysiologic feature of asthma. It has been proposed that although there is no direct variable capable of assessing the small airways, a forced expiratory flow of between 25 and 75 percent ($FEF_{25-75}$) might be considered a more sensitive early marker of small airway obstruction than the forced expiratory volume in 1 second ($FEV_1$). Thus, we proposed that the presence and degree of positive responses to bronchial methacholine testing were related to the difference (DFF) and ratio (RFF) between $FEV_1$ and $FEF_{25-75}$ in asthmatic children. Methods : The subjects were 583 symptomatic children, including 324 children with BHR and 259 controls. Pulmonary function tests, methacholine challenge tests, and skin prick tests were performed, and the total eosinophil count, total serum IgE, and serum eosinophil cationic protein level were measured in all subjects. From a concentration-response curve, the methacholine concentration required to produce a decrease of 20% from post-saline $FEV_1$ was calculated ($PC_{20}$). Results : The median DFF and RFF values decreased in controls compared to subjects with bronchial hyperresponsiveness, and this trend was found in groups ranked by its severity. $PC_{20}$ had a negative correlation with DFF and RFF. Cutoff values of 0.5 for DFF and 1.042 for RFF were identified, and sensitivity and specificity were calculated. Conclusion : This study revealed that DFF and RFF might be predictive of bronchial hyperresponsiveness in the context of normal $FEV_1$ in children.

The Significances of Chest X-ray Findings and Serum IFN-γ, IL-5, ECP as Predictors of Prognosis of Bronchiolitis and Bronchopneumonia in Infants (영아의 세기관지염과 기관지 폐렴의 예후 지표로서 혈청 IFN-γ, IL-5 및 ECP치와 흉부 방사선 소견의 의의)

  • Cho, Su Hyun;Lee, Hyun Seung;Lee, Mi Hee;Lee, Joon Sung
    • Clinical and Experimental Pediatrics
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    • v.46 no.3
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    • pp.230-235
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    • 2003
  • Purpose : Lower respiratory tract infections in infant and young children are often due to a virus, especially the Respiratory syncytial(RS) virus. Chest X-ray findings in bronchiolitis and bronchopneumonia are different. The radiographic hallmark of bronchiolitis is pulmonary hyperinflation and similar to that of bronchial asthma. Bronchiolitis is predisposed to later development of bronchial asthma. To evaluate the difference of immuno-pathophysiology between bronchiolitis and bronchopneumonia, we measured $IFN-{\gamma}$(Th1 cytokine), IL-5(Th2 cytokine) and ECP. We also investigated whether X-ray findings in infants with viral infected respiratory disease are useful in predicting the development of asthma. Methods : We measured IL-5, ECP, $IFN-{\gamma}$ levels in serum from 21 infants with bronchiolitis and 21 infants with bronchopneumonia and 16 infants without pulmonary viral diseases. Results : IL-5 levels of bronchiolitis and bronchopneumonia were significantly higher than those of the control(P=0.02, P=0.042). IL-5 levels of bronchiolitis were higher than those of bronchopneumonia but there was no significant difference. $IFN-{\gamma}$ levels of bronchopneumonia were higher than those of bronchiolitis but there was no significant difference. ECP levels of bronchiolitis and bronchopneumonia were higher than those of the control but only those of bronchiolitis were significantly higher than those of the control(P=0.045). IL-5 and ECP levels did not show any significant correlation in bronchiolitis, bronchopneumonia and control groups. Conclusion : We cannot prove the distinct differences in serum Th1/Th2 cytokine profiles between bronchiolitis and bronchopneumonia in infants. These results suggest that the different findings on chest X-ray between bronchiolitis and bronchopneumonia could not be a predictor of later development of asthma.

Asthma predictive index in children with recurrent wheezing (반복성 천명을 가진 소아에서 천식 발생 예측 지표의 적용)

  • Jang, Joo Young;Kim, Hyo Bin;Lee, So Yeon;Kim, Ja Hyung;Kim, Bong Seong;Seo, Hee Jung;Hong, Soo-Jong
    • Clinical and Experimental Pediatrics
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    • v.49 no.3
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    • pp.298-304
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    • 2006
  • Purpose : We compared the asthma predictive index(API) and the modified asthma predictive index (mAPI) of the Tuscon Children's Respiratory Study Group in Korean children with recurrent wheezing. We investigated the atopic profiles and presence of allergen sensitization of each risk group, and ascertained the significant clinical risk factors. Methods : Two hundred and sixty two children, who visited for recurrent wheezing from 1998 to 2005, were enrolled and divided into groups by API and mAPI. We investigated the history of the patients and their families, atopic profiles, and sensitization to aeroallergen and food allergens. Twenty nine children were followed up to 6 years of age and we evaluated the sensitivity, specificity and positive and negative predictive value of both indices. Results : The high risk group of API were of older age, were more likely to be sensitized to aeroallergen(P=0.001) and food allergen(P=0.034) and had higher levels of total eosinophil count, eosinophil percent, serum ECP, total IgE, and D.p-, D.f-specific IgE. High risk group of mAPI showed higher levels of atopic markers such as egg-, milk-, D.p- and D.f-specific IgE. Even though API did not include allergen sensitization, the high risk group was more significantly sensitized to common allergens than the low risk group. Twenty nine children were followed up until 6 years of age; therefore 15 children were diagnosed as asthma, clinically. The sensitivity, specificity, positive and negative predictive values of mAPI were higher than API. Conclusion : Both high risk groups of API and mAPI had higher levels of atopic markers and were more sensitized to common allergens. These findings suggest that sensitization to aeroallergens and food allergens are more objective markers as asthma predictive indices. In addition, mAPI is a more reliable index in predicting asthma in Korean children with recurrent wheezing than is API. But only 29 patients were followed until the age of 6, so we need to include more children with long term follow up for future study.