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Growth Inhibition of Diatom, Chaetoceros calcitrans by Marine Bacteria, Shewanella sp. SR-14 - Effects of Marine Bacteria on the Changes of Fatty Acid Composition of Diatoms - (해양세균, Shewanella sp. SR-14에 의한 규조류 Chaetoceros calcitrans의 증식저해 - 해양세균이 규조류의 지방산 조성변화에 미치는 영향 -)

  • KIM Ji Hoe;YOON Ho Dong;PARK Hee Yun;LEE Hee Jung;CHANG Dong Suck
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.1
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    • pp.24-29
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    • 2003
  • In the previous reports, the authors isolated two strains of marine bacteria, Shewanella sp. SR-14, which has Chaetonros sp. growth inhibition activity, and Vibrio alginolyticus, that did not affect growth of the alga. In the present study, fatty acid compositions of diatoms, Chaetoceros calcitrans and Skeletonema costatum, and marine bacteria, Shewanella sp. SR-14 and V. alginolyticus, were analyzed. Changes of fatty acid composition in the diatoms grown with the marine bacteria were also determined. Major fatty acids of Sbewanella sp. SR-14 were 16:1n-7 $(29.4\%)$ and 16:0 $(19.2\%)$ during incubation in peptone broth at $20^{\circ}C$ for 3 days. The compositions of V. alginolyticus detected were 16:0 $(23.7\%),$ 16:1n-7 $(27.7\%)$ and 18:1n-7 $(21.0\%).$ C. calcitrans consisted of 16:1n-7 $(33.3\%),$ 16:0 $(27.1\%)$ and 14:0 $(12.1\%).$ S. costatum mainly contained 16:1n-7 $(28.9\%),$ 16:0 $(21.6\%)$ and 20:5 $(19.8\%).$ When halves of cell numbers of C. calcitrans were moribund cells by Shewanella sp. SR-14, the C. calcitrans and S. costatum simultaneously cultured with the bacteria were harvested by filtration with GE/D glass microfibre filter. In the fatty acid composition of both diatoms, saturated fatty acid contents in both diatoms grown with Shewanella sp. SR-14 were decreased, but unsaturated fatty acid contents were increased. The differences were greater in C. calcitrans than those in S. costatum. During the growth of diatoms with V. alginolyticus, C. calcitrans showed increase of saturated fatty acid contents and decrease of unsaturated fatty acid contents; however, S. costatum did not show sharp difference in fatty acid content. In this study, Shewanella sp. SR-14, which showed growth inhibition activity against C. calcitrans, influenced on the changes of fatty acid contents in the diatom. It was suggested that increased unsaturated fatty acid was synergistically activated algal growth inhibition activity of Shewanella sp. SR-14.

Comparison of Stress Response in Diallel Crossed Korean Domestic Chicken Breeds (토종 종계를 이용한 이면 교배조합 계통 간 스트레스 반응정도 비교 분석)

  • Cho, Eun Jung;Park, Ji Ae;Choi, Eun Sik;Sohn, Sea Hwan
    • 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.

Esophagectomy for Esophageal Cancer in Elderly Patients Over 70 Years of Age (70세 이상의 식도암 환자에시 식도절제술 및 재건술)

  • Song Suk-Won;Lee Hyun-Sung;Kim Moon Soo;Lee Jong Mog;Zo Zae Ill
    • Journal of Chest Surgery
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    • v.38 no.6 s.251
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    • pp.428-433
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    • 2005
  • Advanced age in Esophagectomy increases the risk of postoperative morbidity and mortality. However, the recent development of operative technique and perioperative care might have decreased the postoperative morbidity and mortality after esophagectomy. Material and Method: From March 2001 to July 2004, 174 patients underwent esophageal resection for esophageal cancer in the Center for Lung Cancer, National Cancer Center. The patients were divided into two groups : group 1 consisted of 27 patients aged 70 years or more, and group 2 consisted of 147 patients under 70 years of age. The two groups were compared according to preoperative risk factors, postoperative morbidity, operative mortality and survival. Result: The mean age was 63_4. There were 159 men. On histopathological examination, $93.1\%$ had squamous cell carcinoma. On the locations, $78.7\%$ were in mid and lower esophagus. Curative resections for esophageal cancer were possible in $162(93.1\%)$ patients. Mean hospital stay was 19.4 days with out difference between the groups. The overall postoperative morbidity were occurred in 61 patients $(35.1\%)$. The most frequent morbidity was pulmonary complication in $30(17.2\%)$. Preoperative incidence of hypertension, cardiac and pulmonary dysfunction were more common in Group I. However, there was no difference in overall postoperative morbidity, operative mortality and survival rate between the two groups. Conclusion: Esophagectomy for esophageal cancer could be carried out safely in patients over 70 years of age with satisfactory short-term results. Advanced age is no longer a risk factor for esophagectomy.

Feasibility of Reflecting Improvement of Tumor Hypoxia by Mild Hyperthermia in Experimental Mouse Tumors with $^18F-Fluoromisonidazole$ (저온온열치료에 의한 종양 내 저산소상태 개선효과를 $^18F$-Fluoromisonidazole의 섭취 변화를 이용한 평가)

  • Lee Sang-wook;Ryu Jin Sook;Oh Seung Joon;Im Ki Chun;Chen Gi Jeong;Lee So Ryung;Song Do Young;Im Soo Jeong;Moon Eun Sook;Kim Jong Hoon;Ahn Seung Do;Shin Seong Soo;Lee Kyeong Ryong
    • Radiation Oncology Journal
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    • v.22 no.4
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    • pp.288-297
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    • 2004
  • Puporse: The aims of this study were to evaluate the change of $[^18F]fluoromisonidazole$($[^18F]FMISO$) uptake in C3H mouse squamous cell carcinoma-VII (SCC-VII) treated with mild hyperthermia ($42^{circ}C$) and nicotinamide and to assess the biodistribution of the markers in normal tissues under similar conditions. Methods and Materials: $[^18F]FMISO$ was producedby our hospital. Female C3H mice with a C3H SCC-VII tumor grown on their extremities were used. Tumors were size matched. Non-anaesthetized, tumor-bearing mice underwent control or mild hyperthermia at $42^{circ}C$ for 60 min with nicotinamide (50 mg/kg i.p. injected) and were examined by gamma counter, autoradiography and animal PET scan 3 hours after tracer i.v. injected with breathing room air, The biodistribution of these agents were obtained at 3 h after $[^18F]FMISO$ injection. Blood, tumor, muscle, heart, lung, liver, kidney, brain, bone, spleen, and intestine were removed, counted for radioactivity and weighed. The tumor and liver were frozen and cut with a cryomicrotome into 10- um sections. The spatial distribution of radioactivity from the tissue sections was determined with digital autoradiography. Results: The mild hyperthermia with nicotinamide treatment had only slight effects on the biodistribution of either marker in normal tissues. We observed that the whole tumor radioactivity uptake ratios were higher in the control mice than in the mild hyperthermia with nicotinamide treated mice for $[^18F]FMISO$ ($1.56{\pm}1.03$ vs. $0.67{\pm}0.30$; p=0.063). In addition, autoradiography and animal PET scan demonstrated that the area and intensity of $[^18F]FMISO$ uptake was significantly decreased. Conclusion: Mild hyperthermla and nicotinamide significantly improved tumor hypoxia using $[^18F]FMISO$ and this uptake reflected tumor hypoxic status.

The Results of Postoperative Radiotherapy for Hypopharyngeal Carcinoma (하인두암 환자에서의 수술 후 방사선치료의 결과)

  • Kim Won Taek;Ki Yong Kan;Nam Ji Ho;Kim Dong Won;Lee Byung Ju;Wang Su Gun;Kyuon Byung Hyun
    • Radiation Oncology Journal
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    • v.22 no.4
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    • pp.254-264
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    • 2004
  • Purpose: This study was carried out to confirm clinical values and limitations of postoperative radiotherapy for hypopharyngeal carcinoma, to evaluate various prognostic factors which may affect to the treatment results and to use these results as fundamental data for making a new treatment strategy. Methods and Materials:. A retrospective analysis was peformed on 64 previously untreated patients with squamous cell carcinoma of the hypopharynx, seen between 1988 and 1999 at Pusan National University Hospital. Most of patients were treated by laryngopharyngectomy and neck dissection followed by conventional fractionated postoperative radiotherapy on surgical bed and cervical nodal areas. Results: The five-year overall survival rate and cause-specific survival rate were 42.2 percent and 51.6 percent, respectively. Univariate analysis of various clinical and pathologic factors confirmed the overall stage, TN-stage, secondary primary cancers, surgical positive margin, nodal extracapsular extension, total radiation doses as significant prognostic factors of hypopharyngeal carcinomas. But in multivariate analysis, TN-stage, surgical positive margin and extracapsular extesion were only statistically significant. Conclusion: In resectable cases of hypopharyngeal carcinoma, combined surgery and postoperative radio-therapy obtained good treatement results, even though sacrificing the function of larynx and pharynx. But in advanced and unresectable cases, with respect to survivals and qualify of life issues, we were able to confirm some limitations of combined therapy. So we recommend that comparative studies of recent various chemo-radiotherapy methods and advanced radiotherapy techniques with these data should be needed.

Immunogenicity of Synthetic Peptide Specific for Major Immunogenic Determinat of Hepatitis B Surface Antigen (B형간염(型肝炎) 표면항원(表面抗原)의 주면역원(主免疫原) 결정기(決定基)에 특이(特異)한 합성(合成) Peptide의 면역원성(免疫原性)에 관한 연구(硏究))

  • Shin, Kwang-soon;Han, Su-nam
    • Korean Journal of Veterinary Research
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    • v.25 no.1
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    • pp.7-17
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    • 1985
  • Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

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Analysis of Clonorchis sinensis antigens and diagnosis of clonorchiasis using monoclonal antibodies (단세포군 항체를 이용한 간흡충 항원의 분석 및 간흡충증의 진단)

  • Yong, Tae-Sun;Im, Gyeong-Il;Jeong, Pyeong-Rim
    • Parasites, Hosts and Diseases
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    • v.29 no.3
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    • pp.293-310
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    • 1991
  • Clonorchis sinensis is a common parasite of man in Korea. Researches on the specific antigens of C. sinensis would be valuable not only because those elucidate the molecular characteristics of this fluke but also because it is applicable to immunodiagnosis. Although many monoclonal antibodies have been used in the field of parasite immunology, few articles on monoclonal antibodies against C. sinensis have been published so far. The aim of this study was to analyse C. sinensis antigens recognized by monoclonal antibodies, and to set up ELISA-inhibition test using C. sinensis specific monoclonal antibodies for improved specificity of immunodiagnostic tests. By fusion between spleen cells of the mice immunized with C. sinensis water-soluble crude adult worm antigens and plasmacytoma cells of mouse origin, 29 hybridoma clones secreting anti-C. sinensis monoclonal antibodies were made, and 8 clones among those were found specific. After cell cloning, isotypes of 6 selected specific monoclonal anti- bodies were determined to be IgGl, IgG2b and IgA. Four exposed antigenic determinants of natural infection were recognized by different specific monoclonal antibodies. By enzyme-immunoelectrotransfer blot, 10 KD, 34 KD antigenic determinants were found to be reacted with CsHyb 0714-20, CsHyb 0605-10 monoclonal antibodies, respectively, The antigenic determinant recognized by CsHyb 0714-20 monoclonal antibody was revealed to be located at the surface and parenchyme of a parasite by indirect immunoauorescent antibody technique, and those reacted with CsHyb 0605-10, CsHyb 0714-25 monoclonal antibodies were found at the parenchyme and intestine. The antigenic determinant reacted with CsHyb 0605-23 monoclonal antibody was found mainly around the uterine eggs. Four antigenic determinants recognized by specific monoclonal antibodies were all found to be present in the early eluted fractions of C. sinensis antigens separated by Sephadex G-200 gel filtration. By conventional ELISA, 75% of clonorchiasis cases were found positive, but 7.1% of normal controls and 37.5% of paragonimiasis cases showed false positives. However, by ELISA-inhibition test using C. sinensis specific monoclonal antibody (CsHyb 0605-23), 77.1% of clonorchiasis cases were found positive, and there were no false positives in normal controls or paragonimiasis cases, indicating 100% specificity. The ELISA- inhibition test using monoclonal antibodies was found to have same sensitivity and definitely high specificity in comparison with conventional ELISA for serodiagnosis of human clonorchiasis.

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Cytotoxicity of lymphokine activated peritoneal macrophages against Trichomonas vaginalis (질트리코모나스에 대한 림포카인황성대식세포의 세포독성능)

  • Yoon, Kyong;Ryu, Jae-Sook;Min, Duk-Young
    • Parasites, Hosts and Diseases
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    • v.29 no.4
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    • pp.381-388
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    • 1991
  • Trichomonas vaginalis is a parasitic nagellate in the urogenital tract of human. Innate cytotonicity of macrophages against T. vaginalis has been recognized, but any report on the cytotoxicity of Iymphokine-activated macrophages to T vaginalis is not yet available. The present study aimed to elucidate the Iymphokine-activated cell mediated cytotoxic effect against T. vaginalis by mouse peritoneal macrophages. Cytotoxicity was measured by counting the release of $^3H-thymidine$ from prelabeled protozoa, and tested in U-bottom microtiter plates. Nitrite concentration in culture supernatants was measured by standard Griess reaction. The results obtained are as follows: 1, The cytotoxicity of macrophages was increased by addition of rIL-2 or $rIFN-{\gamma}$$. 2, Cytotoxicity of macrophages was reduced by addition of rIL-4 to rOM-CSV, rIL-2 or $rIFN-{\gamma}$. 3. Crude Iymphokine mixed with anti-lL-2 decreased the cytotoxity of macrophages. 4. In case of macrophages cultured with $rIFN-{\gamma}$ or rIL-4, the concentration of nitrite was related with cytotokity of macrophages against T. vaginalis, but the cytotoxicity of macrophages cultured with rIL-2 and $rIFN-{\gamma}$ was decreased in spite of its high production of llitrite. From the results obtained, it is assumed that rIL-2 and $rIFN-{\gamma}$ enhance the cytotoxicity of macrophages while rIL-4 inhibits the cytotoxicity against T. vaginalis, and that the production of nitrite does not relate with the cytotoxicity of macrophages, but nitric oxide may play a role as an inhibitory factor on the proliferation of T. vaginalis.

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Inhibitory effects of extracts from Smilacina japonica on lipopolysaccharide induced nitric oxide and prostaglandin E2 production in RAW264.7 macrophages (RAW264.7 대식세포에서 풀솜대 추출물의 nitric oxide 및 prostaglandin E2생성 저해효과)

  • Nam, Jung-Hwan;Seo, Jong-Taek;Kim, Yul-Ho;Kim, Ki-Deog;Yoo, Dong-Lim;Lee, Jong-Nam;Hong, Su-Young;Kim, Su-Jeong;Sohn, Hwang-Bae;Kim, Hyun-Sam;Kim, Bo-Sung;Lee, Kyung-Tea;Park, Hee-Jhun
    • Journal of Plant Biotechnology
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    • v.41 no.4
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    • pp.201-205
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    • 2014
  • Smilacina japonica is a localized common rhizomatous flowering plant, This plant is often used in Korean traditional systems of medicine as a remedy for migrain, diplegia, physical impurity, blood circulation, abscess and contusion. Generally drugs that are used for arthritis have antinociceptive and anti-inflammatory properties. However, validity of the anti-inflammatory activity has not been scientifically investigated so far. Therefore, the aim of this study was to investigate the anti-inflammatory potential of S. japonica using the ethanolic extract and its subfractions. To evaluate the anti-inflammatory effects, we examined the inflammatory mediators such as nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) on RAW 264.7 macrophages. Our results indicated that hexane fraction significantly inhibited the LPS induced NO and $PGE_2$ production in the cells. The hexane fractions inhibitory activity for NO tests with $IC_{50}$ values showed in $53.3{\mu}g/ml$ and $PGE_2$ tests with $IC_{50}$ values showed at $32.5{\mu}g/ml$. Theseis result suggest a potential role of hexane fraction from S. japonica as source of anti-inflammatory agent.

The Characteristics of IgA Nephropathy when Detected early in Mass School Urine Screening (학교 집단 요검사로 조기 진단된 IgA 신증 환아의 임상적 특징)

  • Kim, Sae Yoon;Lee, Sang Su;Lee, Jae Min;Kang, Seok Jeong;Kim, Yong Jin;Park, Yong Hoon
    • Childhood Kidney Diseases
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    • v.17 no.2
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    • pp.49-56
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    • 2013
  • Purpose: IgA nephropathy (IgAN) is one of the major causes of end-stage renal disease. Mass school urine screening (SUS) has been performed to enable early detection of chronic renal diseases, including IgAN. We wanted to evaluate the patients with IgAN, including those diagnosed through SUS. Methods: Between 1998 and 2010, 64 children were diagnosed with IgAN based on renal biopsy results obtained at the Pediatric Nephrology Department, ${\bigcirc\bigcirc}$ University Hospital. We divided these patients into the SUS group (37 cases), diagnosed through SUS, and the symptomatic (Sx) group (27 cases), diagnosed clinically. The medical records of both groups were analyzed retrospectively. Results: The mean age of the SUS and Sx groups was $10.8{\pm}2.7$ and $9.5{\pm}3.4$ years (P >0.05), respectively. Both groups had a higher proportion of male patients. The time from the notification of an abnormal urinary finding to a hospital visit or renal biopsy was shorter in the Sx group than in the SUS group. Regarding clinical manifestations, there were fewer cases with gross hematuria (P <0.001) and edema (P =0.008) in the SUS group, but there were no differences in terms of the therapeutic regimen and treatment duration. Regarding laboratory parameters, the Sx group had a higher white blood cell count (P =0.007) and lower hemoglobin (P =0.007) and albumin (P =0.000) levels. There were no differences in the renal biopsy findings in both groups, based on the history of gross hematuria or the severity of proteinuria. However, in all 64 patients with IgAN, the light microscopy findings (Hass classification) were related to a history of gross hematuria or the severity of proteinuria. Conclusion: There were no significant clinical and histological differences between the groups, as both had early stage IgAN. Although SUS facilitates the early detection of IgAN, long-term, large-scale prospective controlled studies are needed to assess the benefits of early diagnosis and treatment in chronic renal disease progression.