• Title/Summary/Keyword: Indirect Immunofluorescence

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Hematological Change of Korean Native Goats Experimentally Infected with Bovine Leukemia Virus (Bovine Leukemia Virus에 실험감염된 한국재래산양의 혈액상의 변동)

  • 이필돈;김종호;전무형
    • Korean Journal of Veterinary Service
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    • v.18 no.1
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    • pp.1-21
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    • 1995
  • To elucidate pathogenesis of bovine leukemia virus(BLV) in Korean native goats, the goats experimentally infected with BLV were studied especially for the aspects of infectivity and hematological changes. The experimental goats were examined for 27 months by agar-gel immunodiffusion(AGID) test and syncytium formation assay. During this period, changes of total leucocyte, absolute Iymphocyte and atypical Iymphocyte were examined, and the distribution of surface immunoglobulin ( sIg ) -bearing cells and rosette forming cell (RFC) in the peripheral Iymphocyte were also investigated. By indirect immunofluorescence (IFA) and complement dependent antibody cytotoxicity (CDAC) assay using monoclonal antibody(Mab) against bovine leukosis tumor-associated anti-gen(BL-TAA), changes of BL-TAA positive Iymphocyte in peripheral blood were measured. The results obtained through the experiment were summarized as follows. 1. Antibody titers were measured by AGID using gP51 and P24 antigens. The animals were serologically converted at 2 months post-inoculation(pi) in gP51 antigen, whereas sero-converted at 4 months pi in P24 antigen. In comparison with antibody titers for gP51, P24 antigen showed lower titers throughout the trial period. 2. The peripheral lymphocytes from all of the infected goats, as co-cultivated with F8l cells manifested syncytial formation at 4 months pi. 3. On counting total leucocyte, Iymphocyte and atypical Iymphocyte, two out of four infected goats showed normal distribution, while No 2 of the remaining two revealed temporal and No 3, Persistant increasing number of the cells. 4. The optimal condition of rosette formation of the peripheral Iymphocyte of normal Korean native goats was shown in the sheep erythrocyte treated with 0.1M AET for 30 nun at $37^{\circ}C$. When the Iymphocytes were treated in nylon wool column, the number of sIg-bear-ing cell were increased in the nylon wool adherent cells, but RFC was increased in the non-adherent cells. Of the infected goats, No 2 and No 3 showed significantly increasing number of sIg-bearing cells at 18 months pi. 5. The Iymphocytes of No 2 and No 3 goats reacted positively in IFA using Mab against BL-TAA at 12 months pi and 18 months pi, respectively. In CDAC test, all of four infected goats revealed positive reaction at 24 months pi. The higher positive rates were observed in No 2 and No 3 as compared with the remainders.

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Effects of Trichostatin A on In vitro Development of Porcine Embryos Derived from Somatic Cell Nuclear Transfer

  • Jeong, Yeon Ik;Park, Chi Hun;Kim, Huen Suk;Jeong, Yeon Woo;Lee, Jong Yun;Park, Sun Woo;Lee, Se Yeong;Hyun, Sang Hwan;Kim, Yeun Wook;Shin, Taeyoung;Hwang, Woo Suk
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.12
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    • pp.1680-1688
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    • 2013
  • Many different approaches have been developed to improve the efficiency of animal cloning by somatic cell nuclear transfer (SCNT), one of which is to modify histone acetylation levels using histone deacetylase inhibitors (HDACi) such as trichostatin A (TSA). In the present study, we examined the effect of TSA on in vitro development of porcine embryos derived from SCNT. We found that TSA treatment (50 nM) for 24 h following oocyte activation improved blastocyst formation rates (to 22.0%) compared with 8.9% in the non-treatment group and total cell number of the blastocysts for determining embryo quality also increased significantly ($88.9{\rightarrow}114.4$). Changes in histone acetylation levels as a result of TSA treatment were examined using indirect immunofluorescence and confocal microscopy scanning. Results showed that the histone acetylation level in TSA-treated embryos was higher than that in controls at both acetylated histone H3 lysine 9 (AcH3K9) and acetylated histone H4 lysine 12 (AcH4K12). Next, we compared the expression patterns of seven genes (OCT4, ID1; the pluripotent genes, H19, NNAT, PEG1; the imprinting genes, cytokeratin 8 and 18; the trophoblast marker genes). The SCNT blastocysts both with and without TSA treatment showed lower levels of OCT4, ID1, cytokeratin 8 and 18 than those of the in vivo blastocysts. In the case of the imprinting genes H19 and NNAT, except PEG1, the SCNT blastocysts both with and without TSA treatment showed higher levels than those of the in vivo blastocysts. Although the gene expression patterns between cloned blastocysts and their in vivo counterparts were different regardless of TSA treatment, it appears that several genes in NT blastocysts after TSA treatment showed a slight tendency toward expression patterns of in vivo blastocysts. Our results suggest that TSA treatment may improve preimplantation porcine embryo development following SCNT.

Expression of the Proto-oncogene Pokemon in Colorectal Cancer - Inhibitory Effects of an siRNA

  • Zhao, Gan-Ting;Yang, Li-Juan;Li, Xi-Xia;Cui, Hui-Lin;Guo, Rui
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.9
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    • pp.4999-5005
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    • 2013
  • Objective: This study aimed to investigate expression of the proto-oncogene POK erythroid myeloid ontogenic factor (Pokemon) in colorectal cancer (CRC), and assess inhibitory effects of a small interference RNA (siRNA) expression vector in SW480 and SW620 cells. Methods: Semi-quantitative reverse transcription-polymerase chain reaction (PCR) and immunohistochemistry were performed to determine mRNA and protein expression levels of Pokemon in CRC tissues. Indirect immunofluorescence staining was applied to investigate the location of Pokemon in SW480 and SW620 cells. The siRNA expression vectors that were constructed to express a short hairpin RNA against Pokemon were transfected to the SW480 and SW620 cells with a liposome. Expression levels of Pokemon mRNA and protein were examined by real-time quantitative-fluorescent PCR and western blot analysis. The effects of Pokemon silencing on proliferation of SW480 and SW620 cells were evaluated with reference to growth curves with MTT assays. Results: The mRNA expression level of Pokemon in tumor tissues ($0.845{\pm}0.344$) was significantly higher than that in adjacent tumor specimens ($0.321{\pm}0.197$). The positive expression ratio of Pokemon protein in CRC (87.0%) was significantly higher than that in the adjacent tissues (19.6%). Strong fluorescence staining of Pokemon protein was observed in the cytoplasm of the SW480 and SW620 cells. The inhibition ratios of Pokemon mRNA and protein in the SW480 cells were 83.1% and 73.5% at 48 and 72 h, respectively, compared with those of the negative control cells with the siRNA. In the SW620 cells, the inhibition ratios of Pokemon mRNA and protein were 76.3% and 68.7% at 48 and 72 h, respectively. MTT showed that Pokemon gene silencing inhibited the proliferation of SW480 and SW620 cells. Conclusion: Overexpression of Pokemon in CRC may have a function in carcinogenesis and progression. siRNA expression vectors could effectively inhibit mRNA and protein expression of Pokemon in SW480 and SW620 cells, thereby reducing malignant cell proliferation.

Seroreactivity to Q Fever Among Slaughterhouse Workers in South Korea

  • Chu, Hyuk;Yoo, Seok-Ju;Hwang, Kyu-Jam;Lim, Hyun-Sul;Lee, Kwan;Park, Mi-Yeoun
    • Journal of Preventive Medicine and Public Health
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    • v.50 no.3
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    • pp.195-200
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    • 2017
  • Objectives: Q fever is a zoonotic disease that occurs worldwide; however, little is known about its prevalence in South Korea. We attempted to determine the prevalence of Q fever seroreactivity among Korean slaughterhouse workers and the risk factors for seroreactivity according to the type of work. Methods: The study was conducted among 1503 workers at a total of 73 slaughterhouses and 62 residual-product disposal plants. During the study period, sites were visited and surveys were administered to employees involved in slaughterhouse work, and serological tests were performed on blood samples by indirect immunofluorescence assays. Serological samples were grouped by job classification into those of slaughter workers, residual-product handlers, inspectors and inspection assistants, and grading testers and testing assistants. Employee risk factors were analyzed according to the type of work. Results: Out of 1481 study subjects who provided a blood sample, 151 (10.2%) showed reactive antibodies. When these results were analyzed in accordance with the type of work, the result of slaughter workers (11.3%) was similar to the result of residual-product handlers (11.4%), and the result of inspectors and assistants (5.3%) was similar to the result of grading testers and assistants (5.4%). Among those who answered in the affirmative to the survey question, "Has there been frequent contact between cattle blood and your mouth while working?" the proportions were 13.4 and 4.6%, respectively, and this was identified as a risk factor that significantly varied between job categories among slaughterhouse workers. Conclusions: This study found a Q fever seroreactivity rate of 10.2% for slaughterhouse workers, who are known to be a high-risk population. Contact with cattle blood around the mouth while working was the differential risk factor between job categories among slaughterhouse workers.

Experimental infection of piglets with a field isolate of Aujeszky's disease virus in Korea: Pathogenecity, excretion, distribution and immunogenicity of virus (국내분리 Aujeszky's disease virus의 실험적 감염 자돈에 대한 바이러스학적 연구)

  • Park, Jeong-woo;Jun, Moo-hyung;An, Soo-hwan
    • Korean Journal of Veterinary Research
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    • v.30 no.2
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    • pp.177-186
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    • 1990
  • To investigate the etiology, pathogenicity and virological properties of NYJ-1-87 strain of Aujeszky's disease virus (ADV) that was isolated from the diseased piglet in Korea, the virus at $10^{6.0}TCID_{50}/0.1ml$ was inoculated intranasally and subcutaneously into 30 to 35 days-old piglets. Results obtained through the experiments were summarized as follows. 1. Ten of the infected piglets were clinically observed for 15 days. On the 2nd day post-inoculation(pi), the signs of pyrexia, anorexia and convulsion were noted. On the 4th to 7th days pi, nervous signs of incoordination and intermittent spasm were shown in the most of piglets, and one out of 5 piglets infected intranasally was died with severe nervous signs at the 7th day pi. The signs became relieved on the 8th day pi and all of remainder were completely recovered on the 13th to 14th days pi. 2. In hematological study, prominent decrease in the number of total leukocyte and lymphocyte was shown in the ADV-infected piglets on the 6th day pi. On the 8th day pi, the cell numbers were slightly increased and returned to normal level on the 10th day pi. 3. Viral excretion of the ADV-inoculated piglets was examined by swabbing of nasal and oral cavities, and rectal feces. During the periods of the 3rd to 11th days pi, the virus was excreted intermittently from nasal and oral cavities, and rectal feces. The nasal excretions were shown the highest virus concentration of $10^{5.2}TCID_{50}/0.1ml$ at the 5th day pi. 4. Recovery of the inoculated virus from various organs of the piglets that were died or experimentally slaughtered was attempted, and the virus was isolated from the tissues of brain and tonsil by the cultured cell-inoculation method. The highest recovery rate was noted in the tonsil. By indirect immunofluorescence antibody assay using ADV-monoclonal antibody, the viral antigens were detected in tissues of spleen and liver as well as brain and tonsil on the 7th to 9th days pi. The virus was not isolated from blood and the tissues of lung and kidney throughout the experiments. 5. Titers of virus neutralizing antibody in the piglets experimentally infected with ADV became increased after the 6th to 9th days pi in both of intranasal and subcutaneous inoculation showing the highest titers of 64 to 128 on the 29th day pi. When the antibody levels were measured by radial immunodiffusion enzyme assay, the reactive diameter was enlarged to be positive after the 4th to 6th days pi in both of intranasal and subcutaneous inoculation showing the largest diameter of 13 to 14mm on the 29th day pi.

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Seroprevalence and risk factors of Lyme disease among Korean deer farmers (대한민국 사슴농가 종사자의 라임병 혈청유병률 및 위험요인)

  • Acharya, Dilaram;Cho, Jae-Hak;Yoo, Seok-Ju;Park, Ji-Hyuk
    • Journal of agricultural medicine and community health
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    • v.44 no.2
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    • pp.82-89
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    • 2019
  • This study was undertaken to determine the seroprevalence and risk factors associated with contracting Lyme disease (LD) among Korean deer farmers. This cross-sectional study devised questionnaire that addressed farm activities, was devised and the blood samples of 516 Korean deer farmers were tested. LD seroprevalence was determined by Western blot test. Fisher's exact test, the chi-squared test, and the chi-squared test for trend analysis were performed to assess the risk associated with LD. Of total 516 study participants recruited, only 12 (2.3%) were seropositive for LD. The result of the study revealed that only deer farmers raising Elk (Cervus Canadensis) were found to be at significantly higher risk of contracting LD than other deer farmers (p=0.033). In addition, a male sex, an age of < 40 years, and those that had raised deer for < 19 years had higher rates of LD infection than their counterparts. Similarly, those that managed livestock sheds, fed deer, processed industry and prepared compost had higher rates of LD infection. In terms of protective factors, those who did not use protective measures such as wear glasses and masks, or protective gloves and aprons, and those that did not disinfect work instruments and did not shower after work had higher rates of LD than those that used protective measures. In conclusion, preventive health strategies should take into account the profiles of deer farmers at greater risk based on considerations of personal, type of work, and the use of personal protective measures.

Surface Marker Analysis in Acute Leukemias (백혈병의 면역학적 표지자검사의 결과분석)

  • Moon, Jin-Young;Lee, Chae-Hoon;Kim, Kyung-Dong;Kim, Chung-Sook
    • Journal of Yeungnam Medical Science
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    • v.14 no.2
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    • pp.359-369
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    • 1997
  • We studied the expression of the cell surface antigen associated with myeloid and lymphoid leukemias on bone marrow or peripheral blood blast cells from 153 leukemic patients including 61 cases of acute myelogenous leukemias(AML), 46 of acute lymphocytic leukemias(ALL) and 12 of acute leukemias. They were analyzed by direct or indirect immunofluorescence method for reactivity with the monoclonal antibodies to B cells(CD10, CD19, SmIg), T cells(CD2, CD5, CD7, CD3, CD4, CD8), myeloid antigen(CD13, CD14, CD33, CD61) and a nonspecific antigen, HLA-DR. Lymphoid associated markers detected on AML is CD7 32.8%, CD10 14.8%, CD5 13.1%, CD2 6.6% and CD19 1.6%. TdT was positive in 4.9% of AMLs. Hybrid leukemias were 8 cases out 61 AML cases and were mainly composed of monocytic lineage, M4 and M5a. Myeloid markers detected in ALL were CD13 2.2% and CD33 2.2%. In this study, immunologically classified ALLs were composed of 65.2% of CALLA (+) B precursor type, 10.9% of CALLA (-) B precursor pattern, 8.7% of T cell type, 2.2% of B cell type, 4.5% of mixed lymphoid lineage(B&T), 2.2% of undifferentiated leukemia, and 6.5% of hybrid leukemia. Twelve cases of acute leukemias ware finally diagnosed to be 5 cases of hybrid leukemia, 3 cases of B lineage, 3 case of T lineage and 1 case of mixed lymphoid(B&T) leukemia. In summary, we think the best method for typing acute leukemias is by using a combination of FAB classification and immunophenotying.

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Adenovirus Pneumonia with Respiratory Failure in Pediatric Patients (소아에서 발생한 호흡부전이 동반된 아데노 바이러스 폐렴)

  • Do, Sung Suk;Ma, Sang Hyeok;Park, Jae Sun;Lee, Young Ho;Lee, Hwan Jong;Lee, Gyu Man
    • Pediatric Infection and Vaccine
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    • v.5 no.2
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    • pp.258-266
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    • 1998
  • Purpose : Cases of adenoviral penumonia with rapidly progressive clinical course were experienced. We reviewed these patients in viewpoint of clinical manifestation and adenoviral serotypes. Methods : Culture and indirect immunofluorescence for respiratory viruses including respiratory syncytial virus, influenza virus, parainfluenza virus, adenovirus was done with nasopharyngeal aspirates from patients who admitted due to respiratory infections in Fatima Hospital, Masan from Nov. 1996 to Jul. 1997. Cultured adenovirus was serotyped by both neutralization and hemagglutination inhibition test. Medical records were reviewed for 5 patients with respiratory failure from adenovirus was isolated and serotyped. Results : The total number of examined patients was 460 patients. We isolated respiratory viruses in 143(30.9%) patients. Adenovirus was isolated from 66 out of 143(46.2%) patients. During Jan 1997 to May 1997, five patients with ages of 18 days to 11 months who were infected by adenovirus and had high fever with dyspnea and required assisted mechanical ventilation. One patients discharged against doctor's advice then died. Two of four patients had complications of disseminated intravascular coagulation; two had bronchiolitis obliterans. Two isolates were serotype 7, and one was serotype 5, and two were untyped. Conclusion : Severe pneumonia caused by serotype 7 continued to occur in 1997 following the epidemic in 1996, and severe pneumonia may also be caused by serotype 5 and other serotypes.

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Epidemics of Aseptic Meningitis in Kyoungsangnamdo from March to October, 1997 (1997년 경상남도 중부지방에서 유행한 무균성 뇌막염의 임상적 고찰)

  • Park, Sun Young;Kwon, Oh Su;Kim, Won Youb;Jung, Won Jo;Ma, Sang Hyouk;Kim, Sang Ki;Nam, Sung Jin;Jo, Sung Rae;Gu, Bon Chun;Lee, Kyu Man
    • Pediatric Infection and Vaccine
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    • v.5 no.1
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    • pp.104-114
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    • 1998
  • Purpose : Enteroviruses are the most common cause of aseptic meningitis. The epidemics of aseptic meningitis in 1993 and 1996 were mostly caused by echovirus type 9. Identification of the causative virus of aseptic meningitis in epidemics, is very important not only for diagnosis but also for epidemiologic purpose. The purpose of this study was to identify the causative virus and investigate the relationship between aseptic meningitis, prevailed in Masan and surrounding areas in Kyoungsangnamdo in 1997, and its clinical manifestations. Methods : One hundred twenty eight cerebrospinal fluid(CSF) and 239 stool specimens were obtained from 239 patients(213 children and 26 adult patients) with aseptic meningitis were admitted to Masan Fatima Hospitals from March to October 1997. Viral isolation and serotype identification was performed by cell culture and immunofluorescent test. Enteroviruses not typed by immunofluorescent test was confirmed by reverse transcription-polymerase chain reaction(RT-PCR). Results : 1) The peak incidence was noted in June. 2) The age of 239 patients(pediatrics-213 cases, internal medicine-26 cases) that were diagnosed ranged from neonate to 35 years, the age of the patients of pediatrics ranged from neonate to 15years(mean 4.9 years), the age of the patients of internal medicine (above 16 years) ranged from 16 years to 35 years(mean 24.2 years). 3) Fifty-three(41.4%) of 128 CSF specimens were positive for enteroviruses, and 163(68.2%) of 239 stool specimens were positive for enteroviruses respectively. 4) Serotypes of 53 enteroviruses isolated from CSF were 16(30.2%) of echovirus type 30, 6(11.3%) of echovirus type 6, 1 of echovirus type 4, 4 of untyped echovirus, 1 of coxsackievirus type B5, and 24 isolates of untyped enteroviruses. Of 163 enterovirus isolated from stool were 72(44.2%) of echovirus type 30, 21(12.9%) of echovirus type 6, 1 of echovirus type 4, 17(10.4%) of undetermined subtyped echovirus, 1 of coxsackievirus type B5, 2 of A24, 3 of undetermined subtyped coxsackievirus type B, and 46 isolates of untyped enterovirus. Conclusion : There were epidemics of aseptic meningitis in the central areas of Kyoungsangnamdo from March to October 1997. The main causative organism was thought to be the echovirus type 30, and echovirus type 4, 6, coxsackievirus B5 and A24 were also thought to contribute to the epidemics.

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Antineutrophil Cytoplasmic Antibody in Korean Children with Henoch-$Sch{\ddot{o}}nlein$ Purpura (소아 Henoch-$Sch{\ddot{o}}nlein$ 자반증에서의 항호중구 항체)

  • Choi, Ji-Na;Shin, Jae-Il;Lee, Jae-Seung;Kim, Hyon-Suk
    • Childhood Kidney Diseases
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    • v.12 no.2
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    • pp.164-169
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    • 2008
  • Purpose : There have been some reports on the prevalence of positive antineutrophil cytoplasmic antibody(ANCA) in Henoch-$Sch{\ddot{o}}nlein$ purpura(HSP), but the results were conflicting. We performed this study to evaluate the clinical significance of ANCA(c-ANCA and p-ANCA) in Korean children with HSP. Methods : The medical records of 30 patients(13 boys and 17 girls) aged 6.0$\pm$1.9(5-12) years with a clinical diagnosis of HSP based on the EULAR/PReS criteria were reviewed retrospectively. From the years 2007 to 2008, the sera from children with acute HSP were tested for antineutrophil cytoplasmic antibodies(ANCA). The target antigens of these autoantibodies are proteinase 3(c-ANCA) or myeloperoxidase(p-ANCA). Results : Palpable purpura was seen in all 30 patients(100%), abdominal pain in 20(67%), arthralgia in 17(57%), and renal involvement in 11(37%). Laboratory findings showed leukocytosis in 4 patients(13%), thrombocytosis 18 in(60%), and elevated erythrocyte sedimentation rate in 18(60%). Anti-streptolysin O titers were elevated in 7% of the patients and no patient showed elevation of serum IgA level. The sera from 29 patients were negative for c-ANCA and p-ANCA by indirect immunofluorescence, but only one patient had weakly positive results, which became negative at follow-up. Conclusions : We conclude that c-ANCA or p-ANCA is not an important serologic marker in children with HSP, because it was neither diagnostically nor immunologically specific in children with HSP. These results suggest that ANCA are not involved in the pathogenesis of HSP in children.