key: cord-0041884-p8e8r6z8
authors: nan
title: The 3rd Congress of Asian Society for Pediatric Research
date: 2008-09-22
journal: Pediatr Int
DOI: 10.1111/j.1442-200x.2008.02727.x
sha: 3ef709d285cbdc7196f2e315894c0f9a526f7f14
doc_id: 41884
cord_uid: p8e8r6z8

nan

[Background] Rhabdomyosarcoma (RMS) is the most common soft-tissue sarcoma in children and classified into two major histological subgroups, embryonal and alveolar types that are presented with different clinical pictures and prognosis. Although distinct genetic mechanisms are implicated in their pathogenesis, little has been known about those genetic changes that characterize these distinctive histology types, except for common PAX3 and PAX7 rearrangements in alveolar types. [Methods] In order to investigate the genetic changes that underlie the pathogenesis of both alveolar and embryonal RMSs, we performed comprehensive analysis of copy number alterations in RMS genomes obtained from a total of 30 RMS samples including RMSderived cell lines using high-density SNP-genotyping microarrays (Affymetirx GeneChip 100K/500K arrays).

[Results] A number of gains and losses loci were identified that involves not only previously-described regions such as 1p, 2p, 9p and 11p, but also novel regions including 3p, 11q, 15p and 16q. High-grade amplifications of PAX3 and FKHR as well as MYCN were observed in the subset of patients with alveolar histology. Furthermore, we found a number of additional genetic changes characteristic to the alveolar RMS but not observed in the embryobal RMS. [Conclusion] The SNP-genotyping microarray platform is a powerful tool for genome-wide detection of genetic abnormalities in RMS and expected to facilitate identification of novel genes relevant to the development of RMS.

Introduction: Recombinant factor VIIa (rFVIIa) has been first used for controlling massive bleeding in patients with DHF in Thailand since 1997. Methods: Thirty-eight patients (17 males, 21 females) with DHF grade II (n=4), III (n=19) and IV (n=15) who received rFVIIa between July 1997 to June 2007 were retrospectively studied. They were admitted at Ramathibodi Hospital (n=17) and other hospitals (n=21). Their median age was 9.0 years. The patients were divided into two groups: group 1 included 23 patients waiting for platelet concentrate and group 2 included 15 patients who had already received platelet concentrate with unresponsiveness. The dose of rFVIIa was 100 µg/kg at 15-30 min interval for 1-4 doses until the bleeding significantly reduced followed by 2-4 h interval until the bleeding was completely ceased. Results: Each patient exhibited bleeding at 2-5 sites including gastrointestinal tract 48.0%, epistaxis 16.4%, oral cavity 11.0%, hemothorax 8.2%, lungs 4.1%, hypermenorrhea 4.1%, hematuria 2.7% and other sites 5.5%. An effective response was found in 25 patients (65.8%), including 18 patients in group 1 and seven patients in group 2. The case-fatality rate in this study was 23.6% (9/38) found in group 2 only. They were in the condition of prolonged shock for more than 24-48 h before receiving rFVIIa. Conclusion: The use of rFVIIa, a life-saving medicine, is effective in restoring hemostasis to control massive bleeding in patients with DHF. De La Salle University Medical Center All throughout the year, dengue fever has always been present in pediatric census in different hospitals in our country. It still has been a health problem. However, supportive treatment has always been the conventional management in dengue. For some pediatricians, the incorporation of Carbazochrome sodium sulfonate or Adenogen has been part of their management in dengue cases. But there are no studies yet to prove the efficacy of it in dengue. This is a descriptive study of pediatric dengue patients admitted at De La Salle University-Medical Center from January-December 2004 given and not given Adenogen. There were 249 pediatric patients diagnosed with dengue fever/dengue hemorrhagic fever admitted. Nineteen had other concomitant illness, thus were excluded in the study. A total of 230 pediatric patients were included in the study, and 83 of which were given Adenogen and the rest were subjected to conventional dengue management. For dengue patients given Adenogen, most of them live in Dasmarinas, Cavite and were female aged 6-9 years old. Out of 83 patients, only 7 developed complication (8.4%), all of which had hemoconcentration, and no morbidities or mortalities were noted among them. Compared with the group who were not given Adenogen, 17.0% had complications such as pleural effusion, hemoconcentration, bleeding and mortalities.

Assessment of influenza vaccine effectiveness in schoolchildren using rapid test results obtained from a questionnaire survey Shinya Yamaguchi

Background: Virtually all the Japanese schoolchildren who developed a sudden onset of high fever during the influenza epidemic season are taken to a clinic and most of them are tested with an influenza rapid test. Such a circumstance has enabled us to estimate the vaccine effectiveness (VE) without conducting an expensive study. Methods: Prospective cohort study. All the pupils in 4 primary schools in Tsuchiura city, Ibaraki, Japan were included in the study (n=2607). A questionnaire for baseline survey was sent to the parents in January 2007, which inquired about several risk factors to influenza including vaccination status. When a child was diagnosed as influenza with a rapid test during the winter in 2006/2007, the parent was asked to submit an influenza reporting form to the school administration. Adjusted VE was calculated with a logistic regression model. Results: Vaccination rate was 44.8% in total. Incidence of influenza A and B was 5.2% and 11.7%, respectively. The epidemics began in late February 2007, which was markedly delayed compared to other years. The crude VE for influenza A and B was 20% (95%CI: -11 to 43%) and 2% (-21 to 21%), while adjusted VE for influenza A and B was 52.4% (23.1 to 70.7%) and 6.8% (-34.0 to 35.2%), respectively. Disease history of influenza A in the previous year seemed protective to the development of influenza A in the current season (Odds ratio: 0.27, 0.07 to 1.09). Conclusions: Results of rapid tests could be utilized to assess influenza VE without much cost.

1 Department of Pediatric Allergy and Immunology, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung University, 2 Department of Pediatric Cardiology, Chang Gung Memorial Hospital-Kaohsiung Medical Center OBJECTIVE: Kawasaki disease (KD) is a febrile disease with systemic vasculitis, especially coronary artery involvement. Eosinophilia has been found in our and other studies in KD. This study has further investigated whether eosinophil related T helper 2 (Th2) cytokines or eosinophil activation mediator (eosinophil cationic protein, ECP) involved in the KD with coronary artery lesions (CAL). PATIENTS AND METHODS: A total of 95 KD patients were enrolled for this study. Plasma samples were subjected to measurement of interleukin-4 (IL-4), IL-5 and eotaxin by Luminex-Bedalyte multiplex beadmates system and to measurement of ECP by fluoroimmunoassay. CAL was defined as internal diameter greater than 3 mm of coronary artery. RESULTS: Patients with KD had higher C-reactive protein (CRP) and eosinophils than febrile controls. Eosinophil-related mediators: IL-4, IL-5, eotaxin and ECP levels were also higher in KD patients than controls before intravenous immunoglobulin (IVIG) treatment. After IVIG treatment, ECP levels decreased but IL-4, IL-5, and eotaxin increased significantly. The higher the IL-5 and eosinophil levels after IVIG treatment, the lower rate of CAL was found in KD patients. Changes of eosinophils after IVIG treatment were positively correlated to changes of IL-5 levels but not the ECP levels. CONCLUSIONS: The eosinophilia associated with higher Th2 mediator (IL-5) but not ECP level after IVIG treatment was associated with protection of CAL formation.

O-026 IL-10 gene polymorphism, but not TGF-ß1 cytokine gene polymorphisms, is associated with food allergy in a Japanese population.

O-027 16 cases of Food-dependent exercise-induced anaphylaxis (FEIAn) successfully diagnosed by provocation test

Reiko Ito, Norihiro Oku, Tetuharu Manabe, Asako Hujituka, Kazuko Sugai, Shumpei Yokota, Yukoh Aihara

Children's medical center Yokohama city university medical center (Background) Provocation test is required for the diagnosis and identification of food allergens in FEIAn patients. However, provocation tests often fails to provoke symptoms. (Methods) For the diagnosis of 21 patients with FEIAn (17 males and 4 females; 7 to 18 years old), RAST, skin prick test (SPT), and provocation test on the treadmill were performed. Also we measured blood histamine, tryptase, eosinophilic cationic protein (ECP), leukotrieneB4 (LTB4) and,LTC4,D4,E4 levels during provocation test. (Results) Implicated food was identified in 16 patients. Among these 16 patients, eleven patients had more than two suspected food allergens. RAST for all implicated food demonstrated above class 1 in seven patients, negative in four. SPT turned positive in nine patients, negative in four. Both RAST and SPT were negative in three patients. Provocation test was positive in nine patients, negative in seven. There was no significant increase of tryptase and ECP during the tests. (Conclusion) The diagnosis of FEIAn was made by the elevated plasma histamine levels on provocation test in three patients whose RAST, SPT and the symptoms on provocation test were all negative. For clarifying implicated allergen in FEIAn patients, repeated provocation tests with several conditions are required.

Background: Congenital heart defects (CHDs) are the most common birth defects. Women with CHD-affected pregnancies may be genetically susceptible to alterations in folate-dependent metabolic pathways and these susceptibilities may be modified by maternal pro-oxidant lifestyle factors. Methods: We conducted a population-based case-control study of 620 women who had pregnancies affected by nonsyndromic congenital heart defects, and 391 women without such a history. All pregnancies ended between January 1, 1999 and March 1, 2005. Maternal interviews were conducted on cases and controls to determine lifestyle factors such as smoking and alcohol intake. The Genomic DNA was extracted from maternal buccal cell samples. Using ABI PRISM 7900HT Sequence Detection System, DNA was genotyped for methylenetetrahy drofolate reductase (MTHFR) 677 C>T, transcobalamin II (TCII) 776 C>G, and betaine homocysteine methyltransferase (BHMT) 742 G>A polymorphisms. Results: Women who smoked or consumed alcohol periconceptionally had an increased risk of having an infant with a CHD if they carried the TCII 776 G allele. Women who were obese periconceptionally had an increased risk of having an infant with a CHD if they were homozygous for the MTHFR 677 C>T polymorphism or if they carried the BHMT 742 A allele.

Our findings indicate that maternal pro-oxidant lifestyle factors influence the association between CHD-affected pregnancies and genetic susceptibilities. Our findings are in accord with evidence from previous studies that indicate folate-dependent genetic and metabolic susceptibilities increase the risk of CHD-affected pregnancies.

[Background] Caudal regression represents a pattern of malformation defined by hypoplasia of the caudal end of the embryo: aplasia or hypoplasia of the spinal cord, vertebrae, the urogenital system, the hindgut, and the lower extremities. It has been hypothesized that pathogenesis of caudal regression is associated with abnormality within the primitive streak. However, direct evidences for the hypothesis have been lacking.

[Methods] Increasing amount of adriamycin was injected in the yolk of 2-day old embryo. The distribution of the adriamycin within the embryonic body was monitored by detecting fluorescence from adriamycin and morphological changes of the embryo were followed in a serial manner.

[Results] When 5 μg of adriamycin was injected at stage 10-11, signal from adriamycin was detected most intensely in the caudal eminence 6 hours after injection. Twenty-four hours after injection, cells at the level of upper limb bud underwent massive cell death. Caudal regression was evident 48 hours after injection. Administration of 2μg lead to caudal regression whereas that of 1 μg did not. Administration of 2-5 μg at stage 13 did not lead to caudal regression.

[Discussion] We have developed a novel animal model of caudal regression. The induction of caudal regression was developmental stage-and dose-dependent. Admriamycin was accumulated to caudal eminence, a structure that evolves from primitive streeak. We speculate that the primitive streak, a tissue consisiting of expanding population of pluripotent cells, is vulnerable to adriamycin which intercalates to double-stranded DNA of undifferentiated cells undergoing rapid cell division.

Transcriptome analysis of Down syndrome and the discovery of perturbagen patterns in the developing fetus.

Cheng-Hsien Tsai 1 , Yie-Kun Mao 2 , D. Frank Hsu 3 , Fon-Jou Hsieh 4 , Tsu-F Yeh 5 1 National Taiwan Univ. Hospital Yun-Lin Branch, Dept. Pediatr., 2 National Taiwan Univ. Hospital Yun-Lin Branch, Dept. Obstetrics and Gynecology, 3 Dept. of Computer and Information Science, Fordham Univ., New York, USA, 4 Angiogenesis Research Center, Natl. Taiwan Univ., Taipei, Taiwan, 5 Dept. of Ped., China Med. Univ., Taichung, Taiwan BACKGROUND: Down syndrome (DS) is a model of genomic dosage imbalance. The aim is to investigate the transcriptome difference between DS and euploidy from fetal amniotic fluid (AF). The pertubagen pattern means possible treatment for genomic imbalance. METHODS: Amniocytes cultures were collected from fetuses between 16-22 weeks pregnancy. Gene expression was obtained using oligonucleotide chips (22,572 probes). Significant gene expression was analyzed by two-sample t test, Welch's test and Significance Analysis of Microarrays (SAM), and combination by score and rank function. CMAP-based discretization by up/down tags of genes revealed candidate perturbagens in forms of agonists and antagonists. Pertubargens pattern was plotted by CMAP score versus ATC code (Anatomical Therapeutic Chemical classification), calculated permutation p values. RESULTS: AF were collected from 6 pregnancies with DS fetuses and 6 with normal. One of the DS was partial trisomy. Among 22,572 gene probes investigated, 29 genes were significantly up-regulated by all three tests. Eighteen (63%) of them were on chromosome 21. In data fusion exercises, we found combination of rank scorings was more effective than any single. Using discretization by CMAP, there were 158 agonists and 63 antagonists as candidate pertubagens. Among agonists, ATC-L (immunosuppressive agents) was the most frequent occurrence and two were significant by p value. Among antagonists, ATC-A (oral glucose lowering drugs) were the most frequent, but none was significant. CONCLUSIONS: Our microarray analysis determined expressions of 29 genes potentially important in DS and found immunosuppressive as significant agonist for DS in developing fetus. It is necessary for further study.

The Leu544Ile The biological effects on cells by growth hormone (GH) are mediated directly through the interaction between GH and GH receptor (GHR). We investigated the association between the polymorphisms in the GHR gene and changes in height standard deviation scores(SDSs), or the lipid metabolism during GH treatment for GH deficient children. Design: A one-year study: determining the growth rate and the lipid metabolism under GH treatment. Patients: Eighty-three children (61 boys and 22 girls) with the GH deficiency were treated with GH for one year after the diagnosis. Intervention: The patients were treated with recombinant human GH (0.19 mg/kg/week) for at least one year after the diagnosis. The growth rates and biochemical parameters for lipid metabolism were measured both before and during the treatment. The four types of SNPs in the GHR gene, Cys440Phe, Pro495Thr, Leu544Ile and Pro579Thr, and exon 3 deletion polymorphisms were genotyped by direct-sequencing and multiplex PCR. Results: We found no significant association between the GHR polymorphisms and changes in the height SDSs during GH treatment. The total cholesterol levels of the GH deficient boys with Ile/Ile at codon 544 showed significantly higher cholesterol levels before GH treatment and then maintained those the high levels during the GH treatment than those with other genotypes. No other polymorphisms seemed to have any apparent effects on the lipid metabolism. Conclusion: The Leu544Ile polymorphism of the GHR gene was therefore associated with the cholesterol levels in boys with GH deficiency.

Plasma adiponectin level and sleep structures in children with Prader-Willi syndrome [BACKGROUND] Maple Syrup Urine Disease (MSUD) is a recessive metabolic disorder caused by defective function of the branched chain α-ketoacid dehydrogenase complex (BCKD). Mutation analysis of the dihydrolipoyl transacylase (E2), the alpha and beta subunits of the branched chain α-ketoacid decarboxylase (E1α, E1β) genes of the BCKD complex was done in 33 Filipino patients.

[METHODS] A quick but highly sensitive and specific mutation scanning method, called the COPPER (Condition-Oriented-PCR-primer-Embedded-Reactor) plate system, to analyze the entire coding regions of the E1α, E1β and the E2 genes was used. The coding regions were amplified using 31 primer pairs, all with the same cycling conditions, and aliquoted on a 96-well format polymerase chain reaction (PCR) plate. This method allowed simultaneous amplification of all coding regions of the 3 genes using a single block in a thermal cycler.

[RESULTS] Using this method, 7 novel mutations were identified -2 missense mutations (G132S, M348K) in the E2 gene, 1 missense mutation (S339L) in the E1β gene and 2 nonsense mutations (Q157X, Q190X) in the E1α gene. We also identified an A to G nucleotide substitution changing the start codon ATG to GTG in the E1β gene. Another novel deletion involving nt 788-790 (TCT) was identified in exon 6 of the E1α gene.

[CONCLUSIONS] These findings show that the COPPER plate system is an ideal tool for mutation analysis. This study also provides the molecular basis for understanding this genetic disorder among Filipino MSUD patients.

In vivo dilatation of the neonatal and fetal ductus arteriosus by inhibition of Rho-Kinase in rats

Katsuaki Toyoshima, Kazuo Momma, Daiji Takeuchi, Toshio Nakanishi Section of Pediatric Cardiology , Tokyo Women's Medical University, Tokyo, Japan.

Background: Rho-kinase activation is an additional mechanism that sustains ductus arteriosus (DA) constriction. Objectives: To clarify the ductus-dilating effects of Rho-Kinase inhibitor in nearterm neonatal and fetal rat, in in-vivo studies. Methods: The in vivo ductal diameter of neonatal and fetal rats was measured using a rapid whole-body freezing method, by cutting on a freezing microtome and measuring with a microscope and micrometer. Near-term (21st day of pregnancy: term-21.5 days) neonatal rats were incubated at 33 degrees Celsius following caesarean section. The DA constricted quickly after birth, and the DA diameter was 0.80, 0.08 and 0.06 mm at 0, 60 and 90 min after birth. The ductusdilating effect of a Rho-Kinase inhibitor, fasudil, was studied by intraperitoneal injection at 60 min after birth, and the diameter was studied 30 and 60 min later. The differential effect of fasudil on the near-term were studied by orogastric administration of indomethacin (10 mg/kg) and fasudil into pregnant rats and studying the fetal ductus 4 h later.

Results: Fasudil dilated the neonatal DA dose-dependently. The ductus -dilating effect of fasudil was maximal at 30 min. Fasudil (100 mg/kg) dilated the neonatal constricted DA completely to 0.78 mm. Fasudil (1 mg/kg: clinically equivalent dose) dilated the DA to 0.15 mm. Fasudil inhibited fetal ductus constriction induced by a large dose of indomethacin dose-dependently. Conclusions: The Rho-kinase inhibitor (fasudil) dilated the constricted neonatal and fetal DA dose-dependently in rats. Rho-kinase inhibitors may be useful in maintaining ductus arteriosus patency, as a bridge to congenital heart surgery.

Aerosolized Iloprost for Pulmonary Vasoreactivity Testing in Children with Long-Standing Pulmonary Hypertension Related to Congenital Heart Disease Alisa Limsuwan, Anan Khosithseth, Suthep Wanichkul, Pongsak Khowsathit

Mahidol University, Department of Pediatrics Background: In children with congenital heart disease (CHD) and significantly increased pulmonary blood flow and pressure, progressive pulmonary vascular changes can leads to an irreversible stage. Pulmonary hemodynamics are used to determine the stage whether the surgical correction is indicated. In children with pulmonary hypertension (PH) from CHD, the pulmonary vascular resistance (PVR) > 6 Wood U/m 2 or a pulmonary-to-systemic resistance ratio (Rp/Rs) > 0.3 has been shown to be associated with a poor operative outcome. Therefore, assessment of the vasodilator capacity is important. Whether aerosolized iloprost could be used for the vasoreactivity testing in children with a longstanding PH from CHD is needed to determine. Methods: Children with PH from CHD underwent for hemodynamic evaluation and vasoreactivity testing using iloprost 0.5 mcg/kg delivered through jet nebulizer. Hemodynamic calculations, based on the Fick's principle, were obtained. We consider a decrease in both PVR and Rp/Rs of > 10 % as a responder.

Results: Nineteen children aged 7 months to 13 years, mean age 4.8 years, were tested. Eleven children had a positive response, resulting in a mean±SD decrease in PVR of 6.2±4.3 to 3.5±3.1 WU/m 2 and in Rp/Rs of 0.33±0.21 to 0.19±0.17 (p<0.01). Nine of the 11 responder children underwent surgical correction of their CHD. Conclusions: Iloprost induced vasodilator capacity varies among children with PH and elevated PVR. Our study suggested iloprost may be benefit in preoperative identification of children with long-standing PAH secondary to congenital heart disease for surgery. However, additional vasodilator testing may be required.

Does "treating" mild rejection prevent significant rejection in pediatric heart transplantation?

James Y. Coe, Kim Myers, Christine Liberty, Lori West Pediatric Cardiology, University of Alberrta Heart transplantation is an accepted treatment for end stage pediatric heart failure. Life after transplantation include intensive and regular invasive and non-invasive monitoring to preempt acute rejection and the development of transplant coronary artery disease and post transplant lymphoproliferative disease. Compliance is an ongoing challenge in the adolescents. It is generally accepted that mild rejection (ISHLT grade 1R or 1A, 1B & 2) do not require treatment. However, grade 1B was more likely than 1A or 2 to degenerate to moderate 3A rejection. Over the past decade, 57 children (age 0.5-16. 4 years) has been transplanted at Alberta Provincial Pediatric Heart Transplant Program. There were 3 late deaths (compliance n=2, chest infection n=1) and 3 early deaths (high PRAs n=1, multi-organ failure n=2). One teen had a late moderate rejection related to compliance and treated successfully. Excluding the noncompliant patients and early deaths, all remaining patients were monitored with 3-4 biopsies in the first year and annually thereafter if rejection free. All mild rejections (1A & 1B) were "treated" by increasing the immusuppression, balancing the dose against renal function, and re-biopsied monthly until rejection free (ISHLT grade 0). Immunosuppression consists of ATGAM for induction, triple therapy (tacrolimus, mycophenolate, prednisone) for 9-12 months. Steroid wean starts at 6 months if rejection free and the patients were usually off steroid by 9-12 months. Rejections ≥ grade 2 has not been observed in the last decade. We conclude that "treating" mild rejections prevent the development of significant rejection in the pediatric heart transplants.

Noninvasive assessment of pulmonary artery resistance and pressure in patients with congenital heart disease: A new method using M-mode echocardiography Sumito Kimura, Yayoi Nakahata, Natsuko Owada, Hisashi Ando, Masahiro Ishii

Kitasato University, Department of pediatrics Aim: Accurate evaluation of pulmonary artery resistance <Rp> and pulmonary artery mean pressure <PAMP> were important for management of patients with congenital heart disease <CHD>. It is usually required for an accurate measurement by cardiac catheterization. We assessed the severity of pulmonary artery hypertension in patients with CHD using motions of the interventricular septum <IVS> by M-mode echocardiography. Method: We performed echocardiography on 46 patients <2 months to 20 years old> with CHD include in complex CHD. Cardiac catheterization was performed within 1 day of the echocardiography study in all cases. All patients were divided into 2 types by IVS motion evaluated using M-mode echocardiograph: type A marked anterior motion in early systole; type B marked posterior motion in early diastole. Right ventricular (RV) volume overload affects the left ventricular (LV) function in patients after tetralogy of Fallot (TOF) repair. However, the impact of RV dilation on regional LV wall motions was unclear. In this study, we determined the relationships of RV size as derived from cardiovascular magnetic resonance (CMR) with the global and regional LV wall motion. Methods: Echocardiogram was performed on 25 postoperative TOF patients aged 17.8±4.4 years and data were compared with 23 age-matched controls. The longitudinal, radial and circumferential components of LV wall motion were analysed using speckle tracking method. CMR-derived parameters of all TOF patients were used for correlation analysis. Results: When compared with controls, TOF patients had reduced global LV longitudinal strain (p=0.003) and SR (p=0.001), and global LV radial strain (p=0.001). Reduced global LV radial strain was contributed by lower strain in inferior, lateral, posterior, and anterior segments (all p<0.05). TOF patients had lower circumferential strain of LV inferior-septal (p<0.001) and inferior (p=0.001) segments. In TOF patients, global LV circumferential strain and SR correlated negatively with indexed RV end-diastolic volume (r=-0.61, p=0.002 and r=-0.45, p=0.03 respectively) and indexed RV end-systolic volume (r=-0.61, p=0.002 and r=-0.53, p=0.009 respectively). LV ejection fraction decreased with indexed RV end-diastolic (r=-0.67, p=0.002) and end-systolic volumes (r=-0.73, p<0.001), and increased with global LV circumferential SR (r=0.58, p=0.015).

The negative impact of RV dilation on LV function relates to its influence on LV circumferential wall motion in patients after TOF repair. Background: We sought to determine the usefulness of novel 2-dimensional strain indices, based on speckle tracking imaging, for assessment of systemic right ventricular (RV) function in patients after atrial switch operation for transposition of the great arteries. Methods: Twenty-six patients, aged 21.0±3.6 years at 19.9±3.2 years after atrial switch operation, and 27 age-matched controls were studied. Two-dimensional imaging at the 4 chamber view was obtained with tracing of the entire RV endocardial border. The RV global longitudinal strain (GLS) and GLS rate were derived using an automated software (EchoPAC, GE Medical), and correlated with tissue Doppler-derived RV isovolumic acceleration (IVA), and, in patient cohort, with cardiac magnetic resonance-derived RV ejection fraction. Results: Intraobserver and interobserver variability for measurement of GLS, as determined from mean ± SD of differences in 2 consecutive results from 20 studies, were 0.06±1. 93% Ming-Tai Lin 1 , Shyh-Jye Chen 2 , Chun-An Chen 1 , Shuenn-Nan Chiu 1 , En-Ting Wu 1 , Wen-Jeng Lee 2 , Jou-Kou Wang 1 , Mei-Hwan Wu 1 1 Department of Pediatrics, National Taiwan University Hospital, Taipei, Taiwan, 2 Department of Medical Imaging, National Taiwan University Hospital, Taipei, Taiwan Background: MDCT had been applied to congenital heart disease for several years. However, large scale application of this new imaging tool to the patients with PA and VSD had never been reported before. Objective: In a tertiary medical center, 11-year experience of MDCT in PA with VSD will be reviewed with focus on the pulmonary artery morphology and trachea stenosis. Methods: Between 1995 and 2005, 110 patients with PA and VSD visited our hospital. 211 times of MDCT were conducted in 103 of them. Those who received operation three months before or after their MDCT examinations were enrolled. Four patterns of pulmonary artery morphology were defined: native main pulmonary trunk present (group I); confluent central pulmonary arteries without trunk (group II); nonconfluent central pulmonary arteries (group III); absent intrapericardial pulmonary arteries (group IV). Results of pulmonary artery morphology from MDCT were compared with the operation findings. Existence of trachea stenosis was also evaluated. Results: Data of 95 times of MDCT (74 patients) with concurrent operations were collected. Based on the surgical findings, distribution of each group is: 53 (group I, 55.8%), 25 (group II, 26.3%), 12 (group III, 12.6%), 5 (group IV, 5.2%). The overall accuracy for MDCT was 91.6% (87/95). The group specific accuracy was between 88.7% and 100%. In addition, trachea stenosis was detected in 8 patients (10.8%). Conclusion: For PA with VSD, MDCT is a useful tool to define pulmonary artery morphology and possible trachea stenosis, both of which will influence the surgical outcome.

1 Toyama University, Department of Pediatrics, 2 Toyama University, Department of Psycology, 3 Toyama University, Departmento of the first surgery Background: Increased survival in infants with congenital heart disease has raised interest in the neurodevelopmental sequelae, but there is no data regarding neuroanatomical development in infants with critical CHD. Volumetric study of the brain using three dimensional magnetic resonance imaging in infants with CHD can help to identify effects on anatomical development of the brain that may be associated with functional impairment. Methods and Results: Thirty-nine infants with CHD were studied prospectively with brain 3D-MRI several months after heart surgery, and compared to nineteen healthy control infants. Neurodevelopmental assessment was performed using Bayley II infants developmental scales. The global volume of grey matter was significantly reduced in the patients with CHD compared to normal control (P<0.001) while no significant difference of the volume of white matter was observed. Further, decrease of GM volume was more apparent in frontal lobe than that in temporal lobe. Multivariate analysis revealed that preoperative hypoxia is strongly associated with decreased frontal GM volume (P=0.007). Further, hypoxic patients have a trend toward smaller regional brain volumes, most prominently in frontal GM (P=0.017), as well as more delayed psychomotor development (P=0.05) than non-hypoxic patients. Of note, frontal GM volume was positively associated with psychomotor developmental index PDI score (P=0.005) , but not with mental developmental index. Data, such as students' demographics were obtained. Initial developmental quotients and comparing with the developmental quotients at least 6 months after therapy of both groups were gathered. The type of scheduling in which the students were enrolled was done to see if their DQ in the subsequent evaluation were significantly higher than the initial. Improvement is defined by a child's step up in their level of functionality. Those who obtained the same or lower level were labeled as no improvement. Results: Out of 37 autistic children enrolled in DLSU-NDC, only 30 students were included in the study. Fourteen of them received once-a-week therapy session (Group 1), while the rest receive more than once-a-week therapy (Group 2). Twelve of the students in Group 1 gained a higher DQ, but only 5 of them improved to the next level of functionality. Eleven students in Group 2 had increments in their DQs but only half of them stepped up from their initial functional level. Seventy-seven percent (77%) of subjects improved Personal and Social Area, followed by the Performance area of Development (73%). In Group 1, the average difference between the initial DQ and the subsequent DQ is with marginal significance. While in Group 2, their average difference is statistically significant. Conclusion: The frequency of therapy session is not associated with the development or deterioration of their Develepmental Quotients(DQs). Most of the autistic children receiving intervention improved in their performance and personal and social areas, which then helped them to make the most productive use in their advancement in mental skills.

Prevalence of sleep problems among elementary students in southern Thailand Introduction: Although enzyme replacement therapy has been established in mucopolysaccharidosis (MPS), pathogenesis of neurodegeneration still remains to be elusive. We have clarified the involvement of oxidative brain damage and reduction of GABAergic interneurons in the cerebral cortex in MPS autopsy cases. Here we immunohistochemically examined beta-amyloid deposition, tauopathy and synucleinopathy in the brain disorders of MPS. Materials and methods: Subjects were composed of three cases each of Hunter syndrome (MPSII) and Sanfilippo syndrome (MPSIIIB), and four controls without pathological changes in the brain. We performed immunohistochemistry on expressions of beta-amyloid protein, phosphorylated tau protein, phosphorylated alpha-synuclein and ubiquitin in serial sections of the frontal and temporal cortex, striatum, thalamus, cerebellum and midbrain Febrile seizures are the most common seizure disorder in childhood. However, the pathogenesis of febrile seizures has scarcely been resolved. Oxidative stress can be implicated in various diseases of the central nervous system. The aim of the present study has been to further contribute to the understanding of involvement of oxidative stress in febrile seizures.

[Methods] Patients with prolonged/repetitive febrile seizures (n=13) and age-matched controls (n=12) were included in this study. Cerebrospinal fluid levels of 8-hydroxy-2-deoxyguanosine (8-OHdG), hexanoyl lysine adduct (HEL) and acrolein lysine adduct (ACR) were measured using ELISA techniques.

[Results]

In 5 of 13 patients with febrile seizures, CSF levels of 8-OHdG were elevated, although there was no significant difference of the average levels of 8-OHdG between patients with febrile seizures and controls. CSF HEL levels were elevated in 3 of 12 patients with febrile seizures and 2 out of these 3 patients had high levels of CSF 8-OHdG. Another 2 patients with febrile seizures had high levels of CSF ACR. Importantly, elevated 8-OHdG was associated with an increased risk of repeated seizures.

[Conclusions] Evidence shows repeated seizures can lead to the development of mesial temporal sclerosis and our results suggest that oxidative stress may partly contribute to these processes. immunity responses play an important role in viral infections. To identify whether Toll-like receptor (TLR) and related genes (TLR3, TLR7, TLR9, and UNC93B1) contribute to the development of FS, we performed a case-control study using single nucleotide polymorphisms (SNPs). Methods;We genotyped the six SNPs in four genes based from the data of the International HapMap Project in 249 FS patients (186 simple and 63 complex) and 225 control subjects. Genotyping was performed with TaqMan SNP Genotyping Assay. The Haploview was used to construct haplotypes. Results were analyzed using chi-square tests. Results;The frequency of G allele at rs308328 (UNC93B1, intron7) was significantly higher in FS patients than control subjects (p=0.02). In subgroup analysis, the genotype distributions were significantly different between complex FS, but not simple FS, and controls (corrected p=0.039). No associations were detected between SNPs in TLR3, TLR7, TLR9 and FS susceptibility. Conclusion;The present study suggested that UNC93B1 gene contributed to a genetic susceptibility to development of FS, especially complex FS. UNC93B1 interacts with TLR3, TLR7 and TLR9. The congenital deficiency is linked to the etiology of herpes simplex virus-1 encephalitis in humans (Casrouge, 2006) . Inflammatory processes caused by direct viral invasions might be involved in the pathophysiology of complex FS.

Insufficient secretion of aldosterone causes urinary sodium loss in extremely premature infants of less than 26 weeks of gestation Methods: During the past 3 years, we performed a renal Doppler ultrasound in 216 children with macro-(n=40) or microhematuria (n=176). Peak velocity (PV) was measured in the transverse plane at two points in the left renal vein (LRV), one at the hilar portion of the LRV, and the other at the aortomesenteric portion. Anthropometric measurement of height, weight, body surface area (BSA), and body mass index (BMI) were made at the time of ultrasound. Results: They were divided into two groups according to the PV ratios of the LRV: nutcracker group (PV ratio > 4.1, n=72) and non-nutcracker group (PV ratio < 4.1, n=144). There were no differences in the mean age, gender, weight and BSA between the two groups. However, height was significantly higher in nutcracker group than non-nutcracker group (p=0.031), and BMI was significantly lower in nutcracker group than in non-nutcracker group (p=0.006). Furthermore, the PV ratios of the LRV in 216 children with hematuria inversely correlated with BMI (PV ratio = 6.936 -0.17 X BMI, r = -0.188, p=0.006). This inverse correlation was also observed when we selected 113 male patients (r = -0.267, p = 0.004), but not in 103 females (p = 0.478). Conclusions: These results suggest that patients with nutcracker syndrome were leaner than those without nutcracker syndrome, and height and BMI may influence on left renal venous flow velocity in patients with hematuria.

Effective and safe treatment with cyclosporine in nephrotic children: a prospective, randomized, multicenter trial 

Mainstreaming nutrition for improving nutrition in developing countries Tahmeed Ahmed Nutrition Program, ICDDR,B Improved nutrition is critical to improving health of mothers and children in developing countries. Without this improvement in nutrition, the Millennium Development Goals cannot be achieved. Nearly a third of the children in the developing countries are malnourished. About 60% of the deaths in children are associated with malnutrition. There is enough evidence linking nutrition and good maternal and child health (MCH) outcomes; however, nutrition has been poorly integrated with MCH programs in most countries. The Mainstreaming Nutrition Initiative, formed with support from the World Bank, aims to converge the nutrition agenda in selected countries with the MCH agendas towards the achievement of MDGs. This is based on emerging international consensus that convergence of nutrition and MCH is essential. The primary objective of this initiative is to create a partnership between ICDDR,B and global partnerships in health and nutrition to catalyze a convergence of the nutrition and MCH agendas. A conceptual framework for the mainstreaming process as well as tools for assessing the process has been developed. A global review of nutrition has been done with a view to find out evidence for mainstreaming nutrition into MCH programs. The review has contributed substantially to the upcoming Lancet Nutrition Series. Activities related to mainstreaming nutrition have been initiated in several countries including Bangladesh, Viet Nam, Pakistan, Uganda, Ethiopia, Bolivia and Peru. These activities will include assistance in formulating policy on mainstreaming and integration of simple and efficacious nutrition interventions. For more information on the MNI, please visit http://www.icddrb.org/activity/ index.jsp?activityObjectID=2347.

Is Helicobacter pylori infection a cause of gastric hypoacidity in children in developing countries?

Little is known about effect of Helicobacter pylori (Hp) infection on gastric acid secretion (GAS) in children. The aim of this study was to determine whether Hp infection in children is associated with perturbation of GAS. Methods: We conducted this study on 30 asymptomatic Hp-infected (positive urea breath test; UBT) and 30 Hp-non infected children (2-5 years) from a peri-urban community near Dhaka City. The gastric pH and GAS was measured before and 60 days after anti-Hp therapy (omeprazole, clarithromycin and amoxicillin for 2 weeks). Gastric acid outputs (Mmol/h) were quantified during a 1-hr basal period (BAO) and 1-hr stimulated period (SAO) with subcutaneous pentagastrin (6µg/kg). Hp status was reassessed 60 days after anti-Hp therapy. Results: A significantly greater number of infected children had a very high gastric pH (> 5.5) and demonstrated absolutely no acid output (achlorhydria) in basal state compared to the age matched non-infected group. The acid concentration and volume of gastric secretion were also decreased in infected children. Furthermore, the mean BAO and SAO in Hp-infected children were 50% lower than those with non-infected. However, the eradication therapy in infected children resulted in increase in volume of gastric secretion, lowering of gastric pH, reversal of achlorhydria and significant rise of SAO ( [Methods] Totally 441 HBV carrier children followed-up for more than 15 years were recruited. They were grouped based on their initial HBeAg/anti-HBe status. HBeAg (+) group was further divided into HBeAg (+/+) group (n=152) and HBeAg (+/-) group (n=229) depending on whether spontaneous HBeAg seroconversion occurred during follow-up. Another 25 children with HBV-related hepatocellular carcinoma (HCC) were also studied. HLA class I-restricted T cell epitope mutation of HBsAg gene was examined by PCR and direct sequencing at the latest follow-up of these carrier children.

[Results] A lowest T cell epitope mutation rate was found in HBeAg (+/+) group. The overall mutation rate wassinilar among the HBeAg (+/-), HBeAg (-/-), and HCC groups. Those with mutations had a higher peak ALT level than those who did not ( Objectives: Oxidative stress is known to be involved in the pathogenesis of biliary atresia (BA), but the mechanism is yet to be elucidated. We studied 8-hydroxydeoxyguanosine (8-OHdG) and mitochondrial copy number as potential markers for oxidative stress in BA. [Methods] We focused on the known functional SNPs in the MMP genes (MMP-2 -735C>T, MMP-3 -1612 5A/6A, MMP-9 -1562C>T, MMP-12 -82A>G and MMP-13 -77A>G) and performed the association study between these SNPs and the development of CAL in KD. To confirm the genetic association of the MMP-13 gene to CAL formation in KD, we also evaluated the haplotype frequencies of the MMP-13 gene in the promoter region between the groups. The study population consisted of KD patients with CAL (n=44), without CAL (n=92) and controls (n=175). The genotypings of the polymorphisms were performed by PCR-RFLP and TaqMan ® real-time PCR methods.

[Results] Allele and genotype frequencies of -77A>G polymorphism in the MMP-13 gene showed significant differences between KD patients with CAL and without CAL (P = 0.00551 for the genotype and 0.00989 for the allele).

The estimated frequencies of G-C haplotype in the MMP-13 gene promoter were significantly lower in KD patients with CAL than in those without CAL (P = 0.0155). There was no association between other MMP genes and CAL development.

[Conclusions] Our study has demonstrated that the MMP-13 gene appears to be a susceptibility gene to the development of CAL in KD patients.

Polymorphisms of HLA Genes in Korean Children with Kawasaki Disease Purpose: Kawasaki disease (KD) is a leading cause of acquired heart disease in children. The prevalence rate was reported to be varied in different ethnic groups. Recently, with the remarkable development of the genetic studies, polymorphisms of HLA have been defined by DNA analysis. The aim of this study was to assess the influence of the alleles of HLA to susceptibility and complication of KD in Korean children. Methods: DNA was extracted from children diagnosed with KD (n=74). The polymorphisms of the alleles of HLA-A, -B, -C, -DRB1 of the KD patients were determined by utilizing the PCR-ARMS and PCR-SSP analysis. The polymorphisms discovered were compared to those of normal healthy controls (n=159). Results: There was a significant increase in the frequencies of alleles of HLA-B35, -B75, -Cw09 in patients with KD when compared to those of the healthy control group. With subgrouping of KD patients into groups with or without a coronary complication, the frequencies of HLA-DRB1*11, were increased in a group with a coronary complication when compared to a healthy control group. In comparison between subgroups with or without a coronary complication, the frequency of HLA-DRB1*09 was only increased in the group with coronary complication. To examine the efficacy of high-frequency oscillatory ventilation (HFO) combined with partial liquid ventilation (PLV) in a rabbit model of acute lung injury. Methods: 17 animals were treated with repetitive saline lavage to achieve a uniform degree of acute lung injury (PaO 2 below 100 mmHg on a FIO 2 of 1.0), and were randomized to either HFO-PLV or CMV-PLV group. 9 animals in HFO-PLV group received intratracheal administration of 5ml/kg perflubron, and were ventilated with HFO for 3 hrs after the dosing period. 8 animals in CMV-PLV group were provided with 5ml/kg perflubron and ventilated with CMV for 3 hrs. The other 5 animals were not lavaged and served as non-lavaged PLV control. They were ventilated with CMV for 3 hrs after the instillation of 5ml/kg perflubron. After the ventilation, lung lavage fluid of each animal was collected and analyzed. Results: Animals in HFO-PLV group had significantly higher mean PaO 2 and lower PaCO 2 values than animals in CMV-PLV group. The proportion of surfactant in large aggregate (LA) forms to total surfactant was significantly higher in animals of HFO-PLV group and non-lavaged PLV control than CMV-PLV group. SA/LA ratio was significantly lower in HFO-PLV group and nonlavaged PLV control respectively compared with CMV-PLV group. Conclusions: HFO had beneficial effects not only on gas exchange but also on the preservation of active forms of endogenous surfactant during PLV. We conclude that HFO is a superior ventilatory method to CMV in applying PLV for clinical practice from the viewpoint of endogenous surfactant activity.

In vivo Dilatation of the Postnatal Ductus Arteriosus by Atrial Natriuretic Peptide in the Rat.

Katsuaki Toyoshima, Kazuo Momma, Toshio Nakanishi Section of Pediatric Cardiology , Tokyo Women's Medical University, Tokyo, Japan.

Background: Alpha-human atrial natriuretic peptide (hANP) reportedly increases in premature infants with patent ductus arteriosus (PDA). Objectives: To clarify possible hANP effect to reopen the postnatal ductus arteiriosus (DA), we studied in vivo reopening of the postnatal DA by a recombinant hANP (carperitide) in rats. Methods: Near-term neonatal rats were incubated at 33 degrees Celsius following caesarean section. The DA diameter was measured with a microscope and a micrometer following rapid whole-body freezing. The DA constricted quickly after birth, and the DA diameter was 0.80 and 0.08mm at 0 and 60 min after birth. The ductus-dilating effect of carperitide was studied by subcutaneous injection at 60 min after birth, and the diameter was studied 7, 15, 30 and 60 min later. We investigated the serum hANP concentrations following subcutaneous injection of carperitide. Results: Carperitide dilated the postnatal DA dose-dependently.The DA-dilating effect of Carperitide was maximal at 7 min, and disappeared at 60 min after injection. Carperitide (10 mg/kg) dilated the postnatal constricted DA completely to 0.79 mm. Carperitide (1 mg/kg) dilated the DA to 0.55 mm. The serum hANP levels at 7 minutes after the injection of Carperitide (1mg/kg) was 790 pg/ml. Conclusions: hANP reopens the constricted postnatal DA dose dependently in rats. The increased hANP, accompanying symptomatic premature PDA, may delay closure of the DA. Therefore, cycooxygenase inhibitors should be used before the level of hANP becomes high accompanying symptomatic PDA. There existed disparities among the intercultural couples including paternal age, parental educational level, and residential distribution. The rates of low birthweight and preterm births in the neonates born to foreign-born mothers were lower than those of the counterparts. The adjusted odds ratios of these above outcomes were 0.74 and 0.73, respectively. Early neonatal and neonatal mortalities were lower in the former than the latter. The adjusted hazard ratios of being an early neonatal and neonatal death in babies born to foreign-born mothers were 0.69 and 0.75, respectively. Conclusions: Despite lower levels of parental education, advancing paternal age, and other possible socioeconomic disparity, we demonstrated an epidemiologic paradox of favourable neonatal outcomes among immigrant mothers in Taiwan.

Takeshi Morisawa, Ronald J. Wong, Vinod K. Bhutani, Hendrik J. Vreman, David K. Stevenson

Dept. of Pediatrics, Stanford University School of Medicine, Stanford, CA BACKGROUND: Heme oxygenase (HO) catalyzes the degradation of heme to produce bilirubin. Because excess bilirubin production can lead to the development of jaundice, use of HO inhibiting drugs, such as metalloporphyrins, may be a promising treatment strategy for the prevention of neonatal hyperbilirubinemia. We and others have reported that imidazole dioxolanes also can inhibit in vitro HO activity with selectivity for the inducible HO-1 isozyme and do not affect nitric oxide synthase (NOS) or soluble guanylyl cyclase (sGC), in contrast to metalloporphyrins. Therefore, our objective was to investigate the efficacy of the imidazole dioxolane, Azalanstat, towards inhibiting in vivo HO enzyme activity in newborn mice. METHODS: Azalanstat (500 µmol/kg) or vehicle was administered to 7-dayold FVB mice via intraperitoneal injections. At various times, mice were sacrificed and liver, spleen, and brain harvested. Liver, spleen, and brain HO activities were then quantitated by gas chromatography, calculated as pmoles of carbon monoxide (CO) produced/hr/mg fresh weight, and then expressed as % inhibition of tissue HO activity from treated versus controls. RESULTS: HO activity was maximally inhibited in the spleen (54%) and brain (39%) within 0.5h, and in the liver (28%) at 3h after Azalanstat treatment. Liver HO activity returned to control levels within 24h. In contrast, spleen HO activity increased 18%; while brain HO activity was still inhibited (19%).

We conclude that Azalanstat effectively inhibits HO activity, and its action is immediate and tissue-specific. Thus, imidazole dioxolanes may be attractive alternative compounds for use in the treatment of neonatal hyperbilirubinemia. There have been many studies on bilirubin levels in normal newborn infants, the definition of "normal bilirubin level" has not been established. This is because bilirubin levels change rapidly in the first 72 hours and vary considerably depending on gestational age, racial composition of the population, proportion of breastfed infants, and other genetic and epidemiological factors. However, most studies were carried out on infants with white color skin. In Brunei, the majority of newborn infants are Malay. Hospital stays in Brunei are now short, and measurements of predischarge total serum or TcB (JM-102) are used to determine when additional evaluation is necessary and to predict the risk of subsequent hyperbilirubinemia. In this study, we tried to measure bilirubin levels in normal newborn infants using the new Konica-Minolta transcutaneous jaundice device JM-103. We measured bilirubin levels from 0 to 72 hours after birth in 725 newborn infants (gestational age: >35 weeks) who had been admitted to the well-infant nursery of Raja Isteri Pengiran Anak Saleha (RIPAS) Hospital, Brunei Darussalam. Using a daily list of all deliveries, the nurses obtained JM-103 measurements 3 times every day. All infants in the nursery were eligible for the study. This is the first contemporary study, including the velocity of increase in TcB levels, for neonatal bilirubinemia in a predominately breastfed population. In the future, making nomogram of Brunei Darussalam and combining the use of nomogram and JM-103 will be useful for evaluation of neonatal jaundice, especially in newborn infants of Brunei Darussalam. The mean gestational age and birth weight for study and control groups were 38+3 weeks, 3,143gm and 39+1 weeks, 3,228gm, respectively in phase 1 study. The significant factors for readmission included gestational age, ABO incompatibility, breast feeding, jaundice at discharge, oxytocin, discharge within 72 hrs after birth and meconium stain. The formula to predict the readmission among population of phase 2 had sensitivity and negative predictive values of 87%, 75%; 80%, 74%; and 59%, 76% for 85%, 90% and 95% of readmission, respectively. When readmission rate calculated was <85%, 85-94% and ≥95%, then follow-up in 2 weeks, follow-up in 3 days and held discharge, respectively, were done. Following such CP resulted in decreased readmission from 2.2% to 0.7% (p-value 0.013). 4. Conclusion CP based on readmission rate derived by using an equation developed during this study merits further study to verify its accuracy and safety.

Kanokpan Ruangnapa, Prasin Chanvitan

To determine mortality and morbidity rates and selected outcome variables of very low birth weight (VLBW) infants.

A retrospective study of the outcomes of VLBW infants, admitted to the NICU of Songklanagarind Hospital, Thailand, between January 2003 and December 2006. RESULTS: A total of 178 VLBW infants (86 males and 92 females) met the enrollment criteria. Forty-two infants (23.6%) were referred from other hospitals in southern Thailand. The mean birth weight and gestational age were 1,123 ± 273 grams and 29 ± 3 weeks, respectively. The three most common neonatal morbidities were respiratory distress syndrome (64%), hyperbilirubinemia (64%) and apnea of prematurity (42%). The mortality rate was 27%. Among the selected outcomes of survivors evaluated at discharge (N=130), the incidence of bronchopulmonary dysplasia was 34.6%, with an overall incidence for mild cases of 10%; moderate cases of 20%; and severe cases of 4.6%. Six of those infants (13.3%) were still dependent on oxygen at discharge. Retinopathy of prematurity (stages III to V) was found in 9.2%. Intraventricular hemorrhage (IVH ≥ grade 3) and periventricular leukomalacia were encountered in 3.9% and 7.7%, respectively.

The overall mortality and morbidity rates of VLBW infants in this study are similar to other reports from Thailand and some developing countries.

Takeshi Takami, Daisuke Sunohara, Atsushi Kondo, Yuki Kanetaka, Yukito Takei, Tasuku Miyajima, Akinori Hoshika

[Background] Near-infrared spectroscopy (NIRS) is a noninvasive real-time monitoring technique that provides information on both cerebral oxygenation and cerebral blood flow (CBF). The superior vena cava (SVC) flow assessment by echocardiography has been reported to be a useful predictive marker of intraventricular hemorrhage (IVH) in preterm infants. The present study evaluated the relationship between cerebral perfusion and cardiac function in extremely low birth weight (ELBW) infants.

[Methods] The six ELBW infants in this series were stabilized by mechanical ventilation and had a mean gestation of 25 ± 1.7 weeks (range 23-27) and a mean birth weight of 700 ±125 g (range 551-919 g). Serial echocardiographic assessments were performed at 3-6, 12, 18, 24, 36, 48 and 72 hours after birth. The fractional shorthening of left ventricle (LVFS) was measured using M-mode echocardiography. Superior vena cava (SVC) flow and left ventricular output (LVO) were also measured using Doppler echocardiography. The tissue oxygenation index (TOI) was simultaneously monitored in the front-temporal region using NIRS. Heart rate, mean blood pressure, PaO 2 , and PaCO 2 were recorded.

[Results] The LVFS and LVO decreased transiently at 12 to 18 hours, and then increased gradually after 24 hours of life. The SVC flow and TOI showed almost the same changes in cardiac functions.

[Conclusions] Changes in both SVC flow and TOI were reflected by the changes in cardiac functions. To assess early circulatory status using echocardiography may be useful for the management of cerebral circulation in ELBW infants.

Hung-Wen Chen 1 , Peng-Hong Yang 2 , Li-Yi Tsai 2 , Suh-Fang Jeng 3 , Chyong-Hsin Hsu 4 , Hong-Chih Lin 5 , Chien-Yi Chen 2 , Hung-Chieh Chou 2 , Po-Nien Tsao 2 Results: A total of 14 cases (M:F=10:4) of neonatal listeriosis were identified including 11 surveyed from 4 medical centers and another 3 collected from literature review. 11 were found after 2000 and 3 prior to 2000. The mean gestational age was 32.2 weeks (range 26-38 weeks) and 12 cases were preterm infants. Most of their mothers had history of fever or flu-like symptoms before delivery. Chorioamnionitis was noted in 4 mothers and one mother had L. mononcytogenes bacteremia. The age of onset was less than 3 days in all cases. 11 presented with respiratory distress as the initial symptom. 7 had central nervous system involvement. L. mononcytogenes was identified from blood in 13, cerebrospinal fluid in 4 and gastric aspirate in 2. The mortality rate was around 29 %. Conclusions: Our findings have highlighted that listeriosis may emerge as an important health threat among newborn infants in Taiwan.

Najwa Yudhasompop 1 , Pairoj Boonluksiri 1 , Jeerawan Wannaro 1 , Supaporn Disneewate 2 1 Department of Pediatrics , Hatyai Hospital, 2 Department of Pediatrics , Prince of Songkhla University Background: Neonatal meningitis is a disease with high mortality and morbidity. Diagnosing this condition is more difficult in neonates than in children.

To determine the prevalence of neonatal meningitis and to look for any factors that might predispose the patient to increased mortality risk..

The medical records of neonates aged less than 28 days admitted with meningitis in Hat-Yai Hospital, Thailand from January 2002 to December 2006 were reviewed. Results: The records of 61 neonatal meningitis cases were studied, 29 boys and 32 girls. The prevalence was 1.38 per 1,000 live births. Gestational age was 35.5 weeks, birth-weight 2,412.7 g., and the rate of Apgar scores "<" 3 at 1 minute was 6.5%. The age of disease onset was 9.8 days. White blood cell count in cerebrospinal fluid (CSF) was 741 cells/mm3, with 31.8 % neutrophils. CSF sugar/blood sugar ratio was 0.36. Blood and CSF cultures were positive in 9 cases (14.7%) and 4 cases (6.6%) respectively. E. coli and Enterococci each grew in 2 CSF specimens, and 1 group B streptococcus was detected by latex agglutination. Three cases died (4.9%), all prior to 20 days of age. No independent factors could be associated with an increased risk of mortality.

The incidence of neonatal meningitis at Hat-Yai Hospital was 1.38 per 1,000 live births with a mortality rate of 4.9%. No prognostic factors associated with mortality rate were detected.

Background: Procalcitonin (PCT) has largely studied as marker for neonatal sepsis.

Contradictory results were reported. We aimed to derive a definitive estimate of the diagnosis performance of PCT for neonatal sepsis and identify study-level factors that might predict performance. Methods: We undertook a systematic review of diagnostic studies of PCT that were conducted in neonates. All studies without controlled group were excluded. We searched Medline, EMBASE, Digital dissertation databases, and reference tracking without language restriction. Selected QUANDAS tools were used for validity assessment. Publication bias and its impact were assessed by multiple methods. Random effects meta-analysis was used to derive pooled estimates of positive and negative likelihood ratio, diagnostic odds ratio. Subgroup analysis and meta-regression was used to identify study-level covariates that predicted diagnostic performance.

Results: Of 411 studies found (1980 ( -September 2006 , 28 were included: 21 (English), 5 (Non-English) and 2 conference proceedings. Kappa was 0.8. There was evidence of publication bias. Trim & fill suggested an over-estimation of test performance by publication bias. Subgroup analysis and meta-regression showed that quality of studies significantly has inverse association with test performance. Although the estimated overall area under curve was 0.89, the pooled positive likelihood ratio (+LR) was 6.1 (95%CI 4.6, 8.6), the negative LR negative (-LR) was 0.3 (95% CI 0.2, 3.5) adjusted for different cut-off value. PCT tended to perform better for late-onset sepsis. Conclusion: PCT is not an ideal test for neonatal sepsis. Publication bias and study design and reporting quality explains an over-optimism of PCT.

Anti-Annexin A2 IgM Antibody in Preterm Infants: Its Association with Chorioamnionitis [Background] Intrauterine infection is associated with chorioamnionitis (CAM), which can lead to preterm delivery. We previously reported that the levels of IgM and the incidence of CAM were elevated in preterm infants with Wilson-Mikity syndrome. The molecular target of this IgM remains unclear.

[Methods] In order to determine the target proteins, we used Western blot and amino acid sequences. ELISA for annexin A2 was established to titrate the annexin A2 IgM antibody using the recombinant human annexin A2. The plasmin generation inhibition assay was conducted. To examine the correlation between the titer of anti-annexin A2 IgM antibody and CAM, we prospectively titrated anti-annexin A2 IgM antibody in preterm infants who were admitted to the NICU in and after 2005.

[Results] We identified annexin A2 as the target of IgM antibody in preterm infants. About 60% of preterm infants with hyper-IgM showed high titer against annexin A2 in their cord blood. The patients' IgM inhibited plasmin generation. The titer of anti-annexin A2 IgM antibody was significantly higher in patients with high-grade CAM than in patients without CAM and in patients with lowgrade CAM.

[Conclusions] This is a first report about the autoimmune IgM antibody produced by preterm infants at the local area of the fetomaternal interface. The titer of this anti-annexin A2 IgM antibody and/or annexin A2 protein might be able to function as the marker of CAM, i.e. fetal inflammatory response syndrome (FIRS). Further studies are needed to clarify the participation of this IgM antibody in developing CAM.

Hemodynamic changes before and after intravenous gamma globulin infusion in patients with Kawasaki syndrome Kurume University, Department of Pediatrics and Child Health BACKGROUNDS: To determine the difference in cardiac performance before and after intravenous gamma globulin infusion (IVGG) in patients with Kawasaki syndrome (KS). METHODS: Subjects were 17 patients with KS who were treated with 2 g/kg of IVGG and did not have coronary artery lesion and 9 controls. In Doppler and Tissue Doppler echocardiography, peak aortic velocity (D-AV), peak mitral E and A wave velocity, E/A ratio, aortic ejection time (D-ET), mitral annular peak s', e', and a' velocity, isovolumic contraction time, isovolumic relaxation time (T-IRT), heart rate (HR) were determined. In addition, isovolumic acceleration (IVA) were calculated. Echocardigraphic variables were compared between before and after IVGG as well as patients and controls. RESULT: Mean patients' age was 3.5 years old. Pre-and post-IVGG study were performed at 5.4 and at 11.9 days of illness. In pre-IVGG study, there were significant differences in D (N=28), non-CAL group(N=49), responder group(N=70), and the non-responder group requiring the additional IVIG(N=7). Before and after the initial administration of IVIG, serum IgG levels were measured. Initially IVIG therapy was given with single 2g/kg for 1 day, and then additional IVIG (1g/kg for 1day) was treated if the patients had a sustained fever over the 38.0°C after 48hr from starting therapy. Results: The level of IgG at postIVIG and its difference between before and after IVIG were significantly different between CAL & non-CAL group. The levels of IgG at before and after IVIG were significantly different between responder and non-responder group. Conclusion: The levels of serum IgG before or after IVIG treatment may have the predictive value of the occurrence of CAL and response of initial IVIG therapy in KD. So, the pharmacodynamic mechanism shifting to shorten the half-life of IgG may be involved in the pathogenesis and outcome of KD.

Effects of gamma-globulin therapy on cardiac function in Kawasaki disease: evaluation by speckle-tracking and color tissue Doppler echocardiography.

Department of Pediatrics, Nippon Medical School Background: We previously reported that natriuretic peptides are elevated and decline immediately after gamma-globulin therapy (IVIG). This suggests that silent myocardial damage may exist in Kawasaki disease (KD) and recover with IVIG. Purpose: To clarify our hypothesis, we evaluated the myocardial function in patients with KD before and after IVIG by using speckle-tracking and color tissue Doppler echocardiography. Methods: We enrolled 30 children diagnosed as having KD and treated them with high-dose IVIG (2 g/kg/day). Echocardiographic examinations were performed before and at 3-5 days after IVIG therapy. As an index of diastolic function, E/EW was used. Systolic function was evaluated by peak strain values analyzed by EchoPAC and peak systolic values of tissue velocity. Results: Mean values before and after IVIG therapy are given. Radial strain (%) in the anterior LV was 23.2 and 37.6. Longitudinal strain (%) in the free LV wall was 25.0 and 29.2. Longitudinal strain (%) in the free RV wall was 23.2 and 33.5. E/EW in the free LV wall was 16.9 and 14.4. E/EW in the free RV wall was 7.0 and 5.3. BNP (pg/ml) was 88.2 and 28.7. Adrenomedullin (Fmol/ml) was 86.2 and 26.1. Conclusions: Both systolic and diastolic functions were low in the acute phase of KD, but improved immediately after IVIG. These results supported our hypothesis of myocardial dysfunction in the acute phase of KD, which recovers immediately after IVIG.

Clinical characteristics and coronary artery outcomes in Kawasaki disease patients with early intravenous immunoglobulin treatment Suthep Wanitkun, Sandee Santagule

Background: Kawasaki disease (KD) is a syndrome of fever and principal clinical characteristics affecting young children. Some patients may be diagnosed before the 5th day of illness. There are conflicting reports on requirement of additional doses of intravenous immunoglobulin (IVIG) and adverse coronary artery outcomes. The aims of this study were to identify clinical characteristics and assess coronary artery outcomes in this group. Methods: A retrospective case series was performed. All medical records of patients diagnosed with Kawasaki disease at Ramathibodi Hospital during January 1, 1997 to December 31, 2007 were reviewed. The subjects were divided into 2 groups; early IVIG which were diagnosed and treated before day 5, and conventional IVIG which were diagnosed and treated at or after day 5. Demographic data, standard laboratory and echocardiography were analyzed. Results: A total of 150 KD cases were identified. There were 17 cases with early IVIG treatment. Of these, there were incomplete KD 1.3%, and refractory KD 2.7%. The male: female ratio was 1:2. The initial WBC were 12,900 ± 2700/mm 3 in early group compared to 17,600 ± 6200/mm 3 in conventional group (p < 0.01). Echocardiography demonstrated coronary artery abnormalities 2.7%, pericardial effusion 5%, and mitral regurgitation 2%. Conclusions: The prevalence of KD patients with early IVIG treatment was 11%. The patients with early treatment were predominantly female and had lower initial WBC count. There were no significant differences in other clinical characteristics, standard laboratory investigations and coronary artery outcomes between patients with early treatment and those with conventional treatment.

Department of Pediactrics, University of Muenster Background: Kawasaki disease (KD) is an acute systemic vasculitis syndrome of children characterized by extensive activation of innate immunity. The receptor for advanced glycation end products (RAGE) serves as a pattern recognition receptor for a number of endogenous ligands which are potent inducers of inflammation, especially in vasculitis. Soluble RAGE (sRAGE) is supposed to act as a naturally occurring inhibitor of RAGE by neutralizing proinflammatory ligands; S100A12. Objective: The aim of our study was to investigate for the first time circulating sRAGE in with KD and compared these data directly with concentrations of the proinflammatory RAGE-ligand S100A12. Methods: We investigated 50 patients with acute KD, and differentiated by response to intravenous immune globulin (IVIG) treatment, and examined sequential changes in serum levels of RAGE and S100A12 using by ELISA. Results: We found that patients had decreased levels of sRAGE (553±97 pg/ml, P<0.001) during active KD, especially those more severely affected and not responding to treatment. In addition, sRAGE correlated negatively to proinflammatory S100A12 (328 ± 106 ng/ml, P<0.001) which was found elevated in patients with active KD. The initial serum levels of S100A12 were elevated in all patients with KD and decreased within 24 hours of IVIG (P<0.01) in the responders. Conclusions: Our data suggest that endogenously low levels of sRAGE do not only serve as biomarker of inflammation but can also be disease modifying, or even be a susceptibility factor itself. Pharmacologic approaches exploiting the antiinflammatory effects of sRAGE may offer promising therapeutic strategies in the future.

The strategy of immune globulin-resistant Kawasaki disease: a comparative study of additional immune globulin and steroid pulse Thirteen patients who were non-respondor of initial IVIG treatment were treated with additional IVIG therapy (2g/kg/day). Results: There was significant difference in period before fever falling from the 2nd treatment (IVMP puls: 1.5 ± 4days VS IVIG: 3.4 ± 3days p"<".0.05). No significant differences were found in incidence of coronary aneurysm between 2nd IVIG treatment and IVMP pulse therapy (IVIG 2case VS IVMP 0 case). There was no patient who developed giant aneurysms in both therapies.

Our study data stated that IVMP therapy was equal to additional IVIG therapy to prevent the artery aneurysm. Toyama University,Department of pediatrics, 2 Toyama University,1st Department of surgery In recent years, effects of the oral endothelin antagonist bosentan has been reported on patients with pulmonary hypertension (PH) related to congenital heart disease (CHD). We report successful combination therapy of bosentan, beraprost and HOT in three patients who could not undergo right heart bypass surgery because of PH initially. case 1) 1-year-old female with asplenia, SV, DORV, CoA,TAPVR case 2) 8-year-old male with SV, DILV, d-MGA, CoA case 3) 14-years-old female with Down syndrome,TOF, AVSD Patients underwent clinical, exercise and haemodynamic evaluation at baseline and after 6 to 18 moths of chronic oral administration of bosentan. Although we administered initial doses of bosentan adjusting for their body weight, tachypnea and chest discomfort were recognized in two cases. These symptoms disappeard by reduced dosage of bosentan. Bosentan therapy reduced pulmonary arterial pressure (PAP) and pulmonary vascular resistance (PVR) in all three cases. , and miscellaneous conditions. Two diagnoses accounted for 58 % of murmur evaluations, were patent ductus arteriosus (PDA) and innocent murmurs. The most common murmur diagnosis in the neonatal intensive care unit and the well-child nursery was PDA. Patients with recent cardiac surgery typically had hemodynamic instability. The most common basis for evaluation of function was oncologic disease and shock assessment. Although endocarditis was a clinical concern, only 20% of screening cases were identified, involving 2 patients with an indwelling intracardiac catheter. Kawasaki disease was the most common acquired condition leading to consultation. CONCLUSIONS: Although varieties of conditions were consulted, some were encountered more frequently especially the condition that may not required cardiology subspecialty consultation. Future educational curricula developed for hemodynamic assessment and stabilization should appropriately emphasize conditions necessitating consultation.

Objective: It is difficult to predict a risk of sudden death in long QT syndrome (LQTS) before the onset of syncope due to arrhythmia. Subject and Method: Group 1 consists of 15 high risk cases (mean age 13.6 years male/female ratio=8/7) with QT prolongation and with a history of syncope. Group 2 consists of 7 disease controls (mean age 11.1 years, male/female ratio=3/4) with QT prolongation and without syncope. Group 3 consists of 8 controls (mean age 13.1 years, male/female ratio=2/6) without QT prolongation and without syncope. The complex conjugate roots were determined from the characteristic equation of the third order AR model of RR interval from 0 to 5 am. The real part of conjugate root is classified as negative, positive, zero or imaginary root, and is evaluated to discriminate group 1 from the other groups. Results: Absolute negative roots include 13 from group 1 and 2 from group 2. Imaginary roots include 1 from group 1, 1 from group 2, and 5 from group 3. Positive roots include 4 from group 2 and 3 from group 3. Conclusion: As most of the group 1 is characterized a negative root, this position of conjugate root in the complex domain may be useful to predict a risk of syncope and hence a risk of sudden death in LQTS. Sensitivity and specificity of this risk criterion to determine group 1 and 2 were 80% and 40%, respectively. Sensitivity and specificity to determine group 1 and 3 were 82% and 80%, respectively.

Kazuyoshi Saito 1 , Fukiko Ichida 1 , Keijirou Ibuki 1 , Keiichi Hirono 1 , Sayaka Watanabe 1 , Keiichiro Uese 1 , Toshio Miyawaki 1 , Shingo Ohtaka 2 , Naoki Yoshimura 2 , Satoshi Yasukouchi 3 1 Toyama University, Department of pediatrics, 2 Toyama University, Department of the first surgery, 3 Department of pediatric cardiology, Nagano prefectural children hospital We report successful experience of CRT for a 3-year-old girl who has intractable heart failure due to isolated left ventricular noncompaction (LVNC) with narrow QRS complex. She was noticed to have heart failure due to LVNC at 3-months-old. Despite of full doses of several medications including ACE inhibitor, β-blocker, and pimobendan, she had often required to admit to our hospital because of progression of heart failure. At 3-years-old, on her 13th hospitalization, a chest X-ray showed marked cardiomegaly (CTR=0.73) and pulmonary congestion. An ECG showed QRS duration of 103ms. An UCG showed diffuse hypokinetic LV wall motion (EF=29.3%) with obvious dyssynchrony between apex, posterior and lateral walls where numerous prominent trabeculations existed, and severe MR. She could not have clinical improvement even with catecholamine infusion, and CRT was considered for her. Biventricular resynchronization using epicardial pacing leads was performed under general anesthesia. Optimal pacing sites were decided to get best synchronization of the ventricular walls on tissue Doppler imaging of echocardiography, and optimal AV delay was adjusted to achieve maximal systolic blood pressure and maximal cardiac output. Over a follow-up period of 6 months, she has exhibited significant and sustained improvement in LV function and clinical symptoms. BNP level decreased from 1960 to 82 pg/mL following initiation of CRT. QRS duration on ECG did not change after successful CRT. We conclude that pediatric CRT provide a highly useful adjunct for the treatment of heart failure even with narrow QRS duration, and may improve the prognosis of patients with LVNC.

Increased T-wave alternans by modified moving average beat analysis in children with congenital right bundle branch block Shuenn-Nan Chiu, Mei-Hwan Wu, Jou-Kou Wang, Chun-An Chen, Ming-Tai Lin, En-Ting Wu, Hung-Chi Lue National Taiwan University Hospital

Background: Microvolt T wave alternans (MTWA) has been shown as a marker for increased risk of arrhythmia. Its implication in patients with prolonged QRS duration due to right bundle branch block (RBBB) may however be different, but is still unclear. Methods: Twenty-six, aged 10 to 25 years old, children and adolescents with complete RBBB (RBBB group) and normal cardiac structure identified from school EKG survey received Treadmill by Bruce protocol with modified moving average beat analysis of MTWA. Another 26 age-and sexmatched adolescents served as control. Results: The basic characteristics and EKG parameters other than the QRS duration were not different between the two groups. We used 125 bpm as threshold heart rate for those older than 15 years old, and 67% predicted maximal heart rate for those younger than 15 years old. The MTWA was larger in RBBB group than in the control ( [results] The maximal heart rate by double load in the first grade of elementary school was 169bpm, in the fourth grade was 155bpm, and in the first grader of junior high school was 142bpm, in the first grader of high-school was 130bpm. By triple load in the first grade of elementary school was 171bpm, in the fourth grade was 164bpm, in the first grader of junior high school was 149bpm, and the first grader of high-school was 135bpm.

[discussion] As the increased heart rate rapidly falls down to normal range in a short time after exercise, the test seems to be inappropriate to evaluate. The maximal heart rate of double load in the first and fourth grade of elementary school seems to be appropriate. But the maximal heart rate of triple load, in the first grade of the junior high school and of high school seems to be inadequate.

Lipoprotein (a) levels affect both intima-media thickness of the common carotid artery and Achilles tendon thickness in children with mutation of low density lipoprotein receptor gene Mechanical forces, such as shear stress and stretching tension, not only alter the shape and function of various cells but also affect their differentiation. Previously, we have reported that shear-stressed Flk-1-positive (Flk-1 + ) cells derived from ES cells form tube-like structures in collagen gel and shear stress induces their differentiation into endothelial cells (ECs) via ligand-independent phosphorylation of Flk-1.

[Methods] Using standard immunomagnetic techniques, Flk-1 + cells were obtained from ES cells. Flk-1 + cells were cultured on silicon stretch chambers and cyclic stretch was applied by cyclic-stretch loading system. Then we collected mRNA and protein, and examined their expression levels by RT-PCR and Western blotting.

[Results] Subjecting uni-axial cyclic stretching (1Hz, 2~12%) to Flk-1 + cells induced a marked increase in the expression levels of vascular smooth muscle cell markers SM-α-actin, SM-22α-actin and SM-MHC, whereas it had no effect on the expression levels of the ECs, blood cells or epithelial cell markers.

Additionally cyclic stretch phosphorylated platelet-derived growth factor (PDGF) receptor β. But in the presence of AG1296, inhibitor of phosphorylation of PDGFR-β, cyclic stretch-induced increase of smooth muscle cell markers was abolished.

[Conclusion] Cyclic stretch induces the differentiation of Flk-1 + cells derived from ES cells into vascular smooth muscle cells via phosphorylation of PDGFR-β. These results indicate that mechanical forces, such as shear stress and cyclic stretch generated by fluid flow, play an important role in the regulation of early vascular development in embryos.

Endothelin-1 inhibits both L-type Ca 2+ current and ATP-sensitive K + current in neonatal rat ventricular myocytes. 

Objective: Endothelin-1 (ET-1), a potent vasoactive peptide, has wide range biological effects in various organs and tissues. In heart, ET-1 is released by endothelial, vascular smooth muscle and myocardial cells especially in various cardiovascular disorders, including congestive heart failure and ischemic heart disease. In these pathophysiological conditions, intracellular ATP contents decrease and catecholamines are released simultaneously with ET-1 and may contribute to the regulation of cardiac function. In this study, we investigated the effect of ET-1 on L-type Ca 2+ current (I Ca,L ) and ATP-sensitive K + current (I K,ATP ) in immature ventricular myocytes. Methods: Cells were enzymatically isolated from neonatal rat ventricle (aged 3 to 10 days). I Ca,L was measured as the peak inward current at a test potential of + 10 mV from a holding potential of -40 mV. I K,ATP was measured at a test potential of + 60 mV from a holding potential of -70 mV. All experiment was done at 37 °C. Results: ET-1 (10 -100 nM) decreased not only the basal I Ca,L , but also the isoproterenol-stimulated I Ca,L .

On the other hand, ET-1 (10 nM) almost completely inhibited the ATP-sensitive K + channel opener, pinacidil-induced outward current (i.e. I K,ATP ). Furthermore, ET-1 inhibited the shortening of action potential of cardiomyocyte induced by pinacidil. These effects were reduced in the presence of BQ-123, an ET A -receptor antagonist, suggesting that the action of ET-1 is mediated by the ET A receptor.

In neonatal heart, ET-1 will inhibit not only the contractility, but also cardioprotecive action by inhibiting isoproterenol-stimulated I Ca,L and I K,ATP . We have previously shown that SARS-CoV induced low interferons/IL-12 and high chemokine gene expression in dendritic cells (DCs). The chemokine gene expression was significantly higher in cord blood (CB) than adult DCs. We hypothesize that the developmental stages of the host immune system may account for the less severe phenotype of SARS in young children.

To investigate if the chemokines upregulation is implicated in the regulation of DC migration, we further investigated the effect of SARS-CoV on chemokine receptors (CCRs) gene expression in CB and adult DCs.

[Methods] CB and adult CD14+ monocytes were isolated by immunomagnetic separation and differentiated into immature DCs by IL-4+GM-CSF. Mock or SARS-CoV infected DCs were harvested at 3h and 9h post infection. Total RNA was extracted and gene expressions were quantitated by real time PCR based on Taqman fluoresence signals.

[Results] We studied the common receptors that may bind to the chemokines upregulated by SARS-CoV. Basal gene expressions of CCR1, CCR5, CCR7 in CB DCs were 1-2 log higher than that in adult DCs. SARS-CoV induced their expression in adult DCs but not in CB DCs. For CCR3, the basal levels were low in both CB and adult DCs and SARS-CoV induced its expression significantly in some cases.

[Conclusions] The lack of CCRs gene induction in CB by SARS-CoV may reflect the refractoriness of CB DCs to be mobilised in an environment with high chemokines concentration. Further investigation on chemotaxis of CB and adult DCs is needed to substantiate this hypothesis. The cause of influenza to the brain was investigated using the A/NWS/33 influenza virus infected BALB/c mouse model. NOS-2 mRNA levels in the infected mouse brain was greater than in control mice in all brain regions examined, particularly in the olfactory bulb and hippocampus on 1 day p.i. On the contrary, no differences in NOS-1 or NOS-3 mRNA levels were found between infected and control mice. There was also a marked increase in the levels of metabolites of nitric oxide in the olfactory bulb and hippocampus. Immunohistochemistry showed positive staining for anti-NOS-2 primarily in the hippocampus of infected mice. Further, anti-NOS-2 and GFAP staining was mostly found around capillary blood vessels of the hippocampus starting early in the course of the disease. These results indicate that the NWS enhances the activation of astrocytes and NOS-2 expression which in turn enhances NO production and the expansion of capillary blood vessels of the brain. Although IE has been reported for more than a decade, a therapy for IE has not yet been defined. We expect that our results will provide evidence which will aid in the development of a novel IE therapy. Conclusion: The incidence of EM-resistant strains is supposed not to be higher in invasive GAS infection. We could find out possible higher incidence of rheumatogenic strains in the invasive GAS infections. It seems to be important to have an effort on studying of serotyping and genotyping for the monitoring of strains and to know the epidemiologic characteristics.

Experience of three infants with neonatal bacterial meningitis of Escherichia coli and outcome at 18 month old Knowing the defense mechanism of host to organism is important for infection control. In previous study, we had shown that interferon-alpha (IFN-alpha) but not interleukin-12 (IL-12) being produced as human peripheral blood mononuclear cells (PBMCs) infected with varicella-zoster virus (VZV).

Here we investigated what signal molecule(s) and what kind of cell(s) involved in the IFN-alpha production. Methods: PBMCs were isolated and cultured with VZV antigens with/ without specific inhibitors as indicated. IFN-alpha were detected with intracelluar staining and ELISA. Results: We found that Toll-like receptor 9 (TLR9) was involved in the VZV-induced IFN alpha production since inhibitory CpG ODN could inhibit IFN-alpha production. We also found that UV-inactived VZV induced IFN-alpha production insensitive to inhibitory CpG ODN treatment indicating another TLR9-independent pathway. Further studies demonstrated dsRNA-dependent protein kinase (PKR) but not DNA-dependent protein kinase (DNA-PK) was involved in the VZV-induced IFNa production. With intracelluar staining and ELISA detection, we found that plasmacytoid dendritic cells (pDC) were responsible for IFN-alpha production during VZV infection.

Conclusions: Together, these results suggest that pDC play an important role in the IFN-alpha production in VZV infection through TLR9 dependent and independent pathways.

Prolonged acquired neutropenia associated with younger age and cytomegalovirus infection Backgrounds: Neutropenia is defined as a decrease in the circulating or absolute neutrophil count to 1.5 X 10 9 /l. It may be mild (1.0-1.5 X 10 9 /l), moderate (0.5 -1.0 X 10 9 /l), or severe (less than 0.5 X 10 9 /l). Neutropenia is an important cause of morbidity and mortality in infants and children. We investigated whether certain predisposing factors were involved in the prolonged neutropenia in infant and children. To determine whether SP-D gene polymorphisms altering a single amino acid of mature protein is associated with the severity of RSV infection. Subjects and Methods: DNA samples from 47 infants admitted into a tertiary hospital for RSV infection were analyzed. A PCR-RFLP reaction or direct sequencing method was performed for genotyping three biallelic polymorphisms at the SP-D codons for the amino acids 11, 160, and 270 in the mature SP-D (Met11Thr: rs721917, Ala160Thr: rs2243639, Ser270Thr: rs3088308). The duration of hospital stay, oxygen supplement, endotracheal intubation and laboratory data on admission were compaired among the genotypes. Results: No significant difference was found among the genotypes and allele frequencies of Met11Thr, Ala160Thr and Ser270Thr. In the subgroup analysis of infants less than 1 year old, the hospital stay of the infants with 11Met/Met was longer than the other genotypes (mean 7.0 vs 9.0 days; p=0.002).

The 11Met/Met genotype may increase the severity of RSV infection in Japanese infants less than 1 year old.

Kazuhiro Mori, Ryusuke Sugiyama, Katsuhiro Maeyama Saitama Municipal Hospital, Department of Pediatrics Background: Surfactant protein D (SP-D) is a member of collagenous host defense lectins. Intrauterine infection and cytokines contribute to the development of bronchopulmonary dysplasia (BPD). Genetic variability of SP-D that nonspecifically participates in the immune and inflammatory regulation within the lung may influence the risk of BPD. Purpose: To determine whether SP-D gene polymorphisms altering a single amino acid of mature protein is associated with a predisposition to BPD. Subjects and Methods: DNA samples from 35 premature infants hospitalized in a newborn intensive care unit were analyzed. A PCR-RFLP reaction or direct sequencing method was performed for genotyping three biallelic polymorphisms at the SP-D codons for the amino acids 11, 160, and 270 in the mature SP-D (Met11Thr: rs721917, Ala160Thr: rs2243639, Ser270Thr: rs3088308). Results: Of the 35 subjects, 15 had respiratory distress syndrome (RDS) and 6 had BPD. Three of the BPD patients were further classified with Wilson-Mikity syndrome (WMS). The frequency of the allele coding for 11 Thr was 0.58 in the BPD group and 0.76 in the non-BPD group. The 11 Thr allele was 0.89, 0.67, and 0.50 in the patients of RDS+/BPD-, RDS+/BPD+/WMS-, and RDS+/BPD+/ WMS+, respectively. Although the allele for 11 Thr tended to be less frequent among the BPD group, no statistically significant difference was observed. No significant differences were found in the allele frequencies for amino acids 160 or 270.

The SP-D allele coding for threonine 11 may diminish a predisposition to developing BPD.

High expression of interleukin-10 and plateletderived growth factor-B genes in tracheal aspirate fluid cells of low birth weight infants with bronchopulmonary dysplasia (BPD) is a late critical effect, occasionally occurring in low birth weight infants with and without the history of respiratory distress. Methods: The gene expression of cytokines and associated inflammatory molecules in tracheal aspirate fluid (TAF) cells was quantified by real-time polymerase chain reaction. Results: Interleukin (IL)-10 and interferon (IFN)-γ mRNA levels in TAF cells within the first 24 hours of birth, were higher in the neonates who developed BPD than in those who did not. Plateletderived growth factor (PDGF)-B was exclusively transcribed at high levels in the neonates who developed BPD but not respiratory distress syndrome (RDS). The morbidity rate of BPD was higher in high expression group of TGF-β1, IL-10 and IFN-γ than in the each low expression group. On the other hand, the morbidity rate of RDS in BPD patients was lower in high expression group of PDGF-B than in the low expression group. The immunohistochemistry of lung tissue revealed that PDGF-BB-positive cells were dominantly found in a patient with Wilson-Mikity syndrome. Conclusions: These results suggested that IL-10 expression was involved in the damage of lung tissue irrespective of the presence or absence of RDS, and that PDGF-B in pulmonary inflammatory cells was associated with the development of BPD in the neonates without RDS. IL-10 and PDGF-B levels in TAF cells could be a prognostic marker for the long-term outcome of the newborns with early respiratory distress. Conclusions: New BPD has a characteristically early bubble lung, requires a longer duration of ventilator, and may not be adequately prevented by antenatal steroid therapy. Understanding these differences between new and classical BPD is important to predict infants at risk of developing severe BPD and to design appropriate strategies for the prevention.

Objective: The aim of the study was to explore the feasibility of using venous catheter in substitution of chest tube in the treatment of pneumothorax in preterm neonates. Patients and Methods: 16 preterm neonates treated with chest tube insertion (n=9) or venous catheter placement (n=7) for pneumothorax in the neonatal intensive care unit of a tertiary medical center were enrolled retrospectively over a 6-year period. The duration of the procedure, and the chest tube or catheter in place were analyzed. The complication rate of the two groups was also compared. Mann-Whitney U test, and Fisher's exact test were used for analysis. Results: High reinsertion rate (42.9%) due to catheter malfunction was noted in the venous catheter group. The duration of the procedure and the duration of the device in place were no longer in the venous catheter group. The complication rate of both groups showed no statistical differences. Obstruction of venous catheters due to blood clots resulted in the only failure of using the venous catheter in the treatment of pneumothorax in a premature infant with underlying coagulopathy.

We concluded that using venous catheter in the treatment of pneumothorax in preterm neonates with underlying coagulopathy is not recommended. Further investigation involving more cases is mandatory for testing the feasibility.

The Effect of Surfactant Replacement Therapy in the Neonates with Massive Amniotic Fluid Aspiration Pneumonia (AFAP).

Bo Young Lee, In Kug Bang, Chun-Soo Kim, Sang-Lak Lee, Tae-Chan Kwon Deparment of Pediatrics, Dongsan Medical Center Keimyung University, Daegu, Korea Background: The massive aspiration of amniotic fluid was known as one cause of a fatal postpartal asphyxia and respiratory distress. We conducted this study to evaluate the effect of surfactant replacement therapy (SRT) in neonates who were required ventilatory care due to massive amniotic fluid aspiration pneumonia (AFAP). Methods: 28 patients received ventilatory care due to massive AFAP who were admitted in NICU of Dongsan Medical Center, Keimyung University from Jan. 2000 to Dec. 2006 were enrolled. It was excluded the patients whose amniotic fluid was contaminated by meconium. All patients were under artificial ventilator care and received SRT, the mean doses of surfactant (modified bovine surfactant) was 120 mg/kg. They were evaluated several clinical data (gestational age, delivery type, duration of ventilator care) and outcome (complications and mortality rate). They were also analyzed the change of oxygenation index (OI) between pre-SRT and post-SRT. Results: A large number of cases were term infants (82%) and born by caesarean section (85.7%). Radiological improvement was evident in 27 patients 6 hours post-treatment except one who was expired immediately after birth. Compared with pre-SRT, the mean OI was significant improvement at 6 and 12 hours after SRT (p<0.05 The overall neonatal mortality rates for male and female were 3.39‰ and 2.80‰, respectively. Birth weight/gestational age-specific neonatal mortality rates chart was plotted with curves representing 10th and 90th birth weight percentile.

There was a gradual increase in the prevalence of low birth weight and preterm birth together with the percentage of infants born to immigrant mothers during this period. Conclusion: We provide an easy to use birth weight/gestational age-specific neonatal mortality rates chart as a reference to facilitate decision making for physicians and parents. The normative data is crucial in further investigation of socioeconomic issues and public health policies in Taiwan.

Two cases of ultra-micropremie of birth weight less than 300 g HIDEKAZU HOMMA, TAKESHI ARIMITSU, MASAYUKI MIWA, GEN KURATSUJI, ISAMU HOKUTO, KAZUSHIGE IKEDA

Background: We experienced ultra-premature babies of birth weight less than 300g in 1999 and 2006. We herein report the difference of these two cases. Case reports Case 1: The baby girl was born by cesarean section at 23 weeks of gestation because of pregnancy induced hypertension. Her birth weight was 289 g. We used an umbilical vein catheter for 21 days. We did not give intravenous amino acids until 17th days of age. Although we supplemented calcium, phosphorus and vitamin D to maintain the serum calcium, phosphorus level 9.0 mg/dl and 5.5 mg/dl respectively, rickets of prematurity was exacerbated resulting in pathological bone fractures. Case 2: The baby girl was delivered by cesarean section at 25 weeks of gestation because of pregnancy induced hypertension. Her birth weight was 265 g. We used an umbilical vein catheter for 24 days. We started early aggressive nutritional support using intravenous amino acids immediately after birth. Her weight gain was much better than Case 1. Although we supplemented calcium, phosphorus and vitamin D as case 1, rickets of prematurity was exacerbated resulting in a thigh-bone fracture. Conclusions 1. It was useful to use an umbilical vein catheter for long days. 2. Aggressive nutrition was useful even for the infant who was born less than 300 g. 3. Not only supplements of calcium and phosphorus more than usual dose but also other nutritional device might be required to prevent pathological fracture of ultra-micropremie of birth weight less than 300 g.

Risk factors for rehospitalization within one year after discharge from NICU . Results: At normothermia smaller body-weight led to lower cerebral temperatures at 5, 10 and 15mm from the surface but not at 20mm. During systemic hypothermia, a lower rectal temperature, smaller body-weight and a shorter distance from the cerebral surface all led to a lower regional cerebral temperature (all p<0.01). Systemic hypothermia did not affect the relationships between regional temperatures. During selective head cooling, a lower cap temperature, smaller BW and a smaller sensor depth all resulted in a lower regional cerebral temperature (all p<0.001). Discussion: Brain cooling was more efficient with lower body-weight due to greater head surface-area to volume ratios. In the CoolCap Trial, low hypothbody-weight infants may have been excessively cooled. Therapeutic ermia may require body-weight adjustment to accomplish consistent regional temperatures and optimal neuroprotection.

Why dose immature myocardium have high tolerance against hypoxia.

Tomoyuki Sato 1 , Makino Watanabe 2 , Masahiko Kishiro 1 , Okada Takao 2 , Toshiaki Shimizu 1 1 Juntendo University School of Medicine, Department of Pediatrics and Adolescent Medicine, 2 Juntendo University School of Medicine, Department of Physiology (Background) It is widely known that neonatal hearts show a greater resistance against hypoxia than adult hearts, but its mechanism is not clear yet. We examined the effects and kinetics of magnesium ion which is known to have myocardial protective effect. (Methods) Rat hearts of adult (10weeks old) and neonate (1week old) were Langendorff-perfused and subjected to a 45 min hypoxia /30 min reoxygenation. Heart rate, ventricular tension and GOT release were monitored throughout the experiment. The magnesium content of myocardium after the perfusion was quantified by atomic absorption spectrophotometry. (Results) Heart rate and ventricular contraction in neonatal hearts were maintained higher level by hypoxia and were recovered faster by reoxygenation than those of adult. The recovery level of tension-rate product was not significantly different between neonates and adults after reoxygenation. Although GOT release increased by reoxygenation in adults, it was not the case in neonates. The magnesium content of adult and neonatal heart after perfusion without hypoxia was 793.9 ± 16.7 µg/g and 819.2 ± 39.1 µg/g, respectively.

After hypoxia-reoxygenation, the magnesium content of adult hearts decreased to 607.0 ± 22.8 µg/g; significantly (p<0.01) lower than that after aerobic perfusion. However, in neonatal hearts, magnesium content (853.0 ± 13.8 µg/g) was not significantly difference from that without hypoxia. (Conclusion) We suggest that the high tolerance against hypoxia-reoxygenation injury in neonatal hearts may be derived from the ability of maintaining intracellular magnesium. Purpose; Several studies suggest an association of eNOS gene polymorphisms to cerebrovascular diseases in adults. There are three clinically significant polymorphisms identified in the eNOS gene. We analysed the association of eNOS gene polymorphisms to perinatal hypoxic ischemic encephalopathy(HIE) in neonates. Method; Thirty seven term newborn infants with moderate to severe perinatal HIE(HIE group) and fifity four normal term infants without any perinatal problems(control group) were genotyped by PCR for two single-nucleotide polymorphisms (SNPs): 786 T→C in promoter region and 894 G→T in exon 7 (Glu298Asp). A variable number of tandem repeats (VNTR) polymorphism in intron 4 was also investigated. Results; No significant difference was found in the distribution of genotype frequencies of the three eNOS polymorphisms between HIE and control groups. However the increased frequency of the G allele of Glu298Asp polymorphism was found in HIE group(odds ratio=8.3, P=0.018). Conclusion; G allele of Glu298Asp polymorphism of eNOS gene might have a weak association with the occurrence of perinatal HIE.

New optical brain imaging in infants by using near-infrared diffuse optical tomography Hypoxic-ischemic encephalopathy remains a major cause of permanent neurodevelopmental disability and infant mortality. The late metabolic deterioration indicates that there is metabolic stress and implies that there may be a therapeutic window during which appropriate therapy could markedly improve outcome. However, the limitation in the 31 P-MRS method related to infants is the inability to estimate cerebral metabolism at the bedside. Recently, we have demonstrated that near-infrared time-resolved spectroscopy (TRS) can be used for non-invasive measurements of cerebral blood volume (CBV) and cerebral Hb oxygenation (ScO2) in infants at the bedside. In this study, we investigated changes in cerebral hemodynamics in asphyxiated infants by using TRS. Measurements by TRS were performed in 5 asphyxiated infants (3 with cerebral palsy and 2 with no developmental abnormalities) to estimate the values of CBV and ScO2 during a period of 80 hours after birth, and these values were compared with the values in 12 infants without asphyxia. In asphyxiated infants who had cerebral palsy, CBV and ScO2 increased transiently after birth and reached maximum levels after 36 and 48 hours, respectively. However, asphyxiated infants with good prognosis and infants without asphyxia did not show transient increases in CBV and ScO2. The increase in CBV is due to loss of cerebral autoregulation and, the increase in ScO2 is due to a decrease in oxygen consumption that leads to a decrease in venous Hb oxygen saturation.

The results have demonstrated that the TRS method is useful for monitoring cerebral hemodynamics in asphyxiated infants. AIM: This is a retrospective analytic study which aims to establish that the administration of hepatitis b vaccine at birth promotes compliance on the completion for the hepatitis b vaccine series. MATERIAL: A review of charts for a period of six months, from July to December 2004 was done. This was supplemented by follow on the patient's records with their attending physicians and personal call on the caregiver to obtain the needed information regarding the patient's hepatitis b vaccination status and acceptability of hepatitis b vaccine birth dose. RESULTS: Maternal occupation affects administration of birth dose and its completion, that is greater in employed mothers . The compliance for completion of the vaccine series among those given and not given the birth dose is not significant. Completion of the hepatitis b vaccine for the whole population is 90.2% however, the acceptability of receiving the vaccine at birth is only 50% of the population. CONCLUSION: The administration of the birth dose of hepatitis b vaccine does not predict the likelihood of completion of the series, The compliance rate on the completion of the vaccine series were the same with those who were given and not given the birth dose. (range) ). Number of patients with CRT (number of examinations) was 2 (134), 5 (60), and 3 (10), for first, second, and third week of insertion, respectively. The majority of patients with CRT were with extremely low birth weight (ELBW) (8 of 10). The number of catheter tip position with CRT (the ratio to the number of PICC: %) were 0 (0), 1 (6), 9 (17.6), and 0 (0) Background: ApoE-rich HDL is essential for the central nerve system neuron growth. Recently, it was found that large sized high density lipoprotein particles were identified predominance in cord blood. Purpose: The aim of this study is to investigate the relationship between subclasses of HDL particles and ApoE levels in cord blood, and the change of this association in postnatal period. Subjects and Methods: Sixty-five healthy, term, appropriate for gestational age neonates (38 males, 27 females) were included. Anthropometric parameters and serum lipids and apolipoproteins were determined at birth and at 1 month. Serum lipoprotein analyses were performed by high-performance liquid chromatography (HPLC) with gel permeation columns, which could classify HDL into 5 subgroups on the basis of particle size. ApoE levels were measured by turbidimetric immunoassay. Results: Levels of very large (VL) HDL were 5.4±2.6, and 7.0±4.1 (male, and female, Average±SD), which were higher than those reported in adults. One month after birth, the levels of VLHDL-C increased to 8.0±4.0, and 9.3±5.3. Cord blood apoE levels correlated positively with very large HDL cholesterol levels at birth (male, r= 0.548, p<0.01; female, r= 0.631, p<0.01), but in male this association was not significant at 1 month. Recently it has been shown that venipuncture was less painful than heel prick for blood taking requiring minimally four drops of blood. The aim of this study was to determine whether there was a difference in the pain indicators and effectiveness between venipuncture and heel prick if only one drop of blood is required (e.g. blood glucose testing) in term neonates. Methods: Sixty six term neonates undergoing blood glucose monitoring in the neonatal intensive care unit of Hospital Universiti Sains Malaysia during June and July 2006 were included in this study. The neonates underwent venipuncture (VP) or heel prick(HP). Primary outcome measures included were the Neonatal Facial Scoring System(NFCS) score, duration of the first cry, total duration of cry and duration of procedure. Secondary outcome measured was the number of skin puncture needed to obtain blood. Results: The NFCS score was not significantly different between the two groups and the duration of the procedure was significantly longer for the VP than the HP group (median 27.5sec versus 7sec; P < 0.001). The differences between the two groups for duration of first cry, total duration of cry and number of skin punctures needed to obtain blood were not statistically significant but these parameters displayed a trend, favouring the heel prick. Conclusions: Heel prick is still the preferred method of blood taking in neonates if only one drop of blood is required as in determination of blood glucose level.

A change of the bilirubin photo isomer with New Green LED in vitro.

Takashi Iwase, Toru Kuboi, Kenichi Isobe, Susumu Itoh Kagawa University, Department of Pediatrics LED (light emitted diode) has been used as one of the sources of light of new phototherapy for neonatal jaundice. We developed purely green LED before and gave a presentation. However, a peak wavelength is 525nm and generation rate of cyclobilirubin (cycloBR) was low level. New Green LED (a bed type) (Toitu) (by about 500nm peak wavelengths) was developed and we studied a change of the bilirubin photoisomer in vitro. <methods> We made complex solution (HAS 2g/dl, bilirubin 10mg/dl, 0.1M phosphate buffer pH 7.4) using bilirubin (Tokyo formation) and HAS (Sigma) and entered 10ml's Pyrex pipe by 1ml. It was put on a New Green LED horizontally, and irradiated light for 0, 5, 15, 20, 30 minutes. We measured bilirubin and the photoisomer for high performance liquid chromatography (HPLC). And we calculated ((ZE) -bilirubin/(ZZ) -bilirubin) and quantity of generation of CycloBR (a total of (EZ) -cyclobilirubin and (EE) -cyclobilirubin)). <results> Light energy is 60µ W/cm2/nm with MinoltaFluoro-LiteMeter451. The quantity of initial generation of CycloBR was about 0.30mg/ dl/min. (ZE)-bilirubin/(ZZ) -bilirubin was around 0.15. <conclusion > We showed a low value when we compared quantity of of CycloBR of green LED initial generation with BiliBlanket Plus. However, (ZE) -bilirubin/(ZZ) -bilirubin showed the value that was similar to the green light which assumed around 510nm a peak. Ultraviolet rays and light of 400-450nm that a DNA strand break, a survival fall of a cell are noted do not include new green LED. And a peak of the wavelength area has the greatest effect of phototherapy.

Yoko Yamakawa, Yoshikazu Ohtsuka, Tohru Fujii, Kiyotaka Ohtani, Satoru Nagata, Toshiaki Shimizu Juntendo University School of Medicine AIM: Although we know the clinical efficacy of leukotriene receptor antagonist (LTRA) in bronchial asthma, the effects of LTRA in food allergy still remains unclear. In this study, we investigated the benefits of early intervention of LTRA in food allergy children. METHODS: 50 children aged between 0 to 3 years of age with food allergy who experienced adverse responses, such as diarrhea, vomiting, bloody stool, rush, eczema, cough, and wheezing, against food antigens were enrolled for this study. Children were randomly divided in two groups. First 22 children were treated only with dietary control as a control group, and the rest of 28 children were treated with LTRA in addition to dietary control for one year. Peripheral blood eosinophil counts and serum IgE, IL-4, -5, -6, ECP, and PAF levels between pre and post-treatment were analyzed. RESULTS: Peripheral blood eosinophil counts and serum IgE, IL-4, -5, -6, ECP, and PAF levels were increased in children with food allergy. Although both groups showed clinical benefits with decreased eosinophil counts, only children treated with LTRA showed decreased serum IgE, IL-5, and ECP levels in addition to eosinophil counts. CONCLUSION: Our findings may indicate the possible anti-allergic effects of LTRA, by regulating peripheral blood eosinophil counts and serum IgE, IL-5, ECP levels especially in young children, aged between 0 to 3 years, with food allergy.

Nonlinear relationship between the levels of house dust mite allergen exposure and hospitalized wheeze in children aged 18 months Department of Pediatrics, Yokohama City University Background: Childhood-onset MCTD present with features of more than one rheumatic diseases and speckled ANA pattern. However, some patients have difficulty to make differential diagnosis from SLE. Objective: Clinical symptoms and laboratory findings were evaluated referred to profile of anti-ds-DNA and anti-RNP to figure out the differences between SLE and MCTD. Methods: Eighty children (7 male and 73 female) suspected as having as SLE or MCTD were enrolled in this study. They were classified into three groups: 48 with anti-ds-DNA and no anti-RNP (group A), 22 with both anti-ds-DNA and anti-RNP (group B), and 10 with anti-RNP and no anti-ds-DNA (group C). WBC counts, ESR, CRP, C3, C4, CH50, and serum IgG were examined. Renal pathology, ANA pattern, anti-ds-DNA and anti-RNP were followed (mean 8.0± 6.4 years). Results: Differences between Group A and C Incidence of Raynaud's phenomenon and speckled ANA and level of IgG showed significant differences between two groups. Course of children in Group B Anti-RNP disappeared in eight patients (36.3%), whereas other (63.7%) kept positive. However, four of the eight changed their ANA pattern from homogeneous or homogenous/speckled to speckled alone. Incidence of speckled ANA in group B increased from 45.5 to 73.7% during follow-up. Glomerular basement membrane abnormality was recognized in only 10% of anti-RNP positive children. However, the incidence of membranous nephritis increased by 18.6% on re-biopsies. Conclusion: Presence of speckled ANA and anti-RNP and renal pathology were changeable during long term follow-up. It is important to repeat these findings to verify diagnosis.

Cellular-level effect of interleukin-6 on growth plate chondrocytes Shoko Nakajima, Takuya Naruto, Takako Miyamae, Tomoyuki Imagawa, Masaaki Mori, Shumpei Yokota Yokohama City University, Department of Pediatrics Background: Growth impairment severely complicates systemic juvenile idiopathic arthritis (sJIA). Recent NSE/IL-6 transgenic mouse studies have indicated the pathogenic role of IL-6 in growth impairment of sJIA through systemic mechanisms that alter the GH/IGF-I axis. We investigated how IL-6 affects growth plate chondrocyte differentiation and proliferation. Materials and Methods: IL-6 receptor and gp130 expression by the murine chondrocyte embryonal carcinoma cell line ATDC5 was investigated by FACS analysis. Cells were incubated in 5% CO2 at 37°C in culture medium containing 10 μg/ml of bovine insulin (culture day 0), and were consecutively harvested for 14 full days. Recombinant murine IL-6 was added to the cultures on all the days between days 2 and 14 to observe cell proliferation, and, useing a quantitative RT-PCR (qRT-PCR) to three chondrogenic differentiation markers, collagen type II, aggrecan, and collagen type X, the effect on cell differentiation. To test IL-6 function blocking, the anti-mouse IL-6 receptor monoclonal antibody MR16-1, was added from day 2. Findings: The cells expressed IL-6 receptor and gp130. The expression of chondrogenic differentiation marker genes was reduced by IL-6, but this was abrogated by MR16-1. Interpretation: IL-6 directly inhibits early growth plate chondrocyte differentiation. Relevance to practice: Growth impairment in sJIA is brought about in part through the direct inhibitory effect of IL-6 on early differentiation of growth plate chondrocytes. [Background] Weber-Christian disease is also known as a relapsing febrile nonsuppurative nodular panniculitis.

[CaseReport] A 13-year-old girl was admitted to our hospital for close investigation and therapy against fever lasting over 2 months and erythema nodosum. Two kilograms of body weight loss was shown in 2 months. Laboratory testing may reveal elevation of ESR and CRP. No specific pathogen, autoantibody, and hormonal abnormality were detected. Remittent fever was also continued after admission. In the gallium scintigram, accumulation image was shown on the erythema nodosum of lower thigh. Biopsy of the skin nodules showed lobular rather than septal panniculitis. We diagnosed her disease as Weber-Christian disease, and introduced corticosteroid therapy. It resulted in speedy decline of fever and disappearance of erythema. Since then, her condition was fairly good, so we performed dose reduction of predonisolone. Suddenly, severe pain of coccygeal region occurred to her, and she could hardly walk. In addition, urinary disturbance became clinically evident, so we placed urethral catheter and/or introduced clean intermittent self catheterization. No abnormal findings such as infection, tumor, demyelination, and degenerative disease were detected on MRI or CSF/blood examination. Such symptoms were subsided in 2 weeks. Afterwards, even though subcutaneous nodules on the lower limbs were presented several times, these lesions were vanished in 2-3 days. In each time, her general condition was good without inflammation signs.

[Conclusion] Even though we did not clarify the definite cause of urinary disturbance and pain of coccygeal region, we thought it might be due to some peripheral neuritis.

A case of possible Neuro-Behcet's disease presented following minor head trauma

In Ok Hwang, Young Hwan Lee

Introduction: Behcet's disease(BD) is characterized by recurrent aphthous oral and genital ulcerations, uveitis, and it is recognized as being multisystemic.

Nervous system involvement is one of the most serious manifestation of BD.

We present a case of child, who show typical features of Neuro-Behcet's disease following minor head trauma. Case: A 9-year-old boy was transferred with diplopia and left sided weakness following minor head trauma. Neuroophthalmologic examination showed impairment of abduction of left eye. The muscle strength of left upper and lower extremities is grade 3+ and 4, respectively. T2-weighted images on MRI showed abnormal high signal intensity at right mesodiencephalic lesion. On diffusion weighted images, slightly high signal change was revealed, and apparent diffusion coefficient (ADC) maps had high signal intensity. Despite a negative pathergy test and absence of typical genito-ocular lesion, we diagnosed him as having NBD, based on history of recurrent oral aphthous ulcer, erythema nodosum, joint pain, absence of autoantibodies and characteristic findings of brain MRI. Subsequently, pulse methylprednisolone was initiated, followed by daily oral prednisolone. After 9 days of therapy, second MRI showed complete improvement of the brain lesion accompanied by improvement of neurological manifestations. During 2-months of follow up, he was complete recovery from diplopia and left sided weakness. Conclusion: Although NBD is rare in children, it should be considered when brain MRI reveals parenchymal inflammatory lesions that include mesodiencephalic area. And it should be further evaluated that causal relationship between minor head trauma and the onset of NBD. Accumulation of information on patients with various diseases, intervention trials based on the accumulated case information, and analyses of disease information are required for the practice of evidence-based medicine (EBM) and evidence-based policy making. A research project to develop "Integrated DB for Pediatric Diseases" in the National Center for Child Health and Development (NCCHD) has been under way since 2005. A case registration method to achieve two goals that are different in character, exhaustive registration of case information on many types of diseases and improving accuracy of individual case information, needs to be developed for the integrated DB. In order to achieve the almost conflicting goals, development of the integrated DB to accumulate information on multiple cases was thought to be necessary. "Case Information DB", a prototype of "Integrated DB for Pediatric Diseases", is comprised of information accumulated in the internal server of the NCCHD. The information sources for the DB include data from Research Project for Specific Chronic Pediatric Diseases and Follow-Up Study of Neonatal Screening Programs. We decided to develop an automatic cross-check program for the two information sources. The automatic cross-check program compared the two information sources and automatically calculated the estimated number of cases of specific diseases by using the capture-recapture method merely based on date of birth, place of residence, and sex of patients. We will conduct further demonstration experiments on the program by using more information sources.

The relationship between insulin-like growth factor binding protein-3 promoter polymorphism and growth Sae Min Lee 1 , Seung Yang 2 , Sae Min Lee 1 , Jeh Hoon Shin 1 1 Department of pediatrics, Hanyang university, 2 Department of pediatrics, Hallym university Background: The variability in insulinlike growth factor binding protein-3(IGFBP-3) levels is related to polymorphic variants of the promoter region of the gene. The most common single nucleotide polymorphism of IGFBP promoter region is -202 locus. In vitro, significantly higher promoter activity of the A allele at the -202 locus compared with the C allele, consistent with the relationship observed between genotype and circulating IGFBP-3 was documented. The polymorphic variation occurs frequently and may influence GH responsiveness, somatic growth, but the effects of IGFBP-3 promoter polymorphism on growth in children are unknown. Methods: RFLP genotyping of the -202 single nucleotide polymorphism was performed in 33 Korean children who visited our clinic for evaluation of their growth. The serum levels of IGF-1 and IGFBP-3 were compared according to genotyping. Results: Height SDS of subjects was -1.48±1.10. 11 of 33 children (33%) were heterozygous AC and the others are homozygous AA in genetic distribution, but no homozygous CC was detected. C allele frequency was 16.7%. There were no significant differences in height, body mass index, serum IGF-1 and IGFBP-3 levels between AC and AA groups. Conclusion: The effects of IGFBP-3 promoter polymorphism on growth at the -202 locus deserve investigation because there is no CC and the children were relatively short in this study. [Background] It has been reported that infants borne with small for gestational age may be accompanied with obesity and type 2 diabetes leading causes of cardiovascular disease (CVD) in later life, which is supposed to be a link with catch-up mechanism on their growth. Our aim of present study is to examine the characteristics of changes on their growth in SGA babies by measuring skinfold thickness (SFT).

[Methods] Forty-two neonates (birth weight varied from 1800g to 2300g) were studied. Anthropometry including SFT measurements were performed from birth to 1 year of age. Infants are divided into two groups according to birth weight: SGA was defined that the birth weight was lower than 1.3SD (<-1.3SD), and the other was appropriate for gestational age (AGA)(>-1.3SD). And SGA group was more divided into two groups whether or not those did show catch-up in body weight in this period.

[Results] It was found that SFT in SGA group gained faster than that in AGA group, and that the SFT in catch-up group was thicker than that in non catch-up group.

[Conclusions] Present study suggested that SGA babies have a nature of gaining more body fat than that in AGA. It seemed that the fact may be reasonable alterations in SGA for adapting to their late gestation and after birth, but there may be the mechanism intrigued with deteriorating health condition in later life, especially obesity related type 2 diabetes and CVD

Tomohiro Kitamura, Natsuki Ohkawa, Tsubasa Lee, Ken Hisata, Hiromichi Shoji, Kyoko Tanaka, Koichi Shinohara, Toshiaki Shimizu Juntendo University,Department of Pediatrics Background: Ghrelin plays an important role in the growth of infants through its effects on growth hormone secretion, feeding, and metabolism. Yet no study has evaluated the levels of circulating active ghrelin in very low birth weight (VLBW) infants from immediately after birth. Aim: To investigate the relationship between plasma ghrelin levels and postnatal growth in preterm infants, this study was performed. Methods-We measured the active ghrelin concentrations in the cord blood and in the plasma of 25 VLBW infants immediately, 2, 4, 6, and 8weeks after birth and examined the relationship between those levels and several anthropometric and biochemical parameters.

Results: The plasma immediately after birth in VLBW infants were significantly lower than those at 2, 4, 6 and 8 weeks of age. There were no significant correlations between the plasma active ghrelin levels and gestational age, birth weight, body mass index(BMI), weight gain rate, feeding volume, or plasma levels of leptin and insulin-like growth factor(IGF)-1; at any time point during the study period. Conclusion: These results suggest that the level of circulating ghrelin markedly increases after birth through the stimulation of food intake in VLBW infants, and indicate that the effects of ghrelin on postnatal growth should be further investigated.

A case of newborn with activating mutation of calcium-sensing receptor-should we use Thiazide or not?- [Background] Calcium-sensing receptor (CASR) is a G-protein-coupled receptor that is highly expressed in the parathyroid gland and the kidney. Activating mutations of the CASR can cause hypoparathyroidism with hypercalciuria. Treatment with 1-hydroxylated vitamin D3 derivatives can worsen hypercalciuria and nephrocalcinosis. In some papers it is empathized that thiazide can alleviate hypercalciuria but long-term safety and benefit of thiazide is unknown.

[Case] A male baby (2947g) was born in 36 gestational weeks. His 32-yearold mother has had hypocalcaemia from infancy and took alphacalcidol and calcium lactate. She was revealed to have CASR gain-of-function mutation (C131W) at the age of 26. She has mild renal failure and decreased levels of serum potassium and magnesium. The baby's serum levels of calcium and PTH were 5.5 mg/dl and 7 pg/ml, respectively, at day 3. We started treatment with alphacalcidol and calcium lactate without using thiazide.

[Result] We confirmed that he has C131W mutation. We tried to control his serum calcium level around 7.5 mg/dl. His urinary calcium/creatinin ratio was around 1~3 until he was one month old and after that it is around 0.3~0.6. We thought that using thiazide might worsen the condition of him, because his mother showed Bartter-like phenotype.

Characterization of the molecular species of glycerophospholipids in the brain of Zellweger syndrome by FAB-MS Makiko Saitoh 1 , Masashi Mizuguchi 2 , Masayuki Itoh 3 , Takashi Igarashi 1 , Masao Iwamori 4 1 University of Tokyo, Department of Pediatrics, 2 University of Tokyo, Department of Developmental Medical sciences, 3 National Center of Neurology and Psychiatry, 4 Kinki University, Department of Biochemistry Patients with Zellweger syndrome (ZS) exhibit various biochemical abnormalities of lipids, such as accumulation of very long chain fatty acids (VLFA) and defect of plasmalogens. The relation between these biochemical abnormalities and impaired differentiations and migration of neural cells, which are the major pathological findings in the central nervous system of ZS, remains to be clarified. In order to characterize the molecular events in the pathogenesis of ZS, we analyzed molecular species of glycerophospholipids in the brain of ZS by fast atom bombardment mass spectrometry (FAB-MS). At first, purified phosphatidylethanolamine (PE) and phosphayidylcholine (PC) were treated with 0.05 M HCl to estimate the amounts of plasmalogen, and were found that plasmalogen-type PE was comprised 30% of total PE in control brain, but was not detected in ZS brain, while plasmalogen-type PC was not contained in both control and ZS brain. FAB-MS revealed that plasmalogen-type PE in control gray matter were contained arachidonic (20:4) and docosapentaenoic (22:5) acids, all of which were completely absent in ZS gray matter. Diacyl PE with polyunsaturated fatty acids were compensatory elevated in ZS brain in comparison to those in control brain. Thus, altered molecular species of PE might affect neural membrane properties such as membrane fluidity, thereby playing a crucial role in the pathogenesis of ZS.

Identification of ATP7A mutations and carrier detection associated with Menkes disease Chie Fujisawa, Katsuaki Shiga, Ayafumi Kaga, YanHong Gu, Hiroko Kodama

Menkes disease (MD) is an X-linked recessive disorder of copper metabolism characterized by progressive neurological degeneration and connective tissue abnormalities. The disease is caused by mutations in the ATP7A gene, which encodes a copper-transporting P-type ATPase. ATP7A is a large gene, and consists of 23exons spanning a genomic region of about 150 kbp. We examined mutations in the ATP7A gene in 46 unrelated patients and 1 related patient with MD. Furthermore, carrier detection was performed in 22 mothers of the patients, and prenatal/neonatal diagnosis was performed in 7 male siblings. Genomic DNA was prepared from peripheral blood lymphocytes, cultured fibroblasts or amniocytes, and amplified by PCR. The direct sequencing of exons was performed with a 3700 DNA analyzer (Applied Biosystems). Forty different mutations were identified in the 43 patients; 10 nonsense mutations, 6 missense mutation, 10 splice-site mutations, and 14 insertion/deletion mutations. Most ATP7A mutations in these patients were identified in important functional/ structural domains of protein products such as copper binding domains, transmembrane domains, CPC motif, and ATP binding domains. Four patients were not found mutations. Eighteen of 22 mothers of patients (77.3%) were carriers of Menkes disease, while 5 mothers (22.7%) were not carriers. Four of the 7 siblings (57.1%) that underwent prenatal/neonatal diagnosis suffered from Menkes disease. These results indicate that gene analysis is useful for the early diagnosis of MD and for quality of life of patients and families.

Tetsuya Ito 1 , Yasuhiro Maeda 2 , Hazuki Kobayashi 2 , Mai Nagao 2 , Yukihisa Kurono 2 , Akihito Ueta 1 , Kyoko Yokoi 1 , Yoko Nakajima 1 , Hajime Togari 1 , Naruji Sugiyama 3 1 Nagoya City University, Department of Pediatrics, 2 Nagoya City University, Laboratory of Hospital Pharmacy, 3 Aichi-Gakuin University, School of Dentistry, Department of Pediatrics Methylmalonic aciduria (MMA) is one of the inborn errors of metabolism, caused by the defect of methylmalonyl-CoA mutase or vitamin B12 insufficiency. In this disease, methylmalonyl-CoA and propionyl-CoA accumulate and conjugate with internal carnitine by carnitine acetyltransferase, and then methylmalonylcarnitine (MMC) and propionylcarnitine (PC) are excreted into urine. Thus, the administration of L-carnitine is essential for the patients of this disease, and monitoring of carnitine and acylcarnitine (AC) levels is useful for the diagnosis and assessment of the metabolic state. We have developed the method of detailed determination of ACs including MMC and PC using HPLC-ESI/MS-MS (Maeda Y, Ito T, et al, Rapid Commun Mass Spectrom, 21: 799-806, 2007) . Here we applied this method to tissue AC analysis and measured tissue ACs in two patients with MMA. Cases: Two girls were chemically diagnosed by GC/MS and had genetic mutations of methylmalonyl-CoA mutase gene. Both were dead because of acute metabolic decompensation and tissue samples were collected by autopsy after informed consent. Method: ACs that were extracted from the lyophilized tissues were purified by solid phase extraction and applied to the HPLC-ESI/MS-MS system with multiple reaction monitoring. Result: In the samples of urine and serum, PC concentrations were much higher than the MMC. The same result was seen in the kidney samples, whereas, high concentrations of MMC were determined in the liver and skeletal muscle. This discrepancy of the AC levels in the tissues should be elucidated.

Novel mutation of the OXCT1 gene in the first Asian patient diagnosed during neonate as having succinyl-CoA:3-oxoacid CoA-transferase (SCOT) deficiency Department of Neonatalogy, Aiiku Hospital, 4 Department of Pediatrics, Graduate School of Medicine,Gihu University SCOT deficiency is one of rare and potentially fatal inborn errors of metabolism. Early diagnosis could improve prognosis of patients with SCOT deficiency. We report a Japanese boy with SCOT deficiency diagnosed by critical samples in acidotic crisis during neonate. The patient was born through uneventful vaginal delivery with Apgar 9/10. At 2 days of age, tachypnea and poor sucking were noted. The blood gas analysis showed metabolic acidosis (pH 7.072, HCO3-5.8 mmol/L). The urine ketone bodies were positive (+++). He was treated with intravenous infusion of glucose and sodium bicarbonate. When transferred to our institution at 4 days of age, he no longer represents tachypnea and metabolic acidosis. The urine ketone bodies remained positive (+). Urine organic acid analysis during the metabolic acidosis showed only massive excretion of metabolites from ketone bodies. Enzymatic assay showed the significant decrease in the activity of SCOT from the patient (0.25 nmol/min/mg protein; control, 3.9). Direct sequence of genomic DNA identified compound heterozygous mutations in the OXCT1 gene (c.658_666dup, p.Asn220_Ile222dup; c.1304 C>A, p.Thr435Asn). The 658_666dup has not described yet. The three amino acid residue (Asn220_Ile222) of SCOT protein was reported to be important for dimerization of the SCOT. Although he suffered from ketoacidotic crises twice since the discharge at 30 days of age, he overcome these crises after early intervention of intravenous glucose administration. These data confirm that Asn220_Ile222 is important residue for the activity of SCOT and suggest that early diagnosis with critical samples is beneficial for children with SCOT deficiency.

Naoaki Hori, Naoko Amano, Mikako Inokuchi, Tomohiro Ishii, Tomonobu Hasegawa

Keio University School of Medicine, Department of Pediatrics Background: Adult females with classical 21-hydroxylase deficiency (21OHD) commonly have poor adult height and impaired gonadal function. The long-term outcome in adult males with 21OHD, however, has not been well described. Subjects and Methods: Subjects were 13 21OHD males with adult heights, aged 17.3-36.6 (median 27.3) years. Twelve of 13 were salt wasting and one simple virilizing. We assessed the following parameters for long-term outcome; 1) SD scores of adult height (AH) and AH-target height (TH) (N=13), 2) the average of bilateral testicular volume using an orchidometer (N=10), 3) presence or absence of testicular adrenal rest tumors by ultrasonography (N=10), 4) semen analysis in 4 subjects having testicular adrenal rest tumorsand Results: 1) AH was -3.4 to 0.6 (median -1.7) SD, and AH&#65293;TH was -3.6 to 1.5 (median -1.8) SD.

2) Testicular volume was 10 to 25 (median 13.9) mL, being <12 mL (10th percentile in Japanese healthy 16 yr boy) in 3 subjects.

3) Testicular adrenal rest tumors were present in 7 subjects. There was no significant difference in testicular volume between presence and absence of testicular adrenal rest tumors. 4) Semen analysis showed azoospermia in 1, and oligozoospermia in 3 subjects. Discussion: 1. Adult height was not normalized, as previously reported. 2. We reconfirmed that testicular volume was small and testicular adrenal rest tumors were present in some subjects. 3. Although preliminary, the presence of testicular adrenal rest tumors may indicate azoospermia or oligozoospermia.

Reference Values for Serum Steroids in Term Newborn Infants Using Liquid Chromatography / Mass Spectrometry / Mass Spectrometry.

Gen Kuratsuji 1 , Takeshi Arimitsu 1 , Masayuki Miwa 1 , Hidekazu Homma 1 , Isamu Hokuto 1 , Keiko Homma 2 , Kazushige Ikeda 1 , Tomonobu Hassegawa 1 1 Keio University, Department of Pediatrics, 2 Keio University Hospital, Department of Laboratory Medicine BACKGROUND: Large quantity of fetal cortex steroids in serum exist in newborn infants, leading to false positive errors by commercially available measurements. Serum steroid profile analysis using Liquid Chromatography / Mass Spectrometry / Mass Spectrometry (LCMSMS) has not been reported in newborn infants.

The goal of the current study is to obtain the reference values of steroid hormones in neonates of term delivery in this study. DESIGN/METHODS: Ninety-seven full-term Japanese newborn infants (50 males, 47 females; birth weight 2532-3804 gram) without endocrinological abnormality were enrolled. Ten steroid hormones were measured simultaneously by LCMSMS using 100 µl of serum, such as cortisol, cortisone, 17-hydroxypregnenolone, progesterone, 17-hydroxyprogesterone, 21-deoxycortisone, dedydroepiandrosterone, androstenedione, testosterone, dihydrotestostone. Serum testosterone levels were measured using both radioimmunoassay(RIA) and LCMSMS in two males and two females.

We set forth the reference values of 10 steroids. The serum levels of testosterone using LCMSMS were lower than those using RIA.

We have established reference values of 10 steroid hormones in full-term newborn infants by LCMSMS. Our reference values are clinically relevant to the biochemical diagnosis of various disorders in steroidogenesis in newborn infants. It offers the following advantages over commercially available measurements such as RIA: 1. More than 10 steroid hormones can be simultaneously measured with 100 µl of serum. 2. False positive errors are theoretically unlikely.

We are planning to determine reference values at various weeks of gestation, birth weights, and days of age.

Iodine supplementation with seaweed Laminaria japonica drink for iodine deficiency due to long term gastric tube feeding Kazuko Yoshimura, Soutaro Itou, Yukako Yoshikane, Naomi Morishima, Takahito Inoue, Shinichi Hirose Department of Pediatrics, Fukuoka University, School of Medicine Hypothyroidism due to insufficient iodine intake is a rare nutritional problem in Japan, where marine products containing plenty iodine are consumed. We encountered a 29-year-old disable man who had been on gastric tube feeding for seven years and presented a diffuse goiter and myxedema. His thyroid function was low <fT4 <, 0.4ng/dl > with a high thyrotropin level<TSH 56.288µU/ml>, while both anti-thyroglobulin and anti-thyroid peroxidase antibodies were negative. The liquid formula used for 7 years was found to provide him with 20µg iodine per day. Hypothyroidism due to iodine deficiency was suspected as daily requirement of iodine is suggested to be from 150 to 200µg while the average iodine intake in Japan is 1mg. Levo-throxine <25µg/day>and supplementation of iodine were initiated. 500µg/day of iodine was supplemented with a seaweed drink via the gastric tube. His diffuse goiter was improved and euthyroid function was achieved immediately after the treatment. These days the number of cases on long term gastric tube feeding is increasing. Insufficient iodine intake during a gastric feeding should be considered and appropriate amount of iodine may be supplemented if necessary. The drink used for the present case is a Japanese traditional drink obtained as a preservable and soluble powder made from seaweeds or Laminaria japonica and salt. Since over intake of iodine may also result in thyroid dysfunction, the seaweed drink seems to be a safe, effective and economical remedy to supply iodine during long term gastric feeding.

Han-Ping Wu 1 , Kai-Wei Chien 2 1 Division of Pediatric Emergency Medicine, Department of Pediatrics, 2 Department of Emergency Medicine Introduction: Bronchial asthma is a common cause of respiratory distress in the pediatric emergency unit. However, some other extrathoracic causes may cause respiratory distress, such as diabetic ketoacidosis (DKA). We presented a very uncommon case of asthma concurrent with the first onset of DKA for intending to alert the emergency physician to the additional respiratory distress imposed by DKA Case Report: An eight-year-old girl with a history of asthma presented to pediatric emergency unit with shortness of breath and dyspnea. An asthma attack was considered and therapies were initiated. She also denied any history of other underlying diseases, including DM. Because of the poor control of respiratory distress, we intended to find whether any other diseases causing respiratory distress may coexist with asthma, and arranged some imaging and laboratory examinations. The blood glucose concentration by using the glucometry strip was found exceeding the measurable capacity. Furthermore, the urinalysis showed glycosuria exceeding 1000 mg/dL and ketonuria of 3+. The definite serum glucose level showed 398 mg/dL and serum ketone body was 40 mg/dL. Therefore, the presumption of DKA was made. The patient was transferred to the pediatric intensive care unit. Biochemical laboratory tests about DM performed and showed serum glycosylated hemoglobin of 12.4% Discussion: Asthma attack with concurrent first-onset DKA is rare but critical. Prompt use of glucometry for determination of glucose concentration is of use in rapidly distinguishing DKA from other concurrent respiratory emergencies. When confronted with an intractable asthma exacerbation in the pediatric emergency unit, 1 Gyeongsang National Univ. of Hosp. Depart. of Pediatrics, 2 Gyeongsang National Univ. of Hosp. Depart. of Pathology, 3 Gyeongsang National University College of Medicine. Depart. of Microbiology Objectives: The purpose of this study was whether it is possible to culture of H. pylori from cryopreserved gastric mucosal biopsies frozen with or without OCT compound for more than 10 years. Method: Forty-six cryopreserved antral biopsy specimens with OCT compound and 31 ones without OCT compound sampled from 46 pediatric patients were used. The samples were collected in either 1990 (15, OCT only) or 1992 (31) and deeply frozen in eppedndorf tube without other preservatives (-70°C). Frozen gastric biopsies were cultured at 37°C for 7 days. Results: H. pylori were recovered from 29 (63%) among 46 biopsies. Histopathologic examination of 42 patients revealed bacteria infiltration on gastric antral mucosa. The recovery rate from biopsies cryopreseved in OCT compound was 93% at 1990 years and 13% at 1992 years. The recovery rate was higher in antral specimens stored in OCT compound at 1990 years than 1992 years. And the recovery rate from biopsies cryopreseved in OCT compound was 39%, and the rate of positive culture utilizing without OCT compound was 48%. There were no differences in histopathologic findings between biopsies cryopreserved with OCT compound and without that. The recovery rate would be higher if the gastric mucosal biopsy specimens were kept at -70°C with tight seals and were kept from being freeze-dried. Conclusion: Gastric mucosa per se is the best cryostorage media as well as the best transport media if it were kept in tight sealing and from being freeze-dried.

Cut-off volume of dietary fiber in the relief of constipation in children Purpose: Allergic proctocolitis is a major cause of bloody stool in early infancy, but pathophysiology, progress, treatment and outcome have not been fully identified yet. Methods: We reviewed 26 infants retrospectively, presented with bloody stool in early infancy with diagnosis of allergic proctocolitis. Result: Ten male and sixteen female infants were included. The mean age at onset of symptom was 15.0±13.0 weeks. The mean age at diagnosis was 17.4 ± 12.5 weeks.Fifteen infants(57.6%) were exclusively breast-fed and one infant(3.8%) was formula-fed and six infants(23%) were fed combined formula. Four infants(15%) fed weaning diet. Mean hemoglobin level was 12.4±2.1 g/dL and peripheral eosinophil count was 516.5±473.9/mm 3 .Sigmoidoscopic findings showed multiple focal erythema(76.9%), erosion(61.5%), mucosal hemorrhage(19.2%) and lymphoid hyperplasia (65.3%). Histologic findings in 25 infants showed focal infiltration of eosinophils in lamina propria(96%), muscularis mucosa(8%) and crypt epithelium(96%). In twenty four infants(96%) number of eosinophils in mucosa was increased more than 60/10HPF. Among twelve infants whose mother eliminated the four major food groups from diet, bloody stool was diappeared in ten(83%).Three infants fed exclusively breast milk were improved spontaneously.Cow's milk was changed with HA milk with impro vement. Four infants who were fed weaning diet were all improved after food restriction such as milk, soy, egg and nuts. Conclusion: Allergic proctocolitis should be considered as major cause of bloody stool in healthy appearing infants. Sigmoidoscopy with biopsy is the most effective diagnostic methods. Most infants experience a benign courses and recover with elimination of causative foods.

Diagnostic accuracy of serum biomarkers in predicting pediatric appendicitis, compared with the Alvarado score and abdominopelvic computerized tomography scan Laboratory of Epidemiology and Biostatistics, 3 Evidence Base Medicine Background: The aim of this prospective study was to determine the cutoff values of serum biomarkers, including white blood cell (WBC) count and C-reactive protein (CRP) in predicting pediatric appendicitis based on how long the patients' symptoms were present. Methods: A prospective study comprised 594 pediatric patients with suspected appendicitis at a medical center from 2004 to 2006. Receiver operating characteristic (ROC) curves were used to establish the best cutoff values of serum biomarkers for discriminating pediatric appendicitis. We further analyzed the diagnostic values of performing the Alvarado score, and abdominopelvic computerized tomography scan (CT) in predicting appendicitis, and then compared with our selected serum biomarkers.

Results: ROC analysis showed that the best cutoff values of WBC counts on the first day after onset of symptoms (Day 1), and CRP concentration on Day 2-3 in diagnosing acute appendicitis; CRP cutoff values which indicated perforated appendicitis were also determined. The diagnostic accuracies of the serum biomarkers, the Alvarado score of 7 or more, and CTs in predicting acute appendicitis were all favorable, but in predicting perforated appendicitis, the serum biomarkers had the highest diagnostic value. Conclusion: The kit of selected cutoff serum biomarker values may offer a clinical aid in diagnosing pediatric appendicitis, and we propose the addition of the serum biomarkers in children with clinically suspected appendicitis.

Han-Ping Wu 1 , Chin-Yi Huang 2 , Yu-Jun Chang 3 , Ching-Yuang Lin 1 1 Division of pediatric Emergency Medicine, 2 Evidence Base Medicine Center, 3 Laboratory of Epidemiology and Biostatistics Aims: To determine the change between primary and repeated serum laboratory tests may improve diagnostic accuracy in children with equivocal findings of abdominal computerized tomography (CT) scan in predicting appendicitis Methods: We prospectively collected 129 patients aged from 4 to 18 years with equivocal findings of abdominal CTs suspected which intended to diagnose acute appendicitis in the pediatric emergency department. We repeated serum laboratory tests measures 8 hours later to evaluate the change in serum inflammatory biomarkers. The significantly changes in serum parameters between primary and repeated serum laboratory examinations in patients with appendicitis were selected as the discriminating variables, and receiver operating characteristic (ROC) curves were used to determine the cutoff values of the changes between 2 laboratory examinations in predicting appendicitis.

Results: ROC analysis revealed that the cutoff values for the change in total neutrophils (3.2%) on the first day after onset of symptoms (day 1), the changes in CRP concentration (4.5 mg/L ) on day 2, and the change in CRP concentration (17.0 mg/L) on day 3 were taken as significant serum parameters for acute appendicitis,. Conclusions: Significant change between primary and repeated serum laboratory tests during in-hospital observation may serve as a useful method in predicting pediatric appendicitis in children with equivocal findings of abdominal CT scans P-107 Acute appendicitis in neurologically impaired children In total, 239 severely malnourished children, aged 6-36 months were studied. They received nutritional supplements, and were discharged when the nutritional status improved to either oedema-free weight-for-age >50% or weight-for-length >80%. Results: 56% of the study children (n=239) were male. Median age of the mothers was 22 years, and 18% of them had chronic energy deficiency. Half of the mothers were illiterate, and 24% of them had some form of employment, 16% had no sanitary toilets, and 34% had no access to safe drinking water. About one-third (n=85, 36%) of the children had admission pedal edema, 84% were severely underweight, 62% were severely stunted and 19% had severe wasting. The mean weight gain of children with non-pedal oedema was 8 g/kg/day; weight gain of the severely underweight were significantly higher than those of non-severely underweight (8 g/kg/day vs. 5 g/kg/day, p=0.02); severely stunted had better weight gain than non-severely stunted (9 g/kg/day vs. 5 g/kg/day, p=0.02); and severely wasted gained better weight than non-severely wasted (11 g/kg/day vs. 6 g/kg/day, p=0.001). Conclusions: Improvement of nutrition was more marked in severely malnourished children who are more eligible for nutrition rehabilitation interventions which may be directed towards such population. Risperidone is an antipsychotic in wide use for psychiatric disorders such as ADHD and autism in paediatric patients. It is converted by P450 enzymes into 9-hydroxyrisperidone, which has a pharmacological activity comparable to its parent compound. Substantial inter-individual variability has been observed in the dose-normalized concentrations of risperidone and the 9-hydroxy metabolite. This variability has been partly accounted for by polymorphisms in CYP2D6, which has more than 10 mutant alleles.

[Methods] In the present study, 90 adult Japanese patients with schizophrenia receiving risperidone were evaluated to see whether their CYP2D6 genotypes were associated with the altered metabolism of risperidone and its metabolite. A P450 microarray system was used, by which most major mutant alleles of CYP2D6 can be scored.

[Results] The dose-normalized concentrations of risperidone exhibited significant differences among the three CYP2D6 genotypes (*1/*1, *1/*10 and *10/*10; p=0.025). The concentrations were 2.8 times higher in the homozygous *10 (n=7) than in the wild type (n=16). Among the patients who were given concomitant phenothiazines (n=44), the total active moiety (i.e. risperidone and 9-hydroxyrisperidone) was higher in those with the *10 allele than in those with the wild type (p=0.014).

[Conclusions] The CYP2D6 genotypes and the concomitant phenothiazine administrations were identified as the major determinants of the steady-state concentrations of the total active moiety of risperidone. The present finding is particularly relevant for Asian populations, among whom the *10 allele is quite prevalent. -array) to detect the precise deletion region. To confirm array data, fluorescence in situ hybridization (FISH) analyses were performed using BAC clones.

[Results] CGH-array analysis identified the deletion of 9pter-p23.3 in this patient. FISH analyses refined breakpoint of the deletion region.

[Conclusion] CGH-array was effective to detect the region of deletion for this patient. FISH analyses agreed the result of CGH-array. The deletion region in this patient was not common with the previously reported region responsible for trigonocephaly, but the physical distance is not so far away. Thus, we still believe that the critical region for trigonocephaly is in this narrow region.

[Background] Fabry disease is an X-linked disorder caused by the deficiency of lysosomal enzyme, α-galactosidase A (α-gal A). Accumulation of glycosphingolipids in systemic organs results in pain, hypohidrosis and angiokeratoma during adolescence and renal and cardiac failure at the later stage. Enzyme-replacement therapy is now available, but its efficacy is often inadequate for adult patients with advanced clinical symptoms. AAV vector mediated continued and systemic supply of α-gal A is an important option for treatment of Fabry disease.

[Methods & Results] We compared therapeutic efficacies of AAV vector delivery to neonatal and adult Fabry mice. When adult male Fabry mice (12 weeks old) were treated by intravenous injection of AAV1 vector carrying the α-gal A cDNA, sustained high levels of α-gal A activity were detected in plasma, liver and heart. AAV1 mediated in vivo transduction was significantly suppressed in female adult mice. Injection of AAV1 vector into neonatal mice (3 days old) resulted in long-term cardiac specific expression of α-gal A irrespective of sex of mice. Accumulation of globotriaosylceramide was efficiently inhibited in the liver and heart by a single injection at the neonatal period.

[Conclusions] AAV1 mediated gene therapy at the non-symptomatic early stage of the disease should be beneficial for prevention of major organ failure in adulthood. 

Moyamoya disease is a rare cerebrovascular disorder mainly involving the middle cerebral arteries, is a steno-occlusive disorder which cause ischemic events in the childhood period. Etiological aspects of moyamoya disease are somewhat similar to that of idiopathic pulmonary hypertension. Because, both diseases are caused by narrowing of arteries and their incidence is higher in female than in male. Some responsible genes have been identified in idiopathic pulmonary hypertension, i.e. endoglin (ENG) and activin A receptor type II-like 1(ACVRL1). In this study, we analyzed these genes in patients with moyamoya disease.

MATERIALS AND METHOD Materials were DNA purified from blood of six typical moyamoya patients. All coding regions of ENG and ACVRL1 were analyzed

There was no mutation in the candidate genes.

Because moyamoya disease is popular only in east Asian people, we believe that there should be a common mutation in patients with moyamoya disease, and that the number of materials with six samples would be enough. However, we identified no mutation in ENG and ACVRL1, and failed to confirm positive relationships between them. Since pathways of signal transduction in vascular system are suspected to be involved in moyamoya disease, other molecules should be studied in the future. Acute myelogenous leukemia (AML) is a heterogeneous disease with a variety of genetic alterations including the MLL gene rearrangement. Microarray-based gene expression profiling has been applied to diagnose AML patients and to explore their underlying molecular pathology. In this study, we analyzed gene expression of 54 (14 adult and 40 pediatric) myelo-monocytic AML patients with FAB M4 and M5 subtypes (excluding inv(16) and t(8;21) cases), and found striking differences among the patients in an age-associated manner. Especially most of the infant patients (less than 1y) displayed very distinct gene expression.

With the use of this distinctiveness, we were able to divide the pediatric patients into three age-associated subgroups with different outcomes. In addition, we found that gene expression signature of MLL rearrangement was different among these pediatric subgroups as well. These results suggest that age is an important factor contributing to the biology of myelo-monocytic AML. Recently, it was found that rhabdomyosarcoma (RMS) was induced with extremely high penetrance in Ink4a/Arf -/mice transgenic for MET protooncogene. PTPN11 encodes SH2 domain-containing protein tyrosine phosphatase, which relaying signals from activated growth factors and cytokine receptors to RAS and other intracellular signaling molecules. To address the roles of these genes in RMS, the most common soft-tissue sarcoma in children, we performed mutational analysis in 39 samples by PCR-SSCP. No mutations of MET were detected whereas a nonsense mutation of p16INK4A was identified in an alveolar RMS. Also, we identified a missense mutation of PTPN11 in an embryonal RMS (ERMS) and missense mutations of NRAS in 2 ERMSs having no PTPN11 mutations. By real-time quantitative PCR, expression of MET was found varied more than 10-fold among these samples, despite a normal DNA copy number. The protein expression level was consistent with that of mRNA, and in cell lines with a higher expression level, MET was constitutively activated. Notably, the expression level of MET was significantly higher in patients who died (P = 0.02), in patients with stage IV (P = 0.04), as well as in patients with PAX3-FKHR chimeric transcript (P = 0.04). On the other hand, reduced or absent expression of p16INK4A/p14ARF only showed significant correlation with the age at diagnosis. These data suggests that p16INK4A/ p14ARF, PTPN11 and RAS may involve in the pathogenesis of some RMS whereas MET may play a role in the progression of RMS. Before treatment,their immune function, including NK cell activityand CD4(Th1/Th2) were tested.HUVEC were isolated from an umbilical cord at cesarean section,and were cultured and expanded in vitro.After fixation with glutaraldehyde, they were stored in a refrigerator until vaccination. A vaccine mixed with HUVEC and PPD was injected intradermally to our patients every 2 weeks interval, and 1500 JRU/kg ofIL-2 was administered intravenously by drip infusion for 4-5 consecutive days.

[Results] In case 1, NSE was within normal range after 3 months of treatnent. She has been still tumor free for more than 2years. Case 2 became a stable condition without elevating tumor marker after 3 months of treatment. Major sideeffects were not seen. Before immunotherapy, activated T and NK cells were remarkably increased in both cases.Skin test for HUVEC was positive in both cases.

[Conclisions] Anti-angiogenic therapy, using HUVEC and IL-2 is safe and presumably effective in our cases.Additional trils formany patients are required, and at the same time the mechanism should be analyzed in detail. Department of Pediatrics, Ramathibodi Hospital, Mahidol University Background: Osteoporosis represents an important cause of morbidity in patients with severe thalassemia. The pathogenesis of osteoporosis in severe thalassemia is multifactorial, and includes bone marrow expansion, endocrine dysfunction, and iron overload. However, the mechanism through which these factors lead to bone loss have not been complete clarified. Osteoblasts are the bone-forming cells and arise from mesenchymal stem cell (MSCs). We therefore would like to study the MSC function in severe thalassemia patients to elucidate the bone formation dysfunction mechanism in these patients. Patients and Methods: Twenty ml of heparinized bone marrow was recovered from 10 severe thalassemia patients and 12 healthy volunteer donors. CD105+ cells were purified from each marrow sample by immunomagnetic isolation. Osteoblast differentiation genes expressions by Quantitative Real-Time RT -PCR were studied from these CD105+ cells. The osteoblast differentiation genes were consisted with cbfa1, osterix, osteocalcin, alkaline phosphatase, bone morphogenic protein-2, MSX-2, and type I collagen Results: The medians of fold difference of the most of osteoblast differentiation genes expressions from thalassemia patients were lower than healthy controls (genes expression = 1). Background: Ligneous conjunctivitis is a rare form of membranous conjunctivitis characterized by fibrin-rich, wood-like membranes mainly on tarsal conjunctivae. The disease usually affects children, girls more than boy, but may occur at any age. It appears to be the ocular manifestation of a systemic disease, which might be accompanied by formation of pseudomembrane or membrane in multiple organs such as the mouth, the tracheobroncial tree, and the female genital tracts. Most treatment attempts have so far been unsatisfactory. This report describes a case of ligneous conjunctivitis presented at early infancy whose investigation revealed a plasminogen deficiency. Method: Case report Result: A 4 year-old girl presented with bilateral recurrent membranous conjunctivitis since 1 month of age. Plasma plasminogen functional activity from the patient and mother was 15.2% and 78% respectively (normal 80-120%). Multimodality treatment was given including membrane excision, topical corticosteroid, topical cyclosporine, topical heparin, topical serum and fresh frozen plasma, with improvement. Histology of the excised membranes revealed thick amorphous material composed predominantly of mononuclear inflammatory cells, scattered polymorphonuclear cells and areas of fibrin, consistent with ligneous conjunctivitis. A patient was improved with nearly resolved of the membrane in her right eye and thin layer of membrane in her left eye. Conclusion: Ligneous conjunctivitis associated with plasminogen deficiency should be considered in differential diagnosis of cases with recurrent membranous conjunctivitis.

Hematopoiesis in regenerated bone marrow on the hydroxyapatite scaffold 

Insulin-like growth factor-II: a novel autocrine growth factor promoting the apoptosis, proliferation, and maturation of umbilical cord blood erythroid progenitors Human CBderived CD36+ EPCs were subjected to cDNA microarray. The gene expression and biological property of CB-EPC and adult PB-EPC were compared by using real-time PCR and serum-free culture system with erythropoietin (EPO).

[Results] The microarray revealed 124-fold higher levels of insulin-like growth factor-II (IGF-II) gene expression in CB-EPCs than in stimulated-lymphocytes of adult PB. Real-time PCR verified that IGF-II mRNA levels were highest in CB-EPCs compared to other CB-or adult PB-fractionated cells. When CB-EPCs were cultured with EPO in serum-free medium, anti-IGF-II-antibody (Ab) reduced the number of erythroid colonies. When CB-and adult PB-derived erythroid colony-forming cells (ECFCs) were cultured with interleukin-3, stem cell factor and EPO, mRNA levels of IGF-II, type 1 and type 2 receptors increased with both ECFCs maturation. The maturation rate by IGF-II was higher in CB-ECFCs than in adult PB-ECFCs. The majority of CB-ECFCs expressed IGF-II protein. Anti-IGF-II-Ab, but not anti-IGF-I-Ab, reduced the number of CB-ECFCs in liquid culture with EPO. Anti-IGF-II-Ab inhibited proliferation, and accelerated apoptosis of ECFCs, assessed by MTT and BrdU assays. ECFCs failed to attain full maturity in the presence of anti-IGF-II-Ab.

These results suggest that IGF-II is produced by EPCs themselves, and has a crucial role in fetal erythropoiesis by modulating apoptosis, proliferation and maturation in an autocrine fashion.

Clinical course and immunological study of an infant with Wiskott-Aldrich syndrome (WAS) Wikott-Aldrich syndrome (WAS), a primary human immunodeficiency, results from defective expression of the hematopoietic-specific cytoskeletal regulator protein (WASp). Children with WAS are compromised immunologically and also exhibit a high prevalence of autoimmune disease. We experienced severe phynotypes of WAS infant who presented life-threatening inflammatory bowel disease very early in life with recurrent ulcer which resemble to Bechet's disease. <Case history> A 13-day-old Japanese boy, presented with fever, bloody stool with raised serum CRP. At 1 month, he developed recurrent oral aphtha, genital ulcer. Intermittent fever continued with persistent leukocytosis and thrombocytopenia. Examination of bone marrow was compatible with the myelodysplastic syndrome (MDS). Colonoscopy at 2 months showed numerous small ulcers in the sigmoid and descending colon. A tentative diagnosis of BD was made because the manifestations were sufficient to meet the Criteria for BD despite the early age of presentation. The efficacy of treatment with systemic steroids, mesarazine, additional colchicine was unsatisfactory. At 8 months, a massive colonic resection was done. Because of persistent thrombocytopenia, eczyma with high IgE, we analyzed WASp gene. Exon4, G431A, (E→K) No expression WASp by western blot. <Discussion> Patient with WAS exhibit in both immunodeficiency and marked susceptibility to systemic autoimmunity. The impairment of regulatory T cell functions might explain these paradoxical findings. Our current goal is to perform more profound immunogenetic studies in an attempt to discover the pathogenesis of BD-like auto inflammatory disorders in WAS. [Background] Recent investigation revealed that slit diaphragm protein complex not only serves as a filtration barrier, it also conducts phosphorylation-mediated signals to integrate podocyte intercellular junction and cytoskeletal dynamics. Our previous biochemical studies revealed that slit diaphragm component Neph1 is tyrosine phosphorylated by Src-family tyrosine kinase Fyn. To extend our knowledge of the mechanism regulated by phosphorylation of slit diaphragm, we investigated phosphorylation of Neph1 in vivo.

[Method] To characterize the phosphorylation of tyrosine residues, we produced several antibodies which specifically recognize the phosphorylation of tyrosine residues of Neph1 along with an antibody against C-terminus of Neph1. Using these antibodies, tyrosine phosphorylation was evaluated by western blot analysis using glomerular lysates or by immunohistological analysis using cryosection from protamine-sulfate (PS) perfused rats (podocyte injury model).

[Results] Western blot analysis using Neph1 precipitates from glomerular lysate with anti-phosphotyrosine antibody revealed that tyrosine phosphorylation of Neph1 was significantly increased in PS-treated glomeruli. Affinity-purified rabbit polyclonal antibodies against Neph1 phosphopeptides specifically recognized phosphorylated tyrosine residues of Neph1 in vitro. Western blot analysis with phospho-Neph1 antibody revealed that Y637 was phosphorylated in vivo in PS model, but not in control. Biochemical analysis revealed that phosphorylation of Y637 was responsible for binding of Neph1 with Grb2, and also had a negative effect on ERK activation.

[Discussion] Neph1 is transiently phosphorylated by Fyn in response to podocyte injury which may serve as a link between junctional structure with intracellular signaling.

Functional Role of Na+-Independent System L Amino Acid Transporter 3 (LAT3) in Podocyte We previously reported that human Na+-Independent System L Amino Acid Transporter 3 (LAT3) is already expressed at S-shaped stage of the fetal kidney co-expressed with podocyte marker, synaptopodin.The aim of the present study is to determine the functional role of LAT3 in podocyte biogenesis. Immnofluoresence and confocal microscopy showed the specific distribution of LAT3 as a linear pattern on the plasma membrane of mice podocyte foot processes (FP) as well as human.When mice were starved for 24 h and 48 h, this linear distribution pattern of LAT3 on the plasma membrane was gradually but apparently diminished compared to the fed-mice.Moreover both transmission electron microscopy and scanning electron microscopy showed that the nutrient starvation induced apparent morphological change of podocytes as 1) foot processes became thin and elongated, 2) podocyte cell body became flat and wide.Finally, to determine the functional conservation of lat3 in the development of podocytes, we targeted the zebrafish lat3 ortholog using antisense morpholino oligonucleotide.Interestingly embryos injected with lat3 exon1 donor morpholino showed apparent cell number loss of glomerulus and extended bowman's space in addition to severe cardiac edema at day 5 development.

Recently identified study suggests that amino acids could activate new signaling pathway, rapamycin-insensitive pathway (mTOR2), regulating the cytoskeleton. We speculate that LAT3 likely transports branched chain amino acids into the podocyte, activating mTOR2 pathway and regulating actin-cytoskeletn system, which may play an essential role in the podocyte development and differentiation and maintenance of FP structure in the mature podocyte.

Implication of Na+-Independent System L Amino Acid Transporter 2 (LAT2) in the pathomechanism of the glomerular crescent formation Ryota Kurayama 1 , Yuji Sekine 1 , Yoshikatsu Kanai 2 , Kunimasa Yan 1 1 Kyorin university departments of pediatrics, 2 Kyorin university departments of toxicology and pharmacology Glomerular crescent formed mainly by proliferated Bowman's cells (BC) leads to rapidly progressive glomerular injury and requires early stage therapy. We reported that LAT2, specifically expressed in the basolateral membrane of proximal tubules. The aim of the present study is to determine whether mammalian target of rapamycin (mTOR) 1 is implicated in the cell proliferation process of the glomerular crescent. First we tried immunohistochemitry with LAT2 and its functional ligand, 4F2 hevay chain (4F2hc) in the specimens of crescentic glomerulonephritis (GN). Both LAT2 and 4F2hc were strongly observed in the cellular crescent lesion of IgA nephropathy and ANCA related GN. Rat glomeruli injected with anti-GBM antibody revealed apparent plasma membrane pattern of LAT2 and 4F2hc expression in the cellular crescent. RT-PCR and Western blot study revealed time dependent increase of LAT2 and 4F2hc at both mRNA and protein levels in the samples from isolated glomeruli at day 0, 2, 5, and 7 after injection of anti-GBM antibody. Finally to determine whether LAT2 is directly involved in the cell proliferation machinery, we established S2 cell line expressing full-length human LAT2 (S2hLAT2). Starvation experiment revealed significant increase of phosphorylation with eIF4E binding protein and ribosomal protein S6 (p70) kinases in S2hLAT2 cells compared to wild S2 cells. Taken together, we conclude that LAT2 may participate in the crescent formation through activating mTOR1 pathway via the transport of branched-chain amino acid into BC in various glomerular diseases. Rapamycin may be a candidate of new therapeutic tools for rapid progressive GN. On multivariate analysis, GU anomalies, particularly vesicoureteral reflux (VUR), were the most significant risk factors. Children aged greater than 5 years had a slightly higher risk of recurrence, irrespective of gender.

Comparison of organisms associated with recurrent UTI with those associated with first UTI showed that the prevalence of E. coli decreased from 76.9% to 56.2% but was still the major causative agent. Klebsiella pneumoniae and unusual or mixed organisms significantly increased from 7.8% to 15.0% and 6.2% to 16.3%, respectively. Conclusion: One-fifth of UTI children had a recurrence, with a similar rate for males and females. Independent risk factors for recurrent UTI were found to be age > 5 years, and underlying disease of either GU anomalies or VUR.

Association of 5HTR2A polymorphism with nonmonosymptomatic primary nocturnal enuresis Background. Tricyclic antidepressants(TCA) was used for treatment of NE for decades of years. Recently, some case studies demonstrated the efficacy of some serotonin reuptake inhibitors (SSRI) in the treatment of enuresis. Both TCA and SSRI have the similar influence on serotonin neurotransmission. Serotonin participates many physiologic processes such as sleep, thermoregulation, pain perception and hormone secretion. Animal studies also exhibits TCA exerts an inhibitory action on the micturition reflex by a central cholinergic mechanism. This study aimed to investigate 5-hydroxytryptamine (serotonin) receptor 2A ( 5-HTR2A ) gene polymorphisms in children with primary nocturnal enuresis (PNE). Methods. The 5-HTR2A gene polymorphism was investigated in 151 Taiwanese children (99 PNE cases and 62 healthy, non-enuretic controls). The 5-HTR2A genes polymorphism (rs6313)of the 5-HTR2A genes were genotyped using PCR. Results. There were no significant differences when comparing the allelic frequencies and genotypes of 5HTR2A polymorphisms between NE with and controls. We subsequently compared in turns: arousal score, diurnal voiding symptoms (DVS), and constipation according to 5HTR2A genotypes and allelic frequencies. A significantly differences in genotype distribution in NE patients with DVS or constipation compared with mono-symptomatic NE patients were observed (p=0.037 and 0.010 ) and allele T was also increased in non-monosymptomatic NE patients. Conclusions. This study is the first to search the 5-HTR2A gene polymorphisms in children with PNE. It was determined that 5-HTR2A gene polymorphism, a predominantly TT genotype, may be associated with non-mono-symptomatic PNE in Taiwanese children.

Pulse cyclophosphamide induction treatment in Thai children with diffuse proliferative lupus nephritis Sauwalak Opastirakul, Wattana Chartapisak Chiang Mai University, Department of Pediatrics Background: Intravenous cyclophosphamide has been the standard treatment for diffuse proliferative lupus nephritis for more than 20 years. Recently, this recommendation is questioned regarding its effectiveness and side effects. We reported the effectiveness of pulse cyclophosphamide induction therapy and identified predictors for unresponsiveness to treatment in Thai children. Methods: Children under 15 years of age with biopsy-proven diffuse proliferative lupus nephritis who were admitted to Chiang Mai University hospital between 2001and 2006 were retrospectively studied. Responsiveness to treatment, defined as urinary protein to creatinine ratio of less than 0.3, was assessed at the end of induction period. The clinical characteristics and laboratory data including gender, age at diagnosis of SLE, duration of disease before treatment, hypertension, clinical nephrotic syndrome, amount of proteinuria, serum creatinine, creatinine clearance, serum C3 level and presence of crescentic formation were compared between responsive and nonresponsive groups. Results: A total of 29 patients (90% female) with the mean age of 10.3 ± 2.6 years were studied. Hypertension, microscopic hematuria and nephroticrange proteinuria were seen in 66%, 86% and 60% of the patient respectively. Forty one percent of biopsy showed cellular or fibrocellular crescents. Twenty patients (69%) achieved remission at the end of induction therapy. There were no significant differences in all parameters studied between responsive and nonresponsive groups. Conclusions: Pulse cyclophosphamide is an effective regimen for induction therapy in children with diffuse proliferative glomerulonephritis. No definite predictor for unresponsiveness was detected in this study.

We reported the preliminary results of clinical analysis on cardiomyopathy in autopsy cases of severe motor and intellectual disabilities (SMID) (2006, 2nd ASPR). To further explore pathomechanisms, we immunohistochemically examined accumulation of oxidative products and expressions of antioxidant enzymes.

The subjects consisted of 5 SMID autopsy cases of cardiomyopathy aged 3-59 years, and 4 controls aged 3-66 years. The cause of cardiomyopathy was speculated to be chronic carditis of unknown origin, cor pulmonale, suspicious selenium deficiency, subacute carditis related to hepatitis C virus (HCV) infection and acute selenium deficiency, respectively. Serial sections of cardiac tissue were treated with monoclonal antibodies against mitochondria and oxidative stress markers to DNA, lipid and protein, in addition to polyclonal antibodies against cytochrome c oxidase (COX) and superoxide dismutases (Cu/ ZnSOD and MnSOD). RESULTS: Remnant cardiac tissues were immunoreactive for oxidative product to protein in cases but not controls, whereas neither cases nor controls showed accumulation of oxidative products to DNA or lipid. The remaining cardiac tissues demonstrated patchy or reduced expressions of mitochondria, COX and SODs in three cases of cardiomyopathy, being related to selenium deficiency and HCV infection. CONCLUSION: Data indicate the possibility that oxidative damage to protein can be involved in cardiomyopathy in SMID patients. Disturbed mitochondrial functions with reduced antioxidant capacity seem to make an additional contribution to generation of cardiomyopathy, being related to selenium deficiency and HCV infection.

Abnormal white matter appearance on term FLAIR predicts neuro-developmental outcome at 6 years old consequential to preterm birth Division of Rehabilitation, Nagano Children's Hospital, 5 Department of Pediatrics, Saitama University School of Medicine Purpose: Preterm infants are at significant risk of minor neuro-developmental disorders and learning disabilities at school age. Early reliable screening tools to identify high-risk infants are urgently required. The predictive value of term MRI with the minor neuro-developmental outcome at school age is still unknown. Materials and methods: 210 preterm-infants (<36 weeks) were studied to investigate clinical factors associated with white matter (WM) appearance on MRI. Results: Low-intensity signal in WM on fluid-attenuated inversion recovery (FLAIR) imaging was commonly observed (69%) at less than 2 months corrected-age. Its incidence correlated with corrected-age at scan and incidences of maternal pyrexia and cystic periventricular leukomalacia. Lowintensity signal in WM on FLAIR significantly correlated with physical and total developmental quotients, whereas diffuse high-intensity signal in WM on T2-weighted imaging correlated only with the total developmental quotient at 6 years (n=75, WISC-R). FLAIR imaging, but not T2-weighted imaging, predicted mild neuro-developmental delay at this age. Conclusion: FLAIR appeared to detect subtle WM injury related with neuro-developmental disorders at schoolage, whereas T2-weighted imaging seemed to identify relatively more severe WM injury. FLAIR is a potentially sensitive screening tool for many infants that is readily available and easily interpretable.

Oxidative stress in neurodegeneration in dentatorubral-pallidoluysian atrophy Background: Dentatorubral-pallidoluysian atrophy (DRPLA) is one of CAG-repeat diseases, and it is classified into juvenile, showing progressive myoclonus epilepsy (PME), early adult and late adult types. The common neuropathological findings in DRPLA include degenerations of the basal ganglia and dentate nucleus. It has been discussed in Huntinton's disease that expanded polyglutamine can lead to oxidative neurodegeneration, and we examined involvement of oxidative stress in brain lesions in DRPLA. Method: We performed immunohistochemistry for accumulation of oxidative products and expression of superoxide dismutase (SOD), a radical scavenger, in serial brain sections in autopsy cases of DRPLA. Results: Oxidative products of nucleosides, 8-hydroxy-2'-deoxyguanosine and 8-hydroxyguanosine, were accumulated in the lenticulate nucleus predominantly in DRPLA cases having PME. Mild neuronal accumulation of 4-hydroxy nonenal, a reactive lipid aldehyde, was found in the hippocampus, globus pallidus and cerebellar dentate nucleus in the adult DRPLA cases and controls. Cytoplasmic immunoreactivity for Cu/ZnSOD was reduced in the external segment of globus pallidus, dentate nucleus and cerebellar cortex in most DRPLA cases. Mitochondrial immunoreactivity for MnSOD was reduced in the lenticulate nucleus and cerebellum in DRPLA cases having PME. Some DRPLA cases also showed reduced immunoreactivity for MnSOD in the cerebral cortex. Coexistence of reduced SOD expression and expanded polyglutamine was observed in the frontal cortex or lenticulate nucleus in a few cases. Conclusion: It is likely that oxidative stress can be involved also in DRPLA, although the exact relationship with expanded polyglutamine remains to be elusive.

Sawa Yasumoto 1 , Takahito Inoue 1 , Toshiko Mori 1 , Hiroko Ogata 2 , Akihisa Mitsudome 3 , Shinchi Hirose 1 1 Fukuoka University, Department of Pediatrics, 2 Aoba Clinic, 3 International University of Health and Welfare

Background: There are several studies on the maturation of proprioceptive nerves in neonates and a few in preterm infants. Purpose: To evaluate the maturation of efferent motor nerve and afferent sensory nerve in comparison between full-term (FT) and preterm infants (PT), M waves and H-reflex were examined at a similar post-conceptional age. Subjects and Methods: Eleven FT (gestational age 38~41weeks) and PT infants (gestational age 26~34weeks) were recruited for the study. Informed consents were obtained from the parents. M-wave and H-reflex were recorded within the first week of life for the FT infants while the same parameters were measured for premature babies when they reached the expected time of birth using four channel evoked potential examination apparatus (Signal processor-7S12; NEC Medical Systems). The median nerve was stimulated at the wrist and elbow and M-wave and H-reflex were recorded from the abductor pollicis previs muscle. The motor and proprioceptive nerves conduction velocities (MCV and HCV, respectively) were determined by dividing the difference in latency between the onset of the proximal and distal responses by that distance. Results: The MCV of PT (25.0±4.5m/s) was comparable to those of FT infants (25.5±4.5m/s). In contrast, the HCV of PT (20.1±5.6m/s) was significantly lower than those of FT infants (25.0±10.0m/s). Conclusion: These findings suggest that myelination in the afferent sensory nerves may delay in the extrauterine environment for PT infants. Maturation of proprioceptive nerve of PT infants hence may be influenced by nutrition or medication after birth. To be able to determine the prevalence of developmental concerns in apparently well children aged 1-3 years old by using the Filipino Denver Developmental Screening Tool. STUDY DESIGN: cross sectional. STUDY POPULATION: All children ages 1-3 years old (63) at Brgy Minantok, Amadeo, Cavite who fulfilled the inclusion criteria. METHODOLOGY: All children aged 1-3 years old were interviewed. Parents were asked to fill-out a questionnaire regarding socio-demographics. A scheduled interview regarding the socio-demographic, maternal factors, and neonatal factors was conducted. Afterwards, Filipino Denver Developmental Screening Test was administered. Background data of subjects who have been identified with developmental concern were revisited and risk factors present in their histories were determined and reported for correlation. RESULT: Almost 63 subjects screened had unremarkable demographic, maternal, paternal and neonatal histories which may contribute to the child's development. Two of the population screened were noted to have developmental concern, 1 as language delay and the other as fine motor delay. Back ground: Olmsted syndrome is a rare disorder which was first described by Olmsted HC in 1927, and is dermatologically characterized with sharply marginated keratoderma in the palms and soles, perioral keratoderma and alopecia. We reported a 23-year-old male patient with Olmsted syndrome showing rare neurological and immunological complications. Case presentation: There was no family history of keratosis or neurological disorders. In infancy, he suffered from marginated keratoderma, and acrodermatitis entheropathica was suspected, which was temporally controlled by replacement therapy of zinc sulfate. Gradual development of periorificial and bilateral palmoplantar keratoderma with alopecia led to the diagnosis of Olmsted syndrome, and hyperkeratosis in skin biopsy specimen supported the diagnosis. He also demonstrated motor clumsiness, mental retardation, optic nerve atrophy, spastic gait, and hyporeflexia with numbness at the ends of extremities. He was suspected as having primary peripheral neuropathy because of delayed conduction velocity and reduced myelinated fibers in biopsied peroneal nerve. He also developed multiple demyelinated lesions and asymptomatic signal change in the corpus callosum on magnetic resonance imaging in addition to having sepsis repetitively. At the age of 23, he was complicated with myelodysplastic syndrome (MDS), and he died of sepsis one year after. Conclusion: The occurrence of neurological and immunological complications itself is rare in Olmsted syndrome, but that may suggest the possible linkage between hyperkeratotic syndrome and neurological and/or immunological disturbances.

Expectant mothers of premature infants often have psychological problems. When pregnant women admit to the mother-fetus intensive care unit (MFICU), they would feel to be isolated and would have mood disturbances. The purpose of this study is to evaluate the effects of prenatal visit on mood status of expectant mothers of infants.

Participants consisted of 27 women who were treated at the MFICU in the Kansai Medical University Hospital and have delivered from December 2006 to May 2007. Prenatal visit sessions were held once a week by a neonatologist and a clinical psychologist of the neonatal intensive care unit (NICU). In the sessions, the neonatologist ask the expectant mothers about their anxieties, complaints and requests while clinical psychologists observed and helped them to open their thoughts freely. Before and after the sessions, the participants were asked to complete the Japanese version of the Profiles of Mood Status (POMS).

Ages of participants ranged 24 to 42 years, average of gestational age was 31 weeks, and average length of the stay in MFICU was 52 days. Compared the scores of POMS subscales after the prenatal visit with those of before the sessions, average score in regard to "depression" among POMS subscales was significantly decreased after the sessions. The results indicated the mood status of the participants improved after the prenatal visit sessions.

Prenatal visits by a neonatologist and a clinical psychologist would relieve psychological stress and improve mood status of the expectant mothers of premature infants. Objectives: Adnexal torsion is an acute abdominal condition often confused with other diseases in children. The objective of this study is to evaluate the diagnosis, characteristics and treatment of adnexal torsion in children to reduce morbidity. Methods: We reviewed the medical records of all cases of children whose adnexal torsion diagnosis was proven by surgery from 1992 to 2005. Prenatal cases were excluded. Results: A total of 49 cases were enrolled; the mean age was 12.5 years. Sixteen (32.6%) cases were premenarchal girls. This group was significantly more likely to have adnexal problems missed at first clinical diagnosis compared to postmenarchal girls (P=0.032). Of patients, 32% had severe abdominal pain and 14.2% had a history of recurrent pain over 2 weeks. Of abdominal gray scale ultrasound evaluated in 43 patients, 41 were suspicious for ovarian pathology. Seventeen patients had both ultrasound (US) and abdominal computerized tomography (CT). These had relatively high infarction or necrosis compared with US alone (82.3% versus 61.5%). Fever was significantly associated with infarction or necrosis (P=0.004).

Conclusions: Adnexal torsion should be considered in pediatrics girls with abdominal mass and any degree of abdominal pain. Most pediatric adnexal torsion occurs in postmenarchal patients but should not be overlooked in premenarchal girls with abdominal pain. Ultrasound plays an important, but not definitive role in diagnosis. Multiple radiographic studies with combined CT and US did not adversely affect viability or infarction. An emergency operation should be performed, especially in suspected adnexal torsion with fever to avoid further ovarian damage.

Two adolescent-boy of influenza B virus infection complicated with abnormal behaviors.

Shoji Yoshimatsu, Yuki Enomoto, Kazunori Nishimura, Makoto Hasegawa

Introduction: In last March, the Japanese Ministry for Health, Labor and Welfare issued a warning to doctors not to prescribe oseltamivir phosphate to adolescents aged 10-19 years. This announcement was in response to unusual suicide after taking oseltamivir phosphate in Japan. We present two adolescent-boys who manifested abnormal behavior during the each clinical course of influenza B virus infection. Case1: A 14-year-old Japanese boy was diagnosed influenza B and prescribed oseltamivir phosphate by another hospital. 3.5 hours after taking oseltamivir phosphate, he suddenly manifested abnormal behavior (He was walking around the room, defying his mother and his older brother) and arrived to our hospital. Case2: A 13-year-old Japanese boy was diagnosed influenza B and not prescribed oseltamivir phosphate by another hospital. His temperature became normal on 6th day, but he manifested drowsiness and abnormal behavior(e.g. he could answer the question, but could not stay awake. He suddenly called his mother on the phone and told her what had never happened.), then his family took him to our hospital. Discussion: Both cases are completely recovered without any medication. It is very difficult to diagnose case1 as side effect of oseltamivir phosphate because influenza infection, itself, manifests abnormal behavior, which was seen in case 2. Although the relationship between oseltamivir phosphate and suicide was not clarified, we must take these events extremely serious. And also we need to distinguish between influenza encephalopathy and these cases very carefully.

Effectiveness of Government-sponsored Disease Management Program in Improving Severity of Pediatric Asthma in Taiwan

On the other hand, [ 3 H]-leucine incorporation was significantly increased by leptin in a dose dependent manner. The protein/DNA ratio increased 1.7-fold by leptin 10 ng/mL. Leptin 10 ng/mL activated both ERK and p38. The levels of phosphorylated-ERK (P-ERK) and phosphorylated-p38 (P-p38) peaked at 10-30 min (1.6-and 1.7-fold increase, respectively) remaining elevated for 1-6 h, and returned to the baseline level at 24 h. Leptin increased P-ERK and P-p38 at 30 min in a dose dependent manner. The levels of total ERK and p38 remained unchanged. Increase in [ 3 H]-leucine incorporation stimulated by leptin 10 ng/mL was completely inhibited by coincubation with a MEK inhibitor PD98059 or a p38 inhibitor SB203580. These results demonstrate that leptin induces hypertrophy of metanephric mesenchymal cells via ERK and p38

P-157 CSF d-ROM levels in Japanese pediatric patients with CNS diseases

Akinori Hoshika Tokyo Medical University, Department of Pediatrics [Background] Free radicals have been suggested to play a pathogenic role in a variety of central nervous system (CNS) diseases based on a growing body of evidence obtained from experimental models and the human brain

adrenoleukodystrophy (ALD; n=2) and multiple sclerosis (MS; n=1). [Results] An analysis of the infection-associated group (BM, VM, FS, RC) showed that the CSF d-ROM levels in the BM group (average 119.4±76.2 U.CARR) were significantly higher than those in the VM (average 33±8.1 U.CARR), FS (average 9

As for demyelinating disease such as ALD (n=2) and MS (n=1), patients with progressive ALD or MS showed high d-ROM levels (39 and 74 U.CARR, respectively) compared of the other ALD patient at early stage (2 U.CARR). [Conclusion] In patients with CNS diseases, the CSF d-ROM levels may correlate with their clinical severities or symptoms

P-165 13q partial monosomy presenting with Arima syndrome and bilateral retinoblastoma

The present study assessed the role of GABRG2 gene in idiopathic generalized epilepsies(IGEs) of Korean population. Methods: 23 IGEs and 94 healthy control subjects were selected throughout a collaborative study of Catholic Child Neurology Research Group. The SNP211037 of GABRG2 were screened by DHPLC. DNA fragments showing variant chromatograms were subsequently sequenced. Genotypes and allelic frequencies for GABRG2 gene polymorphism in three groups were compared. Results: Genotypes and allelic frequencies of the γ2 subunit of the GABA receptor gene(SNP211037) in both groups were not significantly different. The most common genotype for GABRG2 (SNP211037) gene in both groups were T/C heterozygote. The allele C and T frequencies for GABRG2 (SNP211037) in IGEs group were 45.7% and 54.3%, respectively; and in healthy control group, 42.6% and 57.4%, respectively. The number of individuals with the GABRG2 (SNP211037)-C/C genotype in IGEs group greater compared with that in healthy control group(21.7% versus 12.8%). The odds ratio for developing

A Randomized Clinical Trial on the Effectivenes of Icegel Compress versus Lidocaine-prilocaine Cream in Reducing Pain Associated with Diphtheria-Pertussis-Tetanus Vaccination

Methods: One-hundred twenty-five students scheduled for BDPT/ B booster were randomized into lidocaine-prilocaine, 5-minute icegel compress, 10-minute icegel compress and placebo group prior to BDPT/B injection. Effectiveness of icegel was compared with lidocaine-prilocaine and placebo in terms of pain reduction using Wong-Baker Faces and BFLACC/B (Face-Legs-Activity-Cry-Consolability) pain scales. Increase in heart rate and decrease in O2 saturation during vaccination were measured and compared with the baseline value taken before injection. Results: The mean pain scores of all

Conclusion: Icegel compress is as effective as topical lidocaine-prilocaine with regards to pain reduction. Tenminute application is more effective than 5-minute application. There is lower increase in heart rates and lower decrease in O2 saturation when icegel is applied

Method: This is a retrospective study by pediatric pulmonologists and asthma educators in a regional teaching hospital. The children from an outpatient pulmonology clinic were expected to receive 5 times of comprehensive evaluation visit (CEV) to classify asthma severity and receive asthma education in 1 year. We analysis the asthma severity and the severity change. Result: 700 asthma children (0-17y/o, mean 5

Natural history of trisomy 18 fetus diagnosed in the third trimester Trisomy 18 is the second most common autosomal trisomy in liveborn infants, and occurs at a frequency of 1 per 3,000-5,000 live births. Conbined with the ultrasonographic examination, multiple marker screening test involving the maternal serum AFP, uE3, and hCG in second trimester was reported to identify about 60% of trisomy 18 with a 0.4% false positive rate. The life expectancy of trisomy 18 after live born reveals the high infant motality rate. About 90% children with trisomy 18 die before their first birthday. Recently the natural history of trisomy 18 of live born was investigated and reviewed extensively.To delineate the natural history of 18 trisomy fetus prenally diagnosed in the third trimester, we reviewed the 50 cases diagnosed after 27 weeks of gestation in our institute between 1994 and 2003. 57% of the cases were live births, and 43% were stillbirths. Prenatally diagnosed cases reached to live-born showed no difference in the prognosis and life expectancy to the control group. The sex ratio was about 1 at that time of prenatally diagnosed, but rapidly declined at term. These information was valuable for counseling to families with the prenatally diagnosed cases and will provide insight into the sex ratio of the disorder.

Gene therapy of neonatal Fabry mice to prevent disease progressionKoichi Ogawa 1 , Yukihiko Hirai 1 , Masamichi Ishizaki 3 , Hideki Hanawa 1 , Yoshitaka Fukunaga 2 , Takashi Shimada 1Background: The purposes of the study were to determine β-cell function and insulin sensitivity and the association of variants in β-cell differentiation genes with insulin resistance (β-cell dysfunction) in childhood ALL survivors. Methods: Childhood ALL patients who had been diagnosed during [1987] [1988] [1989] [1990] [1991] [1992] [1993] [1994] [1995] [1996] [1997] [1998] [1999] [2000] [2001] [2002] [2003] [2004] and completion of treatment for at least 6 months. The oral glucose tolerance test and lipid screening were performed. Impaired glucose tolerance and diabetes were defined according to WHO criteria. Common polymorphisms of TCF7L2 were genotyped and assessed their effect on β-cell function and insulin sensitivity. Results: 110 children were included in this study (45 females and 65 males; age 4-20 yr). 101 children (91.8%) had normal glucose tolerance (NGT). The others (8.2%) had impaired glucose tolerance (IGT) which associated to older age at test (13.7 and 10.3 yrs, p = 0.006), longer period of completion of treatment (47 and 26 months, p = 0.003), and lower value of insulin sensitivity indicies (WBISI) (5.82 and 9.56, p = 0.001) than children in NGT group. The typical signs of metabolic syndromes (hypertriglyceridemia, low HDL-C, obesity) were found up to 20%. The genotypes of TCF7L2 were not difference between NGT and IGT group (p=1). Conclusion: Impaired glucose tolerance (IGT) related to longer period of completion of treatment and older age group. Long-term survivors of childhood ALL had potential risk of impaired β-cell function and impaired glucose tolerance (IGT), even in non-obese children. Our findings did not support association of TCF7L2 variants with insulin resistance (β-cell dysfunction) in childhood ALL survivors These enzymes may also be potentially important for dissemination and tissue invasion of malignant lymphocytes. We examine, in this study, the correlation of MMP and TIMP gene expression in leukemia cells with the clinical characteristics of infants with acute lymphoblastic leukemia (ALL), especially with the extramedullary organ involvement. Methods: The mRNA contents of MMP-2, MMP-9, TIMP-1 and TIMP-2 in leukemia cells from 33 infants with infant ALL were quantified at initial presentation by the real time quantitative RT-PCR (TaqMan methods). The association between their expression and patient clinical characteristics was examined, including patients' age and white blood cell count at diagnosis, MLL gene rearrangement in leukemia cells, and presence of hepatosplenomegaly or central nervous system (CNS) involvement. Results: The mRNA contents of MMP-2 and MMP-9 were not associated with any patient characteristics. The MMP-2/TIMP-1 and MMP-2/TIMP-2 ratios were significantly higher in patients with hepatosplenomegaly than those without it (p=0.005 and 0.009). The MMP-2/TIMP-2 ratio was also higher in patients with CNS involvement (p=0.012). The MMP-9/TIMP-1 and MMP-9/TIMP-2 ratios did not associate with the organ involvement. Conclusions: The MMP/TIMP balance, but not the mRNA amounts of a single gene, may play an important role in dissemination and organ invasion of leukemia cells in infant ALL.

Age-associated difference in gene expression of myelo-monocytic AML Recently, a number of methods have been devised for detection of mutations in the field of molecular genetics. The newly available LightCycler ® system has been used for rapid PCR, while simultaneously quantifying and analyzing the amplification results. We tried to apply the LightCycler ® system to the diagnosis of Japanese-type (type 2) adenine phosphoribosyltransferase (APRT) deficiency in two families with 2,8-dihydroxyadenine urolithiasis.[Cases] The first patient was a 3-year-old girl who presented with left flank pain. The second patient was a 2-year-old girl who presented with complaints of sudden dysuria.The spectrophotometric analysis of the stone fragments revealed an absorption spectrum for 2,8-DHA. Results of Gene Analysis: We used the LightCycler ® system to detect APRT*J and the PCR-SSCP method to detect APRT*Q0. The first patient was homozygous for APRT*J/APRT*J and the second patient was compound heterozygous for APRT*J/APRT*Q0.[Disccusion] The analysis of the APRT gene is the most reliable diagnostic method in infants. Major problems with the present PCR-SSCP method, however, are that it reguire multiple manual step, and it takes a very long time before the detection can be confirmed. Meanwhile, the analysis of the APRT gene using the LightCycler ® system can provide mutation results within approximately 30 minutes after DNA isolation, and the results are easily and clearly visualized. The genetic diagnosis of APRT deficiency using this system may be useful not only as a diagnostic test for infants with known 2,8-DHA, but also as a screening of infants with a suspicion of urolithiasis.

Immunohistochemical analysis of cardiomyopathy in autopsy cases of sever motor and intellectual disabilities Figure 1 . Majority (90%) was febrile seizure (FS) and only 5 (0.6%) were encephalopathy. Two patients died; 1 with underlying methylmalonic aciduria while the other was a previously healthy 6 y.o. girl who developed shock, DIC and died on D2 fever. CT brain showed generalized cerebral edema and hypodense lesions in thalamus, brainstem and basal ganglia. The picture is compatible with acute necrotizing encephalopathy reference 3 frequently reported in Japan. Conclusion: Combining previous findings, incidence of INCs was ~240 per 100,000 person-years in HK. The discrepancy between HK and USA is at least partly attributable to different admission threshold for influenza-related illnesses and FSs. INC data from Japan will be of great interest if inter-ethnic difference in the neurotropic effect of influenza is to be explored.P-163 CYP2C19 polymorphisms as a predictor of adverse effects of clobazam Background: Clobazam (CLB) is effective in treating intractable epilepsy, but with occasional adverse effects that often require dose decrease or termination. CLB is metabolized to N-desmethyl CLB (N-CLB) by CYP3A4, and then to an inactive metabolite by CYP2C19. We have previously shown that 1) the ratio of serum levels of N-CLB to CLB was increased in patients with two mutated CYP2C19 alleles, 2) patients co-medicated with CYP3A4 inducer showed lower CLB concentration/dose. Purpose: To examine effects on occurrence of adverse effects of CLB of the number of mutated alleles of CYP2C19 and co-medication with CYP3A4 inducer(s). Methods: 41 patients with intractable epilepsy who had been on multiple AED; between the age of 3 to 38 years; 22 males and 19 females. Median CLB administration duration was 36.6 months (range; 8 to 58 months). Results: In 11 patients it was necessary to reduce the dose of or discontinue CLB due to severe adverse effects. Of those 11 patients, 3 are among 17 with no mutant alleles (3/17 or 18%), 3 among 16 with one mutant allele (19%), and 5 among 8 with two mutant alleles (63%). The most sever adverse effects were observed in 3 patients; those patients had either one or two mutant alleles, and all were co-medicated with CYP3A4 inducer. Conclusion: Patents with two mutant alleles of CYP2C19 are more likely to develop sever adverse effects, particularly when co-medicated with CYP3A4 inducer(s), which may lead to discontinuation or reduction in dosage of CLB.

Autosomal dominant hyperkalemic periodic paralysis with biallelic mutations of the SCN4A gene Yoshito Ishizaki, Ryutaro Kira, Hiroyuki Torisu, Masafumi Sanefuji, Naoko Yukaya, Toshiro Hara Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University Introduction: More than 20 missense but no truncation mutations in the skeletal muscle voltage-gated sodium channel α subunit (SCN4A) gene have been identified in various forms of skeletal muscle sodium channelopathies including hyperkalemic periodic paralysis (PP), paramyotonia congenita, potassiumaggravated myotonia, hypokalemic PP and congenital myasthenic syndrome. Method: The proband was a Japanese 12-year-old boy, who experienced his first attack of paralysis at the age of one year. Thereafter, paralytic attacks intermittently recurred a few times per day, each lasting for several minutes to an hour. The weakness most often appeared during a rest after strenuous exercise, and repeated exercise alleviated the weakness. After informed consent was obtained, we analyzed exons 9, 13, 14, 19, 22, 23 and 24 of SCN4A, the regions of SCN4A containing mutations in hyperkalemic PP patients reported previously, using PCR and direct sequencing. Results: We detected a missense mutation in exon 13 (c.2375T→G, V792G) of the maternal origin and a one-base deletion in exon 24 (c.4677delC, P1560fsX1650) of the paternal origin in the heterozygous form in the proband and his younger brother. Discussion: We have identified biallelic heterozygous novel muations V792G and P1560fsX1650 in the SCN4A gene in siblings with hyperkalemic PP. Their mother and five her relatives had the same symptoms, while their father had severe stiff shoulders but no paralytic attack. The phenotype of the siblings appeared to be caused by not the P1560fsX1650 mutation, but the V792G mutation, located at the sixth transmembrane segment of domain 2 of SCN4A. There are only a few study in Thailand and lack of complete data collection system. Objectives: To study the epidemiologic data of child abuse case for creating an optimized prototype for data collection system at the emergency room. Methods: The medical records of child abuse cases attended in the emergency room at Ramathibodi Hospital between Jan 2001 to Dec 2004, were retrospectively reviewed. The demographic data included age, sex, type of abuse, abuser, sender, education, sibling and order, family member, parent's career, family relationship, substance abuse in family and socioeconomic status. Then the multivariate analyses were used for determining the correlation of each variable. Results: One hundred child abuse cases (84 females, 16 males) with a mean age of 9 years, were enrolled in the study. The child abuse was defined as sexual abuse (60%) and physical abuse (40%). The prevalence of sexual abuse was gradually increased form 6 cases in the year 2001 to 9, 22 and 23 in the year 2002, 2003 and 2004 respectively. The multivariate analyses revealed that there was no statistically significant correlation between the sexual/physical abuse and all variables. This may be explained by a rather small sample size. However, the computerized data collection system for child abuse has been created.

The prevalence of sexual abuse gradually increased. A further cohort study in a large sample size is warranted.

Early detection of anorexia nervosa in school tetsuya tanaka 1 , takashi esaki 2 , mikako inokuchi 1 , tomonobu hasegawa 2 , mitsuaki tokumura 1 , seiichiro nanri 1 , hisako watanabe 2 , takao takahashi 2 1 Health center of Keio University, 2 Keio University, Department of pediatrics Background: In spite of the fact that anorexia nervosa is refractory disease with a high mortality rate, many patients and their parents are in strong denial about this disease and therefore early detection is considered to be difficult. We are trying to prevent and detect anorexia nervosa early by using growth curves and pulse rates in school. Methods: Since 2001, based on the physical measurement values of the girl students in the health examinations conducted twice a year in two private junior high schools, we have collected those who fall under the following conditions: (1) downward shift of weight by 1 channel and more on a growth curve and the degree of obesity ≤-15%;(2) weight loss of ≥3 kg. Among these students, when bradycardia (heart rate ≤60/min) or amenorrhea is detected, we refer them to medical institutions. Results: Twenty students were referred to medical institutions, and 19 of them except one consulted a doctor. Among these 19 students, 2 were diagnosed as underweight, 17 as anorexia nervosa. In these two schools, we have performed early interventions before the level of emaciation reaches to a severe condition. Therefore, progressive anorexia nervosa has not been detected since our screening program was launched. Conclusions: When weight loss or poor weight gain on a growth curve are observed based on the physical measurement values, and bradycardia or amenorrhea is detected, the possibility of a diagnosis of anorexia nervosa should be considered, and early close examination is required.

Takashi Esaki, Tetsuya Tanaka, Hisako Watanabe, Takao Takahashi

Background: Against the backdrop of ever increasing number of anorexia nervosa(AN)among early teen-age girls in Japan, training of inpatient treatment by a full-time child psychiatrist has become an essential component of the postgraduate training programme at the Department of Pediatrics, Keio University Hospital in Tokyo. Over the past 12 years within our pediatric ward, we developed ANICU (anorexia nervosa intensive care unit), a comprehensive step-by-step program for recovery from extreme emaciation. Methods: 70 children(64 girls,6 boys) who underwent inpatient treatment of ANICU at Keio Hospital over the past 12 years. Results: All patients were seriously ill cases with multiple organ failures. With sincere empathic attendance and care including systematic feeding session and firm emotional containment at times of children's distress, all patients steadily recovered physically and emotionally to their healthy weight, and endocrinological state, feeding behaviors and enhanced self-esteem. Conclusions: AN is a serious psychosomatic disease with a high risk of death, recurrence and life-long suffering. With ever increasing numbers of young teenage AN in Japan today,combined team of child psychiatrist and pediaticians play a vital role in rescuing children with AN.

An urgent need for a 24 hour crisis center for sexually-abused young children and disabled people in Japan.Michiko Sakai 1 , Takashi Esaki 1 , Tetsuya Tanaka 2 , Hisako Watanabe 1 , Takao  Takahashi 1 1 Keio University, Department of Paediatrics, 2 Health Center of Keio University Background: Recently, many sexual crimes are reported by mass media, which include murders, kidnaps and sexual abuse by pedophiles. Although Japan approved ratification the Treaty of Children's Right in 1994, pedophiles go unpunished in the society. Methods: We study the present system for sexually-abused children in Japan through following case studies of young and disabled children which became court cases. Subjects: A 3-year-old girl who was sexually abused by the male teacher in the kindergarten. A 9-year-old and a 11-year-old girls who were sexually abused by the teacher in the special education class of the elementary school. A 16-year-girl who was raped by a elder student in the high school for handicapped children. Results: Our study revealed that there are no 24 hour crisis center for sexually-abused children and disabled people in Japan. These cases revealed that the victims had no access to medical after care, making it impossible to produce any evidence. Conclusions: We should create a 24 hour crisis center to help sexually-abused children and disabled people.

Voraluck Phatarakijnirund 1 , Suphachai Janjindamai 1 , Jaturaporn Sangkul 2 1 Prince of Songkla university, department of Pediatrics, 2 Prince of Songkla university, Department of Psychiatry Background: Attention deficit-hyperactivity disorder (ADHD) is a disorder in which those affected display symptoms of hyperactivity, inattention and impulsivity. Its prevalence has been reported as 3-20% in other countries but its prevalence has not been studied in our region. Objectives: To study the prevalence and distribution of subtypes of ADHD in primary school children in Hat Yai, southern Thailand. Method: A cross-sectional study was performed in first grade Hat Yai students from 15 primary schools using the Conners' Abbreviated Parent/Teacher Questionnaires. Children with positive scores ( > 12) and their parents were randomized and interviewed by a pediatric psychiatrist using DSM IV. Result: 2,539 questionnaires were sent out of which 2,293 (90.3%) were returned. 1,479(90.7%) of the students were from private schools and 814(89.5%) from public schools. The age range was 4.0-9.9 years with a mean of 7.8+0.5 years. 794 children (34.6%) tested positive for ADHD, of whom 198 were interviewed with their parents to confirm the ADHD diagnosis. 46 were proven to have ADHD, giving a prevalence of 8.0%. The prevalences in private and public schools were not significantly different (7.6% vs. 8.1%, p=0.3). The male to female ratio was 2.5:1. Hyperactive, inattentive, and combined subtypes were found to be 2.1%, 3.1% and 2.8%, respectively. (p=0.9). Conclusions:. The prevalence of ADHD among school children in southern Thailand was 8% and males predominated. The distribution of the 3 major subtypes was similar. Results: Of the total sample, 2,236/3,692 (60.6%) shared a bed with their parents, and 39.3% shared a room but slept in a different bed. Sixty percent of the parents place their infants to sleep in a supine position, 32.2% place on their side and 4.9% in a prone position. Bed sharing was associated with older maternal age, higher parental education and with professional careers. Places the infants to sleep in a non-supine position was associated with older maternal age, higher parental education, higher income, professional careers, and with infant birth weight of > 2,500 gm. Conclusions: Bed sharing is more common in Thai neonates than has been reported from other countries, but rates of sleeping in the non-supine positions are comparable. The main factor associated with bed sharing as well as putting infants to sleep in the non-supine positions was a higher parental socioeconomic status (SES), in contrast to previous studies in which both are associated with low maternal SES.

Preliminary study of visual reliance and target size effects on reach-to-grasp movement in children with high functioning autism Hsiu-Ching Yang, I-Ching Lee National Cheng Kung University, Department of Occupational TherapyMotor coordination problems have been reported in children with autism, however there were little studies addressing whose sensory reliance in fine motor tasks. In this study, there were four conditions of reach-to-grasp movement: large target with visual feedback (VL), small target with visual feedback (VS), large target with non-visual feedback (NL), and small target with non-visual feedback (NS) conditions to investigate the effects of visual condition and target size. A counterbalanced repeated measure was designed. Ten children with high functioning autism (HFA) and 10 normal subjects were matched according to age and handedness. Qualisys motion analysis system was used to capture the reachto-grasp movement. Kinematic data such as movement time (MT), peak velocity (PV), normalized jerk score (NJS), movement unit (MU), maximal grip aperture (MGA), normalized maximal grip aperture (NMGA) and so on were analyzed. The results of kinematic performance showed that children with HFA have larger MU (which represented for motor facileness) than controls in VS condition, but no differences between two groups in VL condition were found. In non-visual feedback condition, children with HFA also demonstrated significantly longer MT, larger NJS (which represented for motor smoothness) and more MU than controls, especially while the target was small.This study suggested that children with HFA have problems on motor smoothness and coordination in the process of the reach-to-grasp movement, especially in non-visual feedback and small target condition which suggested that children with HFA depend more visual cuing on performing prehension tasks and demonstrate poor coordination in high accuracy tasks. Intervention: Subjects were randomized into refrigerated and room temperature needle group prior to injection of DPT Outcome Measures: The effectiveness of refrigerated needles and room temperature needles were compared in terms of pain reduction using the Wong Baker Faces Pain Scale. The heart rate, blood pressure and oxygen saturation before, during and after the vaccination were the secondary outcome measures. Results: The increase in heart rate, increase in blood pressure and decrease in oxygen saturation from baseline, during and after vaccination among treatment groups were statistically insignificant (p-values 0.128, 0.209, and 1.00, respectively). The pain scores from the two groups showed marginally statistically significant result (p-value 0.055). Fifty-two percent of subjects in room temperature needle group have 0-1 pain score compared to forty-two percent with refrigerated needles. However, it is significant to note that with pain scores of 4-5, the room temperature needle group showed a higher percentage of 36.5% compared to 26.9% in refrigerated needles. Reported adverse events were redness for both needle groups. Blanching was noted with room temperature needle group. Conclusion: The use of refrigerated needles does not decrease the pain associated with DPT vaccination in 4-6 year-old children as compared with room temperature needles. Jikei University School of medicine, Department of Pediatrics, 2 Mie National Hospital, 3 Inje University Background: Use of aminophylline is an additional treatment for acute exacerbation of asthma is controversial .IV aminophylline is a "classical" bronchodilator but may have therapeutic advantages in its putative antiinflammatory effect and availability to peripheral airways where inhaled drugs may inadequately reach at. Objective: To assess if the addition of aminophylline infusion to standard treatment for acute exacerbation of asthma would enhance the recovery without serious adverse events. Methods: Children aged 2 to 15 years with acute asthma who did not respond to repeated inhalations of β2 agonists were enrolled. All subjects were treated with inhaled salbutamol and methylprednisolone/hydrocortisone infusion and were randomized to receive additional aminophylline infusion(Group A) or none (Group B).Asthma symptom score and wheezing on auscultation were compared. Results: Fifty subjects were enrolled with 26 randomly allocated to group A and 24 to group B. The groups were well matched at baseline. One in group A and 7 in B dropped out because of exacerbation or non-compliance and outcome was analyzed in 24 from A and 17 from B. Faster improvement in symptoms were seen in Group A and there was a significant difference in symptom score at 24 hours after treatment between the groups(p<0.05).The time to resolution of wheezing was significantly shorter in Group A than in B(p<0.01). Four minor adverse events were observed in Group A but all completed the treatment. Conclusion: Our results showed that an addition of aminophylline may be a beneficial therapeutic option for children with acute asthma.

The prognosis of acquired vocal cord paralysis after the open-heart surgery [Results] The cause of VCP was identified in 20 patients; open-heart surgery in 11 patients, extra-low-birth-weight at delivery in 4 patients, other disorders in 5 patients, and unidentified cause (idiopathic condition) in 3 patients. VCP after open-heart surgery subsided spontaneously in 8 of 11 patients (6 were fully, 2 were partially) by 6 months after the surgery. However in 3 patients, VCP did not subside by 14 to 53 months after the surgery; a case received tracheostomy at 5 months after surgery as the severity of symptoms.[Conclusions] Our observation suggests that we can predict the prognosis of VCP after open-heart surgery at 6 months after the surgery. Tracheostomy is not recommended for the mildly symptomatic VCP patients within 6 months, as most acquired VCA who had received open-heart surgery subsided spontaneously by 6 months after the surgery. However, the VCP that did not subside by 6 months after the surgery did not subside later.