Keijiro Yabuta

Serum levels of tumor necrosis factor, interleukin 2 receptor, and interferon-γ in Kawasaki disease involved coronary-artery lesions☆

We investigated 45 patients with Kawasaki disease (KD) and report the first simultaneous determination of tumor necrosis factor (TNF), interleukin 2 receptor (IL-2R) and interferon-gamma (IFN-gamma) in the serum during acute phase. Serum levels of TNF were measured by a sandwich enzyme-linked immunosorbent assay. Serum levels of soluble IL-2R and IFN-gamma were measured by a sandwich enzyme immunoassay and radioimmunoassay, respectively. Serum levels of TNF, IL-2R, and IFN-gamma were seen to increase during the acute phase of KD. In KD patients with coronary-artery lesions (CAL), the percentage of positive cases for TNF (greater than or equal to 10 U/ml), IL-2R (greater than or equal to 1056 U/ml), and IFN-gamma (greater than or equal to 0.3 U/ml) was higher than that in patients without CAL. Our results suggest that aggressive activation of immunocompetent cells develops in KD with CAL.

Peripheral blood monocyte/macrophages and serum tumor necrosis factor in Kawasaki disease

We analyzed the populations of peripheral blood monocyte/macrophages in 27 patients using a fluorescence-activated cell sorter, and investigated the possibility, in another 30 patients, that tumor necrosis factor (TNF) might be detectable in serum during the acute phase of Kawasaki disease (KD). Percentages of peripheral blood monocyte/macrophages among mononuclear cells and serum TNF levels were both seen to increase during the acute phase of the illness in patients with KD. The percentage of TNF positive cases in KD patients with coronary involvement was higher than that of patients without coronary involvement. These results suggest the possibility that immunological activation, accompanied by the secretion of TNF from monocyte/macrophages, is an important predisposing condition for the exacerbation of vascular damage in KD.

Four Novel KVLQT1 and Four Novel HERG Mutations in Familial Long-QT Syndrome

BACKGROUND Familial long-QT syndrome (LQTS) is characterized by prolonged ventricular repolarization. Clinical symptoms include recurrent syncopal attacks, and sudden death may occur due to ventricular tachyarrhythmias. Three genes responsible for this syndrome (KVLQT1, HERG, and SCN5A) have been identified so far. We investigated mutations of these genes in LQTS families. METHODS AND RESULTS Thirty-two Japanese families with LQTS were brought together for screening for mutations. Genomic DNA from each proband was examined by the polymerase chain reaction-single-strand conformation polymorphism technique followed by direct DNA sequencing. In four of the families, comprising 16 patients, mutations were identified in KVLQT1; five other families (9 patients) segregated mutant alleles of HERG. All 25 of these patients carried the specific mutations present in their respective families, and none of 80 normal individuals carried these alleles. Mutations were confirmed by endonuclease digestion or hybridization of mutant allele-specific oligonucleotides. No mutation in SCN5A was found in any family. CONCLUSIONS We identified nine different mutations among 32 families with LQTS. Eight of these were novel and account for 25% of all types of mutations reported to date. Such a variety of mutations makes it difficult to screen high-risk groups using simple methods such as endonuclease digestion or mutant allele-specific amplification.

Application of Percutaneous Transluminal Coronary Angioplasty to Coronary Arterial Stenosis in Kawasaki Disease

BACKGROUND Percutaneous transluminal coronary angioplasty (PTCA) has rarely been performed on patients with coronary lesions that result from Kawasaki disease. In this study, we retrospectively evaluated the effectiveness of PTCA in five patients with coronary arterial stenosis that resulted from Kawasaki disease and reviewed previous reports for possible indicators of PTCA effectiveness. METHODS AND RESULTS Five patients, ranging in age from 2 to 16 years (median 8 years) underwent conventional PTCA for localized stenosis. The lesion targeted for PTCA was located in the middle right coronary artery of three patients and in the left anterior descending artery in two patients. In four of the five patients, PTCA was angiographically effective, with stenosis rates improving from 84 +/- 10% to 33 +/- 11% (P<.05). When the previously reported cases of six similar patients were taken into consideration, the only predictor of successful PTCA seemed to be the time elapsed between the onset of Kawasaki disease and performance of this procedure. CONCLUSIONS In cases in which patients show significant localized stenosis as a result of Kawasaki disease, PTCA should be attempted within 6 to 8 years of the onset of the disease. Additionally, intravascular ultrasound imaging was found to be a useful tool for evaluating internal morphology before and after PTCA. In older patients with coronary calcification, other alternatives to PTCA, such as the use of a rotablator or an atherectomy catheter, should be considered.

Early detection of specific IgE antibody against house dust mite in children at risk of allergic disease

OBJECTIVES House dust mite (HDM) is a representative inhalant allergen that triggers allergic disease in childhood. The aim of this study is early detection of HDM-specific IgE antibody and prediction of the risk of a positive reaction to this antibody by in vitro parameters in infants with allergic manifestations. STUDY DESIGN Levels of HDM IgE in a range below the standard cutoff point of 0.35 U/ml, serum concentrations of IgE, and specific IgE antibodies against egg white, cow milk, and soybeans were determined in 108 infants with allergic manifestations at 6 months of age, and these infants were monitored for conversion of HDM IgE to positive levels greater than 0.35 U/ml up to 5 years of age. The presence of active allergic disease at 5 years of age in relation to HDM-specific IgE was also examined. RESULTS We were able to determine reliably the HDM IgE values between 0.23 and 0.35 U/ml, using a fluorescent enzyme immunoassay that measured the intensity of fluorescence. The HDM IgE levels increased, resulting in positive values, in 54 of 108 subjects during the first 5 years of life. In multiple regression analysis, an HDM IgE value between 0.23 and 0.35 U/ml, a high serum IgE level, and a positive reaction to specific IgE antibody against egg white in infants at 6 months of age proved to be significant predictors of the future positive reaction to HDM IgE (p = 0.0006, 0.0043, and 0.0001). In particular, the sensitivity and specificity of specific IgE antibody against egg white for the conversion of HDM IgE to positive values were the best among these indicators. Moreover, active allergic diseases were observed significantly more often in children with positive HDM IgE values than in children with negative HDM IgE values at 5 years of age (p < 0.001 for each). CONCLUSIONS A determination of these predictors in infants at 6 months of age can be used for early detection of HDM IgE and would be valuable in a screening test for later allergic disease among infants with allergic manifestations.

Serum levels of p60 soluble tumor necrosis factor receptor during acute Kawasaki disease

To evaluate the role of tumor necrosis factor alpha (TNF-alpha) during acute Kawasaki disease, we measured p60 soluble tumor necrosis factor receptor (sT-NF-R) shedding into the circulation in 48 patients with acute Kawasaki disease, all of whom received intravenous infusions of gamma-globulin. Of the 48 patients, 5 had coronary artery lesions. Serum concentrations of p60 sTNF-R and TNF-alpha were measured by a sandwich enzyme immunoassay. Patients with Kawasaki disease had increased serum levels of p60 sTNF-R. We found a positive correlation between serum levels of p60 sTNF-R and levels of TNF alpha during acute Kawasaki disease. Moreover, patients with coronary artery lesions had higher levels of sTNF-R than did those without coronary artery lesions. Our findings indicate that p60 sTNF-R levels in serum may be useful for determining the severity of vascular damage during acute Kawasaki disease, and that patients with Kawasaki disease and high sTNF-R levels seem to be susceptible to coronary artery lesions even if they receive therapy with intravenous infusions of gamma-globulin.

Effect of testosterone on bone density and bone metabolism in adolescent male hypogonadism

To assess the influence of gonadal steroid testosterone (T) on bone mineral status in males during puberty, we observed the response of cortical bone density and serum biochemical parameters of bone metabolism to T treatment in 12 adolescent patients with hypogonadotropic hypogonadism (11 with both gonadotropin and growth hormone deficiency and one with isolated gonadotropin deficiency). The 12 patients aged 15 to 21 years (Tanner stage I to II) were divided into two groups: group 1 (n = 6) given T treatment for 2 consecutive years, and group 2 (n = 6) without T treatment for the first year and then with T treatment for the second year. Cortical bone density measured in the radius was less than the age-matched mean value for normal subjects in all 12 patients (groups 1 and 2) at the start of the study. Bone density in group 1 increased significantly during the 2-year T treatment period, but did not increase in group 2 during the first year without T treatment, although an increase was observed during the subsequent year with T treatment. Among circulating biochemical factors such as osteocalcin, parathyroid hormone (PTH), 25-hydroxyvitamin D (25-OHD), and 1,25-dihydroxyvitamin D [1,25-(OH)2D], only osteocalcin showed an increase in response to T treatment in both groups. Levels of insulin-like growth factor-I (IGF-I) remained consistently low and did not change in any patients except one with isolated gonadotropin deficiency.(ABSTRACT TRUNCATED AT 250 WORDS)

Problems of mass screening for neuroblastoma: Analysis of false-negative cases

The Japanese mass screening (MS) system for neuroblastoma at 6 months of age has resulted in the earlier diagnosis of the tumor with excellent therapeutic results. However, some problems are involved in the present MS system. We present six false-negative cases, ages ranging from 1 year 11 months to 3 years 11 months. Neuroblastoma cell taken from four of these patients were studied biologically. These patients had advanced disease (one was stage III; five were stage IV). Three of the patients have died and one is terminally ill despite undergoing surgery combined with intensive chemotherapy. Cytogenetic analysis performed in three cases showed that all the cases had diploid chromosome mode associated with 1P-, double minutes (DMs), or marker chromosomes. N-myc oncogene analysis, performed in four cases, showed amplification in two; one patient had diploid chromosomes, but the other was not examined cytogenetically. These findings were strikingly different biologically from those of cases found by MS. The majority of neuroblastomas detected by MS were found to be triploid tumors without N-myc amplification. These findings suggest that the main reason for the false-negative results in the patients we examined is that they were tumor-free or the tumors were so small in size that they were unable to produce urinary vanillylmandelic acid and or homovanillic acid levels high enough to be detected at the time of MS. Therefore, we conclude that MS at 6 months of age is too early to detect neuroblastoma with a diploid chromosome mode and/or amplified N-myc oncogene. We propose that MS at the age of 1 year 6 months would be more effective to pick up these cases, because treatment strategies depend on the different biological characteristics of tumor cells.

Apoptosis as a Possible Cause of Wall Thinning in End-Stage Hypertrophic Cardiomyopathy

A 15-year-old boy with hypertrophic cardiomyopathy died of congestive heart failure with progressive left ventricular wall thinning with poor systolic function. Microscopic examination revealed patchy fibrosis in the ventricular myocardium with wall thinning, and immunohistochemical evaluation of apoptosis showed apoptotic cells and bodies in the destroyed myocytes along the border between the fibrotic area and myofibril.

Immunohistochemical Studies on Small Intestinal Mucosa in Kawasaki Disease

ABSTRACT: To investigate the etiology of Kawasaki disease (KD), the cell surface phenotypes of mononuclear cells and enterocytes in the jejunal mucosa of KD were investigated in a case-control study. Sixteen Japanese patients with KD were enrolled in the study. As disease controls, jejunal tissues from 10 patients with diarrhea due to cows milk protein intolerance were used. The numbers of cells stained by an immunofluorescent technique were counted and analyzed statistically by t test. Both HLA-DR+CD3+ and DR+CD4+ cells were significantly increased in the lamina propria of KD patients in the acute phase compared with numbers in controls and patients with cows milk protein intolerance (p ≤ 0.01). CD8+ cells were significantly reduced in both the epithelium and the lamina propria of KD patients in the acute phase in comparison with numbers in both controls (p ≤ 0.05) and patients with cows milk protein intolerance (p ≤ 0.01). HLA-DR+ cells were significantly increased in both the enterocytes and the lamina propria of KD patients in the acute phase compared with numbers in controls (p ≤ 0.01). These cell patterns returned to normal in the convalescent phase of KD. Differences between these cell patterns in KD patients with and without diarrhea were not significant, although these immunohistochemical features tended to be more marked in patients with diarrhea than in those who lacked diarrhea. These results are consistent with what one would expect to find if a delayed-type hypersensitivity reaction had occurred in the small intestinal mucosa of KD patients. Although other explanations cannot be ruled out, it is conceivable that causative antigens may have invaded the body by breaching the barrier of the intestinal mucosa.