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Background There have been few reports on Helicobacter pylori (H. pylori) infection in asymptomatic Japanese children and adolescents. We hypothesized that the prevalence of H. pylori infection is very low among Japanese children and that clinical variables such as serum pepsinogen and iron levels are associated with H. pylori infection. Materials and Methods We conducted a cross-sectional analysis of a sample of 454 junior high school students aged 12-15 years in four areas in Nagano Prefecture. A commercial ELISA kit (E-plate Eiken H. pylori antibody) was used to measure IgG antibody against H. pylori. Serum pepsinogen and iron levels were also measured using standard methods. A urea breath test was performed for seropositive students. Results The overall prevalence of H. pylori was 3.1% (14/454). There were no significant differences in H. pylori prevalence among mountain, rural, and urban areas. The mean level of both serum pepsinogen (PG I) and PG II was significantly increased in the seropositive subjects compared with the seronegative subjects. When the cutoff values for adults (PG I: 70 ng/mL and PG I/II ratio: 3) were used, 4 of 14 subjects had PG I ≤70 ng/mL and PG I/II ratio ≤3. The results of a logistic regression analysis showed that low serum iron levels were significantly associated with H. pylori infection (P=.02). Conclusions The prevalence of H. pylori infection is as low as 3% among junior high school students aged 12-15 years in Japan. The disappearance of H. pylori is accelerating in Japanese children.

Background: The present study was designed to examine whether serum uric acid (SUA) levels were associated with cardiometabolic risk factors and to determine optimal cut-offs for SUA to identify multiple risk factors among Japanese junior high school students. Methods and Results: A total of 958 students (518 boys and 440 girls, aged 12.1-15.0 years) who were enrolled between April 2005 and June 2008 were divided into 4 groups according to SUA quartiles. Compared with the lowest quartile of SUA, prevalence of abdominal obesity, hypertension, and dyslipidemia was significantly increased in the highest quartile in boys and that of abdominal obesity was increased in the highest quartile in girls. The adjusted odds ratios (95% confidence interval) of the highest quartile of SUA for 2 or more cardiometabolic risk factors were 2.59 (1.16-5.79) for boys and 1.54 (0.43-5.56) for girls. Receiver operating characteristic curve analysis demonstrated that the most appropriate cut-offs for SUA to identify multiple cardiometabolic risk factors were 6.4 mg/dl for boys and 4.9 mg/dl for girls. Conclusions: SUA was strongly associated with the prevalence of cardiometabolic risk factors among male Japanese junior high school students. The present study may provide insights into the role of SUA in the school screening system for the development of educational programs on prevention of lifestyle-related diseases among school children. (Circ J 2010; 74: 1570 - 1577)

Serum cystatin C (CysC), a novel marker of renal function, is associated with the components of metabolic syndrome in adults. Little is known about the utility of CysC and its association with cardiometabolic risks in young subjects.In a cohort of 454 Japanese junior high school students, the distribution of serum CysC levels and associated variables were analyzed. CysC levels were significantly higher in boys than in girls (0.92±0.10mg/L vs. 0.77±0.08mg/L, p0.001). CysC was significantly correlated with serum creatinine (r=0.473, p0.001), and serum uric acid (SUA) (r=0.546, p0.001). Multivariable regression analysis revealed significant associations between CysC and SUA in all subjects (β=0.241, p0.001), and in boys and girls separately (β=0.264 and 0.240, respectively, both p0.001). Importantly, subjects with elevation of both serum CysC and SUA levels had the highest ratio of triglyceride to high-density lipoprotein cholesterol.CysC had significant associations with both creatinine and SUA in Japanese junior high school students. The concomitant elevation of serum CysC and SUA levels was associated with subclinical lipid metabolism dysregulation, and suggested the presence of cardiometabolic risk accumulation.

A variation of the fatty acid composition is closely associated with the clinical state of inflammatory disorder and metabolic syndrome. The analysis of serum fatty acid composition of neonates and infants has been measured hardly at the laboratory, because a large quantity of serum was required for an analysis and the measurement procedure was cumbersome. We examined the rapid and easy analysis in a small amount of serum using the combination methods of the gas chromatography mass spectrometry (GC MS) and the quick transmethylation. The serum fatty acid composition of neonates and infants were compared with the young people. The serum fatty acids with the internal standard material were performed transmethylation using the microwave, and then the lipids were extracted. The fatty acid esters were analyzed by GC MS with capillary column, and the statistical procedure used nonparametric method. The repeatability of each fatty acid concentration was CV = 5-11.3% (n = 5) with serum 50 μl. The lowest quantity of sample was possible to measure with 13 μl of serum. The total serum fatty acid, saturated fatty acid and monounsaturated fatty acid levels did not show a significant difference at all age, but the polyunsaturated fatty acid (PUFA) level of neonates and infants was significantly lower than young people, p = 0.007. The four main PUFA exclusive of α-linolenic acid showed a significant difference. The fatty acid composition with small quantities serum was measured by the rapid and accurate method using the GC MS and the microwave. The serum PUFA concentrations have fluctuated according to growth, therefore it was necessary to evaluate serum fatty acid composition in each age category.

Background Sphingomyelin (SM) is a key component of extracellular membranes and lipoproteins, and plays roles in cell signaling and as a component of lipoproteins. SM species differ in terms of fatty acid (FA) composition. However, no simple, rapid, quantitative assay for identifying different SM species has yet been reported. In this study, lipid hydrolase treatment and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) were used to identify serum SM species. Methods Sera were collected from healthy young individuals. To identify SM species, sera were treated with phospholipase A2 and lipoprotein lipase, and lipids were extracted using the standard chloroform/methanol (2/1 v/v) method. Results We detected 15 peaks from serum using MALDI-TOF MS, which were assigned to SM species bound with FA components ranging from C15:0 to C24:2. The most prominent serum SM species was SM [C16:0], which accounted for approximately 26% of serum SM. Some SM species contained an odd-carbon FA (C15, C21, and C23), and these accounted for approximately 4% of serum SM. The reproducibility of major SM species within and between application positions on MS-sample plate was CV = 3.0%–7.9% and CV = 3.1%–6.8%, respectively. The concentration and dilution ratio were linearly related. The SM species composition of 10 healthy young subjects showed a similar profile. Conclusions We developed a rapid, and quantitative method for identifying serum SM species using lipid hydrolase treatment and MALDI-TOF MS. This method will be suitable for clinical laboratory studies to examine the associations between SM species and disease states.

We investigated the exact prevalence of dementia of Alzheimer type (DAT) and apolipoprotein E (ApoE) phenotypes in 106 Japanese Down's syndrome (DS) patients. Among these patients 16 were diagnosed as having DAT. The prevalence of DAT was 0% in the 30- to 39-year-old group, 16% in the 40- to 49-year-old group, and 38% in those over 50 years old. The frequency of the epsilon4 allele in DS patients with DAT was 18.8%, which was considerably higher than that of nondemented DS patients (4.5%) and Japanese nondemented controls (6.7%). Especially, the frequency of the epsilon4 allele in DS patients who developed DAT under 50 years was significantly higher (28.6%). DS patients certainly develop DAT at earlier ages but the prevalence of DAT in each group of patients was lower than previously recognized. It is very likely that the ApoE epsilon4 is a risk factor for DAT even in DS patients with a genetic predisposition to Alzheimer's disease.

Background: An increase in plasma low-density lipoprotein (LDL) is a well-known risk factor in the development of atherosclerosis. Dairy consumption may lower the risk of atherosclerosis; however, studies on the effects of milk on cardiovascular risk factors are still scarce. We were interested in investigating whether the intake of milk improves the atherogenic lipoprotein profile. Aims: We investigated the effects of consuming whole or nonfat milk on plasma lipoprotein composition in healthy Japanese subjects as a pilot study. Methods: Normolipidemic subjects consumed 500 ml of whole milk (whole milk group; n = 7) or nonfat milk (nonfat milk group; n = 7) every day for 2 weeks. Results: The consumption of nonfat milk resulted in a lowering of plasma triglyceride (TG) and phospholipid levels and TG level in high-density lipoprotein (HDL) and increased the plasma apolipoprotein (apo) C-III level. In addition, the TG/cholesterol ratios in HDL and LDL were significantly decreased, and LDL particles became larger. In contrast, the only changes observed following whole milk consumption were increases in the plasma levels of apoC-III and apoE. Conclusions: These findings suggest that consumption of nonfat milk, but not whole milk, may result in a less atherogenic lipoprotein profile, and that the constituents of nonfat milk may improve lipid metabolism.

The treatment effects of rosuvastatin on arterial stiffness were assessed and compared to those of fluvastatin in high-risk Japanese patients with dyslipidemia in a primary prevention group.Patients were randomly assigned to either 2.5-5 mg/day of rosuvastatin (Group A) or 20-40 mg/day of fluvastatin (Group B) and followed up for 12 months. In Group A (n=38), there was a progressive reduction in brachial-ankle pulse wave velocity (baPWV) along with a decrease in the low-density lipoprotein cholesterol/high-density lipoprotein cholesterol (L/H) ratio and high-sensitivity C-reactive protein (hsCRP), and the change in baPWV correlated significantly with that of the L/H ratio and that of hsCRP after rosuvastatin treatment. In Group B (n=37), although fluvastatin achieved a significant improvement in baPWV, L/H ratio, and hsCRP, baPWV was significantly greater than that in Group A and showed a significant correlation with that of hsCRP alone after fluvastatin treatment. In a subgroup of patients (n=26), switching from fluvastatin to rosuvastatin further improved baPWV and the L/H ratio without altering hsCRP after 12 months.Low-dose rosuvastatin would be more effective than fluvastatin in improving arterial stiffness in high-risk Japanese patients with dyslipidemia. The results suggest that improvement in arterial stiffness by rosuvastatin mainly depends on its strong lipid-lowering effects, whereas that by fluvastatin is strongly dependent on the pleiotropic effects, especially an anti-inflammatory action.

There is accumulating evidence that soluble amyloid-beta (Abeta) oligomers, rather than amyloid fibrils, are the principal pathogenic species in Alzheimer disease (AD). Here, we have developed a novel enzyme-linked immunosorbent assay (ELISA) specific for high-molecular-weight (HMW) Abeta oligomers. Analysis of Abeta oligomers derived from synthetic Abeta 1-42, by size-exclusion chromatography (SEC), revealed that our ELISA specifically detected HMW Abeta oligomers of 40-200 kDa. Using this ELISA, we detected significantly higher (P

The present study was designed to test the hypothesis that fluvastatin might improve arterial stiffness, as assessed with pulse wave velocity (PWV), in patients with coronary artery disease (CAD) and hyperlipidemia over the long term.Ninety-three patients were randomly assigned to either fluvastatin (group A, n=50) or bezafibrate (group B, n=43) and followed for 5 years. There was no difference in the clinical findings between the 2 groups. In group A, there was a progressive reduction in the brachial-ankle PWV along with a decrease in serum low-density lipoprotein-cholesterol (LDL-C) and C-reactive protein (CRP) by 12 months after fluvastatin, and the improvement was maintained until 5 years after treatment. In group B, despite identical lowering of the serum lipid, PWV was progressively increased. In group A, the percentage change in PWV correlated significantly with that of the serum CRP (r=0.49, p0.001), but not with that of the serum LDL-C after treatment.The beneficial vascular effects of fluvastatin persisted for a long period in patients with CAD and hyperlipidemia. Its anti-inflammatory action might contribute to the favorable effects on arterial stiffness.

Two cases with abnormal laboratory data due to interaction between immunoglobulins and other serum proteins are described. Case 1 was a patient with lactate dehydrogenase (LD) -IgG3 complex whose serum LD value was moderately elevated. Case 2 were nondiabetic patients with IgA type M-proteinemia who had significantly increased serum fructosamine (FRA). The IgG3 in Case 1 was found to be conjugated to LD by immunoprecipitation assay. The LD-IgG3 complex was easily dissociated by affinity chromatography on 5'-AMP or Cibacron Blue F3G-A. The relative molecular weights of the patient's gamma3 chains and light chains were 67,000 and 28,000, respectively, by Western blotting, which corresponded to the expected values. However, the patient's IgG3 did not react to the anti-kappa and anti-lambda light chain antibodies in Immunofixation electrophoresis. Serum FRA concentrations were higher in patients (Case 2) with IgA type M-proteinemia or polyclonal hyper-IgA than those with the IgG type or IgM type. The sera from the patients with IgG or IgM type M-proteinemia had FRA only at the position of albumin, but 11 of 13 sera with IgA type M-proteinemia stained for glycoprotein at the position of the M-protein band as well as the albumin band. The abnormal precipitin arcs of IgA-albumin complex were observed in 11 of 13 sera from patients with IgA type M-proteinemia that were glycosylated at the position of M-protein band.

Functional heterogeneity of pancreatic islets was systematically analyzed for the first time using freshly isolated single rat pancreatic islets. First, 60 islets were sequentially exposed to 3, 9.4, 15.6, and 24.1 mM glucose for 30 min each in incubation experiments: 36 (60%) responded in a concentration-dependent and 19 (32%) in an all-or-none manner, and 5 (8%) islets did not respond to high glucose. As a group, the larger the islet, the higher the beta cell glucose sensitivity. However, glucose-stimulated elevation of [Ca2+]i in the beta cell. insulin/glucagon ratio in the islet, and expression of glucose transporter 2, glucokinase, and pancreatic duodenal homeobox factor-1 in the beta cell were not significantly related to islet size. Second, 50 islets were stimulated with 16.7 mM glucose in perifusion. A biphasic insulin release was found in 39 (78%), and no or little first phase response in 11 (22%) islets, irrespective of the islet size. Nevertheless, when the response was plotted as a group, it was clearly biphasic. Islet size, insulin content and the amount of insulin release were positively correlated with each other. In conclusion, there are size-related and size-unrelated functional diversity among pancreatic islets. The reason for such heterogeneity remained to be determined.

BACKGROUND AND AIMS—Helicobacter pylori locate not only on the apical surface of surface mucous cells but also in the mucous gel layer covering the gastric mucosa. The present study was undertaken to observe the mucous gel layer itself and any H pylori in this layer at the electron microscopic level, and to determine whether H pylori proliferate in this layer. METHODS—We examined resected human stomachs (five cases, fixed in Carnoy's solution, paraffin embedded) under the light microscope, and gastric biopsy specimens (10 cases, fixed in glutaraldehyde with or without osmium, epoxy embedded) under the electron microscope. We performed histochemical staining for gastric mucins and immunostaining for H pylori, gastric gland mucous type mucins, and intestinal mucins. RESULTS—Under the electron microscope, surface mucous cell type mucins and gland mucous cell type mucins in the mucous gel layer covering gastric mucosa without intestinal metaplasia showed reticular and band like structures, respectively. H pylori were frequently found as small aggregates within the mucous gel layer of surface mucous cell type mucins, and H pylori within these aggregates were seen dividing. H pylori were frequently found in the mucous gel layer of the surface mucous cell type mucins along the border with the layer of gland mucous cell mucins. Occasionally, H pylori were trapped by frayed thin threads of the gland mucous cell type mucins. CONCLUSIONS—The two types of gastric mucins in the mucous gel layer differ in ultrastructure. H pylori preferentially colonise and form microcolonies within the mucous gel layer of surface mucous cell type mucins. Mucins from gland mucous cells may disturb the movement of H pylori within the mucous gel layer. Keywords: gastric mucin; Helicobacter pylori; immunohistochemistry; surface mucous gel layer

The specificity of monoclonal antibodies against gastric mucins (designated as HIK1083, PGM 36, and PGM 37) was studied immunohistochemically in normal, metaplastic, and neoplastic human tissues. These antibodies labeled class III mucin-producing cells identified by paradoxical concanavalin A staining in normal stomach, duodenum (Brunner gland), biliary tract, and main pancreatic duct; in mucinous metaplasia of pancreas and gallbladder; and in adenocarcinomas of stomach (90%), bile duct (80%), gallbladder (100%), pancreas (80%), lung (100% of goblet cell type adenocarcinomas), ovary (67% of mucinous carcinomas), and uterine cervix (100% of adenoma malignum tumors). Normal and neoplastic cells of esophagus, colon, salivary gland, kidney, endometrium, breast, prostate, and liver, as well as normal small intestine, lung, and uterine cervix, were all negative. The antibodies used should be valuable for the detection of class III mucin and class III mucin-producing cells in normal, metaplastic, and neoplastic tissues.

To analyze both hemoglobin (Hb) and globin chain variants, we modified a commonly used method, capillary isoelectric focusing (CIEF), with detection at 280 nm. The samples were hemolysates prepared from red blood cells, and globin chains obtained from the hemolysates by treatment with cold acidified acetone. When the migration time for the internal reference, carbonic anhydrase I (isoelectric point, pI 6.60), was taken as 1.0, the migration ratio for Hb A0 in normal human blood was 0.877 +/- 0.004 (mean +/- SD, n = 9), and those of the alpha- and beta-globin chains were 0.673 +/- 0.004 and 0.847 +/- 0.005 (mean +/- SD, n = 4), respectively. The ratio of peak heights between the beta- and alpha-globin chains (beta/alpha) in the normal Hbs obtained from four subjects was almost constant at 2.5 +/- 0.1 (mean +/- SD). This ratio indicates which of the globin chains includes a mutation (if one exists). When an Hb variant, Hb Hoshida (in which Gln is substituted for Glu at residue 43 in the beta-globin chain), was analyzed by this method, two main peaks were observed (migration ratios 0.836 and 0.877, corresponding to an abnormal and the normal Hb, respectively). An additional peak with an abnormal migration ratio of 0.788 was also detected in the globin chain profiles. The ratio of peak heights between normal beta- and alpha-globin chains was 1.57, indicating that a mutation exists in the beta-globin chain. We thus established a convenient system using CIEF that provides a rapid and reproducible method for the random analysis of both Hb and globin chain variants.

This is the first systematic study on the pancreatic beta cell function in the heterozygous mouse with targeted disruption of the beta cell glucokinase gene. The heterozygotes' beta cell displayed the following characteristics: (1) impaired glucose sensitivity with normal glucose responsiveness, (2) poor discrimination of alpha and beta glucose anomers, and (3) normal response to glucose in the presence of 25 mM K+ and 150 microM diazoxide. Both the first and the second phases of glucose-stimulated insulin release were depressed. Although the heterozygotes were mildly hyperglycemic, insulin treatment further suppressed beta cell function, implying the beta cell glucose toxicity is not the cause of impaired glucose sensitivity. The data are compatible with the glucokinase glucose sensor concept inasmuch as glucose sensitivity is reduced in the heterozygotes' beta cell. The anomeric malaise and preservation of the ATP-sensitive K+ channel-independent glucose action were considered due to chronic hyperglycemia.