Serum levels of vault RNA significantly varied in patients with haematological malignancies.

Among extracellular non­coding RNAs, serum levels of microRNAs have been extensively investigated in cancers. In contrast, the serum levels of vault RNAs (vtRNAs) in relation to various disease conditions remain poorly understood. The present study evaluated the clinical significance of serum vtRNA1­1 levels in patients with blood diseases. The stability and sub­localisation of serum vtRNA1­1 was assessed and a reverse transcription­quantitative PCR method using spiked RNA to quantify serum vtRNA1­1 was developed. Serum vtRNA1­1 levels were assessed in 102 individuals with blood diseases. Serum vtRNA1­1 was demonstrated to be stable for three weeks at 4˚C and was not confined to the exosome fractions. Spiking RNA was used to correct for the inconsistency in RNA extraction. The serum vtRNA1­1 levels ranged between 7.28 and 8.76 log10 cps/ml (median 8.05) in control individuals (n=46). Serum vtRNA1­1 levels correlated with leukocyte counts and increased to a maximum of 10.01 log10 cps/ml in patients with bulky leukaemia and lymphoma and decreased to 6.52 log10 cps/ml during intensive chemotherapy. The serum vtRNA1­1 levels varied significantly in patients with haematological malignancies. Serum vtRNA1­1 may originate from haematological cells and are a potential biomarker of normal and malignant haematological activities.

Differences in the Composition of Activated Partial Thromboplastin Time (APTT) Reagents Affect Clot Waveform Analysis.

Background: Clot waveform analysis (CWA) based on activated partial thromboplastin time (APTT) is a useful assay for hemostasis. However, the effects of activators and phospholipid conditions on CWA have not been adequately investigated. Therefore, we characterized CWA using four different APTT reagents. Methods: We used 39 archived plasma samples from patients with hemophilia A (HA), 16 samples from patients with HA under emicizumab treatment, and 10 samples from healthy individuals for CWA with four different types of APTT reagents (reagents A, B, C, and D). We then compared Ad|min1|, Ad|min2|, and Ad|max2| from the CWA, which reflect the maximum velocity, maximum acceleration, and maximum deceleration, respectively, among the four reagents. Results: Similar clot waveform shapes were observed for each reagent in the healthy donor group, HA group, and HA under emicizumab group, and the waveform was different for each target group. Significant changes were found in clotting time (CT) (s), Ad|min1| (%/s), Ad|min2| (%/s2), and Ad|max2| (%/s2). The waveform pattern for the coagulation reaction by reagent D, comprising silica and synthetic phospholipids, was the fastest among the reagents examined. Further, the difference in Ad|min1| (%/s) and Ad|min2| (%/s2) was larger than that in CT depending on the reagent used(s), indicating that the measured value of CWA was affected by the reagent composition. Conclusion: Our results showed a significant difference among reagents with varying composition and concentration; this was found to affect the parameters obtained from CWA. Thus, the differences between reagents hinder standardization of quantitative evaluation using these parameters; further, this highlights the necessity of understanding the characteristics of APTT reagents and determining the reference range in individual facilities.

Free advanced glycation end product distribution in blood components and the effect of genetic polymorphisms.

One hypothesis regarding the cause of diabetic complications is that advanced glycation end products (AGEs) bind to the AGE receptor and induce changes in gene expression. However, what AGEs exist in vivo and how individual AGEs are produced and impact body metabolic process to cause diabetes complications are not understood. We developed a new precise method to measure AGEs using LC-MS/MS with a new column and measured 7 free AGEs, including N(6)-carboxymethyllysine (CML), N(6)-(1-carboxyethyl)-l-lysine (CEL) and N5-(5-hydro-5-methyl-4-imidazolon-2-yl)L-ornithine (MG-H1), in human blood components. Blood was obtained from 9 people, and free AGEs were measured in individual blood components with LC-MS/MS before and after a meal. Free CML and CEL were abundant in erythrocytes, with 92% of free CML and 85% of free CEL localized in erythrocytes. In contrast, 60% of free MG-H1 was distributed in the serum. After the meal, free serum MG-H1 increased, but CML and CEL did not. CML and CEL are mainly distributed in erythrocytes and were not affected by the meal, indicating that they are produced in vivo. However, the main source of MG-H1 is the meal. The effect of genetic polymorphisms on AGEs was also investigated. Low activity type aldehyde dehydrogenase 2 (ALDH2) increased the CML concentration in the blood. This is the first observation that shows that the metabolic process of CML and CEL is different from that of MG-H1 and the effect of ALDH2 SNPs on CML.

A Randomized Placebo-controlled Trial of an Oral Preparation of High Molecular Weight Fucoidan in Patients with Type 2 Diabetes with Evaluation of Taste Sensitivity.

BACKGROUND: Fucoidan is derived from seaweed widely used in Japanese cuisine, but little is known about its influence on glucose metabolism. To obtain information about the physiological effects of fucoidan on glucose metabolism, the digestive system, and the gustatory system controlling taste sensation in patients with type 2 diabetes, we conducted a randomized, double-blind, placebo-controlled study. METHODS: Thirty patients with type 2 diabetes on diet therapy were recruited from an outpatient clinic (22 men and 8 women aged 59.10 ± 13.24 years, body mass index: 25.18 ± 3.88, hemoglobin A1c: 7.04 ± 1.24%). They were divided into 2 groups and underwent 2 interventions with a 4-week interval. One group received fucoidan for 12 weeks (a daily 60 mL test beverage containing 1,620 mg of fucoidan) and then placebo (60 mL) for the subsequent 12-week period, while the order was reversed in the other group. Evaluation was performed just before and after each intervention. Taste sensitivity was measured for 5 basic tastes by the filter paper disk method and food intake was evaluated with a validated diet questionnaire. RESULTS: No adverse events occurred during the study period. Despite no change of the diet, stool frequency increased during fucoidan intake (from 7.78 ± 4.64/week in Week 1 to 9.15 ± 5.03/week in Week 5, P < 0.001), and it increased more in lean subjects. In 11 subjects whose stool frequency exceeded the mean value, the thresholds for sweet, salty, bitter and umami tastes were significantly reduced (enhancement of sensitivity) after fucoidan intake. In 14 subjects with normal HOMA-IR (homeostatic model assessment of insulin resistance, < 2.5), hemoglobin A1c decreased after fucoidan intake (from 6.73 ± 1.00 to 6.59 ± 1.00%, P < 0.05), as did the fasting plasma level of GLP-1 (glucagon-like peptide-1, from 6.42 ± 3.52 to 4.93 ± 1.88 pmol/L, P < 0.05). CONCLUSION: Sustained fucoidan intake led to alterations of gastrointestinal function, including increased stool frequency and enhanced taste sensitivity, which could contribute to better control of diabetes.

Evaluation of antigen-positive toxin-negative enzyme immunoassay results for the diagnosis of toxigenic Clostridium difficile infection.

Clostridium difficile (C. difficile)-associated diarrhea (CDAD) is a challenging nosocomial infectious disease. C. DIFF Quik Chek Complete assay is widely used to detect glutamate dehydrogenase (GDH) antigen and toxin A/B of C. difficile simultaneously. However, the interpretation of GDH positive/toxin negative results is problematic. We performed a retrospective study of patients with GDH positive/toxin negative results to determine the probability of detecting toxigenic C. difficile and its risk factors. Between April 2012 and March 2017, we investigated cultures of fecal specimens followed by toxin detection tests. The clinical histories of patients with and without toxigenic C. difficile were compared using univariate- and multivariate-analyses. In total, 2675 patients were examined using C. Diff Quik Chek Complete assay. Among 356 GDH positive/toxin negative patients, cultures were performed in 220 cases and toxigenic C. difficile was recovered from 139 (63.2%) specimens. Patients with toxigenic C. difficile had significantly lower body mass index than those without. Over half the GDH positive/toxin negative patients were infected with toxigenic C. difficile. Lower BMI was a CDAD risk factor in this patient population. These data can be utilized to initiate isolation and clinical interventions before confirmatory test results are available. J. Med. Invest. 65:131-135, February, 2018.

Association between serum soluble low-density lipoprotein receptor levels and metabolic factors in healthy Japanese individuals.

BACKGROUND: Soluble low-density lipoprotein receptor (sLDL-R) is formed by cleavage of the extracellular domain of low-density lipoprotein receptor (LDL-R). It is unclear whether serum sLDL-R is a marker of diseases associated with triglyceride (TG) metabolism. We investigated the association between serum sLDL-R concentrations and other biochemical parameters in healthy Japanese individuals. METHODS: Study subjects consisted of 102 healthy adult Japanese volunteers (42 men, 60 women) with body mass index (BMI) < 30 kg/m(2) and serum TGs, LDL cholesterol (LDL-C), aspartate aminotransferase, alanine aminotransferase, γ-glutamyl transpeptidase, and glucose concentrations within normal ranges. Serum sLDL-R concentrations were determined by enzyme-linked immunosorbent assay and their correlations with biochemical parameters were analyzed. RESULTS: Mean serum sLDL-R concentration was 120.9 ± 39.9 ng/ml. Serum sLDL-R levels were significantly and positively correlated with BMI (rs = 0.252) and TG (rs = 0.408) and LDL-C (rs = 0.325) concentrations. Multiple regression analysis adjusted for age, gender, and smoking showed that BMI (ß = 0.274), TG (ß = 0.328), and LDL-C (ß = 0.224) were factors independently correlated with sLDL-R levels. CONCLUSION: Serum sLDL-R concentration may be a marker of diseases associated with TG metabolism. This is the first report to date describing the clinical relevance of sLDL-R.

Factor XII Osaka: abnormal factor XII with partially defective prekallikrein cleavage activity.

A healthy Japanese male had reduced factor XII (FXII) activity (35%) in contrast to normal antigen levels (81%). The F12 of this proband had a 9775G to C mutation in exon 10 and an 11276G to A mutation in exon 13 that resulted in two amino acid substitutions of Ala324Pro (GCG→CCG) in the proline-rich connecting region and Gly531Glu (GGG→GAG) near the active Ser544 in the catalytic domain. His father had the nucleotide 46T/T and a heterozygous 9775G/C mutation. The FXII activity (32%) and antigen level (38%) of the father were about half that of normal individuals with 46T/T, suggesting a heterozygous cross reacting material (CRM)-negative deficiency. His mother had a 46C/T and heterozygous 11276G/A mutation, and 80% FXII activity was incompatible with the corresponding antigen level (125%), suggesting a heterozygous CRM-positive deficiency. The substitution of Ala324Pro probably caused the CRM-negative mutation and the Gly531Glu caused the CRM-positive mutation. We developed three methods based on chromogenic substrates to assay the distinct functions of FXII, namely its autoactivation on a negatively charged surface, activation by kallikrein cleavage and the prekallikrein cleavage activity of FXIIa. The ratios of autoactivated FXIIa/FXII antigen (0.80-1.10) and of kallikrein-induced FXIIa/FXII antigen (0.86-1.00) in plasma from the proband were within normal ranges, whereas those of FXIIa-induced kallikrein/FXII antigen were reduced to 0.41-0.45. In conclusion, the 9775G to C and 11276G to A mutations of F12 led to a CRM-negative and -positive FXII deficiency, and the F12 with 11276A produced a dysfunctional type of FXII with a partial defect (0.41-0.45) in prekallikrein cleavage activity.

Relationship between lipoprotein(a), metabolic syndrome, and carotid atherosclerosis in older Japanese people.

BACKGROUND: Lipoprotein(a) [Lp(a)] or metabolic syndrome (MS) is individually considered an atherosclerotic factor. Serum Lp(a) may reportedly show the additive effects on atherosclerosis under certain particular pathologies. We do not know the association between serum Lp(a) and carotid artery intima-media thickness (CIMT) with relation to MS in older people. OBJECTIVE: The present study aims at investigating the relationship between Lp(a) and CIMT levels in relation to MS among older subjects. METHODS: We studied 182 Japanese subjects of >or=60 years (mean 72.5 years), free of cardiovascular/cerebrovascular disease. MS was based on the NCEP-ATPIII criteria with a minor modification for Japanese. The CIMT was ultrasonographically measured. RESULTS: The CIMT levels were significantly greater in the MS group (n = 60, 1.03 +/- 0.22 mm) than the non-MS group (n = 122, 0.96 +/- 0.22 mm). Multivariate analysis, using Lp(a) levels or the product term for interaction between Lp(a) and MS, showed that age significantly and independently correlated to CIMT, along with male gender. DISCUSSION: Even when Lp(a) and MS were simultaneously considered, age was the best determinant of CIMT in this population. The mechanism of our results including weak additive effects of Lp(a) and MS among older subjects may be partly throughout aging.

The association between an increased level of gamma-glutamyl transferase and systolic blood pressure in diabetic subjects.

Gamma-glutamyl transferase (GGT) is an enzyme present in serum and on most cell surfaces and serves as an oxidative stress marker. Although serum GGT is associated with hypertension development, little data are available on the associations between GGT and hypertension among populations with diabetes mellitus (DM). Our aim was to investigate the potential association between the changes in systolic or diastolic blood pressure (SBP/DBP) and the GGT level in type 2 DM subjects, in comparison with non-DM subjects. In 179 non-DM and 177 DM subjects, SBP/DBP, body mass index (BMI), fasting plasma glucose, serum asparate aminotransferase, alanine aminotransferase and GGT were measured at the baseline and after a 1-year period. Between these 2-measurement points, in non-DM subjects, SBP and DBP levels were significantly increased, while GGT tended to increase. In contrast, in DM subjects, the mean levels of SBP, DBP and GGT remained unchanged. Multivariate analysis revealed that in non-DM subjects the degree of increase in SBP was significantly and positively correlated to that of GGT (beta = 0.165), along with age and BMI. Likewise, the increase in DBP was correlated to that of GGT in non-DM subjects (beta = 0.170). In contrast, in DM subjects, the degree of increase in SBP was significantly correlated to that of only GGT (beta = 0.166). These results suggest that the presence of DM may attenuate the effects of GGT on DBP.

Non-high-density lipoprotein cholesterol levels and recent involuntary weight gain among asymptomatic female subjects.

BACKGROUND: Serum non-high-density lipoprotein cholesterol (HDL-C), an easily identifiable atherogenic index, has attracted attention within a clinical meaning different from that of other lipid indexes. The link between body weight gain and the occurrence of cardiovascular diseases may be mediated through non-HDL-C. However, there have been few reports examining the independent associations between weight gain and non-HDL-C, over a period of at least 1 year, especially in females. METHODS: We examined data on 200 asymptomatic Japanese females (mean 49.1 years) with an involuntary weight gain of at least > or = 0.1 kg/m2 as a body mass index (BMI) 1 year after a baseline check-up. At baseline and after the 1-year period, we measured BMI, blood pressure (BP) and blood metabolic variables, including non-HDL-C. RESULTS: The mean BMI levels rose from 22.9 to 23.5 kg/m2 during this period. Non-HDL-C levels had a significant increase (from 3.87 to 3.93 mmol/L), and a partial correlation test, adjusted for age and all measured metabolic variables, revealed that BMI change was significantly and independently correlated to non-HDL-C levels (r=0.25, p<0.0001), along with systolic BP. The subgroup with an age of <50 years had a clear correlation between BMI change and non-HDL-C levels (r=0.34, p=0.001), contrary to those of > or = 50 years. CONCLUSIONS: A short-term involuntary weight gain was significantly and independently correlated to an increase in non-HDL-C levels among asymptomatic Japanese females, particularly in relatively young subjects. In achieving a more favorable lipid profile of non-HDL-C, even a modest weight gain may need attention.

Relationship between A-3826G polymorphism in the promoter of the uncoupling protein-1 gene and high-density lipoprotein cholesterol in Japanese individuals: a cross-sectional study.

BACKGROUND: A-3826G polymorphism within the promoter region of the uncoupling protein-1 (UCP-1) gene is possibly involved in the pathophysiology of obesity and metabolic disorders. However, the effects of UCP-1 A-3826G polymorphism on high-density lipoprotein cholesterol (HDL-C), a major contributor to atherosclerotic disease, still have not been established. METHODS: A total of 298 healthy Japanese subjects (144 males and 154 females, mean age: 45.2 years) with a body mass index (BMI) of 20.0-30.0 kg/m(2), regular lifestyles, and receiving no medication were enrolled in the cross-sectional study to estimate the relationship of serum HDL-C levels with UCP-1 A-3826G polymorphism by genomic PCR and Bcl1-restriction fragment length polymorphism analysis. We used 1.04 mmol/L of HDL-C in Japanese males and 1.29 mmol/L in Japanese females as cut-off values of low HDL-cholesterolemia. RESULTS: The genotype and allele frequencies of UCP-1 A-3826G polymorphism were similar to those previously reported in the Japanese population. In males, HDL-C levels of the GG genotype (1.75+/-0.49 mmol/L) were significantly higher than those found in the AA genotype (1.45+/-0.34 mmol/L, p=0.015). In females, the occurrence rate of low HDL-cholesterolemia was significantly different by genotype: a low prevalence in the GG genotype (15.4% in the AA, 4.8% in the AG, 15.4% in the GG genotype, p=0.022). Logistic regression analysis was used to identify risk factors for low HDL-cholesterolemia, with adjustments for age, gender, smoking, alcohol intake, BMI, hypertriglyceridemia, and genotype. The GG genotype was detected as being a significant associated factor (odds ratio =0.11 [95% confidence interval =0.01-0.90], p=0.01), in addition to BMI and the presence of hypertriglyceridemia. CONCLUSIONS: These results suggest that the GG genotype may be an independent protective factor associated with low HDL-cholesterolemia in this population, although the role of the UCP-1 A-3826G polymorphism in HDL-C is complex and remains controversial. This hypothesis needs further investigation.

Mood change tendency and fasting plasma glucose levels in a Japanese female population.

Internal stress can modify values in blood examinations such as glucose. Mood change releases us from the stress state, and the mood change tendency (MCT) may show individual differences. However, little is known about whether individual mood change tendencies in daily life affect fasting plasma glucose (FPG) levels. We investigated the effects of clinical characteristics including age, body mass index (BMI), smoking, alcohol habits and self-reported MCT (an answer to the inquiry on their daily MCT: good, average, or poor) on FPG values among 272 Japanese females (mean age 48.4 +/- 9.3 years). Subjects with normal to impaired fasting glucose levels (less than 7.0 mmol/L) were included in this study. The mean FPG levels in subjects with good, average and poor MCT were 5.32 +/- 0.48, 5.36 +/- 0.50 and 5.58 +/- 0.69 mmol/L, respectively. A significant difference was noted between subjects with good and poor MCT (p = 0.02). There was no significant difference in BMI levels among MCT-based groups. Pearson's rank correlation and multiple regression analysis, using FPG levels and other variables, demonstrated a significant relationship between FPG levels and MCT (p < or =0.01), along with age and BMI. These results suggest slight but significant effects of individual MCT on FPG, and that a consideration of MCT may occasionally be needed in the interpretation and management of FPG levels in the Japanese female population.

The uncoupling protein-1 gene -3826A/G polymorphism and hypertension in Japanese subjects.

BACKGROUND: The possible effects of the uncoupling protein-1 (UCP-1) gene -3826A/G polymorphism on hypertension (HT) have yet to be elucidated. METHODS: A total of 578 Japanese subjects (231 males and 347 females, mean age 58.4 years) were enrolled in the study to investigate the association between HT and the -3826A/G polymorphism by genomic PCR and Bcl1-restriction fragment length polymorphism methods. RESULTS: Multivariate logistic regression analysis for HT, adjusted for genotype (recessive model, AA+AG vs. GG) and other covariates such as cardiovascular risk factors [e.g., smoking, body mass index (BMI), dyslipidemia and diabetes] showed age [odds ratio (OR) 1.11 (95% confidence interval 1.08-1.13)] and BMI [OR 1.13 (1.06-1.21)] as independent significant factors. In the subgroup analysis, as well as age and BMI, GG genotype [OR 2.32 (1.08-4.99)] was also an independent significant factor for HT in males. Similarly, as well as age and BMI, GG genotype [OR 1.89 (1.00-3.57)] was also an independent significant factor for HT in the relatively older subgroup (> or =60 years). CONCLUSIONS: The results suggest that the GG genotype may be associated with the presence of HT in Japanese males and older subjects. Further investigation is needed to confirm our hypothesis.