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BACKGROUND Type 2 diabetes is a risk factor for atherosclerosis. Oxidative stress, which is a causative factor in insulin resistance, leads to atherosclerosis in patients with diabetes. Xanthine oxidoreductase (XOR) is an enzyme that catalyzes the oxidation of hypoxanthine to xanthine and xanthine to uric acid and is related to oxidative stress. We aimed to examine the influence of plasma XOR activity on arterial stiffness in patients with type 2 diabetes. METHODS In total, 458 patients with type 2 diabetes not receiving antihyperuricemic agents were enrolled and their clinical parameters including plasma XOR activity and the cardio-ankle vascular index (CAVI) were measured. Patients were divided into the liver dysfunction and absence of liver dysfunction groups. Multiple regression analysis was performed. RESULTS The median plasma XOR activity level was 64.3 pmol/h/mL (33.3-147.3 pmol/h/mL). Plasma XOR activity was correlated significantly and positively with aspartate transaminase and alanine transaminase (ρ > 0.5). The level of plasma XOR activity in the liver dysfunction group was eight-fold higher than that in the absence of liver dysfunction group. A significant positive correlation was observed between plasma XOR activity and the CAVI only in the liver dysfunction group (ρ = 0.3968, P < 0.0043). Multiple regression models demonstrated that plasma XOR activity was an independent predictor of the CAVI in the liver dysfunction group (P = 0.0055). CONCLUSIONS Our results suggest that plasma XOR activity is associated with arterial stiffness and may have a role in atherosclerosis development in patients with type 2 diabetes and liver dysfunction.

Aim We aimed to investigate the effect of resveratrol (Rsv) on expression of genes regulating triglyceride (TG) accumulation and consumption in differentiated 3T3-L1 preadipocytes. Methods 3T3-L1 preadipocytes were cultured in DMEM supplemented with 10% fetal calf serum. Upon reaching confluence, cells were induced to differentiate for 4 days, cultured for 10 days for TG accumulation, and then incubated with Rsv (0, 25 or 50 μM) for 3 days. TG accumulation was analyzed by Oil Red-O staining. To understand how Rsv regulates TG accumulation and consumption, changes in gene and protein expressions of several factors associated with free fatty acid (FFA) uptake and β-oxidation were investigated by real-time RT-PCR and Western blot. For further elucidation of underlying mechanisms, we also investigated gene expressions using Sirtuin1 (Sirt1) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α) siRNA. Results Rsv dose dependently enhanced Sirt1 expression and reduced TG accumulation. Rsv-induced reduction of TG accumulation was abolished by inhibition of Sirt1 and PGC1α. Rsv also enhanced expressions of genes involved in FFA uptake [peroxisome proliferator-activated receptor-gamma (PPARγ) and lipoprotein lipase] and in β-oxidation regulation [PGC1-α and carnitine palmitoyl-transferase 1a (CPT1a)]. All these effects were abolished by Sirt1 inhibition. Conclusion The present results suggest that Rsv may augment synthesis and oxidation of fatty acid, and possibly increases energy utilization efficiency in adipocytes through activation of Sirt1. The present study may provide meaningful evidence supporting the efficacy of Rsv in the treatment of obesity.

An increased risk of arteriosclerosis has been noted in cancer survivors. Currently, there are only a few reports available that consider the risk of arteriosclerosis in patients treated with chemotherapy. Patients with an advanced stage B-cell malignant lymphoma are typically treated with a combination therapy of rituximab and cyclophosphamide, doxorubicin, vincristine, and prednisolone (R-CHOP). Complications such as diabetes mellitus (DM), hyperlipidemia (HL), and osteoporosis due to prednisolone and cardiotoxicity due to anthracyclines are well known. However, there are no studies that have investigated the link between R-CHOP therapy and arteriosclerosis. We discussed a patient with follicular lymphoma who was evaluated using cardio-ankle vascular index (CAVI) as an arterial stiffness parameter during R-CHOP therapy in this report. She achived complete remission after the eighth course therapy without complications such as hypertension (HT), HL, DM, and infection. This patient showed elevated CAVI with new plaque formation in the carotid arteries after the end of chemotherapy. These data indicate that R-CHOP therapy may progress the arteriosclerosis.

Measurement of arterial stiffness in routine medical practice is important to assess the progression of arteriosclerosis. So far, many parameters have been proposed to quantitatively represent arterial stiffness. Among these, pulse wave velocity (PWV) has been most frequently applied to clinical medicine because those could be measured simply and non-invasively. PWV had established the usefulness of measuring arterial wall stiffness. However, PWV essentially depends on blood pressure at the time of measurement. Therefore, PWV is not appropriate as a parameter for the evaluation of arterial stiffness, particularly for the studies involving blood pressure changes.On the other hand, stiffness parameter β is an index reflecting arterial stiffness without the influence of blood pressure. Recently, this parameter has been applied to develop a new arterial stiffness index called cardio-ankle vascular index (CAVI). Therefore, CAVI does not depend on blood pressure changes during the measurements; CAVI could represent the stiffness of the arterial tree from the origin of the aorta to the ankle.Many clinical studies obtained from CAVI are being accumulated. CAVI showed high value in arteriosclerotic diseases, such as coronary artery diseases, cerebral infarction, and chronic kidney diseases, and also in majority of people with various coronary risk factors. The improvement of those risk factors decreased CAVI. Furthermore, the role of CAVI as a predictor of cardio-vascular events was reported recently.We review the clinical studies on CAVI and discuss the clinical usefulness of CAVI as a candidate surrogate end-point marker for cardiovascular disease.

AIM We investigated whether cardio-ankle vascular index (CAVI), an arterial stiffness marker, independently predicts future cardiovascular events in subjects with metabolic disorders. METHODS 1562 outpatients underwent CAVI between April 2004 and March 2006 at Toho University, Sakura Medical Center in Chiba, Japan. Patients who already had cardiovascular events at baseline, patients with low ankle brachial index (＜0.9), and patients with atrial fibrillation were excluded. After exclusion, 1080 subjects with metabolic disorders including diabetes mellitus, hypertension and dyslipidemia were screened and followed prospectively. RESULTS Eventually, 1003 subjects (92.9% of 1,080 subjects) followed until March 2012 (follow-up duration 6.7±1.6 years) were analyzed. During the observation period, 90 subjects had new-onset myocardial infarction or angina pectoris confirmed by angiography. All subjects were stratified into quartiles by baseline CAVI (Q1: CAVI ≤8.27, Q2: CAVI 8.28-9.19, Q3: CAVI 9.20-10.08, Q4: CAVI ≥10.09). Age, male ratio and future cardiovascular events increased as CAVI quartile became higher. In Cox proportional hazards regression analysis, the factors independently associated with higher risk of future cardiovascular events were every 1.0 increment of CAVI [hazard ratio (HR) 1.126, p= 0.039], male gender (HR 2.276, p=0.001), smoking (HR 1.846, p=0.007), diabetes mellitus (HR 1.702,p=0.020), and hypertension (HR 1.682, p=0.023). CONCLUSION In individuals with metabolic disorders, CAVI was a predictor of future cardiovascular events, independent of traditional coronary risk factors. CAVI is a potentially valuable tool to identify persons likely to benefit from more intensive therapeutic approaches.

7-Ketocholesterol (7-KCHO) is a highly proinflammatory oxysterol and plays an important role in the pathophysiology of diabetic nephropathy (DN). Lipoxygenases (LOXs) and cyclooxygenases (COXs) are also involved in the development of DN. The aim of this study was to clarify the effects of 7-KCHO on mRNA expression of LOXs and COXs as well as pro-inflammatory cytokines in human mesangial cells (HMC). We evaluated cell viability by WST-8 assay and measured mRNA expression by reverse transcription-polymerase chain reaction. Intracellular reactive oxygen species (ROS) production was evaluated by flow cytometry. Although 7-KCHO did not affect cell viability of HMC, 7-KCHO stimulated significant increases in mRNA expression of 12-LOX, COX-2 and pro-inflammatory cytokines. 7-KCHO also induced an increase in ROS production, while N-acetylcysteine partially suppressed the increase. The 12-LOX and COX-2 inhibitors also suppressed mRNA expression of cytokines. These findings may contribute to the elucidation of the molecular mechanism of the pathophysiology of DN.

Daiji Nagayama,1,2 Haruki Imamura,2 Yuta Sato,2 Takashi Yamaguchi,2 Noriko Ban,2 Hidetoshi Kawana,2 Masahiro Ohira,2 Atsuhito Saiki,2 Kohji Shirai,3 Ichiro Tatsuno2 1Center of Endocrinology and Metabolism, Shin-Oyama City Hospital, Oyama-City, 2Center of Diabetes, Endocrinology and Metabolism, 3Department of Vascular Function, Sakura Medical Center, Toho University, Sakura-City, Japan Objective: The objective of this study is to investigate the association of body mass index (BMI) with arterial stiffness assessed by cardioankle vascular index (CAVI). Subjects and methods: A retrospective cross-sectional study was conducted in 23,257healthy Japanese subjects (12,729 men and 10,528 women, aged 47.1 ± 12.5years, BMI 22.9 ± 3.4kg/m2) who underwent health screening between 2004 and 2006 in Japan. Exclusion criteria were current medication use and a past history of cardiovascular disease, hypertension, stroke, diabetes, and nephritis. Results: Male subjects showed significantly higher BMI, CAVI, and triglycerides and lower high-density lipoprotein (HDL)-cholesterol compared with female subjects. Next, the subjects were divided into tertiles of BMI: lower, middle, and upper, in a gender-specific manner. After adjusting for confounders including age, systolic blood pressure, and HDL-cholesterol identified by multiple regression analysis, the mean CAVI decreased progressively as BMI tertile increased in both genders. Furthermore, a negative inverse relationship between BMI and adjusted CAVI was observed throughout the BMI distribution. Multivariate logistic regression model for contributors of high CAVI (≥90th percentile) identified obesity (odds ratios (95% confidence interval): 0.804 (0.720–0.899)], older age [15.6 (14.0–17.4)], male gender [2.26(2.03–2.51)], hypertension [2.28 (2.06–2.54)], impaired fasting glucose [1.17 (1.01–1.37)], and low HDL-cholesterol [0.843 (0.669–1.06)] as independent factors. Conclusion: We demonstrated an inverse relationship between CAVI and BMI in healthy Japanese subjects, suggesting that systemic accumulation of adipose tissue per se may lead to a linear decrease of arterial stiffness in nonobese and obese subjects without metabolic disorders. Keywords: BMI, cardioankle vascular index, arterial stiffness

Summary Objective: Obesity is associated with type 2 diabetes, dyslipidemia and hypertension, contributing to atherogenesis. Weight reduction is the fundamental therapy for obesity. Recently, a novel arterial stiffness parameter called cardio-ankle vascular index (CAVI) has been developed. We hypothesized that CAVI may be a candidate marker of increased vascular stiffness in obese patients. The aim of this study is to investigate the effect of weight reduction on CAVI. Subjects and methods: Using CAVI as an indicator, we assessed the changes in arterial stiffness in 47 obese Japanese subjects (aged 46 ± 13 years) who underwent a 12-week weight reduction program consisting of a calorie restriction diet (20—25 kcal/day) and exercise therapy. Visceral fat area (VFA) was evaluated by CT. Results: At baseline, CAVI correlated positively with age (r = 0.70), blood pressure (r = 0.23), VFA (r = 0.26) and HbA1c (r = 0.39). After 12 weeks of weight reduction, mean BMI decreased from 33.3 ± 7.5 to 30.7 ± 6.4 kg/m 2 (p < 0.0001), and mean CAVI decreased from 8.3 to 7.9 (p < 0.01). The change in VFA correlated positively with change in CAVI in subjects with decrease in CAVI (r = 0.47). Furthermore, change in VFA was a significant independent predictor for change in CAVI. No significant correlation was observed between change in CAVI and clinical variables such as BMI, HbA1c and lipids. Conclusion: This study demonstrated that CAVI decreased after weight reduction, and was associated with a decrease in VFA. CAVI reduction maybe a marker of improved vascular stiffness after weight reduction in subjects with visceral adiposity. © 2011 Asian Oceanian Association for the Study of Obesity. Published by Elsevier Ltd. All rights reserved.

Aim Probucol has antioxidant as well as cholesterol-lowering effects. We examined the effect of probucol on the progression of diabetic nephropathy. We named this study 'Sakura Study' after our hospital and city. Methods We performed a randomized, open trial on 162 type 2 diabetic patients with clinical albuminuria (urinary albumin excretion >300 mg/g creatinine). Eighty patients were assigned to probucol treatment (500 mg/day) and 82 patients to no probucol treatment. All patients were followed for five years. The primary outcome was the time to renal dysfunction events, defined as the initiation of chronic hemodialysis therapy and renal dysfunction-related death. Results Probucol decreased total cholesterol, HDL-cholesterol, and LDL-cholesterol compared to the control group. The serum creatinine increase rate was significantly lower (p= 0.015) in the probucol group (0.066 mg/dL/month) than in the non-probucol group (0.116 mg/dL/month). Renal dysfunction events occurred in 72 patients during this study. The 69 patients who were initiated on chronic hemodialysis comprised 42 in the non-probucol group and 27 in the probucol group. Three patients in the non-probucol group, but no patients in the probucol group died of renal dysfunction. The renal dysfunction event-free survival rate was significantly higher (log-rank: p= 0.02) in the probucol group than in the non-probucol group. Conclusion Probucol suppressed the progression of diabetic nephropathy and renal dysfunction events.

Postprandial hyperglycemia is known to be associated with increasing cardiovascular mortality in type 2 diabetes mellitus patients. Cardio-ankle vascular index (CAVI) reflects arterial stiffness and is more useful for predicting coronary atherosclerosis than intima-media thickness. Premixed human insulin 30/70 (BHI30) containing rapid-acting insulin has been used conventionally as a biphasic insulin. Recently, a biphasic insulin analogue preparation, biphasic insulin aspart 30/70 (BIAsp30), containing ultrarapid-acting insulin has been approved and expected to improve postprandial hyperglycemia. The aim of this study was to clarify the effects of switching the biphasic insulin from BHI30 to BIAsp30 on arterial stiffness in type 2 diabetes mellitus patients. Twenty-six type 2 diabetes mellitus patients (glycosylated hemoglobin >6.5%) who were already receiving biphasic insulin therapy with BHI30 twice daily were observed for 3 months. Afterward, BHI30 was switched to BIAsp30. At 3 months after switching, relative mobility of the peak of LDL fraction decreased significantly (from 0.3462 ± 0.041 to 0.3356 ± 0.035, P < .01); and CAVI also decreased significantly (from 9.77 ± 1.11 to 9.35 ± 1.17 m/s, P < .005). A significant negative correlation was observed between the change in CAVI and change in 1,5-anhydroglucitol (1,5-AG) (r = -0.3929, P < .05). A stronger correlation between change in CAVI and change in 1,5-AG was observed in the subgroup of patients whose 1,5-AG levels were elevated after switching (r = -0.6261, P < .05) compared with all subjects. These results suggest that switching biphasic insulin from BHI30 to BIAsp30 improves arterial stiffness, and the improvement of arterial stiffness may be associated with improvement of postprandial hyperglycemia.

SUMMARY Aims: Glimepiride, a third generation sulfonylurea (SU), is known to have extrapancreatic effects, but its vascular effect is unclear. We investigated the efficacy of glimepiride in improving arterial stiffness assessed by cardio-ankle vascular index (CAVI) in type 2 diabetic patients, compared with glibenclamide, a conventional SU. Methods: Forty type 2 diabetic patients were randomly assigned to two groups. One group was administered glimepiride 1.5 mg ⁄day, and the other group was administered glibenclamide 1.25 mg ⁄day for 6 months. Results: No significant difference in hypoglycaemic effect was observed between two groups. CAVI significantly decreased only in glimepiride group (9.4 ± 1.4 fi 8.9 ± 0.8, p < 0.05). Decrease in CAVI was greater in glimepiride group than in glibenclamide group ()0.50 ± 0.98 vs. )0.04 ± 0.57, p = 0.048). Urinary 8-hydroxy-2¢-deoxyguanosine (8-OHdG) decreased in glimepiride group and increased in glibenclamide group, and the changes were significantly different between groups ()1.5 ± 3.5 vs. + 1.8 ± 3.6, p = 0.009); whereas serum lipoprotein lipase mass increased in glibenclamide group and decreased in glibenclamide group, and the changes tended to be different between groups (+ 2.1 ± 19.1 vs. )7.4 ± 19.2, p = 0.096). Change in urinary 8-OHdG was a significant independent predictor for change in CAVI in all subjects. Conclusions: These results suggest that glimepiride improves CAVI compared with glibenclamide. Reduced oxidative stress and improved insulin resistance may contribute to the improvement of CAVI by glimerpiride. What’s known Diabetes mellitus is a risk factor for all manifestations of atherosclerotic vascular disease.

Aim: Recently, a novel device for measuring the cardio-ankle vascular index (CAVI) as an arterial stiffness parameter has been developed. In this study, we evaluated the effect of angiotensin II receptor blocker (ARB) and calcium channel (Ca) blocker on CAVI in type 2 diabetic patients with hypertension.Methods: Seventy type 2 diabetes mellitus patients with hypertension were enrolled and randomly divided into two groups. One group was administered olmesartan medoxomil 10 mg/day for 12 months (ARB group), and the other group was administered amlodipine besilate 5 mg/day for 12 months (Ca blocker group).Results: In the ARB group, a significant decrease in CAVI was observed after 12 months; however, no significant change in CAVI was observed in the Ca blocker group although changes in blood pressure were almost the same. By simple regression analyses, CAVI changes correlated positively with 8-OHdG changes.Conclusions: Olmesartan, an ARB, improved arterial stiffness more than amlodipine, and this effect might be due to not only the blood pressure-lowering effect but also to reducing the potential of oxidative stress recognized in olmesartan.

Aim A novel device has been developed for measuring the cardio-ankle vascular index (CAVI) as an indicator of arterial stiffness. In this study, we evaluated the effect of pitavastatin on CAVI in type 2 diabetic patients. Methods Forty-five type 2 diabetes mellitus patients with low-density lipoprotein cholesterolemia were enrolled and treated with pitavastatin 2 mg/day for 12 months. Before and after pitavastatin administration, HbA1c, serum lipids, serum malondialdehyde-LDL (MDA-LDL), urinary 8-hydroxy- 2'-deoxyguanosine (8-OHdG) and CAVI were measured. Results After pitavastatin treatment for 12 months, significant decreases in 8-OHdG, MDA-LDL and CAVI were observed. DeltaCAVI significantly correlated with DeltaMDA-LDL. Conclusions In type 2 diabetic patients, pitavastatin may have an oxidative stress-reducing effect, especially in a state of enhanced oxidative stress, and CAVI may be useful as a routine test for the diagnosis and therapeutic monitoring of atherosclerosis.

Aim: The three types of calcium channel blocker (CCB), L-, T- and N-type, possess heterogeneous actions on endothelial function and renal microvascular function. In the present study, we evaluated the effects of two CCBs, efonidipine and amlodipine, on renal function and arterial stiffness.Methods: Forty type 2 diabetic patients with hypertension and nephropathy receiving angiotensin receptor II blockers were enrolled and randomly divided into two groups: the efonidipine group was administered efonidipine hydrochloride ethanolate 40 mg/day and the amlodipine group was admin-istered amlodipine besilate 5 mg/day for 12 months. Arterial stiffness was evaluated by the cardio-ankle vascular index (CAVI).Results: Changes in blood pressure during the study were almost the same in the two groups. Sig-nificant increases in serum creatinine and urinary albumin and a significant decrease in the esti-mated glomerular filtration rate were observed in the amlodipine group, but not in the efonidipine group. On the other hand, significant decreases in plasma aldosterone, urinary 8-hydroxy-2'-deoxy-guanosine and CAVI were observed after 12 months in the efonidipine group, but not in the amlo-dipine group.Conclusions: These results suggest that efonidipine, which is both a T-type and L-type calcium chan-nel blocker, has more favorable effects on renal function, oxidative stress and arterial stiffness than amlodipine, an L-type calcium channel blocker.

Oxysterols have cytotoxic effects and contribute to the development of atherosclerosis. To examine association between 7-ketocholesterol and diabetes mellitus, and other coronary risk factors, we developed a reliable quantitative method to measure serum 7-ketocholesterol (s-7KCHO) and studied s-7KCHO in patients with type 2 diabetes mellitus (T2DM). The s-7KCHO was detected by gas chromatography-mass spectrometry assay. The s-7KCHO was significantly higher in patients with T2DM (n=137, 33.8 ng/ml) compared to non-diabetic healthy subjects (n=89, 16.1 ng/ml). Patients with T2DM were divided into two groups with two or more than two risk factors (defined as multiple risk factors group) and with zero or one risk factor (non-multiple risk factors group). The s-7KCHO was significantly higher in multiple risk factors group (39.5 ng/ml) compared to non-multiple risk factors (30.1 ng/ml). Among patients with multiple risk factors group, s-7KCHO was significantly higher in patients with high low-density lipoprotein cholesterol (LDL-C) levels (45.1+/-5.9 ng/ml) compared to those with normal LDL-C levels (35.3+/-7.0 ng/ml). Furthermore, s-7KCHO increased according to the number of concurrent coronary risk factors. These results suggest that serum 7-ketocholesterol levels may depend on the multiple risk factors and serum LDL-C levels.

Aim: Previous reports indicate that serum lipoprotein lipase mass levels (LPL mass) and common carotid artery intima-media thickness (CCA-IMT) are independent predictors of atherosclerotic diseases. The aim of this study was to examine the effects of combination therapy of sulfonylurea and acarbose on LPL mass and CCA-IMT.Methods: Eighty-four patients with type 2 diabetes mellitus, who were treated with only sulfonylureas and showed CCA-IMT of more than 0.9 mm at baseline, were selected and randomly divided into two groups. One group was administered acarbose 300 mg/day for 12 months (acarbose group, n=41), and the other group was not administered acarbose (non-acarbose group, n=43).Results: After 12 months, a significant increase in LPL mass and a significant decrease in CCA-IMT were observed in the acarbose group (1.024 to 0.964 mm), but no significant changes were observed in the non-acarbose group. In a subgroup analysis of patients with HbA1c improved by 0.5% or more, the increase of LPL mass and decrease of CCA-IMT was significantly greater in the acarbose group than in the non-acarbose group although the changes in HbA1c were similar in two groups.Conclusions: We concluded that reducing postprandial hyperglycemia might increase LPL mass levels and might be useful to prevent macroangiopathy in type 2 diabetic patients treated by sulfonylurea.

Objective To investigate the association of serum uric acid (SUA) with arterial stiffness assessed by cardio-ankle vascular index (CAVI). Methods We analyzed the cross-sectional data from 27,360 healthy Japanese subjects (12,910 males and 14,450 females) aged between 20 and 74 years without a past history of heart disease, stroke, hypertension, diabetes, nephritis or gout. We investigated whether SUA was independently associated with CAVI in a gender-specific manner. Results BMI, CAVI, systolic/diastolic BP, GOT, GPT, γ-GTP, triglyceride (TG), creatinine and SUA were higher and HDL-C was lower in males than in females. Next, they were stratified by SUA into 3 groups: lower tertile (T1), middle tertile (T2) and upper tertile (T3) and by gender. CAVI increased progressive with increasing SUA tertile, after adjusting for age, BMI and systolic BP (sBP) identified in multiple regression analysis for CAVI. Multivariate analysis showed that the odds ratios (95% CI) relative to T1 for high CAVI (≥90 th percentile) were 1.233 (0.928–1.638) in T2 and 1.352 (1.031–1.773) in T3 for males, and 1.133 (0.984–1.303) in T2 and 1.361 (1.098–1.687) in T3 for females, after adjusting for confounders. Furthermore, increase in adjusted CAVI was observed in a lower SUA range in females compared to that observed in males. Conclusion We demonstrated an independent correlation between SUA and CAVI, and observed gender difference in the SUA range for increase in CAVI. These results may suggest the need to set different target SUA levels for men and women in anti-hyperuricemic treatment for atherosclerosis prevention.

We propose a novel single stranded DNA (ssDNA) immobilization process for improving sensitivity and reproducibility of DNA microarrays. We immobilized 50 mer 5'-amino-modified oligonucleotides by forming a covalent bond between the oligonucleotides, and phenylisothiocyanate groups tethered on the surface. We used a solution containing a phase-transfer catalyst, cetyltrimethylammonium bromide (CTAB). The amount of the immobilized oligonucleotides was estimated from fluorescence intensity of Cy5 dyes, which were attached to the oligonucleotides. The surface density of the immobilized oligonucleotides was also measured using X-ray reflectometry and surface plasmon resonance. We found that the addition of CTAB allowed us to increase the density and uniformity of the immobilized oligonucleotides, as well as to decrease non-specific adsorption of the oligonucleotides. The highest density and uniformity were obtained when the concentration of CTAB was nearly below the critical micelle concentration (CMC). At the concentration level close to the CMC, a CTAB monolayer formed on the surface can attract the negatively charged oligonucleotides effectively because of the positive charge of CTAB. Moreover, the CTAB layer can enhance encounter rates between the hydrophilic oligonucleotides and the hydrophobic surface with the phenylisothiocyanate groups. However, at a higher concentration level above the CMC, the density of the immobilized oligonucleotides decreases because the CTAB micelles can trap the oligonucleotides in the solution. Finally, we obtained higher hybridization signals on oligonucleotides immobilized with the CTAB solution. [DOI: 10.1380/ejssnt.2005.511]

Introduction : Oxidative stress is closely related to iron overload in myelodysplastic syndrome (MDS) and induces DNA damage. A recent study has suggested that oxidative stress leads to increased mutation frequency in a murine model of MDS, and that additional mutations lead to the progression of MDS. We have recently reported that oxidative stress marker reflects not only iron overload but also disease progression in a MDS patient (2016, Shimizu et al). So there is a possibility that the increase in oxidative stress markers may have not only caused the iron overload but also resulted in the disease progression of MDS. To clarify this point, we evaluated the association among oxidative stress marker and disease progression in eight patients with MDS. In this study, we first examined the oxidative stress marker, derivatives of reactive oxygen metabolite (dROM), ferritin in serum and Wilms Tumor 1 gene (WT1) in the peripheral blood throughout the course of treatment. Then, we analyzed the correlations of the levels of dROM, ferritin and WT1. Next, we investigated ROS expression in each fraction of WBC by flow cytometry in a healthy donor and some MDS patients. Materials and methods: Levels of serum dROM were measured by colorimetry, using samples of MDS (RA n=3, RAEB-1 n=4, RAEB-2 n=1). Ferritin in serum and WT1 in peripheral blood were also measured. Statistical analyses were performed by Pearsonfs test. The ROS expression was evaluated during the clinical course by flow cytometry. Results: Table 1 shows the relationship between dROM, ferritin and WT1 in MDS patients throughout the treatment. Case 1 to 3 patients showed disease progression with excess blasts in spite of azacitidine treatment. These patients showed a correlation tendency between dROM and ferritin, dROM and WT1. Case 1 showed close relationship between ferritin and WT1. Case 3 shows slight relation between ferritin and WT1. Case 4 to 8 patients showed no disease progression. They were treated with supportive therapy, including blood transfusion or cytokine therapy. These patients showed no relation between dROM and the elevation of ferritin and WT1. In these patients, there was no relation between ferritin and WT1 except Case 6. We also evaluated the score of WPSS, IPSS and dROM at diagnosis. dROM did not correlate with WPSS and IPSS. Figure 1 shows increased ROS expression during disease progression in a MDS patient compared with healthy control. Considering that other MDS patients with excess blasts showed the similar pattern, the present result suggested that the oxidative stress markers were produced by tumor-derived cells. Conclusions: dROM levels were slightly related with the ferritin and WT1 mRNA expression levels in MDS patients with excess blasts. We speculate that the increase in the oxidative stress markers is a cause of not only iron overload but also the disease progression of MDS. Oxidative stress markers may contribute to evaluate the progression of treatment efficacy for MDS patients. Table 1 Patientfs characteristics and correlation between serum dROM and ferritin, WT1 mRNA in MDS patients. Table 1. Patientfs characteristics and correlation between serum dROM and ferritin, WT1 mRNA in MDS patients. Figure 1 The increment of ROS expression during disease progression in a MDS patient with excess blasts. Figure 1. The increment of ROS expression during disease progression in a MDS patient with excess blasts. Disclosures Nakaseko: BMS: Honoraria, Research Funding; PFIZER: Honoraria, Research Funding; NOVARTIS: Honoraria.

Aim: High cholesterol absorption in the small intestine has been proposed to be a risk factor of atherosclerosis. In this study, we evaluated the effect of ezetimibe monotherapy on arterial stiffness in type 2 diabetic patients.Methods: Forty type 2 diabetes mellitus patients with high serum low-density lipoprotein cholesterol (LDL-C) were enrolled and treated with ezetimibe 10 mg/day for 6 months. HbA1c, serum lipids, remnant-like particle-cholesterol (RLP-C), serum lipoprotein lipase mass (LPL mass) and the cardioankle vascular index (CAVI) were measured before and after ezetimibe treatment.Results: After 6 months of ezetimibe treatment, significant decreases in LDL-C, RLP-C and CAVI were observed. In the group that achieved the LDL-C goal of

Purpose: Type 2 diabetes is known to be associated with increasing cardiovascular mortality. Malondialdehyde-modified LDL (MDA-LDL) is an oxidized LDL and is increased in patients with diabetes or hypertriglyceridemia. Elevated MDA-LDL has been reported to be a risk factor of atherosclerosis or cardiovascular disease. Sitagliptin is a dipeptidyl peptidase-4 inhibitor and a new class of hypoglycemic agents. In this study, the effects of increasing the dose of metformin and add-on sitagliptin on MDA-LDL were examined in type 2 diabetes patients. Methods: Seventy patients with type 2 diabetes, inadequately controlled despite on-going treatment with metformin 500 mg/day, were enrolled in this randomized controlled trial. The patients received additional metformin (500 mg/day) or sitagliptin (50 mg/day) for 6 months, and changes in metabolic parameters including MDA-LDL were evaluated. Results: After 6 months of treatment, add-on sitagliptin (n=35) improved fasting blood glucose (FBG) and hemoglobin A1c (HbA1c) to significantly greater extent than increasing the dose of metformin (n=35). There were no differences in total cholesterol and low-density lipoprotein cholesterol levels between two groups. MDA-LDL levels (mean±S.E.) decreased significantly with increasing the dose of metformin (from 94.40±6.35 to 77.83±4.74 U/L, P < 0.005), but remained unchanged with add-on sitagliptin treatment (from 89.94±5.59 to 98.46±6.78 U/L, p > 0.05). Multiple linear regression analysis identified increasing the dose of metformin treatment as the only independent factor associated with decreased MDA-LDL (β coefficient 0.367, P < 0.0119), and no significant correlation between change in MDA-LDL and fasting blood glucose or HbA1c. Conclusion: These results suggest that increasing the dose of metformin improves serum MDA-LDL levels in type 2 diabetes mellitus.

Masahiro Ohira,1 Takashi Yamaguchi,1 Atsuhito Saiki,1Noriko Ban,1 Hidetoshi Kawana,1 Ayako Nagumo,1 Takeyoshi Murano,2 Kohji Shirai,3 Ichiro Tatsuno1 1Center for Diabetes, Endocrinology and Metabolism, 2Department of Clinical Laboratory Medicine, 3Department of Vascular Function, Sakura Hospital, Toho University Medical Center, Chiba, Japan Background: Type 2 diabetes is known to be associated with elevated cardiovascular mortality. Pioglitazone improves blood pressure (BP) and pulse wave velocity (PWV), which is an arterial stiffness parameter. Arterial stiffness is closely associated with cardiovascular disease. However, PWV is correlated with BP. The cardio-ankle vascular index (CAVI) reflects arterial stiffness independent of BP. Pioglitazone improves PWV but reduces blood pressure. The aim of this study was to re-evaluate the effect of pioglitazone on arterial stiffness with CAVI. Methods: Sixty patients with type 2 diabetes mellitus and already on 500 mg/day of metformin received add-on therapy of pioglitazone 15 mg/day or glimepiride 1 mg/day for 6 months, during which time changes in their metabolic parameters and CAVI were observed. Results: After 6 months of treatment, both pioglitazone (n=30) and glimepiride (n=30) improved fasting blood glucose and glycated hemoglobin. The changes in fasting blood glucose and glycated hemoglobin between the two groups were greater in the pioglitazone group. Systolic and diastolic BP was decreased in both groups, with no significant between-group differences. Only pioglitazone increased serum adiponectin levels, and the change in adiponectin between the pioglitazone and glimepiride groups was significantly different. CAVI was decreased significantly by pioglitazone but remained unchanged after treatment with glimepiride. The change in CAVI between the two groups was significantly different. Conclusion: These results suggest that pioglitazone improves CAVI, a BP-independent arterial stiffness parameter, in patients with type 2 diabetes mellitus treated with metformin. Keywords: biguanide, thiazolidinediones, sulfonylurea, blood pressure, cardiovascular disease, arterial stiffness

Background: Recently, a novel arterial stiffness parameter called cardio-ankle vascular index (CAVI) has been developed. Aim: This study was conducted in order to elucidate the predictive value of CAVI for future cardiovascular events. Methods: A total of 1,000 outpatients (513 males and 487 females, mean age 63±11 years), who had the examination of CAVI between 2004 and 2006 at our Medical center, were included and followed up for 6.7±1.6 years. Those had diabetes mellitus (51.0%), hypertension (52.4%) or/and dyslipidemia (62.5%). Results: New-onset cardiovascular events were observed in 9.0%. In subjects with cardiovascular events, lower HDL-C and higher CAVI (9.9±2.0 vs. 9.2±1.6), mean age, prevalence of hypertension, diabetes and smoking were observed. COX regression model revealed that CAVI independently increased the risk of cardio vascular events with OR of 1.13 (95% CI 1.007-1.269, p=0.038). Male, diabetes, hypertension, low HDL-C and smoking were also significant independent predictors of events. Conclusion: CAVI is an independent predictor of cardiovascular events in subjects with coronary risks.