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Xanthine oxidoreductase exists both intracellularly and extracellularly and induces vascular injury by producing reactive oxygen species (ROS). Here, we investigated the effects and mechanism of action of topiroxostat, a xanthine oxidase inhibitor, on ROS using an animal model of type 1 diabetes with persistent hyperglycemia. Six-week-old male Sprague-Dawley rats were administered 50 mg/kg streptozotocin to induce diabetes; at 8 weeks of age, animals were administered topiroxostat (0.3, 1, or 3 mg/kg) for 2 weeks through mixed feeding after which the aorta was sampled. The production of superoxide, a type of ROS, was measured by chemiluminescence and dihydroethidium staining. Cytotoxicity was evaluated by nitrotyrosine staining. Topiroxostat at 3 mg/kg significantly decreased blood urea nitrogen, e-selectin, urinary malondialdehyde, and the urinary albumin/creatinine ratio compared with the streptozotocin group. Superoxide production by xanthine oxidase anchored to the cell membrane was significantly decreased by topiroxostat at both 1 mg/kg and 3 mg/kg compared with the streptozotocin group. Dihydroethidium staining revealed no significant effect of topiroxostat administration on superoxide production. The fluorescence intensity of nitrotyrosine staining was significantly suppressed by 3 mg/kg topiroxostat. Topiroxostat was found to inhibit the production of ROS in the thoracic aorta and suppress vascular endothelial damage. The antioxidant effect of topiroxostat appears to be exerted via the inhibition of anchored xanthine oxidase.

Nonalcoholic steatohepatitis (NASH), the aggressive form of the most common chronic liver disease nonalcoholic fatty liver disease, is characterized by inflammation and damage in the liver. Although hepatocyte injury and cell death have been identified as cardinal pathological features of NASH, its pathogenesis has not yet been elucidated in detail. Immortalized cell lines and primary cultured cells have been used as in vitro models of NASH. However, these cells have several disadvantages, such as specialized characteristics by immortalization or limited growth potential. To overcome these difficulties and develop a strategy to analyze the pathology of NASH, we employed hepatocyte-like cells differentiated from human induced pluripotent stem cells (hiPSC-HLCs) as an in vitro model of NASH to clarify the intracellular effects of glyceraldehyde-derived advanced glycation end-products (AGEs), also named toxic AGEs (TAGE). The viability of hiPSC-HLCs decreased with the accumulation of TAGE in the cells, which was consistent with previous findings on human hepatocellular carcinoma cells and human primary cultured hepatocytes. In addition, the TAGE accumulation up-regulated the expression of inflammation-related genes (interleukin 6, interleukin 8, and monocyte chemoattractant protein-1) in hiPSC-HLCs. These results indicated that the accumulation of TAGE induced hiPSC-HLC cytotoxicity and inflammation, which are features of the pathology of NASH. Therefore, we suggest the use of hiPSC-HLCs as an important strategy for analyses of the pathology of NASH.

In 2015, Japan's Ministry of Health, Labour and Welfare released a report on the need for pharmacies to support public health, although the details of such a service was left to each pharmacy. Consequently, pharmacists had to determine the nature of such services. We considered the services that pharmacists could offer to improve people's lifestyles. This study tests such a service at a pharmacy. We prepared a lifestyle self-review test. From September 2015 to February 2016, pharmacists interviewed members of the community using the test, and pharmacists at 50 pharmacies in Aichi prefecture, Japan, set goals for lifestyle improvement. We analyzed 289 tests. The number of people who had a dietary goal concerning snacking was reduced from 19 people who snacked every day to 11. The number of people who had a goal to reduce their drinking was reduced from 7 people who drank every day to 4. The number of people who had an exercise goal was reduced from 17 people who did not exercise every day to 7. The people who had a sleep goal increased the number of days in which they got adequate rest. Those who had specific, tangible goals for walking achieved their goal at a higher rate than did those who had a goal but no tangible elements, such as time spent or number of steps achieved. Overall, the lifestyle self-review at the pharmacy was effective for lifestyle improvement. Pharmacists should help people set goals with tangible elements to improve their lifestyle more effectively.

It is reported that statins have inconsistent effects on glycemic status and adiponectin concentrations in patients with type 2 diabetes mellitus (T2DM). We aimed to investigate the effect of statins on these variables in patients with T2DM and hypercholesterolemia. A control group comprising 24 patients with T2DM but without hypercholesterolemia was observed for more than 12 weeks, while 24 patients with T2DM and hypercholesterolemia were treated with statins for the same period (statin group). The percentage changes in the glycemic status [blood glucose and glycated hemoglobin (HbA1c)], and levels of plasma adiponectin [total and high molecular weight (HMW)] were compared between the two groups. The statin group had reduced percentage changes in HbA1c, blood glucose, and total and HMW-adiponectin concentration percentage changes that were similar to those in the control group. However, when matched for sex, age (±5 years) and HbA1c (±0.5%) with the control group, the pravastatin group had reduced percentage changes in the plasma HMW-adiponectin concentrations than the matched controls (p=0.023). However, there were no differences in the percentage changes in the plasma total adiponectin (p=0.137), HbA1c (p=0.202), or blood glucose concentrations (p=0.450) between the two groups. Pravastatin treatment had no effect on the glycemic status of patients with T2DM and hypercholesterolemia, but may reduce the percentage changes in the plasma HMW-adiponectin concentrations. Hence, patients with T2DM and hypercholesterolemia receiving long-term treatment with pravastatin might experience increased insulin resistance.

A complication of diabetes mellitus is the over-production of vascular superoxides, which contribute to the development of arteriosclerosis and peripheral arterial disease (PAD). Hyperglycemia induces the formation and accumulation of advanced glycation end-products (AGEs), which in turn stimulate vascular superoxide production. The mechanism underlying AGE-mediated vascular superoxide production remains to be clarified in lower limb complications associated with diabetes. In the present study, we investigated the role of AGEs and the mitochondrial respiratory complex in superoxide production in femoral arteries using the type 2 diabetes model Otsuka Long-Evans Tokushima Fatty (OLETF) rats [vs. non-diabetic Long-Evans Tokushima Otsuka (LETO) rats]. The effects of in vivo administration of pravastatin on superoxide production in femoral arteries were also examined. Using chemiluminescent assays, luminescence microscopy, and competitive enzyme-linked immunosorbent assay (ELISA), we determined that vascular superoxide production and serum glyceraldehyde-derived AGEs (Glycer-AGEs) increased in OLETF rats. Pravastatin inhibited these responses without changing serum total cholesterol concentrations. The mitochondrial complex II inhibitor thenoyltrifluoroacetone (TTFA) also inhibited vascular superoxide production. Application of Glycer-AGEs in situ increased superoxide production in the vascular wall of femoral arteries from pravastatin-treated OLETF rats, which was then inhibited by TTFA. These results suggest that hyperglycemia increases serum Glycer-AGEs, which subsequently induce superoxide production in the femoral artery of OLETF rats in a mitochondrial complex II-dependent manner. Collectively, our results have partially elucidated the pathological mechanisms leading to diabetes-related PAD, and indicate dual beneficial actions of pravastatin for the prevention of oxidative damage to the vascular wall.

In this study, we took continuous measurements of hemoglobin A1c (HbA1c) levels and conducted lifestyle checks in three cases to determine if these parameters were effective in improving overall wellness. We selected three young men with relatively high HbA1c levels. During the 12-weeks study periods, we regularly measured each participant's HbA1c levels and monitored their lifestyle habits every two weeks at the community pharmacy once every 2 weeks using specific guidelines. The first participant, a 23-year-old man, had a HbA1c level of 5.7% at his first measurement. His HbA1c level decreased to 5.2% at the last measurement. The second participant, a 19-year-old man, had an initial HbA1c level of 5.7% and a final HbA1c level of 5.4%. The third participant was a 22-year-old man with an initial HbA1c level of 5.4%. His HbA1c level had decreased to 5.1% by the last measurement. The lifestyles of all three men improved with respect to exercise and diet. Based on these results, we surmise that continuous measurements of HbA1c and regular lifestyle checks may contribute to reducing the risk of lifestyle-related disease.

One of the Specific Behavioral Objectives (SBOs) of pharmaceutical education model-core curriculum is as follows: "Understand patient's state of mind and be sensitive to patient's feelings". We performed learning through simulation of diabetes drug therapy as a means to achieve the objective and evaluated the educational effects of the learning. The simulation was performed and a questionnaire survey was conducted among the 4th-year students of the 6-year curriculum before and after simulation. The score of "level of understanding patient's feelings" was significantly increased after simulation (p

Pharmacy school students were trained in a program simulating medication administration and giving adherence instructions. Following the training, the educational effects were evaluated. Students were separated into two groups. One group of students played the role of pharmacists and instructed simulated patients on medication adherence. Another group of students played the role of patients receiving simulated drug therapy; they were instructed on medication adherence by the students playing the role of pharmacists. The educational effects were evaluated using a questionnaire. The scores for "recognition of factors that influence medication adherence" tended to increase after the simulation, and they increased significantly after practical training. The scores for "self-evaluation of technique for instructing patients on medication adherence" increased significantly after the simulation, and they increased even more after practical training. The students' understanding of the effects on patients who were being instructed also increased significantly after the simulation, and these changes were maintained after practical training. In particular, students became more aware of the influence of pharmacists' attitudes. In practical training, the simulation training was helpful for bedside practice at hospital pharmacies and over-the-counter service at community pharmacies. Thus, the use of role play and simulated patients was an effective method for training pharmacy students to instruct patients on medication adherence.

Endothelium-derived superoxide induces vascular dysfunctions. The aim of this study was to examine the activity of protein kinase C (PKC) isoforms and endothelial nitric oxide synthase (eNOS), which leads to vascular superoxide production in type 2 diabetes, in addition to the effects of pravastatin. We studied these mechanisms in Otsuka Long-Evans Tokushima Fatty (OLETF) rats (type 2 diabetes model) at the early hyperglycemic stage (vs. non-diabetic Long-Evans Tokushima Otsuka [LETO] rats). Superoxide production and catalase activity were measured in aortas, as were the protein expressions of PKCδ and phospho-Ser(1177) eNOS. Superoxide production was increased in OLETF rats, and this increase was inhibited by the selective conventional PKC (cPKC) inhibitor Go6976 and by the non-selective cPKC and novel PKC inhibitor GF109203X. Phospho-Ser(1177) eNOS was significantly increased in OLETF rats, whereas the protein expressions of PKCδ and phosopho-Thr(505) PKCδ and catalase activity were all greatly reduced. Pravastatin administration to OLETF rats in vivo had normalizing effects on all of these variables. The increment in superoxide production seen in OLETF rats (but not the production in pravastatin-treated OLETF rats) was abolished by high concentration of N(ω)-nitro-L-arginine methyl ester (electron transport inhibitor of eNOS), by sepiapterin (precursor of tetrahydrobiopterin), and by LY294002 (phosphatidylinositol 3-kinase [PI3-kinase] inhibitor). In OLETF rats at the early hyperglycemic stage, aortic superoxide production is increased owing to activation of uncoupled eNOS through phosphorylation by PI3-kinase/Akt. This may be related to the observed reduction in PKCδ/catalase activities. Pravastatin inhibited endothelial superoxide production via normalization of PKCδ/catalase activities.