Development of Simplified in Vitro P-Glycoprotein Substrate Assay and in Silico Prediction Models To Evaluate Transport Potential of P-Glycoprotein.

For efficient drug discovery and screening, it is necessary to simplify P-glycoprotein (P-gp) substrate assays and to provide in silico models that predict the transport potential of P-gp. In this study, we developed a simplified in vitro screening method to evaluate P-gp substrates by unidirectional membrane transport in P-gp-overexpressing cells. The unidirectional flux ratio positively correlated with parameters of the conventional bidirectional P-gp substrate assay ( R2 = 0.941) and in vivo Kp,brain ratio (mdr1a/1b KO/WT) in mice ( R2 = 0.800). Our in vitro P-gp substrate assay had high reproducibility and required approximately half the labor of the conventional method. We also constructed regression models to predict the value of P-gp-mediated flux and three-class classification models to predict P-gp substrate potential (low-, medium-, and high-potential) using 2397 data entries with the largest data set collected under the same experimental conditions. Most compounds in the test set fell within two- and three-fold errors in the random forest regression model (71.3 and 88.5%, respectively). Furthermore, the random forest three-class classification model showed a high balanced accuracy of 0.821 and precision of 0.761 for the low-potential classes in the test set. We concluded that the simplified in vitro P-gp substrate assay was suitable for compound screening in the early stages of drug discovery and that the in silico regression model and three-class classification model using only chemical structure information could identify the transport potential of compounds including P-gp-mediated flux ratios. Our proposed method is expected to be a practical tool to optimize effective central nervous system (CNS) drugs, to avoid CNS side effects, and to improve intestinal absorption.

A Randomized, Double-Blind, Crossover Pilot Trial of Rice Endosperm Protein Supplementation in Maintenance Hemodialysis Patients.

In maintenance hemodialysis (MHD) patients, low protein intake is associated with protein-energy wasting, a risk factor that affects outcome. However, increased protein intake may lead to hyperphosphatemia and hyperkalemia, which are also mortality risk factors. Here, we evaluated the safety and effects of purified rice endosperm protein (REP), which contains less phosphorus and potassium than soy and casein proteins, as a supplemental protein source for MHD patients. This randomized, double-blind, placebo-controlled, crossover pilot study of REP supplementation (5 g/day × 4 weeks) was carried out in 50 Japanese adult MHD patients (1 dropped out); the primary outcome was the change in the urea kinetic-based normalized protein catabolic rate (nPCR), an indicator of protein intake in MHD patients. Intention-to-treat analyses of 24 patients in the REP-first group and 25 in the placebo-first group showed that REP supplementation increased nPCR significantly by 0.07 g/kg/day (95% confidence interval, 0.03-0.11), whereas changes in serum phosphorus and potassium concentrations were not different from the placebo. REP supplementation did not show a significant effect on other nutritional or metabolic parameters and no specific complications. In conclusion, purified REP with efficient bioavailability may be safe and useful for dietary supplementation in MHD patients.

[Secondary osteoporosis or secondary contributors to bone loss in fracture. Vitamin D deficiency and fracture].

Vitamin D plays an important role in maintaining bone and calcium metabolism. Vitamin D deficiency causes not only low bone mineral density via secondary hyperparathyroidism, but also impairment of bone mineralization. Vitamin D deficiency also increases the risk of fall. Thus, vitamin D deficiency results in increased fracture risk. Serum 25 (OH) D level reflects Vitamin D store and more than 30 ng/mL is recommended for bone health, but the measurement of serum 25 (OH) D is not reimbursed by Japanese health insurance system. Vitamin D supplementation to vitamin D deficient osteoporotics reduces the fracture incidence by both increasing bone mineral density and reducing falls. Evaluation of vitamin D status and its adequate supplementation are prerequisite for proper osteoporosis management.

Dietary ratio of n-6 to n-3 polyunsaturated fatty acids and periodontal disease in community-based older Japanese: a 3-year follow-up study.

The longitudinal relationship between dietary n-6 to n-3 PUFAs ratio and periodontal disease in 235 Japanese subjects for whom data were available for the years 2003-2006 was investigated. PUFAs intake was assessed at baseline with a brief-type self-administered diet history questionnaire. Full-mouth periodontal status, measured as the clinical attachment level (CAL), was recorded at baseline and once a year for 3 years. The number of teeth with a change in the loss of CAL ≥3 mm at any site over a year was calculated as 'periodontal disease events'. Poisson regression analysis was conducted, with dietary n-6 to n-3 PUFAs ratio as the main predictor, to estimate its influence on periodontal disease events. A high dietary n-6 to n-3 PUFAs ratio was significantly associated with greater number of periodontal disease events. The findings suggest the dietary n-6 to n-3 PUFAs ratio is associated with periodontal disease among older Japanese.

Longitudinal relationship between dietary ω-3 fatty acids and periodontal disease.

OBJECTIVE: Fish oil has anti-inflammatory actions that may benefit periodontal health. We investigated the longitudinal relation between dietary ω-3 fatty acids (FAs), docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA) to periodontal disease in community-dwelling elderly. METHODS: Fifty-five participants aged 74 y were randomly selected from a longitudinal interdisciplinary study of aging. Dietary intake data were obtained by a 3-d weighed food intake. The dietary intakes of energy, DHA, and EPA were calculated based on the Standard Food Composition Tables in Japan. Dental examinations were carried out at baseline and once a year for 5 y. The number of teeth with periodontal progression over 5 y per person was calculated as "periodontal disease events." Negative binomial regression analysis was conducted, which included DHA, EPA, and other covariates as independent variables to estimate the influence on periodontal disease events. Longitudinal data were analyzed for participants for whom data were available for 5 y (n=36). RESULTS: Low DHA intake was significantly associated with more periodontal disease events. The mean number of periodontal disease events for participants who consumed the lowest tertile of DHA was approximately 1.5 times larger (lowest tertile, incidence rate ratio 1.49, 95% confidence interval 1.01-2.21) than the reference group (highest tertile of DHA consumption), after simultaneously adjusting for possible confounders. CONCLUSION: The findings suggest there may be an inverse, independent relation of dietary DHA intake to the progression of periodontal disease in older people.

De novo CD5+ diffuse large B cell lymphoma with basophilia in the peripheral blood: successful treatment with autologous peripheral blood stem cell transplantation.

Basophils play an important role in allergic inflammation and are pathologically related to hematological disturbances, such as iron deficiency anemia and myeloproliferative disorders; however, they are only rarely encountered in lymphoid malignancies. Here, we report the case of a 33-year-old man with a bulky mass of the small intestine, multiple paraaortic lymphoadenopathy, pleural effusion, and ascites, who was diagnosed as a case of de novo CD5+ diffuse large B cell lymphoma (DLBCL). This patient showed a marked elevation of the basophil count in the peripheral blood, which appeared to run in parallel with the tumor burden. High dose chemotherapy followed by autologous peripheral blood cell transplantation yielded complete remission, and the patient has remained disease free for 5 years. To the best of our knowledge, this is the first report of a case of de novo CD5+ DLBCL showing marked elevation of the PB basophil count.

High serum TNF-alpha level in Type 2 diabetic patients with microangiopathy is associated with eNOS down-regulation and apoptosis in endothelial cells.

A high dose of tumor necrosis factor (TNF)-alpha induces endothelial dysfunction and enhances apoptosis in vitro. The present study was conducted to examine whether incubating human umbilical vein endothelial cells (HUVECs) with serum from Type 2 diabetic patients complicated with retinopathy and/or microalbuminemia demonstrate endothelial dysfunction. Serum levels of TNF-alpha and vascular endothelial growth factor (VEGF) were elevated in diabetic patients. Plasma levels of TNF-alpha, two soluble TNF-alpha receptors (sTNFR), and VEGF were assessed in diabetic patients (CD, n=21) complicated with retinopathy and/or nephropathy, uncomplicated diabetic patients (UD, n=18), and in healthy normal participants (NS, n=16). In HUVECs incubated with patient's serum, endothelial constitutive nitric oxide synthase (eNOS) protein expressions were measured by Western blot analysis. Apoptosis in HUVECs was determined by optical microscopy, DNA fragmentation, and CPP32-like protease activity. Serum TNF-alpha, sTNFR-I, and asymmetric dimethylarginine (ADMA), an endogenous inhibitor of NOS, in CD were significantly higher than in UD or NS. While, serum sTNFR-I and VEGF levels were significantly increased in the both diabetic patients, compared with those of NS, no difference was observed in the serum TNF-alpha, sTNFR-II, and ADMA levels between UD and NS. eNOS down-regulation and apoptosis were seen in HUVECs incubated with serum from CD for 24 h, but those observations were completely counteracted in the incubation by the addition of the antihuman TNF-alpha antibody. These results imply that eNOS down-regulation in CD is associated with high serum TNF-alpha levels despite of high serum of VEGF levels. Therefore, endothelial dysfunction in diabetic patients complicated with microangiopathy may, in part, be attributed to high serum TNF-alpha levels.

Differential increase in natriuretic peptides in elite dynamic and static athletes.

The echocardiographic measures and plasma concentrations of either atrial natriuretic peptide (ANP) or brain natriuretic peptide (BNP) were compared in elite judo practitioners (static athletes), elite marathon runners (dynamic athletes) and healthy controls to investigate the relationship between the different types of left ventricular (LV) hypertrophy and plasma concentrations of natriuretic peptides in athletes. The LV mass and LV wall thickness of marathon runners and judo practitioners were significantly greater than those of controls. The LV end-diastolic dimension index was significantly larger in the marathon group, but smaller in the judo group. The left atrial dimension (LAD) index was significantly larger only in marathon runners. Plasma BNP concentrations were higher in both the judo and marathon groups than in controls, and positively correlated with LV mass as well as with deceleration time. Plasma ANP concentrations were significantly higher in marathon runners than in the controls and judo groups, and positively correlated with the LAD index, but negatively correlated with ejection fraction. Multivariate analyses showed that the type of athlete and LAD index were independent predictors of plasma BNP and ANP concentrations, respectively. Thus, there is an intimate link between plasma concentrations of natriuretic peptides and cardiac morphology in different types of athletes.

[Consideration concerning a care orientated medicine for the patient with terminal stage of cancer].

Home care medicine plays an important role in the care of terminal patients with redundant diseases such as cancer. In this study, we propose a "simple-style medicine" consisting in the participation of both medical and co-medical staffs and the use of Kampo medicine. This system satisfied various and individual requests from terminal patients. We present 2 terminal patients for whom this "simple-style", home care-orientated medicine was effective. This system can contribute to the quality of life of patients who need palliative, home care medicine.