Yukio Arakawa

Effects of Lipid Peroxidation-Derived Products on the Growth of Human Colorectal Cancer Cell Line HT-29

Epidemiologic investigations indicate a close relationship between colorectal cancer and fat intake. However, to date the effects of lipid peroxidation-derived products that are formed from fat (especially free or esterified unsaturated fatty acids) on the initiation or progression of colorectal cancer have not been investigated extensively. Therefore, in the present study, we examined the effects of fatty acids, fatty acid hydroperoxides and aldehydes on the growth of human colorectal cancer cell line HT-29. At concentrations of 1 and 10 µM, linoleic, arachidonic and eicosapentaenoic acids, and 13-hydroperoxyoctadecadienoic and 15-hydroperoxyeicosapentaenoic acids had no significant effects on the growth of HT-29 cells. 4-Hydroxynonenal and 4-hydroxyhexenal had no significant effects on the growth of HT-29 cells up to 10 µM, whereas 4-oxononenal potently inhibited HT-29 cell growth (1–10 µM, 16–85% inhibition). Further experiments concerning DNA fragmentation, expression levels of Bax and Bcl-2 mRNA, expression levels of pro-caspase-3 and caspase-3 proteins, and activity of caspase-3 suggested that 4-oxononenal may increase the sensitivity of HT-29 cells to apoptosis through a decreased expression level of Bcl-2 and then increased formation of caspase-3 from pro-caspase-3.

In-vivo Distribution and Erythrocyte Binding Characteristics of Cyclosporin in Renal Transplant Patients

The pharmacokinetic parameters of cyclosporin, a potent immunosuppressive agent, show large intra‐and inter‐individual variability, possibly because of the different analytical methods used. A recently developed cyclosporin‐specific radioimmunoassay has been used to study the in‐vivo distribution and binding characteristics of cyclosporin in whole blood, plasma and erythrocytes of fifteen renal transplant patients.

Intraocular Concentration of Intravenous Prednisolone in Experimental Autoimmune Uveoretinitis Mice

could be caused by a common primary immune-mediated process affecting both the lung and the eye, although the relationship between COP and intraocular infl ammation is still unknown. To our knowledge, intraocular infl ammation has not been previously reported in a patient with COP. In the future, ophthalmologists might need to accumulate cases of COP and intraocular infl ammation, and to consider the possibility of a new clinical entity.

Immunochemical characterization and measurement of neuronal type nitric oxide synthase in human neuroblastoma NB-OK-1 cell using novel anti-synthetic peptide antibody and specific immunoassay system.

We developed a sensitive and specific immunoassay system for human neuronal nitric oxide synthase (hnNOS) using synthetic hnNOS(998-1024) peptide and anti-hnNOS(998-1024) antibody. The novel antibody and radioimmunoassay system revealed a typical nNOS protein in human neuroblastoma NB-OK-1 cell (160 kDa, 180 fmol/10(6) cells). The kinetic parameters of the enzyme were K(m)=4.88 microM and V(max)=4.34 pmol/min/mg protein for L-arginine. On incubation of NB-OK-1 cell for 24 h, betamethasone phosphate decreased both nNOS-immunoreactivity (nNOS-IR) and enzymatic activity in the cell dose-dependently. On the other hand, pituitary adenylate cyclase activating polypeptide(1-38) (PACAP38) increased both nNOS-IR and enzymatic activity at concentrations of 10(-10) and 10(-9) M, but inversely decreased both at 10(-7) M. These suggest the positive and negative implications of endogenous NO in proliferation and differentiation of the cell, which support mitogenic activity of NO generated by nNOS in the cell. The present findings also provided evidence that the quantitative change of nNOS protein controls the integrated activity of the enzyme in the cell and, in turn, substantiate the validity and reliability of the present immunoassay system for hnNOS and its practical usefulness.

A Preliminary Study about the Relationship between Workload and the Outcomes of Community Pharmacists' Home Visiting Service

Aim : To examine any relations of workload and outcomes of pharmacistsʼ home visiting service for medication management and guidance. Survey Target : Managing pharmacists and home-visiting pharmacists from community pharmacies of the Osaka Pharmaceutical Associationʼs Yao and Toyonaka branches as of March, 2012. Methods : Survey forms regarding drug management and guidance at patientsʼ homes were mailed, requesting mail or online response. Workload indexes were visit frequency and work time on site. The relation of workload and its outcomes was examined using univariate analysis regarding three items : change in unused medication amount ; detection of side effects during visits ; change in prescription. SPSS ver. 20 for Windows was used for statistical analysis. Results : 90 of 201 pharmacies responded (collection rate 44.8%), and 110 home patientsʼ data were analyzed. 5-to-15-minute actual work time scored highest (57.4%), followed by less than 5 minutes and 15 to 30 minutes, both at 21.3%. Visit frequency of twice a month scored highest (70.4%), followed by once a week (19.4%), once a month (7.4%), and once in more than one month (2.8%). 5-minute or longer patient visits had a tendency of higher percentage of patients whose unused medication decreased after visits started than less-than-5-minute visits (P=0.072). “Once a week” visits had a tendency of higher percentage of pharmacists detecting side effects than less frequent visits (P=0.061) and changing in prescription (P=0.085). Conclusion : The results above implied the relationship between workload and outcomes incurred by pharmacistsʼ home visits for medication safety management and guidance.

Change in solubility of insulin in therapeutic mixtures of insulin preparations: HPLC analysis

We systematically and quantitatively observed the changes in solubility when short- and intermediate-acting insulin preparations were mixed together, using high-performance liquid chromatography (HPLC) and centrifugation. We tested 30 combinations of 16 kinds of commercial insulin preparations, including six short-acting and eight intermediate-acting preparations. The soluble insulin component in the short-acting preparations examined was in all cases insolubilized to various extents after mixing. Particularly remarkable was the complete precipitation of the insulin from Actrapid Human when it was mixed with Monotard Human (1:1). This sort of drastic change in insulin solubility after mixture was reflected in the changes in blood glucose level after the subcutaneous injection of the mixture to rabbits. The information provided by pharmaceutical companies on absorption kinetics and action profiles of short-acting insulin preparations may no longer be valid when mixtures with longer-acting preparations are used.

Studies on dissolution behaviors of extended-release articles.

The dissolution behavior of fifteen commercial extended-release articles was examined by the dissolution test in order to characterize their behavior in vitro. The articles were classified into six groups according to the formation types. The paddle method for six hours was used for tests, and the media were the first solution (pH 1.2) and the second solution (pH 6.8) in JP XI.Five articles showed different patterns in the above two media, so that their dissolution seemed to depend on pH. A few articles were also effected by pH on the dissolution although they showed time-dependent behavior. The others showed pH-independent (time-dependent) dissolution. The dissolution curves of time-dependent articles approximated to zero-order or firstorder kinetic model. There was, however, no significant relation between the formation type of articles and their dissolution behavior.In addition, for time-dependent articles, simulation curves of dissolution were calculated by using “Weibull function” with the data obtained from the dissolution test up to six hours. Since most of all curves conformed to the observed curves, the parameters of the simulation curves were re-calculated only with the data up to three hours to obtain the prediction curves. The predicted rate fitted well to the observed rate after six hours in many articles. Therefore the prediction curves could be applicable to shorten time for the dissolution test.