尿中の遊離γ-カルボキシグルタミン酸定量のためのHPLCによる改良法

A rapid and sensitive high performance liquid chromatographic (HPLC) method for the determination of free γ-carboxyglutamic acid (Gla) in urine using precolumn fluorescent derivatization with OPA/ET reagent was previously developed. In the present study, we improved the method for quantitative analysis of urinary free Gla. 1) Fluorescent strength of OPA/ET reagent reached a maximum level after 3 days from the preparation of the reagent. Then the fluorescent strength was maintained at least after 3 weeks by the addition of ethanthiol every 3 days. 2) An urine sample was diluted by 20 to 60-folds and a 2.5μl aliquot of the diluted urine sample containing about 1 to 2 pmol of Gla was injected into the ChemcoPak Liquid Chromatography Columns packed with Nucleosil 5SB. The mobile phase consisted of 0.12 M sodium citrate buffer (pH 5.28) and acetonitrile in the ratio 60:40. Under these conditions, Gla peak appeared in a retention time of about 7 to 10 minutes and was completely resolved from the other amino acids. Since the peak disappeared after the sample was subjected to decarboxylation treatment, the peak was confirmed to contain only Gla. 3) This method gave a linear standard curve in a range of 0.10-150pmol and allowed quantitative analysis of Gla in an amount as low as 0.1pmol. We used the standard curve in a range of 0.10-4.0pmol for urinary Gla analysis. This is a sensitive and simple assay of free Gla in urine which was subjected to only dilution without further treatment

Intake frequency of vegetables or seafoods negatively correlates with disease activity of rheumatoid arthritis

Objective: To clarify the relationship between dietary habit and disease activity of rheumatoid arthritis (RA). Methods: This study enrolled RA patients who met the ACR/EULAR 2010 classification criteria from Kyoto University Rheumatoid Arthritis Management Alliance (KURAMA) cohort in 2015. 22-item food frequency questionnaire (FFQ) was taken for the measurement of dietary habit in a single-institution cohort of RA (Kyoto University Rheumatoid Arthritis Management Alliance: KURAMA) in 2015. The disease activities of RA using the Disease Activity Score calculated based on the erythrocyte sedimentation rate (DAS28-ESR), Simplified Disease Activity Index (SDAI), Health Assessment Questionnaire (HAQ), and serum matrix metalloproteinase-3 (MMP-3) level, the use of disease-modifying anti-rheumatic drugs (DMARDs), disease duration, rheumatoid factor, anti-cyclic citrullinated antibody, and body mass index were also examined. All of them were combined and statistically analyzed. Results: 441 RA patients (81% women; mean age 65 years; mean disease duration 15 years) were enrolled from the KURAMA cohort. Average Disease Activity Score-28 using the erythrocyte sedimentation rate (DAS28-ESR) was 2.7. Univariate analysis showed that intake frequency of vegetables had a statistically significant negative correlation with disease activity markers, such as DAS28-ESR (ρ = −0.11, p<0.01), Simplified Disease Activity Index (SDAI) (ρ = −0.16, p<0.001), matrix metalloproteinase-3 (MMP-3) (ρ = −0.21, p<0.0001), and Health Assessment Questionnaire (HAQ) (ρ = −0.13, p<0.01). Factor analysis with varimax rotation was done to simplify the relevance of disease activity to various food items. 22 foods were categorized into five dietary patterns: “seafoods”, “vegetables/fruits”, “meats/fried foods”, “snacks”, and “processed foods”. The multivariate analysis adjusted for clinically significant confounders showed that “seafoods” had statistically significant negative correlations with DAS28-ESR (β = −0.15, p<0.01), SDAI (β = −0.18, p<0.001), MMP-3 (β = −0.15, p<0.01), and HAQ (β = −0.24, p<0.0001). “Vegetables/fruits” had statistically significant negative correlations with SDAI (β = −0.11 p<0.05), MMP-3 (β = −0.12, p<0.01), and HAQ (β = −0.11, p<0.05) Conclusions: These results suggest that high intake frequency of vegetables/fruits and/or seafoods might correlate with low disease activity

生活習慣病としての骨粗鬆症

Osteoporosis is diagnosed in the presence of increased risk of fracture, but not by the presence of fracture. Osteoporotic fracture is associated with increased mortality and decreased quality of life (QOL) in the elderly. Hip fracture has poor prognosis such that the mortality is high after the fracture and patients quite often need long-term care. Vertebral fracture has high incidence. Although its significance is often overlooked, it is associated with poor QOL, and decreased pulmonary and digestive function. Thus osteoporotic fracture is a preventive disease. Unfortunately, the significance of osteoporosis is not fully acknowledged in the general population. For the prevention of osteoporosis, the collaboration of those with diverse speciality is essential such as the one with medical doctors and dietitians, nurses, and health policy maker

A novel anticancer ribonucleoside, 1-(3-C-ethynyl-β-D-ribo-pentofuranosyl)cytosine, enhances radiation-induced cell death in tumor cells.

1-(3-C-Ethynyl-β-d-ribo-pentofuranosyl)cytosine (ECyd, TAS106) is a newly developed anti-tumor agent that targets RNA synthesis. We report here that a low dose of ECyd induces radiosensitization of caspase-dependent apoptosis and reproductive cell death in cells of the gastric tumor cell lines MKN45 and MKN28 and murine rectum adenocarcinoma Colon26. Flow cytometry demonstrated that TAS106 induced the abrogation of the X-ray-induced G2/M checkpoint. Western blot analysis showed that X rays increased the expression of cyclin B1, phospho-Cdc2 and Wee1, whereas co-treatment with X rays and TAS106 decreased the expression of these cell cycle proteins associated with the G2/M checkpoint. Furthermore, TAS106 was shown to decrease the radiation-induced expression of survivin but not Bcl2 and BclXL regardless of TP53 status and cell type. Overexpression of wild-type survivin in MKN45 cells inhibited the induction of apoptosis induced by co-treatment with X rays and TAS106. These results suggest that TAS106 enhances X-ray-induced cell death through down-regulation of survivin and abrogation of the cell cycle machinery

Counter-facing plasma guns for efficient extreme ultra-violet plasma light source

A plasma focus system composed of a pair of counter-facing coaxial guns was proposed as a long-pulse and/or repetitive high energy density plasma source. We applied Li as the source of plasma for improvement of the conversion efficiency, the spectral purity, and the repetition capability. For operation of the system with ideal counter-facing plasma focus mode, we changed the system from simple coaxial geometry to a multi-channel configuration. We applied a laser trigger to make synchronous multi-channel discharges with low jitter. The results indicated that the configuration is promising to make a high energy density plasma with high spectral efficiency