Restricting type of anorexia nervosa (AN-R) is a serious disorder affecting adolescents and young adults and decreases quality of life over a long period. Successful weight restoration is an important prognostic factor for disease outcome; however, the underlying mechanism of refeeding resistance, a core psychopathology relevant to 'ambivalent' eating behaviors, remains unclear in this disorder. Ghrelin plays an important role in the regulation of growth hormone release, appetite, and energy metabolism. However, the early progress of these patients and changes in the levels of acyl ghrelin and des-acyl ghrelin during treatment were not reported. The purpose of this study was to determine the changes in ghrelin levels (acyl and des-acyl) during early treatment. As a result, des-acyl ghrelin in AN-R patients is higher than in control subjects before the therapy, but it decreases with treatment. The plasma des-acyl ghrelin level in AN-R patients started decreasing more rapidly and in early stage of the hospitalization than ever reported, and after 8 weeks, it is significantly lower than in control subjects. It means that des-acyl ghrelin is sensitive and changeable with their nutrition state. Furthermore, the ratio of the acyl ghrelin to total ghrelin increases with 8 weeks treatment. Eight weeks after, energy intake of the AN-R patients is recovered near the normal range with a daily energy intake of 1 700+/-93.54 kcal. These findings may be valuable for future AN-R treatments in order to increase acyl ghrelin and decrease des-acyl ghrelin, thereby influencing the refeeding outcome.

Quinoline is a chemical with potential pharmaceutical components, such as antimalaria, antiulcer, and antibiotic agents. Quinoline is metabolized by CYP2A6, whose activity is generally shown by coumarin 7-hydroxylation, and the principal product is the 5,6-epoxide of quinoline. We found coumarin 7-hydroxylase activity in bovine liver microsomes and studied the interaction of quinoline and some quinoline derivatives with coumarin 7-hydroxylase activity by fluorometry. Quinoline inhibited coumarin metabolism, and the apparent V max value decreased to 0.39 nmol/ min/nmol cytochrome P-450 (CYP) in the presence of quinoline from the value (V max = 0.63 nmol/min/nmol CYP) in the absence of quinoline. 5-fluoroquinoline (5FQ), 6-fluoroquinoline (6FQ) and 8-fluoroquinoline (8FQ) showed stronger inhibition than quinoline, whereas 3-fluoroquinoline (3FQ) showed weaker inhibition (apparent V max was 0.59 nmol/min/nmol CYP). Almost the same inhibition pattern of fluoroquinolines were found in assays of cDNAexpressed human CYP2A6. The results suggest that bovine CYP2A enzymes (s) as well as human CYP2A6 can interact strongly with monofluoroquinolines such as 5-, 6-, and 8-FQ, but weakly with 3-FQ.

Restricting-type of anorexia nervosa (AN-R) is a serious disorder affecting adolescents and young adults, and decreases quality of life over long period. Successful weight restoration is an important prognostic factor for disease outcome; however, the underlying mechanism of refeeding-resistance, a core psychopathology relevant to 'ambivalent' eating behaviors, remains unclear in this disorder. Obestatin plays an important role in the regulation of growth hormone release, appetite, and energy metabolism. However, the progress of these patients and changes in the levels of obestatin during treatment were not reported. The purpose of this study was to determine the changes in obestatin levels when energy intake increases in AN-R patients. As a result, obestatin was higher in AN-R patients than in control subjects as well as acyl ghrelin and des-acyl ghrelin. An increase in the intake calorie has decreased obestatin as well as des-acyl ghrelin. These findings indicate that the obestatin is an important factor in the diagnosis and treatment of AN-R, similarly to acyl ghrelin and des-acyl ghrelin. In the future, the research on the clinical application of the ghrelin peptide family and the receptor will be expected to progress.

We previously reported that materials leached from bioactive glass (BG) and vitamin D3 induced the formation of nodules on cultured periodontal-ligament fibroblasts (PLF). In this study, we have investigated the relationship between the conditions of the materials and nodule formation, analyzed morphologically, and also studied whether the production of nodules was specific to cultured PLF. PLF and skin fibroblasts were cultured in the presence or absence of BG. The amounts of calcium, phosphate, sodium and silicon in the culture medium and the number of nodules were measured at the 55th day. The nodules were observed microscopically and analyzed using an X-ray microanalyzer. In PLF, nodules were formed regardless of the presence or absence of BG; however, they were more numerous in the presence of BG. In skin fibroblasts, nodules were not observed. The amounts of calcium and silicon were higher in the presence of BG, while the amount of phosphate was lower. The nodules appeared crystalline with a spongy structure and contained calcium and phosphorus. Our results show that the nodules were associated with PLF and precipitated by the materials (higher concentrations of calcium and silicon), and they were spongy crystal composed of calcium and phosphorus.

Previously, the authors reported that bioactive glass promoted formation of nodules in cultured periodontal-ligament fibroblasts and high concentrations of calcium and silicon were detected in the culture medium. Herein, the effects of bioactive glass, calcium alone and calcium and silicon for formation of nodules on the periodontal-ligament fibroblasts are examined. The cells were cultured with bioactive glass, without bioactive glass, calcium and silicon, with calcium alone and with calcium and silicon. The cell growth, alkaline-phosphatase activity, amount of pro-collagen type I and the number of nodules were measured periodically until the 21st day. The cell growth rate with calcium and silicon was the highest, however the rate with bioactive glass and calcium alone were similar to that without them. The alkaline-phosphatase activity and the amount of pro-collagen showed peaks at the 11th day. At the peak, the alkaline-phosphatase activity with bioactive glass or calcium and silicon, and the pro-collagen with bioactive glass was higher than without them. The number of nodules with bioactive glass, calcium alone or calcium and silicon was greater than without them. These findings suggested that the combination of calcium and silicon released from bioactive glass was concerned with the formation of nodules on the periodontal-ligament cells.