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Abstract.We examined the effects of insulin on bone formation including the mineralizing process. Twenty-day fetal rat parietal bones were cultured for 96 h on grids in a serum-free medium. For the precise assessment of bone formation, histomorphometry, with an image analyzing system, was used to measure the areas of mineralized bone and bone matrix, and the numbers of osteoblasts and osteoclasts. In order to confirm the effects of insulin on bone mineralization, the calcium content of the bone and the release of previously incorporated 45Ca into the medium were measured. Insulin, at a concentration of 10-6 M or higher, increased the areas of mineralized bone and bone matrix, and the number of osteoblasts. Osteoclasts were seldom observed in bones on day 0 or in bones treated with insulin. In bones treated with insulin at a concentration of 10-6 M, the calcium content of bone increased. At an insulin concentration of 10-' M or higher, the dry weight of decalcified bone increased. Lactate production in the medium increased dose-dependently. The inhibited release of 45Ca in bones indicated that insulin acts by increasing calcium retention. We demonstrated that insulin has an effect on bone-forming and bone-resorbing cells to enhance the bone forming process from matrix formation to mineralization.

Clostridium difficile is a major causative agent of antimicrobial-associated diarrhea, and the leading cause of nosocomial diarrhea. We clarified intestinal colonization and nosocomial spread of C. difficile in pediatric cancer patients undergoing antineoplastic therapy during long-term hospitalization. Subjects were 10 children with pediatric malignant diseases admitted from November 2005 to December 2006, aged 5 to 15 years, who received antineoplastic agents. Stool specimens were examined at hospitalization, after each course of treatment with antineoplastic chemotherapy, and when symptoms such as diarrhea or fever occurred. While C. difficile was detected from stool specimens of 8 of 10 children during their hospital stay, 6 of these 8 children were negative for C. difficile on the day of their admission. These results demonstrate that the use of antimicrobial agents and antineoplastic agents lead to overgrowth of C. difficile in intestinal tract of pediatric cancer patients. Five of the 8 children carried toxin A-positive, toxin B-positive C. difficle and 2 were diagnosed with C. difficile-associated diarrhea (CDAD). This demonstrates that CDAD is not a rare infection in pediatric cancer patients. Nine C. difficile isolates from 8 children were analyzed by PCR ribotyping. Two isolates from 2 children were typed into the same type;banding patterns of the remaining 7 isolates from 6 children were unique.

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We investigated the bioactivities of the recombinant DNA-derived methionyl 20K hGH (20K-Met-hGH) and methionyl hGH (22K-Met-hGH).The growthpromoting activities in Nb2 cells of 20K-Met-hGH and 22K-Met-hGH were 10.7% and 93.5% of pituitary hGH (P-hGH), respectively.In the IM-9 lymphocyte assay, the binding activities of 20K-Met-hGH and 22K-Met-hGH to hGH receptor were 29.0% and 87.1% of P-hGH, respectively. Our data demonstrate that 20K-Met-hGH may have weaker biological potency than P-hGH.