Measurement of the total purine contents and free nucleosides, nucleotides, and purine bases composition in Japanese anchovies (Engraulis japonicus) using high-performance liquid chromatography with UV detection.

Dietary purine restrictions are recommended for patients with hyperuricemia and gout. While measuring the purine contents of various foods in our laboratory using high-performance liquid chromatography (HPLC), we observed and reported changes in purine composition. In this study, we measured the total purine content and free purine of raw anchovies as well as after fermentation, using two methods by HPLC. Method 1 involved acid hydrolysis of all purines, such as nucleic acids and nucleotides, to form four corresponding purine bases. Method 2, which is a non-hydrolysis method, is used to measure the amount of free purines (nucleotide, nucleoside, purine base). As a result of method 1, after fermentation, adenine-related and hypoxanthine-related purines and the total purine levels decreased significantly. Regardless of being raw or fermented, each anchovy contained mainly hypoxanthine- and guanine-related purines. Among the hypoxanthine-related purines, the results of method 2 revealed that the raw anchovies contained a lot of inosine monophosphate (IMP), while after fermentation contained more inosine. In guanine-related and adenine-related purines, those nucleotides decreased by fermentation and nucleosides and bases increased. Measurements of free purines revealed that those reductions after fermentation observed in method 1 were derived from decreased nucleotides. These results indicate that purines are affected by the fermentation bacteria and period.

The observed variation in the purine composition of food after soaking in sake lees.

In this study, we investigated the alterations in the purine composition of swordfish prepared using a traditional Japanese processing method of soaking in sake lees. These alterations are the byproducts of the yeast fermentation of rice-koji and are renowned for enhancing the umami nature of food. Using a conventional assay method for hydrolyzing all of the purines into four bases and our developed method for simultaneously analyzing purines, we observed the alterations in four purine bases in the soaked sake lees and swordfish. The findings showed that the total purine content, and hypoxanthine-related and guanine-related purines in swordfish decreased after soaking in sake lees. We also analyzed the free purine composition and showed that the ratio of IMP in swordfish was decreased by soaking, while that of inosine in sake lees was increased by soaking swordfish in it.

[Neo-adjuvant therapy in childhood osteogenic sarcoma: a pilot study of selective postoperative chemotherapy based on response to preoperative high-dose methotrexate].

"Neo-adjuvant therapy" with preoperative high-dose methotrexate (HD-MTX) and CF rescue therapy was investigated in four children with osteogenic sarcoma. Immediately after the diagnosis of osteogenic sarcoma from biopsy, the patients were treated with three to five courses of weekly HD-MTX (300 mg/kg) with CF rescue. Three patients had en bloc tumor resection and one patient underwent disarticulation of the hip joint after the pre-operative HD-MTX. The effect of HD-MTX was evaluated on the basis of pathological changes between the specimen of the primary tumor taken at biopsy and that during surgery. Two out of four patients showed marked tumor cell reduction (greater than 50%) of the specimen upon surgery. Two patients who responded to the preoperative HD-MTX were further treated with HD-MTX on a post-operative adjuvant therapy basis for 18 months. Both of these patients survived with no evidence of disease for 35.6+ and 20.9+ months. Two patients who responded poorly to HD-MTX were treated with a multi-drug postoperative adjuvant therapy including cis-platinum, adriamycin, cyclophosphamide, actinomycin D, and bleomycin. One patient had a solitary lung metastasis at 12.2 months after amputation. Wedge resection of the metastatic tumor was performed and adjuvant therapy with cis-platimum has been given for 20 months. He has remained with no evidence of disease for more than 30 months. Another patient has been receiving multi-drug neo-adjuvant therapy without any evidence of disease for 11.9 months after surgery. These data suggest that neo-adjuvant chemotherapy based on the response to preoperative HD-MTX is more useful for increasing the cure rate of childhood osteogenic sarcoma.