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opportunities and challenges in the development of non-obstructive azoospermia options: a way forward for assisted reproductive technology in Japan. With the introduction of intracytoplasmic sperm injection (ICSI) in 1989, this technique has resulted in remarkable improvements in the fertilisation rate and achievement of pregnancy for couples with male infertility. However, ICSI has raised the possibility of spermatogenesis disorders such as azoospermia. A non-obstructive azoospermia (NOA) refers to unexplained azoospermia; it can be of obstructive (OA) or non-obstructive (NOA) azoospermia. We have been conducting basic research and clinical trials on the biological significance and the clinical application of patients with NOA for more than 10 years. In this review, we discuss the recent progress in the development of testis biopsy and sperm retrieval techniques as well as the natural history of NOA. Finally, we discuss the current opportunities and challenges in the development of treatment options for NOA patients.Structural and functional studies of vitamin D-3-26-hydroxylase. Vitamin D-3-26,26-dihydroxyvitamin D-3-26-hydroxylase catalyzes the hydroxylation of vitamin D-3 at the carbon atom at position 26. It is found in rat kidney medulla and in human leukocytes. We previously purified and partially sequenced vitamin D-3-26-hydroxylase from rat kidney. In the present work, cDNAs for human leukocyte and rat kidney vitamin D-3-26-hydroxylase have been cloned and sequenced. The human leukocyte vitamin D-3-26-hydroxylase differs from the rat enzyme only in that it has an insertion of 43 amino acids. The insert contains a bipartite nuclear localization signal and glutamic acid, arginine, valine and leucine rich sequences. In the rat kidney, cDNA clones encoding for the enzyme are composed of four exons and three introns. Two alternate first exons were found in the last intron, one encoding a cDNA for leukocyte vitamin D-3-26-hydroxylase and the other for renal vitamin D-3-26-hydroxylase. The sizes of the respective mRN
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