Five patients with chronic post-operative hypoparathyroidism were treated with 450 \g=m\g/day5,6-trans-25-hydroxyvitamin D3 (5,6-trans-25OHD3) for 14 days, and the treatment was continued with 150 \g=m\g/day for one year. At the end of this period the patients received 450 \g=m\g/day 5,6-cis-25\x=req-\ hydroxyvitamin D3 (5,6-cis-25OHD3) for 14 days. Comparison of the effects of both isomers revealed a similar ability to enhance intestinal calcium absorption and to normalize serum calcium; serum phosphate and alkaline phosphatase, however, remained unaffected. Urinary phosphate and hydroxyproline excretion decreased on the cis-isomer and increased on the trans-isomer. During treatment with the lower dose of 5,6-trans\x=req-\ 25OHD3 intestinal calcium absorption remained in the normal range for one year, whereas the serum calcium decreased to the levels observed before administration of 450 \g=m\g/day within 6 weeks. The results suggest that in hypoparathyroidism 5,6-cis-25OHD3 and 5,6-trans-25OHD3 are equally effective on serum calcium and on intestinal calcium absorption, but that their mode of action on renal phosphate handling and on calcium release from bone is different. Treatment of hypocalcaemia in hypoparathyroidism requires pharmacological doses of vitamin D (Ireland et al. 1968). The refractoriness to the actions of vitamin D has been found to be due to an impaired conversion of the vitamin to its metabolically active form in this condition (Kooh et al. 1975; Russell et al. 1974). Vitamin D3 is hydroxylated in the liver to 25-hydroxyvitamin D3.In a second step this metabolite undergoes renal conversion to 1,25-dihydroxyvitamin D3, the active form of the vitamin. The renal production of 1,25-di-