We have developed a simple, robust assay to measure salivary cortisol using on-line solid-phase extraction to reduce sample clean-up requirements.

Background: Insulin analogues used in the treatment of insulin-dependent diabetes may result in hypoglycaemia; however, these preparations show variable cross-reactivity in many insulin assays. In order to investigate a suspected insulin overdose, it is therefore essential to characterize the cross-reactivity of these preparations in insulin assays. Methods: The cross-reactivity of 12 commercial insulin preparations was assessed on the Beckman DxI analyser, by comparison with the insulin calibration curve corresponding to the midpoint of the dose-response curve. This was achieved by manually plotting the relative light units generated by the analyser for each preparation. Results: All 12 insulin preparations examined showed significant cross-reactivity with the Beckman DxI insulin assay, with 11 of the insulin preparations demonstrating cross-reactivity of 83% or greater. The lowest cross-reactivity was observed with detemir with a value of 47.6%. Conclusion: When investigating hypoglycaemia it is important to be aware of the cross-reactivity of the recombinant insulin analogues in insulin assays. This study has established the cross-reactivity of 12 preparations in the Beckman DxI insulin assay.

Context: Altered hepatic cortisol-cortisone metabolism by type 1 11␤-hydroxysteroid dehydrogenase (11␤HSD1) has previously been linked with polycystic ovary (PCO) syndrome (PCOS).Objectives: Our objectives were to establish whether ovarian 11␤HSD activities are also altered in PCOS and to determine whether any changes in ovarian cortisol metabolism might reflect exposure to elevated concentrations of insulin or androgens.Design: Cortisol and cortisone concentrations were measured in follicular fluid aspirated from size-matched follicles dissected from normal, ovulatory, and anovulatory PCOs. Human granulosalutein cells, recovered during oocyte retrieval for assisted conception, were maintained in primary culture for 4 days, after which 11␤HSD1 activities were measured as the net oxidation of(100 nmol/L) in the absence and presence of insulin (100 nmol/L) with or without metformin (1 mol/L) or a range of androgens/oxy-androgen metabolites (0.01-10 mol/L).Results: Intrafollicular cortisol to cortisone ratios were elevated in anovulatory PCOs (2.1 Ϯ 0.4, P Ͻ .05, n ϭ 13) but did not differ between follicles from ovulatory PCOs (1.6 Ϯ 0.1, n ϭ 24) and normal ovaries (1.2 Ϯ 0.1, n ϭ 14). 11␤HSD1 activities were lower in granulosa-lutein cells recovered from patients with PCOS compared with all other causes of infertility (median ϭ 5.8 vs 14.9 pmol cortisone/4 h, respectively; P Ͻ .05). Cortisol oxidation was unaffected by insulin with or without metformin, dehydroepiandrosterone, and androstenedione, but was inhibited in a concentrationdependent manner by testosterone, 11␤-hydroxyandrostenedione, and 7␣-and 7␤-hydroxy-dehydroepiandrosterone (P Ͻ .01). Conclusions:There is decreased inactivation of cortisol in follicles from anovulatory PCOS. This may reflect inhibition of 11␤HSD1 by androgens and their 7/11-oxy-metabolites, local concentrations of which are increased in PCOS, and may contribute to the block to folliculogenesis seen in PCOS.(J Clin Endocrinol Metab 98: 3375-3383, 2013) P olycystic ovary (PCO) syndrome (PCOS) is the predominant cause of anovulatory infertility (1). This endocrine disorder, which affects up to 10% of premenopausal women (2), is characterized by PCOs accompanied by elevated plasma LH, hyperandrogenism, and peripheral insulin resistance/hyperinsulinemia (3). Recent studies have explored the role of glucocorticoids in the pathophysiology of PCOS. Within potential target cells, the

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