Routes t o hydroxychlorin derivatives of two distinct categories (nuclear hydroxy substituent; sidechain hydroxy substituent) are developed. Such substances, covering a range of amphiphilic character depending on the number of hydroxy groups, are seen as potential sensitisers for photodynamic therapy. Osmylation of octaethylporphyrin gives the dihydroxyoctaethylchlorin 2 and the tetrahydroxybacteriochlorin 3. Pinacol-pinacolone rearrangement of 2 gives the octaethyl-poxochlorin 4, borohydride reduction of which gives the secondary alcohol 6. This is converted via the bromochlorin 7 (prepared using 50% HBr/HOAc at room temperature) into a series of ethers, including those, 10, 11, 12, derived from glycerol, D-glucose, and D-mannitol respectively. Alkylation of these unprotected polyols with the highly hindered bromide 7 occurs preferentially at the primary alcohol functions in each case. Some of these hydroxychlorins are found in animal assays to be highly effective sensitisers of tumour photonecrosis. In this respect the most effective compounds in each category are the most highly hydroxylated. However, the D-glucose derivative 11 proves also to be an effective sensitiser of skin and muscle, i.e. it does not show the selectivity shown by 5,10,15,20-tetra(m-hydroxypheny1)chlorin.

No abstract

The accurate measurement of testosterone remains a challenge. The determination of the blood testosterone concentrations in serum by conventional immunoassays is inaccurate in men and even more so in females and children. A new luminescence enzyme immunoassay (LIA) has been developed and validated. The high analytical (8.7 pmol/L) and functional (17.3 pmol/L) sensitivity allows the quantification of the very low concentration in saliva, as well as in serum, after 1/40 dilution. This study measured salivary testosterone levels and compared the results with the free levels calculated from total testosterone and sex hormone-binding globulin in eugonadal and hypogonadal men. Salivary testosterone concentrations in healthy men in morning hours were 369 pmol/L (mean), range 263-544 pmol/L, which was statistically significantly higher than that in men with androgen deficiency, 215 pmol/L (mean), range 51-249 pmol/L. Repetitive determination of free testosterone concentrations in saliva (once a week for 5 weeks) showed high stability of results over time, with coefficient of variation 9% (range 5-23%). In this study we showed that free salivary testosterone levels in morning samples correlated well with calculated free testosterone in blood, both in healthy men (R = 0.754, P = 0.001), and in patients with androgen deficiency (R = 0.889, P = 0.0001), though in cases with very low testosterone, salivary concentrations were systematically higher than calculated free testosterone levels in blood.

No abstract

Background: Deficiency of 17a-hydroxylase/17,20-lyase is a rare cause of 46,XY disordered sex development. Objective: We characterize in vitro and in vivo effects of two novel CYP17A1 gene mutations identified in a patient with a mild phenotype of CYP17A1 deficiency. Subjects and methods: A 46,XY patient presented with ambiguous genitalia. CYP17A1 deficiency was suspected at 2 months on the basis of steroid analysis performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Mutational analysis of the CYP17A1 gene was performed by PCR and Sanger sequencing. To characterize the effect of CYP17A1 mutation on 17a-hydroxylase and 17,20-lyase activities in vitro, HEK293 cells were transiently transfected with CYP17A1 expression plasmids, incubated with progesterone or 17-OH-pregnenolone and concentrations of 17-OH-progesterone or DHEA were then measured in the cell culture medium by LC-MS/MS. Results: Clinical and hormonal findings in the patient were consistent with partial combined deficiency of 17a-hydroxylase/ 17,20-lyase. The sequencing of the CYP17A1 gene in the patient revealed compound heterozygosity for two novel mutations: c.107delT p.R36fsX107 and p.W121R. After 6-h in vitro culture of transfected HEK293 cells in the presence of 1 mM progesterone, 17a-hydroxylase activity of p.W121R mutant was 60.5G16.3%, while 17,20-lyase activity of mutant measured from the amount of DHEA produced in the presence of 1 mM of 17-OH-pregnenolone was 15.8G2.6% compared with the WT. Conclusions: p.W121R substitution, affecting the first residue in the conserved heme-interacting WXXXR motif of CYP17A1, is associated with partial combined deficiency of 17a-hydroxylase/17,20-lyase.