Aromatase expression in human peripheral blood leucocytes (PBLs) and in various tissues in primates: studies in elderly humans and cynomolgus monkeys.

BACKGROUND: Previous analysis of aromatase gene and protein expression in peripheral blood leucocytes (PBLs), studied in children and adults, was extended to elderly subjects. In addition, we assessed whether aromatase expression in PBLs could be used as a parameter of aromatase expression in other tissues, using the cynomolgus monkey as model. METHODS: Real-time PCR analysis of aromatase gene expression and protein evaluation by Western blot was performed in PBLs of human elderly subjects and in various tissues from cynomolgus monkeys. RESULTS: No gender-related difference in CYP19A1 mRNA and protein expression in PBLs from human elderly women and men was found. In elderly male cynomolgus monkeys, CYP19A1 mRNA and protein were expressed in all cells and tissues analysed, with the lowest levels in PBLs but no clear-cut correlation with other tissues. CONCLUSIONS: Aromatase expression in PBLs in elderly human subjects is not gender-related and cannot be a surrogate of aromatase expression for other tissues.

Oxidative DNA damage in human sperm can be detected by Raman microspectroscopy.

OBJECTIVE: To determine whether Raman microspectroscopy can identify different levels of oxidative sperm nDNA damage and to corroborate the findings using an established method and an alternative but complementary spectroscopic technique. DESIGN: Three-way comparison of Raman profiles, Fourier transform infrared spectroscopy (FTIR) spectra, and flow-cytometric assessments of sperm nDNA damage. SETTING: University-based research laboratory. PATIENT(S): Thirty-eight men attending the infertility clinic at the Centre of Reproductive Medicine and Andrology. INTERVENTION(S): Induction of oxidative damage by Fenton's reaction on semen samples. MAIN OUTCOME MEASURE(S): Raman profiles, FTIR spectra, and flow-cytometric analysis of DNA fragmentation. RESULT(S): Raman and FTIR spectra contained distinctive differences between untreated and fragmented nDNA sperm that were indicative of oxidative attack. The changes in Raman profiles were similar to those previously seen and corresponded to the DNA backbone. The peak attributions were corroborated by the FTIR spectra. Principal component analysis of the entire Raman spectra distinguished samples with varying degrees of damage. After determination of a cutoff value (0.63), estimation of the percentage of sperm with nDNA damage using the intensity ratio of Raman peaks (1,050/1,095 cm(-1)) correlated linearly to the flow-cytometric assessment. CONCLUSION(S): Raman microspectroscopy still requires further validation but may potentially provide a means of assessing the nDNA status of a living sperm.

Pharmacokinetics and pharmacodynamics of injectable testosterone undecanoate in castrated cynomolgus monkeys (Macaca fascicularis) are independent of different oil vehicles.

Testosterone undecanoate (TU) dissolved in soybean oil was developed in China to improve the pharmacokinetics of this testosterone ester in comparison with TU in castor or tea seed oil. As a pre-clinical primate model, three groups of five castrated cynomolgus macaques received either a single intramuscular injection of 10 mg/kg body weight TU in soybean oil, in tea seed oil, or in castor oil (equals 6.3 mg pure T/kg body weight for all preparations). Testosterone, estradiol, luteinizing hormone, and follicle-stimulating hormone as well as prostate volume, body weight and ejaculate weight were evaluated. After injection supraphysiological testosterone levels were induced. There were no significant differences in the pharmacokinetics of the three TU preparations for testosterone and estradiol. The gonadotropin levels showed a high individual variation. Prostate volumes increased equally in all groups after administration and declined to castrate level afterwards. The results suggest that TU in soybean oil produces similar effects as TU in the other vehicles. This study in non-human primates provides no objection to testing of this new preparation in humans.