[25] with minor modifications. Briefly, a 20 μl PCR mixture contained 1 μM each of the primers, 10 μl of FastStart PCR master (Roche), 2 μl of Easymag DNA-extract of the samples JNK-IN-8 concentration and distilled water. Thermal cycling consisted of an initial denaturation of 2 min at 94°C, followed by 35 cycles of 30 sec at 94°C, 30 sec at 60°C and 1 min at 72°C, with a final extension of 10 min at 72°C, and cooling to 10°C. Detection and identification of fungi using fluorescent fragment length analysis of the ITS2-PCR amplicon and sequencing The amplification of the ITS2-region
and subsequent capillary electrophoresis was performed as previously described [26, 27]. Amplicons having a fragment length that was not present in the existing ITS2 library, which contains
most of the clinically Pictilisib concentration important yeast species, were sequenced as previously described [26]. Data analysis Distributions of continuous and discrete variables were summarized as means and standard deviations. Bivariate correlations are represented by Pearson’s R if the observed distribution approximated a normal distribution, either by Spearman’s rank correlation coefficient rho under the non-parametric assumption. Statistical significance was accepted at the conventional two-tailed α = 0.05 significance level. All analyses were performed with the statistical software package SPSS 15.0 (Chicago, IlIinois). Acknowledgements The authors would like to thank Dr. G. De Cuypere, Prof. P. Hoebeke and Dr.
G. T’Sjoen for recruiting the patients. We are of course also greatly indebted to all the patients participating in this study. SW was supported by an unrestricted grant donated by Besins-Healthcare® (Brussels, Belgium). This work was supported through research grant BOF08/GOA/002 of the Bijzonder Onderzoeksfonds of the University Idoxuridine of Gent (UGent). References 1. Fisk N: Gender dysphoria syndrome (the how, what and why of the disease). Proceedings of the second interdisciplinary symposium on gender dysphoria syndrome (Edited by: Laub D, Gandy P). Palo Alto, California: Stanford University Press 1973, 7–14. 2. Meyer W III, Bockting W, Cohen-Kettenis P, Coleman E, DiCeglie D, Devor H, Gooren L, Hage JJ, Kirk S, Kuiper B, Laub D, Lawrence A, LY333531 in vitro Menard Y, Patton J, Schaefer L, Webb A, Wheeler C: The Standards of Care for Gender Identity Disorders – Sixth Version. [http://www.symposion.com/ijt/soc_2001/index.htm]International Journal of Transgenderism 2001., 5: 3. Sohn M, Bosinski HA: Gender Identity Disorders: Diagnostic and surgical aspects. J Sex Med 2007, 4:1193–1208.CrossRefPubMed 4. Marrazzo JM: Evolving issues in understanding and treating bacterial vaginosis. Expert Rev Anti Infect Ther 2004, 2:913–22.CrossRefPubMed 5. Sobel JD: Bacterial vaginosis. Annu Rev Med 2000, 51:349–56.CrossRefPubMed 6.