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Primer sets specific for complement proteins, C3, factor B and factor I, were designed and used to amplify cDNA from cultured human astrocyte mRNA by the polymerase chain reaction. Appropriately sized PCR products of 506 bp, 885 bp and 146 bp, respectively, were generated and specificity was confirmed with Southern blotting using an enhanced chemiluminescence detection system. The sensitivity of detection was high, with amplified product from cDNA of approximately 6250 cells readily visualized. C3 and factor B have previously been reported to be produced by murine astrocytes; however, this is the first report indicating synthesis of C3, factor B and factor I by human astrocytes. These results indicate that PCR is a simple and sensitive technique to detect mRNA transcripts for proteins of the alternative pathway of complement in human astrocytes.

Background/Aim This study aimed to determine the prevalence of Mycoplasma genitalium infection among male patients with dysuria and/or urethral discharge. An analysis of the clinical, demographic and microbiological factors associated with M. genitalium infection was also conducted. Method From May 2007 to June 2011, men presenting to the clinic with self-reported symptoms of dysuria and/or urethral discharge were identified and underwent urethral swab, which was microscopically assessed for objective non-gonococcal urethritis. A first-void urine sample was tested for Chlamydia trachomatis and Neisseria gonorrhoeae using the Aptima Combo-2 assay. A portion of the urine sample was sent for polymerase chain reaction analysis for M. genitalium. Results One thousand, one hundred and eighty-two men with dysuria and/or urethral discharge were tested for M. genitalium. Of those, 96 men (8.1%) were positive for M. genitalium. Men identifying as solely MSM (men who have sex with men) constituted 16.3% ( n = 193) of the sample. Their infection rate was 3.1% ( n = 6). The infection rate for heterosexual and bisexual men was 9.1%. For all men, the M. genitalium co-infection rate was 14.6% ( n = 14) with C. trachomatis and 3.1% ( n = 3) with N. gonorrhoeae. Factors associated with M. genitalium infection were analysed by univariate analysis. We determined that five investigated predictors were significantly associated with M. genitalium infection, urethral discharge, non-gonococcal urethritis on Gram stain of urethral smears, identification as heterosexual or bisexual, and absence of co-infection with C. trachomatis or N. gonorrhoeae. Conclusion In Adelaide, M. genitalium is an important sexually transmitted infection among men with dysuria and/or urethral discharge, and is primarily an infection of heterosexual and bisexual men. [ABSTRACT FROM AUTHOR]