Expression of human telomerase reverse transcriptase, Survivin, DD3 and PCGEM1 messenger RNA in archival prostate carcinoma tissue.

INTRODUCTION: The wide spectrum of biological behavior displayed by prostate cancer (PCa) warrants investigation of potential PCa-specific biomarkers that could identify more aggressive tumor types and therefore provide prognostic value. Upregulation of expression of human telomerase reverse transcriptase (hTERT), Survivin, DD3 and PCGEM1 mRNAs in PCa lesions has recently been described. The purpose of this study was to evaluate the clinical value of detection of over-expression of these biomarkers in the diagnosis and prognosis of PCa. MATERIAL AND METHODS: Archival formalin-fixed, paraffin-embedded (FFPE) prostatectomy tissue from 26 patients with PCa (Gleason score 3-9, mean 7) and 14 patients with benign prostatic hyperplasia (BPH) were analyzed by reverse transcription polymerase chain reaction (RT-PCR) for semiquantitative transcript levels of hTERT, Survivin, DD3 and PCGEM1. In addition, 25 matched normal (MN) tissue samples were examined. The expression of biomarker mRNA relative to b2-microglobulin mRNA was determined using AlphaImager 2200 data analysis software. RESULTS: The biomarkers had sensitivities ranging from 91% to 100%. Clinical specificities evaluated with the BPH tissue were the following: hTERT mRNA (93%), DD3 mRNA (57%), Survivin (29%) and PCGEM1 (14%). Biomarker expressions were up to 13.5-fold higher in PCa tissue as compared to MN tissue. None of the tumor biomarkers showed a positive correlation with pathological stage and Gleason score. CONCLUSIONS: The results of this study indicate potential utility of the hTERT mRNA and DD3 mRNA as diagnostic but not prognostic biomarkers for PCa.

A prospective evaluation of the diagnostic and potential prognostic utility of urinary human telomerase reverse transcriptase mRNA in patients with bladder cancer.

Human telomerase reverse transcriptase (hTERT) mRNA expression has been considered a surrogate marker for telomerase activity based on its parallel detection in urological malignancies, including transitional cell carcinoma (TCC) of the bladder. The objective of this study was to prospectively evaluate the diagnostic performance of urine hTERT mRNA marker and urine cytology in the detection of bladder cancer. The multiplex hTERT/GAPDH (glyceraldehyde-3-phosphate dehydrogenase) reverse transcription polymerase chain reaction (RT-PCR) assay was employed to assess hTERT mRNA expression in urine sediments from 43 patients with clinically apparent TCC undergoing transurethral resection. Tumor grade and pathological stage were determined. The results of urine cytology were compared with urine hTERT mRNA expression. The control group consisted of 46 age-matched healthy volunteers without known urinary tract disease. The sensitivity of hTERT mRNA expression marker in the detection of bladder cancer was significantly better than urine cytology (95% versus 65%, p<0.001). The hTERT mRNA was detected with high sensitivity in both low and high grade tumors, and in superficial and invasive phenotypes. No correlation was seen between hTERT mRNA and the histopathological grade and stage. The specificity of urinary hTERT mRNA marker was 93.5%. The detection of hTERT mRNA expression in urine was a highly sensitive marker for the diagnosis of TCC of the bladder in this study. This urine-based marker shows promise as a non-invasive adjunct to cystoscopy in patients undergoing bladder tumor surveillance.

Adhesion of coagulase-negative staphylococci grouped according to physico-chemical surface properties.

Physico-chemical cell surface properties of 23 coagulase-negative staphylococcal strains, including contact angles, zeta potentials and elemental cell surface composition were measured, together with the adhesion of all strains to hexadecane. The data were employed in a hierarchical cluster analysis, revealing that the 23 strains comprised essentially four different groups. Groups I-III were somewhat similar to each other, but group IV was markedly distinguished from the other strains, predominantly through an elevated acidity of the cell surface. These group distinctions were not related to the presence of a capsule or slime on the strains. Adhesion of the strains to hexadecane depended critically on electrostatic interactions between the hexadecane and the staphylococci, and adhesion only occurred when the electrostatic repulsion between hexadecane and the micro-organisms was less than 500 kT at closest approach. Adhesion of six representative strains from all four groups in a parallel plate flow chamber to silicone rubber, an implant material with similar hydrophobicity to hexadecane, did not show such a critical dependence, nor did it relate with the group distinction. Possibly, microbial adhesion to substratum surfaces like silicone rubber is more complicated than adhesion to an ideally smooth and homogeneous hexadecane surface in an aqueous solution. Adhesion of all six strains to silicone rubber with an adsorbed conditioning film of plasma proteins was less than that to bare silicone rubber: initial deposition rates dropped from 2000-3000 cm-2 s-1 to 100-300 cm-2 s-1 after adsorption of plasma proteins, while the stationary end-point adhesion decreased from 10 x 10(6)-15 x 10(6) cm-2 to 1 x 10(6)-5 x 10(6) cm-2. The adhering staphylococci poorly withstood the passage of an air-bubble through the parallel plate flow chamber, regardless of the presence of a conditioning film, indicating a low affinity of these relatively hydrophilic strains for hydrophobic substratum surfaces.