Inflammation has been suggested to play an important role in onset and progression of prostate cancer (PCa). Histological analysis of prostatectomy specimens has revealed focal inflammation in early stage lesions of this malignancy. We addressed the role of inflammatory stimuli in the release of PCa-specific, tumor-derived soluble factors (PCa-TDSFs) already reported to be mediators of PCa morbidity, such as indoleamine 2,3-dioxygenase (IDO) and interleukin (IL)-6. Inflammation-driven production and functions of PCa-TDFSs were tested “in vitro” by stimulating established cell lines (CA-HPV-10 and PC3) with IFN-γ or TNF-α. Expression of genes encoding IDO, IL-6, IFN-γ, TNF-α, and their receptors was investigated in tumor tissues of PCa patients undergoing radical prostatectomy, in comparison with benign prostatic hyperplasia (BPH) specimens. IFN-γ and TNF-α-treatment resulted in the induction of IDO and IL-6 gene expression and release in established cell lines, suggesting that the elicitation of PCa-TDSFs by these cytokines might contribute to progression of cancer into an untreatable phenotype. An analysis based on timing of biochemical recurrence revealed the prognostic value of IDO but not IL-6 gene expression in predicting recurrence-free survival in patients (RFS) with PCa. In addition, a urine-based mRNA biomarker study revealed the diagnostic potential of IDO gene expression in urines of men at risk of PCa development.

clinical settings, but calculated estimates of GFR appear to provide equally good information at much lower cost. In keeping with current best practice guidelines (12 ), we recommend that calculated estimates of GFR should supplement serum creatinine measurement alone and replace measured creatinine clearance.

This preliminary analysis revealed that the median values of a panel of mRNAs from the maternal blood of PE patients were different from those of the same gestational age control group at the third trimester. If prospective studies at the second trimester could detect a related marker sufficiently able to discriminate between affected and unaffected patients and thus detect the disease before its clinical onset, then a screening project using a panel of mRNAs would be feasible.

The ranges of the RQ values were calculated by use of the equation: RQ ϭ 2 Ϫ⌬⌬Ct , with ⌬⌬Ct ϩ SD and ⌬⌬Ct Ϫ SDs. The Micro Fluidic Cards detect a 2-fold difference in gene expression at the 99.7% confidence level. The efficiencies of all target and reference amplifications were nearly identical as analyzed by serial dilutions using 2, 1, 0.5, 0.25, and 0.125 ng of calibrator cDNA. When we plotted the log input amount of cDNA vs the ⌬Ct values, the slope was Ͻ0.1. The correlation coefficients were always Ͼ0.95, which is comparable to our previously established method (5 ). We assessed the within-run and day-to-day imprecision of our assay by measuring 4 identical samples in duplicate; the CVs were Ͻ1.6% and Ͻ2.9%, respectively.As can be seen in Table 1, 30 ABC transporters displayed detectable mRNA expression in human primary monocytes, based on ⌬Ct values Ͼ25. Twenty-six genes were induced more than 2-fold during macrophage differentiation, and 12 genes were sensitive to LXR/RXR agonist incubation, with 10 up-regulated ABC transporters and only 2 down-regulated genes. Although the differentiation-dependent and sterol-regulated induction of ABCA1 and ABCG1 is well established (7 ), parallel transcript profiling, using our Human ABC Transporter TaqMan Low-Density Array, revealed several additional differentiation-dependent ABC transporters and novel LXR/RXR-regulated ABC transporters, including ABCB1 (MDR1), ABCB9, ABCB11 (BSEP), ABCC2 (MRP2), ABCC5 (MRP5), ABCD1 (ALD), ABCD4, and ABCG2. Despite the large dynamic range of gene regulation (e.g., a 24.5-fold induction of ABCA1 by 9-cis RA/T0901317), the range of differential transcript expression between 3 different donors was quite narrow (21.9 -27.4). This implies that the interindividual differences in response are much smaller than the differentiation-dependent and lipid-regulated effects, allowing screening processes with a limited number of probands.These findings are particularly relevant for lipidomics and cardiovascular research, as these ABC transporters are novel candidates for lipid disorders and pharmacologic targets for lipid-modulating drugs.In summary, we have developed a Human ABC Transporter TaqMan Low-Density Array based on the TaqMan chemistry, the Micro Fluidic Card, and the 7900HT Sequence Detection System. Compared with standard TaqMan reverse transcription-PCR methods, the Micro Fluidic Card requires less sample material (2 ng instead of 50 ng of total RNA-equivalents per gene), only one-tenth the volume of TaqMan Universal Master Mix, and much less hands-on time. This assay could be a useful tool for monitoring dysregulated ABC transporter mRNA profiles in human lipid disorders and cancer-related multidrug resistance. Furthermore, the pharmacologic and metabolic regulation of ABC transporter gene expression important for drug development could be analyzed in large screening approaches using this Human ABC Transporter TaqMan Low-Density Array.We thank Ernst Arnoldi, Astrid Potratz, and Andrea Geiger for support and discussions, and Manfred H...

amount of Sepharose beads can easily be adjusted to the amount of sample available. This has two advantages: (a) To prevent contamination, it is best to allow each sample a fresh batch of Sepharose beads. This can easily be done with our method, whereas with columns, this could be more expensive (in terms of the number of columns and antibodies needed) and requires larger sample volumes. (b) Because of the low-volume elution, proteins remain present at high concentrations. This method is therefore suitable for extracting and concentrating low-abundance (g/L) proteins from small sample quantities without the risk of cross-contamination. For example, using this highly sensitive method, we were the first to be able to detect cTnT fragments in serum samples at concentrations below the 0.010 g/L detection limit of the third-generation cTnT assay (7, 8 ) (see Fig. 2 in the online Data Supplement). Moreover, this method allows us to further investigate cTnT fragmentation and clearance in vivo.We greatly acknowledge Vincent Kleijnen for technical assistance.