Vascular endothelial growth factor A,secreted in response to transforming growth factor-β1 under hypoxic conditions,induces autocrine effects on migration of prostate cancer cells

Hypoxia and transforming growth factor-β1 (TGF-β1) increase vascular endothelial growth factor A (VEGFA) expression in a number of malignancies. This effect of hypoxia and TGF-β1 might be responsible for tumor progression and metastasis of advanced prostate cancer. In the present study, TGF-β1 was shown to induce VEGFA₁₆₅ secretion from both normal cell lines (HPV7 and RWPE1) and prostate cancer cell lines (DU145 and PC3). Conversely, hypoxia-stimulated VEGFA₁₆₅ secretion was observed only in prostate cancer cell lines. Hypoxia induced TGF-β1 expression in PC3 prostate cancer cells, and the TGF-β type I receptor (ALK5) kinase inhibitor partially blocked hypoxia-mediated VEGFA₁₆₅ secretion. This effect of hypoxia provides a novel mechanism to increase VEGFA expression in prostate cancer cells. Although autocrine signaling of VEGFA has been implicated in prostate cancer progression and metastasis, the associated mechanism is poorly characterized. VEGFA activity is mediated via VEGF receptor (VEGFR) 1 (Flt-1) and 2 (Flk-1/KDR). Whereas VEGFR-1 mRNA was detected in normal prostate epithelial cells, VEGFR-2 mRNA and VEGFR protein were expressed only in PC3 cells. VEGFA₁₆₅ treatment induced phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) in PC3 cells but not in HPV7 cells, suggesting that the autocrine function of VEGFA may be uniquely associated with prostate cancer. Activation of VEGFR-2 by VEGFA₁₆₅ was shown to enhance migration of PC3 cells. A similar effect was also observed with endogenous VEGFA induced by TGF-β1 and hypoxia. These findings illustrate that an autocrine loop of VEGFA via VEGFR-2 is critical for the tumorigenic effects of TGF-β1 and hypoxia on metastatic prostate cancers.     

Comparing diagnostic tests: trials in people with discordant test results

Diagnostic tests are traditionally compared for accuracy against a gold standard but can also be compared prospectively in a trial. A conventional trial comparing two tests would randomize each participant to a testing strategy, but a more efficient alternative is to give both tests to all participants and follow up those with discordant results. Participants could be randomized before or after testing. The statistical analysis of such a trial has not previously been described. We investigated two estimates of the risk difference for a binary outcome: one based on analysing outcomes as if from a conventional trial and one combining estimates of different parameters in the manner of a decision analysis. We show that the trial estimate and decision analysis estimate are both unbiased and derive approximate formulae for their standard errors. By using the decision analysis estimate (but not the trial estimate), the same precision can be achieved by randomizing before testing as by randomizing after. To avoid destroying equipoise, and to allow consenting and randomizing to be carried out at the same visit, we recommend randomizing before testing. Giving both tests to all participants means fewer need to be recruited: in one example from the literature, the proposed design was nearly four times more efficient in this sense than a conventional trial design. Copyright © 2012 John Wiley & Sons, Ltd.     

Rasch-based scoring offered more precision in differentiating patient groups in measuring upper limb function

ObjectiveTo compare the discriminatory ability of Rasch-based and summative scoring in the context of assessing upper limb function of patients with stroke.Study Design and SettingData were from a cohort study of 497 adults with stroke undergoing physiotherapy. Upper limb function was assessed at admission and discharge using the upper limb subscale of the Motor Assessment Scale (UL-MAS). Rasch analysis was used to transform raw UL-MAS scores into interval measures. A relative precision (RP) index was used to differentiate patients by discharge destination.ResultsThe analysis confirmed the unidimensional structure of UL-MAS at both admission and discharge and demonstrated the adequate fit of the items. The RP index favored the Rasch-based scoring over the summative scoring in differentiating between the two patient groups, with significant gains in precision at admission (15%) and discharge (11%). When examining patients in the upper or lower quartile of UL-MAS, the gains in precision were statistically significant in favor of the Rasch-based scoring, with 20% precision at admission and 19% precision at discharge.ConclusionRasch-based scoring was more precise in differentiating patient groups by discharge destination than the summative scoring used to measure upper limb function, especially at the extreme range of the scale.