Trace detection of the conformational transition of beta-amyloid peptide (Abeta) from a predominantly alpha-helical structure to beta-sheet could have a large impact in understanding and diagnosing Alzheimer's disease. We demonstrate how a novel nanofluidic biosensor using a controlled, reproducible surface enhanced Raman spectroscopy active site was developed to observe Abeta in different conformational states during the Abeta self-assembly process as well as to distinguish Abeta from confounder proteins commonly found in cerebral spinal fluid.

We have developed an optofluidic device that improves the sensitivity of surface enhanced Raman spectroscopy (SERS) when compared to other SERS approaches. This device has a pinched and step microchannel-nanochannel junction that can trap and assemble nanoparticles/target molecules into optically enhanced SERS active clusters by using capillary force. These SERS active clusters provide an electromagnetic enhancement factor of approximately 10(8). In addition, due to the continuous capillary flow that can transport nanoparticles/target molecules into the junction sites, the numbers of nanoparticles/target molecules and SERS active sites are increased. As a result, the detection limit of SERS for adenine molecules was better than 10 pM.

SignificanceWe describe unexpected cooperation between two cytokines that are important in regulating the growth of cancers, namely, type I interferons (IFNs) and interleukin 6 (IL6). It is well established that IL6 is vital for the ability of many tumor types to prosper, and the work in the current paper reveals that the signaling pathway driven by IFN, which is also evident in many cancers, increases the expression of IL6 through a direct effect on the IL6 gene. The findings may help to identify new antitumor targets for therapy.

Epidermal growth factor receptor (EGFR) is a clinical therapeutic target to treat a subset of non-small cell lung cancer (NSCLC) harboring EGFR mutants. However, some patients with a similar kind of EGFR mutation show intrinsic resistance to tyrosine kinase inhibitors (TKI). It indicates that other key molecules are involved in the survival of these cancer cells. We showed here that 2-[(aminocarbonyl)amino]-5 -(4-fluorophenyl)-3-thiophenecarboxamide (TPCA-1), a previously reported inhibitor of IkB kinases (IKK), blocked STAT3 recruitment to upstream kinases by docking into SH2 domain of STAT3 and attenuated STAT3 activity induced by cytokines and cytoplasmic tyrosine kinases. TPCA-1 is an effective inhibitor of STAT3 phosphorylation, DNA binding, and transactivation in vivo. It selectively repressed proliferation of NSCLC cells with constitutive STAT3 activation. In addition, using pharmacologic and genetic approaches, we found that both NF-kB and STAT3 could regulate the transcripts of interleukin (IL)-6 and COX-2 in NSCLC harboring EGFR mutations. Moreover, gefitinib treatment only did not efficiently suppress NF-kB and STAT3 activity. In contrast, we found that treatment with TKIs increased phosho-STAT3 level in target cells. Inhibiting EGFR, STAT3, and NF-kB by combination of TKIs with TPCA-1 showed increased sensitivity and enhanced apoptosis induced by gefitinib. Collectively, in this work, we identified TPCA-1 as a direct dual inhibitor for both IKKs and STAT3, whereas treatment targeting EGFR only could not sufficiently repress NF-kB and STAT3 pathways for lung cancers harboring mutant EGFR. Therefore, synergistic treatment of TPCA-1 with TKIs has potential to be a more effective strategy for cancers. Mol Cancer Ther; 13(3); 617-29. Ó2014 AACR.