Quantitative Single-Cell Analysis of Isolated Cancer Cells with a Microwell Array.

The heterogeneous nature of tumor-cell populations suggests that quantitative analysis at the single-cell level may provide better insights into cancer biology. Specifically, detection of multiple biomarkers from a single cell offers important initial information about cellular behavior. However, conventional approaches limit biomarker detection at the single-cell level. Here, we fabricated a polymer microwell array to capture single cells from prostate-cancer cell lines and quantitatively analyzed the expression of three different cancer-related biomarkers, CD44, EpCAM, and PSMA, without a membrane protein-extraction step. The resulting information on cell-surface biomarker distributions was compared with that from other standard analytical techniques. Interestingly, a large variation in CD44-expression levels was observed when the cell-proliferation cycle was modulated. On the other hand, the expression levels of EpCAM in three different cell lines are consistent among the different analytical methods with the exception of the microarray, where it has a different substrate material to adhere to. This observation clearly emphasizes that biomarker choice and environmental control are critical for properly understanding the single-cell state.

Sequential assessment via daphnia and zebrafish for systematic toxicity screening of heterogeneous substances.

Environment and organisms are persistently exposed by a mixture of various substances. However, the current evaluation method is mostly based on an individual substance's toxicity. A systematic toxicity evaluation of heterogeneous substances needs to be established. To demonstrate toxicity assessment of mixture, we chose a group of three typical ingredients in cosmetic sunscreen products that frequently enters ecosystems: benzophenone-3 (BP-3), ethylhexyl methoxycinnamate (EHMC), and titanium dioxide nanoparticle (TiO2 NP). We first determined a range of nominal toxic concentration of each ingredient or substance using Daphnia magna, and then for the subsequent organismal level phenotypic assessment, chose the wild-type zebrafish embryos. Any phenotype change, such as body deformation, led to further examinations on the specific organs of transgenic zebrafish embryos. Based on the systematic toxicity assessments of the heterogeneous substances, we offer a sequential environmental toxicity assessment protocol that starts off by utilizing Daphnia magna to determine a nominal concentration range of each substance and finishes by utilizing the zebrafish embryos to detect defects on the embryos caused by the heterogeneous substances. The protocol showed additive toxic effects of the mixtures. We propose a sequential environmental toxicity assessment protocol for the systematic toxicity screening of heterogeneous substances from Daphnia magna to zebrafish embryo in-vivo models.

Diagnosis of prostate cancer via nanotechnological approach.

Prostate cancer is one of the leading causes of cancer-related deaths among the Caucasian adult males in Europe and the USA. Currently available diagnostic strategies for patients with prostate cancer are invasive and unpleasant and have poor accuracy. Many patients have been overly or underly treated resulting in a controversy regarding the reliability of current conventional diagnostic approaches. This review discusses the state-of-the-art research in the development of novel noninvasive prostate cancer diagnostics using nanotechnology coupled with suggested diagnostic strategies for their clinical implication.