A simple, accurate and universal method for quantification of PCR.

BACKGROUND: Research into gene expression enables scientists to decipher the complex regulatory networks that control fundamental biological processes. Quantitative real-time PCR (qPCR) is a powerful and ubiquitous method for interrogation of gene expression. Accurate quantification is essential for correct interpretation of qPCR data. However, conventional relative and absolute quantification methodologies often give erroneous results or are laborious to perform. To overcome these failings, we developed an accurate, simple to use, universal calibrator, AccuCal. RESULTS: Herein, we show that AccuCal quantification can be used with either dye- or probe-based detection methods and is accurate over a dynamic range of ≥10(5) copies, for amplicons up to 500 base pairs (bp). By providing absolute quantification of all genes of interest, AccuCal exposes, and circumvents, the well-known biases of qPCR, thus allowing objective experimental conclusions to be drawn. CONCLUSION: We propose that AccuCal supersedes the traditional quantification methods of PCR.

A Novel Strategy for Softening Gelatin-Bioactive-Glass Hybrids.

The brittle structure of polymer-bioactive-glass hybrids is a hurdle for their biomedical applications. To address this issue here, we developed a novel method to cease the overcondensation of bioactive-glass by polymer cross-linking. Here, an organosilane-functionalized gelatin methacrylate (GelMA) is covalently bonded to a bioactive-glass during the sol-gel process, and the condensation of silica networks is controlled by photo-cross-linking of GelMA. The physicochemical properties and mechanical strength of these hybrids are tunable by the incorporation of secondary cross-linking agents. These hydrogels display elastic properties with ultimate compression strain above 0.2 mm·mm(-1) and tunable compressive modulus in the range of 42-530 kPa. In addition, these hydrogels are bioactive because they promoted the alkaline phosphatase activity of bone progenitor cells. They are also well-tolerated in the mice subcutaneous model. Therefore, our method is efficient for the prevention of overcondensation and allows preparation of soft bioactive hydrogels from organic-inorganic matrices, suitable for soft and hard tissue regeneration.

Androgen receptor actions modify skin structure and chemical carcinogen-induced skin cancer susceptibility in mice.

Men are significantly more susceptible to non-melanoma skin cancers than women, and the androgen receptor (AR) is widely distributed in the skin, suggesting a ro\le for androgens acting via AR. Therefore, we explored the role of androgen action via AR in susceptibility to experimental 7,12-dimethylbenz[a]anthracene (DMBA)-induced skin carcinogenesis and in skin structural development of male and female mice. We demonstrate that both the male gender and androgen action via AR modify the susceptibility to carcinogen-induced skin cancer, but the effect depends on the carcinogenesis model used. Following systemic DMBA exposure, males were significantly (p < 0.05) more susceptible to DMBA-induced experimental skin cancer than females and AR inactivation significantly delayed cancer detection in both male (median time to palpable tumours 19 vs. >35 weeks (wild-type [WT] vs. AR knockout [ARKO], p < 0.001) and female (27 vs. >35 weeks, p = 0.008)) mice. In contrast, following DMBA/12-O-tetradecanoylphorbol-13-acetate (TPA)-induced multistage local skin carcinogenesis, AR inactivation protected against formation of DMBA-induced skin cancers in both male and female mice. The skin structure was also affected by gender effect as well as the AR inactivation and could at least partly explain the different responses between the carcinogenesis models (systemic vs. topical). In addition, AR inactivation modified Cox-1 and Cox-2 expression in the skin, suggesting possible molecular mechanism for the AR effect on skin. Finally, some gender differences are observed also in ARKO mice insensitive to androgens, suggesting that factors other than androgens also play a role in gender-dependent skin carcinogenesis.