A quantitative LumiFluo assay to test inhibitory compounds blocking p53 degradation induced by human papillomavirus oncoprotein E6 in living cells

High-risk human papillomaviruses (HR-HPVs) are the causative agents for the onset of several epithelial cancers in humans. The deregulated expression of the viral oncoproteins E6 and E7 is the driving force sustaining the progression of malignant transformation in pre-neoplastic lesions. Targeting the viral E6 oncoprotein through inhibitory compounds can counteract the survival of cancer cells due to the reactivation of p53-mediated pathways and represents an intriguing strategy to treat HPV-associated neoplasias. Here, we describe the development of a quantitative and easy-to-perform assay to monitor the E6-mediated degradation of p53 in living cells to be used for small-molecule testing. This assay allows to unbiasedly determine whether a compound can protect p53 from the E6-mediated degradation in cells, through a simple 3-step protocol. We validated the assay by testing two small molecules, SAHA and RITA, reported to impair the E6-mediated p53 degradation. Interestingly, we observed that only SAHA efficiently rescued p53, while RITA could not provide the same degree of protection. The possibility to specifically and quantitatively monitor the ability of a selected compound to rescue p53 in a cellular context through our LumiFluo assay could represent an important step towards the successful development of anti-HPV drugs

On the price of morals in markets: an empirical study of the Swedish AP-Funds and the Norwegian Government Pension Fund

This study empirically analyses the exclusion of companies from investors’ investment universe due to a company’s business model (sector-based exclusion) or due to a company’s violations of international norms (normbased exclusion). We conduct a time-series analysis of the performance implications of the exclusion decisions of two leading Nordic investors, Norway’s Government Pension Fund-Global (GPFG) and Sweden’s AP-funds. We find that their portfolios of excluded companies do not generate an abnormal return relative to the funds’ benchmark index. While the exclusion portfolios show higher risk than the respective benchmark, this difference is only statistically significant for the case of GPFG. These findings suggest that the exclusion of the companies generally does not harm funds’ performance. We interpret these findings as indicative that with exclusionary screening, as practiced by the sample funds, asset owners can meet the ethical objectives of their beneficiaries without compromising financial returns

The use of Gompertz models in growth analyses, and new Gompertz-model approach: An addition to the Unified-Richards family

<div><p>The Gompertz model is well known and widely used in many aspects of biology. It has been frequently used to describe the growth of animals and plants, as well as the number or volume of bacteria and cancer cells. Numerous parametrisations and re-parametrisations of varying usefulness are found in the literature, whereof the Gompertz-Laird is one of the more commonly used. Here, we review, present, and discuss the many re-parametrisations and some parameterisations of the Gompertz model, which we divide into <i>T</i>_<i>i</i> (type I)- and <i>W</i>₀ (type II)-forms. In the <i>W</i>₀-form a starting-point parameter, meaning birth or hatching value (<i>W</i>₀), replaces the inflection-time parameter (<i>T</i>_<i>i</i>). We also propose new “unified” versions (U-versions) of both the traditional <i>T</i>_<i>i</i> -form and a simplified <i>W</i>₀-form. In these, the growth-rate constant represents the relative growth rate instead of merely an unspecified growth coefficient. We also present U-versions where the growth-rate parameters return absolute growth rate (instead of relative). The new U-Gompertz models are special cases of the Unified-Richards (U-Richards) model and thus belong to the Richards family of U-models. As U-models, they have a set of parameters, which are comparable across models in the family, without conversion equations. The improvements are simple, and may seem trivial, but are of great importance to those who study organismal growth, as the two new U-Gompertz forms give easy and fast access to all shape parameters needed for describing most types of growth following the shape of the Gompertz model.</p></div

Structure based identification and characterization of flavonoids that disrupt human papillomavirus-16 E6 function.

Expression and function of the human papillomavirus (HPV) early protein 6 (E6) is necessary for viral replication and oncogenesis in cervical cancers. HPV E6 targets the tumor suppressor protein p53 for degradation. To achieve this, "high-risk" HPV E6 proteins bind to and modify the target specificity of the ubiquitin ligase E6AP (E6 associated protein). This E6-dependent loss of p53 enables the virus to bypass host cell defenses and facilitates virally induced activation of the cell cycle progression during viral replication. Disruption of the interaction between E6 and E6AP and stabilization of p53 should decrease viability and proliferation of HPV positive cells. A new in vitro high-throughput binding assay was developed to assay binding between HPV-16 E6 and E6AP and to identify compounds that inhibit this interaction. The compound luteolin emerged from the screen and a library of novel flavones based on its structure was synthesized and characterized using this in vitro binding assay. The compounds identified in this study disrupt the E6/E6AP interaction, increase the levels of p53 and p21(Cip1/Waf1), and decrease proliferation of HPV positive cell lines. The new class of flavonoid E6 inhibitors displays a high degree of specificity for HPV positive cells. Docking analyses suggest that these compounds bind in a hydrophobic pocket at the interface between E6 and E6AP and mimic the leucines in the conserved α-helical motif of E6AP. The activity and specificity of these compounds represent a promising new lead for development as an antiviral therapy in the treatment of HPV infection and cervical cancer