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Mutation-induced activation of WNT-β-catenin signalling is a frequent driver event in human cancer. Sustained WNT-β-catenin pathway activation endows cancer cells with sustained self-renewing growth properties and is associated with therapy resistance. In healthy adult stem cells, WNT pathway activity is carefully controlled by core pathway tumour suppressors as well as negative feedback regulators. Gene inactivation experiments in mouse models unequivocally demonstrated the relevance of WNT tumour suppressor loss-of-function mutations for cancer growth. However, in human cancer, a far more complex picture has emerged in which missense or truncating mutations mediate stable expression of mutant proteins, with distinct functional and phenotypic ramifications. Herein, we review recent advances and challenges in our understanding of how different mutational subsets of WNT tumour suppressor genes link to distinct cancer types, clinical outcomes and treatment strategies.

It has been shown that incorporation of TiO2 nanoparticles into hydrophobic coatings can show self-cleaning performance. Accelerated laboratory testing indicated that the coats retain their hydrophobic nature for an extended time period. In this paper, hydrophobic polydimethylsiloxane (PDMS)/TiO2 nanocomposite coatings with a TiO2 content of 0–40% were fabricated by simple blending of a PDMS dispersion with an aqueous TiO2 nanoparticle dispersion. Their long-term hydrophobicity and self-cleaning performance were investigated both in laboratory and real-world outdoor testing. As expected, TiO2 nanoparticle-based coatings exhibited better self-cleaning relative to the TiO2-free PDMS control coating as measured by methylene blue degradation testing. Excellent long-term hydrophobicity was observed in accelerated weathering testing when they contained the appropriate levels of TiO2 nanoparticles (i.e., 0–30%). However, the same PDMS/TiO2 coatings did not show self-cleaning performance, and instead, exhibited improved dirt pickup resistance, in outdoor exposure testing. Sustained hydrophobicity was observed in outdoor exposure testing for the clear films except when TiO2 levels were at 40%. The hysteresis of water contact angle (HWCA) significantly increased for the PDMS control coating, and water beading was lost as the film surface picked up dirt. In contrast, the TiO2-based coatings with appropriate TiO2 levels maintained a relatively low HWCA after outdoor exposure and no water sheeting on rainy days was observed. This result demonstrates that while photocatalytic TiO2 nanoparticles can maintain coating hydrophobicity upon outdoor exposure, long-term self-cleaning performance in polluted environments has not yet been achieved with this type of coating under real-world conditions.

The evaluation of the dirt resistance of organic coatings according to traditional accelerated methods (e.g., the standard method GB/T 9780-2013 in China) usually provides results which deviate from their real-world performance in seriously polluted environments. As an alternative, a novel adsorption method with methyltrimethoxysilane-modified rice-straw ash as the artificial dirt standard was proposed in this paper. With this method, the ash was dispersed into the air in a testing chamber and then allowed to physically adsorb to the coating, which is believed to more accurately simulate the adsorption behavior of floating particulates outdoors. Two groups of latex coatings from Dow Chemical Company were employed to evaluate the adsorption method by monitoring the changes in color, whiteness, and reflectivity. In parallel, outdoor exposure testing of these coatings was conducted. The influences of charging method, of ash type, and panel position on the testing results were carefully investigated. Multicharging of ash (9 g × 3) and parallel panels at different positions in the testing chamber are designed for the generation of repeatable results. It was demonstrated that the adsorption method basically matched the outdoor exposure results and was more predictive of real-world results than GB/T 9780-2013.

Mammalian Wnt proteins are believed to act as short-range signals(1-4), yet have not been previously visualized in vivo. Selfrenewal, proliferation and differentiation are coordinated along a putative Wnt gradient in the intestinal crypt(5). Wnt3 is produced specifically by Paneth cells(6,7). Here we have generated an epitopetagged, functional Wnt3 knock-in allele. Wnt3 covers basolateral membranes of neighbouring stem cells. In intestinal organoids, Wnt3-transfer involves direct contact between Paneth cells and stem cells. Plasma membrane localization requires surface expression of Frizzled receptors, which in turn is regulated by the transmembrane E3 ligases Rnf43/Znrf3 and their antagonists Lgr4-5/R-spondin. By manipulating Wnt3 secretion and by arresting stem-cell proliferation, we demonstrate that Wnt3 mainly travels away from its source in a cell-bound manner through cell division, and not through diffusion. We conclude that stem-cell membranes constitute a reservoir for Wnt proteins, while Frizzled receptor turnover and ` plasma membrane dilution' through cell division shape the epithelial Wnt3 gradient.

Neuroblastoma (NB) is the most common solid malignancy in childhood and its prognosis is still generally poor. In contrast to many other cancers, mutations of the p53 tumor suppressor are rare. Instead, significant cytosolic sequestration of wtp53 is one of several mechanisms that attenuate p53 function in this cancer. Here, we report that aberrant p53 hyperubiquitylation contributes to p53 cytoplasmic sequestration in NB. NB lines constitutively harbor an elevated portion of wtp53 as stable ubiquitylated species confined to the cytoplasm. p53 hyperubiquitylation is not due to dysregulation by Hdm2 or proteasomal dysfunction. Instead, the defect lies in p53 regulation by HAUSP, a major p53-deubiquitylating enzyme. In contrast to non-NB cancer cells with nuclear p53 and normal ubiquitylation, p53 from NB cells shows impaired HAUSP interaction. Conversely, interference with p53 hyperubiquitylation in NB cells by Nutlin 3a or by a C-terminal p53 peptide (aa 305-393) results in p53 relocalization from the cytoplasm to the nucleus, and in case of Nutlin, in reactivation of p53's transcriptional and apoptotic functions. Moreover, nutlin and camptothecin act synergistically in inducing NB cell apoptosis. Hence, this study strengthens the rationale for targeting p53 deubiquitylation by drugs like Nutlin as a promising new strategy in NB therapy.

FOXO (Forkhead box O) transcription factors are important regulators of cellular metabolism, cell-cycle progression and cell death. FOXO activity is regulated by multiple post-translational modifications, including phosphorylation, acetylation and polyubiquitination. Here, we show that FOXO becomes monoubiquitinated in response to increased cellular oxidative stress, resulting in its re-localization to the nucleus and an increase in its transcriptional activity. Deubiquitination of FOXO requires the deubiquitinating enzyme USP7/HAUSP (herpesvirus-associated ubiquitin-specific protease), which interacts with and deubiquitinates FOXO in response to oxidative stress. Oxidative stress-induced ubiquitination and deubiquitination by USP7 do not influence FOXO protein half-life. However, USP7 does negatively regulate FOXO transcriptional activity towards endogenous promoters. Our results demonstrate a novel mechanism of FOXO regulation and indicate that USP7 has an important role in regulating FOXO-mediated stress responses.

Dishevelled (Dvl) is a key regulator of Wnt signaling both in the canonical and non-canonical pathways. Here we report the identification of a regulatory domain of ubiquitination (RDU) in the C-terminus of Dvl. Mutations in the RDU resulted in accumulation of polyubiquitinated forms of Dvl, which were mainly K63 linked. Small interfering RNA-based screening identified Usp14 as a mediator of Dvl deubiquitination. Genetic and chemical suppression of Usp14 activity caused an increase in Dvl polyubiquitination and significantly impaired downstream Wnt signaling. These data suggest that Usp14 functions as a positive regulator of the Wnt signaling pathway. Consistently, tissue microarray analysis of colon cancer revealed a strong correlation between the levels of Usp14 and b-catenin, which suggests an oncogenic role for Usp14 via enhancement of Wnt/beta-catenin signaling.

A model of impaired oxygen delivery, using an acute surgical preparation in the fetal dog, is described. Fetal heart rate, transcutaneous pO2 and pCO2, and tissue pH were simultaneously recorded during hypoxic episodes produced by a series of successive occlusions of the maternal abdominal aorta. Corresponding values were also determined in the arterial blood of the fetus. The following pathophysiologic sequence of events was observed: 1) a latency period with no changes; 2) a drop in pO2 which stabilized later at a lower pressure; 3) late fetal heart rate deceleration, the pattern of which was not related to the progressively deteriorating fetal condition; 4) a progressive increase in pCO2; 5) a progressive decrease in pH. Abnormal tissue values consistently preceded and were more adversely affected than corresponding blood values. This experimental model demonstrates first that late decelerations of the fetal heart rate are an early sign of fetal hypoxia and second that a fall in fetal blood pH, beyond that level normally seen during labor, occurs relatively late in this pathophysiologic sequence. Between these two, there are intermediary stages that could be continuously monitored in order to identify worsening fetal condition. Continuous tissue pH and transcutaneous pO2 and pCO2 recording may potentially be of significant clinical value.

To investigate the resistance to diffusion between the two phases of the egg, the 34-hr. isotope of bromine (82Br) as bromide was injected into the white in many eggs and its penetration into the yolk measured after various periods of time. About 5 ml. of the white of the egg to be injected was mixed with ½ ml. of the radioactive bromide in isotonic solution, and injected as uniformly as possible throughout that remaining in the shell. The egg was maintained during the experimental period at 1° C., and, at its conclusion, frozen in a mixture of ether and dry ice. The yolk and white could then be separated cleanly with a sharp blow, and were conveniently diluted and their activities measured with a Geiger-Muller counter dipping into the liquids.