E-cadherin can limit the transforming properties of activating β-catenin mutations

Wnt pathway deregulation is a common characteristic of many cancers. But only Colorectal Cancer predominantly harbours mutations in APC, whereas other cancer types (hepatocellular carcinoma, solid pseudopapillary tumours of pancreas) have activating mutations in β-catenin (CTNNB1). We have compared the dynamics and the potency of β-catenin mutations in vivo. Within the murine small intestine (SI), an activating mutation of β-catenin took much longer to achieve a Wnt deregulation and acquire a crypt-progenitor-cell (CPC) phenotype than Apc or Gsk3 loss. Within the colon, a single activating mutation of β-catenin was unable to drive Wnt deregulation or induce the CPC phenotype. This ability of β-catenin mutation to differentially transform the SI versus the colon correlated with significantly higher expression of the β-catenin binding partner E-cadherin. This increased expression is associated with a higher number of E-cadherin:β-catenin complexes at the membrane. Reduction of E-cadherin synergised with an activating mutation of β-catenin so there was now a rapid CPC phenotype within the colon and SI. Thus there is a threshold of β-catenin that is required to drive transformation and E-cadherin can act as a buffer to prevent β-catenin accumulation

Macrophage-derived Wnt opposes Notch signaling to specify hepatic progenitor cell fate in chronic liver disease

During chronic injury a population of bipotent hepatic progenitor cells (HPCs) become activated to regenerate both cholangiocytes and hepatocytes. Here we show in human diseased liver and mouse models of the ductular reaction that Notch and Wnt signaling direct specification of HPCs via their interactions with activated myofibroblasts or macrophages. In particular, we found that during biliary regeneration, expression of Jagged 1 (a Notch ligand) by myofibroblasts promoted Notch signaling in HPCs and thus their biliary specification to cholangiocytes. Alternatively, during hepatocyte regeneration, macrophage engulfment of hepatocyte debris induced Wnt3a expression. This resulted in canonical Wnt signaling in nearby HPCs, thus maintaining expression of Numb (a cell fate determinant) within these cells and the promotion of their specification to hepatocytes. By these two pathways adult parenchymal regeneration during chronic liver injury is promoted

mTORC1-mediated translational elongation limits intestinal tumour initiation and growth.

Inactivation of APC is a strongly predisposing event in the development of colorectal cancer, prompting the search for vulnerabilities specific to cells that have lost APC function. Signalling through the mTOR pathway is known to be required for epithelial cell proliferation and tumour growth, and the current paradigm suggests that a critical function of mTOR activity is to upregulate translational initiation through phosphorylation of 4EBP1 (refs 6, 7). This model predicts that the mTOR inhibitor rapamycin, which does not efficiently inhibit 4EBP1 (ref. 8), would be ineffective in limiting cancer progression in APC-deficient lesions. Here we show in mice that mTOR complex 1 (mTORC1) activity is absolutely required for the proliferation of Apc-deficient (but not wild-type) enterocytes, revealing an unexpected opportunity for therapeutic intervention. Although APC-deficient cells show the expected increases in protein synthesis, our study reveals that it is translation elongation, and not initiation, which is the rate-limiting component. Mechanistically, mTORC1-mediated inhibition of eEF2 kinase is required for the proliferation of APC-deficient cells. Importantly, treatment of established APC-deficient adenomas with rapamycin (which can target eEF2 through the mTORC1-S6K-eEF2K axis) causes tumour cells to undergo growth arrest and differentiation. Taken together, our data suggest that inhibition of translation elongation using existing, clinically approved drugs, such as the rapalogs, would provide clear therapeutic benefit for patients at high risk of developing colorectal cancer

Localization of proteins bound to a replication origin of human DNA along the cell cycle

The proteins bound in vivo at the human lamin B2 DNA replication origin and their precise sites of binding were investigated along the cell cycle utilizing two novel procedures based on immunoprecipitation following UV irradiation with a pulsed laser light source. In G(1), the pre-replicative complex contains CDC6, MCM3, ORC1 and ORC2 proteins; of these, the post-replicative complex in S phase contains only ORC2; in M phase none of them are bound. The precise nucleotide of binding was identified for the two ORC and the CDC6 proteins near the start sites for leading-strand synthesis; the transition from the pre- to the post-replicative complex is accompanied by a 17 bp displacement of the ORC2 protein towards the start site

Phytochemical Statistical Mapping of Red Grape Varieties Cultivated in Romanian Organic and Conventional Vineyards

Red grapes are rich in phytochemicals such as phenolics and flavonoids, which are strongly correlated with their antioxidant activity. Thus, grapes as-harvested and grape extracts, especially those obtained from their seeds and pulp, have been reported to have health benefits, and accordingly, grapes and their derivatives are considered potential functional food ingredients. The total phenolic content, total flavonoid content, and the antioxidant activity of skin, pulp, and seeds of four grape varieties grown both in conventional and organic vineyards were examined in this study. Phytochemical characteristics of one native Romanian variety, Feteasca Neagra, were compared with data measured for three red grape varieties more commonly cultivated worldwide (Merlot, Pinot Noir, and Muscat Hamburg). It was found that the seeds of the Pinot Noir variety grown in an organic system contained the highest total phenolics of 169.53 ± 7.32 mg gallic acid equivalents/g and the highest total flavonoid content of 388.25 ± 10.72 mg quercetin equivalents/g, values corresponding to high antioxidant activity (312.84 ± 12.81 mg ascorbic acid equivalents/g). The total flavonoid content in the hydroalcoholic extracts obtained from seeds of Pinot Noir (organic vineyard) was around 24.5-fold higher than that of the skin of Pinot Noir (conventional vineyard). Experiments showed that seeds of all four tested grape varieties are good sources of total flavonoids, not only of total phenolics. When referring to the organic vineyard, the skin and pulp grapes showed good results for the total phenolic content. The antioxidant activities of the hydroalcoholic extracts were well-correlated with the total phenolic content and total flavonoid content. Lower values of these parameters were found for extracts obtained from skin and pulp than for those obtained from seeds of the same grape variety regardless of the culture management system (organic/conventional). Data mining techniques such as regression analysis, principal component analysis, and clustering analysis were applied to establish the potential correlation between the phytochemical content and the antioxidant activities of the red grapes on the one hand, and grape variety, anatomical parts, and vineyard type (organic/conventional) on the other hand

Chronic Kidney Disease—An Underestimated Risk Factor for Antimicrobial Resistance in Patients with Urinary Tract Infections

(1) Background: Chronic kidney disease (CKD), as well as antimicrobial resistance (AMR) represent major global health problems, with important social and economic implications. It was reported that CKD is a risk factor for antimicrobial resistance, but evidence is scarce. In addition, CKD is recognized to be a risk factor for complicated urinary tract infections (UTIs). (2) Methods: We conducted an observational study on 564 adult in-hospital patients diagnosed with urinary tract infections. The aim of the study was to identify the risk factors for AMR, as well as multiple drug resistance (MDR) and the implicated resistance patterns. (3) Results: The mean age was 68.63 ± 17.2 years. The most frequently isolated uropathogens were Escherichia coli strains (68.3%) followed by Klebsiella species (spp. (11.2%). In 307 cases (54.4%)), the UTIs were determined by antibiotic-resistant bacteria (ARBs) and 169 cases (30%) were UTIs with MDR strains. Increased age (≥65) OR 2.156 (95% CI: 1.404–3.311), upper urinary tract obstruction OR 1.666 (1.083–2.564), indwelling urinary catheters OR 6.066 (3.919–9.390), chronic kidney disease OR 2.696 (1.832–3.969), chronic hemodialysis OR 4.955 (1.828–13.435) and active malignancies OR 1.962 (1.087–3.540) were independent risk factors for MDR UTIs. In a multivariate logistic regression model, only indwelling urinary catheters (OR 5.388, 95% CI: 3.294–8.814, p < 0.001), CKD (OR 1.779, 95% CI: 1.153–2.745, p = 0.009) and chronic hemodialysis (OR 4.068, 95% 1.413–11.715, p = 0.009) were risk factors for UTIs caused by MDR uropathogens. (4) Conclusions: CKD is an important risk factor for overall antimicrobial resistance, but also for multiple-drug resistance

Novel Structures of Functionalized Graphene Oxide with Hydrazide: Characterization and Bioevaluation of Antimicrobial and Cytocompatibility Features

Graphite was oxidized to graphene oxide and activated by thionyl chloride, for further covalently linking three hydrazides with potential biological activity. The obtained materials were characterized by scanning electron microscopy with energy dispersive spectroscopy, Fourier-transform infrared and Raman spectroscopies. The presence of various functional groups specific to graphene oxide (GO) functionalized with different hydrazides was confirmed by spectral data. The ratio between D- and G-bands, observed in Raman spectra, allowed for an evaluation of the disorder degree and the mean crystallite size of the samples. The micrographs highlighted that the samples lead to the occurrence of disorders, probably caused by the sp3 carbons, the formation of oxygen-containing functional groups in the basal planes, and by various structural defects. The new graphene oxide–hydrazide derivatives were tested for their antimicrobial and cytotoxicity activity. Their antimicrobial activity against planktonic and biofilm-embedded cells was inferior to that of free hydrazides, except for GO-3 against planktonic Escherichia coli and GO-2 against Pseudomonas aeruginosa biofilm, demonstrating that further optimization is needed to be able to exploit the huge potential of GO for developing potent antimicrobials