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Pompe Disease (PD) is an inherited metabolic disorder that causes glycogen to accumulate within lysosomes ultimately leading to severe muscle weakness. According to age of onset, it is classified into early and late-onset (EOPD and LOPD). In vitro skeletal muscle 2D models have been reported for EOPD using pluripotent stem cells but concerns exist about their phenotypic and genotypic representation of the disease. To our knowledge there are no reports of a human LOPD skeletal muscle model. We therefore leveraged the MyoScreen platform and our expertise in quantitative image analysis to generate and compare EOPD and LOPD pathological hallmarks. Robust myotube differentiation was obtained from myoblasts from 2 LOPD patients when cultured on micropatterned 96-well plates, while EOPD patient cells differentiate into smaller myotubes and had a lower fusion index (LOPD 40% versus EOPD <20%). Myotubes from both forms of Pompe showed an increase in baseline levels of autophagy compared to Healthy myotubes, as monitored by LC3b intensity (2.3-fold), spot number (1.99-fold) and percent coverage area (2-fold). CQ treatment exacerbated the autophagic build-up further. Only EOPD myotubes presented an accumulation of lysosomes at basal levels (1.7-fold LAMP1 mean intensity versus Healthy). Structurally, the EOPD donor showed large vacuoles filled with LAMP1 positive vesicles that displaced the myofibrillar architecture. Levels of glycogen were quantified biochemically: compared to Healthy myotubes, EOPD and LOPD showed 4 and 1.4-fold increase respectively, and a genotype-phenotype correlation of reduced GAA enzyme activity. Enzyme replacement therapy and liver-directed AAV delivery of the corrective enzyme α-glucosidase have had limited success in improving muscle function due to inefficient internalization. The MyoScreen PD platform provides an unprecedented opportunity for new drug screening, target identification and the assessment of novel emerging RNA and gene therapies. [ABSTRACT FROM AUTHOR]

Recent evidence indicates that the cancer preventive activity of olive oil can be mediated by the presence of minor components, such as antioxidant phenolic compounds. However, their mechanisms of action remain largely unknown. In this study, we investigated the in vitro effects of one of the main olive oil phenols, hydroxytyrosol [3,4-dihydroxyphenylethanol (3,4-DHPEA)], on proliferation, cell cycle progression, apoptosis, and differentiation of HL60 human promyelocytic leukemia cells. 3,4-DHPEA showed a potent inhibitory activity on DNA synthesis, as evidenced by a 92% reduction of [3H]-thymidine incorporation at 100 micromol/L, and an induced apoptosis, as evidenced by the release of cytosolic nucleosomes and flow cytometry. This phenol, 3,4-DHPEA, was also able to inhibit the progression of the cell cycle in synchronized HL60 cells, which accumulated in the G0/G1 phase of the cell cycle after 25 h of treatment. Furthermore, 3,4-DHPEA induced differentiation on HL60 cells with a maximum effect (22% of cells) at 100 micromol/L after 72 h of treatment. Among the different proteins involved in the regulation of the cell cycle, 3,4-DHPEA reduced the level of cyclin-dependent kinase (CDK) 6 and increased that of cyclin D3. With regard to the CDK inhibitors, p15 was not altered by 3,4-DHPEA treatment, whereas the expression of p21(WAF1/Cip1) and p27(Kip1) was increased at both protein and mRNA levels. To our knowledge, these results provide the first evidence that 3,4-DHPEA may effect the expression of genes involved in the regulation of tumor cell proliferation and differentiation. [ABSTRACT FROM AUTHOR]

In the attempt to assess the relationship and interdependency among sediment toxic pollutants, in particular heavy metals, polycyclic aromatic hydrocarbons (PAH), and linear alkyl sulfonates (LAS) and some of the sediment typical components: inorganic carbon (IC), organic material (OM) and acid volatile sulphides (AVS), multivariate techniques of statistical analysis have been applied to a set of chemical data obtained by the analysis of the sediments of the Trasimeno Lake, a central Italy lake characterized by a large surface (128 km2) and a low mean depth (about 4.5 m).The results of principal component analysis (PCA) show interrelationships between: OM content and PAH, Pb, and Cu concentrations of the sediments, LAS and AVS, and AVS and IC. The effect of the different sampling periods on sediment composition and contamination level, and the clustering of the sampling sites as a consequence of pollutant load are also shown.The principal component bi-plot of the variables and samples indicates that PAH have the greatest influence on the separation of samples in the different sampling periods. [Copyright &y& Elsevier]