The Human Melanoma Proteome Atlas—Complementing the melanoma transcriptome

The MM500 meta‐study aims to establish a knowledge basis of the tumor proteome to serve as a complement to genome and transcriptome studies. Somatic mutations and their effect on the transcriptome have been extensively characterized in melanoma. However, the effects of these genetic changes on the proteomic landscape and the impact on cellular processes in melanoma remain poorly understood. In this study, the quantitative mass‐spectrometry‐based proteomic analysis is interfaced with pathological tumor characterization, and associated with clinical data. The melanoma proteome landscape, obtained by the analysis of 505 well‐annotated melanoma tumor samples, is defined based on almost 16 000 proteins, including mutated proteoforms of driver genes. More than 50 million MS/MS spectra were analyzed, resulting in approximately 13,6 million peptide spectrum matches (PSMs). Altogether 13 176 protein‐coding genes, represented by 366 172 peptides, in addition to 52 000 phosphorylation sites, and 4 400 acetylation sites were successfully annotated. This data covers 65% and 74% of the predicted and identified human proteome, respectively. A high degree of correlation (Pearson, up to 0.54) with the melanoma transcriptome of the TCGA repository, with an overlap of 12 751 gene products, was found. Mapping of the expressed proteins with quantitation, spatiotemporal localization, mutations, splice isoforms, and PTM variants was proven not to be predicted by genome sequencing alone. The melanoma tumor molecular map was complemented by analysis of blood protein expression, including data on proteins regulated after immunotherapy. By adding these key proteomic pillars, the MM500 study expands the knowledge on melanoma disease

In vitro evaluation of antimicrobial efficacy of endodontic irrigants.

Purpose: The objective of this study was to compare in vitro, by MTT assay, the antimicrobial efficacy of Niclor 5 (5% NaOCl solution), Cloreximid (0.2% chlorhexidine and 0.2% cetrimide solution), 3% hydrogen peroxide and 17% EDTA against two microorganisms associated with primary endodontic infections. Methods: Enterococcus faecalis and Streptococcus mutans strains were selected for this test. Freshly extracted single-rooted human teeth were endodontically treated, inoculated with bacterial strains and then divided into different groups, each of them rinsed with Niclor 5 (5% NaOCl solution), Cloreximid (0.2% chlorhexidine and 0.2% cetrimide solution), 3% hydrogen peroxide,17% EDTA and with 5% NaOCl solution (positive control). Results: Even though all the tested irrigating solutions demonstrated antibacterial effects against E. faecalis and S. mutans, the results were significantly different between the various groups. The greatest antimicrobial effects were observed in groups treated with 5% NaOCl and 17% EDTA. Interestingly, the effectiveness of EDTA could be ascribed to its capability of detaching biofilm from canal walls

Photoactivated disinfection (PAD) in endodontics: an in vitro microbiological evaluation.

Purpose: The objective of the present study was the in vitro evaluation by MTT test of the antimicrobial effect of photoactivated disinfection (PAD) and, comparatively, of a conventional 5.25% NaOCl irrigating solution. Methods: Enterococcus faecalis, Streptococcus mutans and Streptococcus sanguis strains were selected for the test. Freshly extracted single-rooted human teeth were endodontically treated, inoculated with bacterial strains and then divided into different groups, each of them treated with PAD, with PAD plus 0.5% NaOCl solution, with TBO, with PAD for longer time and with 5% NaOCl solution (positive control). Results: The results were significantly different among the various groups, and for Enterococcus faecalis, Streptococcus mutans and Streptococcus sanguis. PAD applied for a longer time (in respect to manufacturer's instructions) or PAD associated to 5% NaOCl showed the significantly higher antibacterial effects

New evidence of plant food processing in Italy before 40ka

Evidence of plant food processing is a significant indicator of the human ability to exploit environmental resources. The recovery of starch grains associated with use-wear on Palaeolithic grinding tools offers proof of a specific technology for making flour among Pleistocene hunter-gatherers. Here we present the analysis of five grindstones from two Italian sites, Riparo Bombrini and Grotta di Castelcivita, both inhabited during a crucial phase spanning the decline of the Neanderthals and the establishment of Sapiens. The recovery of starch grains on a Mousterian grindstone at Bombrini suggests that the last Neanderthals not only consumed and processed plants but also made flour 43e41,000 years ago. Starch grains attributable to Triticeae on Protoaurignacian grindstones at both sites testify that Sapiens were processing wild cereals at least 41,500e36,500 years ago when they expanded into Eurasia, long before the dawn of agriculture. These new data suggest a profound knowledge of available plant resources in both human groups

Early consequences of the phospholamban mutation PLN-R14del^+/− in a transgenic mouse model

Aims: The heterozygous phospholamban (PLN) mutation R14del (PLN R14del+/−) is associated with a severe arrhythmogenic cardiomyopathy (ACM) developing in the adult. “Superinhibition” of SERCA2a by PLN R14del is widely assumed to underlie the pathogenesis, but alternative mechanisms such abnormal energy metabolism have also been reported. This work aims to (1) to evaluate Ca2+ dynamics and energy metabolism in a transgenic (TG) mouse model of the mutation prior to cardiomyopathy development; (2) to test whether they are causally connected.Methods: Ca2+ dynamics, energy metabolism parameters, reporters of mitochondrial integrity, energy, and redox homeostasis were measured in ventricular myocytes of 8–12 weeks-old, phenotypically silent, TG mice. Mutation effects were compared to pharmacological PLN antagonism and analyzed during modulation of sarcoplasmic reticulum (SR) and cytosolic Ca2+ compartments. Transcripts and proteins of relevant signaling pathways were evaluated.Results: The mutation was characterized by hyperdynamic Ca2+ handling, compatible with a loss of SERCA2a inhibition by PLN. All components of energy metabolism were depressed; myocyte energy charge was preserved under quiescence but reduced during stimulation. Cytosolic Ca2+ buffering or SERCA2a blockade reduced O2 consumption with larger effect in the mutant. Signaling changes suggest cellular adaptation to perturbed Ca2+ dynamics and response to stress.Conclusions: (1) PLN R14del+/− loses its ability to inhibit SERCA2a, which argues against SERCA2a superinhibition as a pathogenetic mechanism; (2) depressed energy metabolism, its enhanced dependency on Ca2+ and activation of signaling responses point to an early involvement of metabolic stress in the pathogenesis of this ACM model.</p