Multiple genome relationships and a complex biogeographic history in the eastern range of Quercus suber L. (Fagaceae) implied by nuclear and chloroplast DNA variation

The complex evolutionary history of Quercus suber has been throughly investigated in many recent works, but the details of its differentiation processes are still largely unknown. In addition, the geographical and evolutionary roles of the eastern parts of the species range have gained much less attention compared to other southern European areas. In order to fill this gap, new insights to infer the species diversification and range establishment of the cork oak in the east-central Mediterranean are here provided by means of inter- and intra-specific plastid DNA and nuclear ribosomal ITS phylogeographic studies. We analyzed 95 natural cork oak populations; 6 closely related, sympatric oaks were included in the study and used for comparisons.Evidence for a clear phylogeographical structure was detected with PCR-RFLP at 5 chloroplast loci, while ITS sequence variation is apparently unrelated with the geographical distribution. Five chloroplast haplotypes and three ITS main lineages were identified. Three haplotypes and all ITS lineages occur in the Italian Peninsula, stressing the importance of these territories for the evolutionary history of the species. Two divergent "Italian" haplotypes are highly shared, and one ITS variant is basal to the ingroup, revealing sister relationships within Cerris taxonomic group. Hypotheses of hybridization, lineage sorting of ancient DNA polymorphisms and of reticulate evolution of the whole species group are presented and discussed

A transcriptome analysis identifies molecular effectors of unconjugated bilirubin in human neuroblastoma SH-SY5Y cells

Background: The deposition of unconjugated bilirubin (UCB) in selected regions of the brain results in irreversible neuronal damage, or Bilirubin Encephalopathy (BE). Although UCB impairs a large number of cellular functions in other tissues, the basic mechanisms of neurotoxicity have not yet been fully clarified. While cells can accumulate UCB by passive diffusion, cell protection may involve multiple mechanisms including the extrusion of the pigment as well as pro-survival homeostatic responses that are still unknown. Results: Transcriptome changes induced by UCB exposure in SH-SY5Y neuroblastoma cell line were examined by high density oligonucleotide microarrays. Two-hundred and thirty genes were induced after 24 hours. A Gene Ontology (GO) analysis showed that at least 50 genes were directly involved in the endoplasmic reticulum (ER) stress response. Validation of selected ER stress genes is shown by quantitative RT-PCR. Analysis of XBP1 splicing and DDIT3/CHOP subcellular localization is presented. Conclusion: These results show for the first time that UCB exposure induces ER stress response as major intracellular homeostasis in surviving neuroblastoma cells in vitro

Aluminium to Carbon Fibre Reinforced Polymer tubes joints produced by magnetic pulse welding

Project SpIM, (2016/10605). ESA (ESA contract 4000111471/14/NL/PA). UID/EMS/00667/2019 (UNIDEMI).In this work, Aluminium AA7075-T6 to Carbon Fibre Reinforced Polymer (CFRP) tube joints using magnetic pulse welding technology were performed. The process was found suitable for joining this material combination when a rigid support is used to constrain the CFRP wall displacement. A minimum discharge energy of 2.5 kJ was seen to produce tight interfaces in tubular geometry for 1 mm of stand-off distance. A larger number of turns in the coil was found to reduce both the damages caused by the impact forces and to increase the joint axial resistance. To protect the CFRP tube from impacts and achieve a weld, an electrolytic Nickel coating was deposited over the CFRP tube. Welding was produced between the coating and the aluminium tube with no intermetallic compounds observed on the interface for the energies tested.authorsversionpublishe

Functional Induction of the Cystine-Glutamate Exchanger System Xc- Activity in SH-SY5Y Cells by Unconjugated Bilirubin

We have previously reported that exposure of SH-SY5Y neuroblastoma cells to unconjugated bilirubin (UCB) resulted in a marked up-regulation of the mRNA encoding for the Na+ -independent cystine∶glutamate exchanger System Xc− (SLC7A11 and SLC3A2 genes). In this study we demonstrate that SH-SY5Y cells treated with UCB showed a higher cystine uptake due to a significant and specific increase in the activity of System Xc−, without the contribution of the others two cystine transporters (XAG− and GGT) reported in neurons. The total intracellular glutathione content was 2 folds higher in the cells exposed to bilirubin as compared to controls, suggesting that the internalized cystine is used for gluthathione synthesis. Interestingly, these cells were significantly less sensitive to an oxidative insult induced by hydrogen peroxide. If System Xc− is silenced the protection is lost. In conclusion, these results suggest that bilirubin can modulate the gluthathione levels in neuroblastoma cells through the induction of the System Xc−, and this renders the cell less prone to oxidative damage

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field