Sex-related differences in death control of somatic cells

In 2001, The United States Institute of Medicine (IOM) Committee on Understanding the Biology of Sex and Gender Differences concluded that ‘Sex…should be considered when designing and analysing studies in all areas and at all levels of biomedical and health-related research…’ and stated an apparent paradox i.e.: ‘every cell has a sex’ 1. Sex is defined as ‘the classification of living things, generally as male or female according to their reproductive organs and functions assigned by chromosomal complement’ whereas gender is defined as ‘a person's self representation as male or female, or how that person is responded to by social institutions based on the individual's gender presentation. Gender is rooted in biology and shaped by environment and experience’ 1. It is unchallenged that there are health differences between males and females and that social and cultural factors could contribute to the observed differences. Anyway, the sex-dependent differences also have a biological base which sometimes has not been deeply investigated. Scientists studying health differences between male and female aim to both considering social/cultural environment and investigating biological/molecular mechanisms different between sexes. Some experimental studies have elucidated important differences in cell death control 2. A sex disparity, in fact, has been shown both in the propensity to apoptosis and in the activation of the autophagic pathway. In the context of cell fate control, hormones represent important regulators of both apoptosis and autophagy. In the cardiovascular system, for example, oestrogens inhibit cardiomyocyte apoptosis by decreasing reactive oxygen species production and increasing intracellular antioxidants 3. Oestrogens may also indirectly control autophagy as they up-regulate urocortin 4, a neuropeptide hormone able to inhibiting autophagy in cardiomyocytes. Conversely, increasing evidence suggests possible adverse effects of androgens on the vasculature showing that androgens, as opposed to oestrogens, may worsen vascular dysfunction in men, thus contributing to sex-based differences in cardiovascular diseases 5. However, it is currently emerging that some cell death programs are differentially controlled by sex-related hormone-independent cellular genetics. Differences in cell death sensitivity in male and female may then occur in the absence of an hormonal context. This is not an immediately obvious finding; Penaloza C et al., 6 have shown that the apoptosis amount differs between the sexes in isolated embryonic cells exposed to similar conditions and this happens at embryonal stages where there are no hormonal influences. Previous studies had reported a sexual dimorphism in embryonic neuronal signal transduction pathways and consequently differences in cell survival 7. Death pathways in XX and XY cells have been poorly investigated as most studies have been performed on established cells lines often irrespective of their male or female origin. Recently, using freshly isolated cells from male and female individuals gave important information on sex disparity in cell fate control. Such sex specificity has been in part clarified thanks to cell culture models where sex steroids can be removed from the media. Even sex-related differences in caspase activation have been found to be independent on hormone exposure. More in detail, cell death occurring in cortical neurons after ischaemia proceeds predominantly via an apoptosis-inducing factor-dependent pathway (a caspase-independent pathway) in male neurons while proceeds via a cytochrome C-dependent pathway (a process mediated by caspase activation) in female neurons 8. In this context, a sex-specific microRNA expression after ischaemia has been described in in vivo studies. In particular, it has been demonstrated that microRNA-23a, by binding the mRNA of the caspase inhibitor named XIAP, induces its translational repression in females, leading to enhanced caspase signalling in the ischaemic female brain. This effect has been shown to be independent of circulating oestrogen levels 9. Sex differences in ischaemic brain injury and cerebrovascular regulation have been observed in clinical and experimental studies and an important determinant of such differences is also represented by the integrity of endothelial cells. In fact, endothelial function is improved in women compared with men, contributing to female cellular higher resistance after ischaemic brain injury. Gupta NC et al. 10 showed that female cerebrovascular endothelial cells express lower level of soluble epoxide hydrolase and consequently have higher levels of vasoprotective epoxyeicosatrienoic acids as compared with male endothelial cells. This study therefore presents a novel additional mechanism underlying differences between male and female cells in apoptotic response after oxygen-glucose deprivation, contributing to explain higher resistance observed in females as compared with males. This study remarks again that differences between male and female cells do not necessarily depend on the hormonal context but may be inherent the cells. We believe that this apparently paradoxical concept has not been sufficiently highlighted in the scientific literature. The present ‘Letter to the Editor’ therefore aims at underlining such an important issue which deserves more attention and discussion in the researchers' community. A practical consequence of sex-dependent discrepancies in cell death control is that cellular response to any stimulus or treatment, in any physiological or pathological context, may well depend on the sex of the cell line used; journals guidelines should therefore require authors to state in any case the sex of the cell lines used in any in vitro study. In addition, at least to some extent, sex-matched or sex-unmatched cell controls may be necessary in many experimental settings. In conclusion, sex-related differences in cell death mechanism may have strong implications for experimental studies and sexual dimorphism dependent on chromosomal rather than hormonal differences have important implications for planning preclinical studies and clinical interventions

The short food supply chains’ phenomenon: a multidisciplinary approach to explore consumer behaviour and preferences

L’odierna sfiducia dei consumatori verso i sistemi agroalimentari industriali e la loro crescente riflessività o “quality turn”, hanno portato negli ultimi anni allo sviluppo delle filiere corte (FC), in alternativa ai mercati convenzionali. Le FC hanno la capacità di aumentare la sostenibilità dei sistemi convenzionali in termini sia di equità socio-economica sia di sviluppo ambientale e locale, incontrando le abitudini e le motivazioni di acquisto del consumatore post-moderno, le quali sono per definizione molto eterogenee. Viepiù, la nuova Politica Agricola Comunitaria 2014-2020 incoraggia oggi la promozione delle FC, attraverso specifici supporti finanziari previsti all’interno del secondo pilastro, al fine di favorire uno sviluppo sostenibile. Questa ricerca indaga, attraverso un approccio multidisciplinare, le preferenze e il comportamento del consumatore verso l’acquisto in questi canali di vendita alternativi. Sulla base di alcuni risultati preliminari di natura qualitativa, tale ricerca esplora più in dettaglio determinati fattori che influenzano il comportamento del consumatore (sostenibilità, fiducia, equità). L’obiettivo più ampio è quello di fornire nuova conoscenza sulle FC, focalizzando l’attenzione in particolar modo all’Italia, al fine di spiegarne il crescente appeal sul consumatore e il continuo sviluppo. Questa ricerca passa da un approccio socio-psicologico (Teoria del Comportamento Pianificato) alla teoria economia, applicando la tecnica dell’esperimento di scelta basato su un mercato ipotetico e relativo ai mercati dei contadini. È stato svolto anche un confronto tra Italia e Brasile e Italia e Germania. I risultati mostrano l’importante ruolo della sostenibilità e della fiducia nell’influenzare gli acquisti alimentari presso le FC, cosi come una rinnovata enfasi su alcuni fattori legati al marketing tradizionale (contatto diretto con il produttore) e la possibilità per i consumatori di contribuire al reddito degli agricoltori.In recent years, the erosion of consumers’ confidence in industrialized agro-food systems and their increasing reflexivity known as “quality turn” have led to the promotion of Short Food Supply Chains (SFSCs) as opposite to conventional markets. SFSCs have the potential to enhance the sustainability of conventional food systems in terms of socio-economic equity and environmental and local development, addressing post-modern consumer’s habits and purchasing motivations that are extremely heterogeneous in natures. In addition, the renewed EU Common Agricultural Policy 2014-2020 encourages the promotion of SFSCs for the first time through a specific financial support within its II pillar, providing a publicly funded stimulus for sustainable development. This research aims at contributing to the growing literature on SFSCs, investigating consumer preferences and behavior towards purchasing food in such alternative schemes through a multidisciplinary approach. Based on some preliminary qualitative findings, this research explores the importance of some major drivers in influencing consumers’ preferences and purchasing behavior (i.e., sustainability, trust, fairness) more in depth. The broader objective is to provide new knowledge around SFSCs’ growing appeal among consumers, focusing especially on Italy, to explain their recent increasing in number. From a socio-psychological approach, i.e. the Theory of Planned Behavior, the research turns to economic theory with a choice experiment (CE) based on an hypothetical market situation and focusing on farmers’ markets. Italian consumers have been also compared with Brazilian and German consumers. Generally speaking, findings show the important role of sustainability and trust in influencing food purchases at SFSCs, as well as consumers’ renewed emphasis on both some traditional marketing patterns (i.e., face-to-face interactions with the producer) and the possibility to contribute to farmers’ income

WIPI1, BAG1 and PEX3 autophagy-related genes are relevant melanoma markers

ROS and oxidative stress may promote autophagy; on the other hand, autophagy may help reduce oxidative damages. According to the known interplay of ROS, autophagy, and melanoma onset, we hypothesized that autophagy-related genes (ARGs) may represent useful melanoma biomarkers. We therefore analyzed the gene and protein expression of 222 ARGs in human melanoma samples, from 5 independent expression databases (overall 572 patients). Gene expression was first evaluated in the GEO database. Forty-two genes showed extremely high ability to discriminate melanoma from nevi (63 samples) according to ROC (AUC ≥ 0.85) and Mann-Whitney (p < 0.0001) analyses. The 9 genes never related to melanoma before were then in silico validated in the IST online database. BAG1, CHMP2B, PEX3, and WIPI1 confirmed a strong differential gene expression, in 355 samples. A second-round validation performed on the Human Protein Atlas database showed strong differential protein expression for BAG1, PEX3, and WIPI1 in melanoma vs control samples, according to the image analysis of 80 human histological sections. WIPI1 gene expression also showed a significant prognostic value (p < 0.0001) according to 102 melanoma patients' survival data. We finally addressed in Oncomine database whether WIPI1 overexpression is melanoma-specific. Within more than 20 cancer types, the most relevant WIPI1 expression change (p = 0.00002; fold change = 3.1) was observed in melanoma. Molecular/functional relationships of the investigated molecules with melanoma and their molecular/functional network were analyzed via Chilibot software, STRING analysis, and gene ontology enrichment analysis. We conclude that WIPI1 (AUC = 0.99), BAG1 (AUC = 1), and PEX3 (AUC = 0.93) are relevant novel melanoma markers at both gene and protein levels

c-Flip KO fibroblasts display lipid accumulation associated with endoplasmic reticulum stress

c-Flip proteins are well-known apoptosis modulators. They generally contribute to tissue homeostasis maintenance by inhibiting death-receptor-mediated cell death. In the present manuscript, we showthat c-Flip knock-out (KO) mouse embryonic fibroblasts (MEFs) kept in culture under starvation conditions gradually modify their phenotype and accumulate vacuoles, becoming progressively larger according to the duration of starvation. Large vacuoles are present in KO MEFs though not in WT MEFs, and are Oil Red-O positive, which indicates that they represent lipid droplets. Western blot experiments reveal that, unlikeWTMEFs, KOMEFs express high levels of the lipogenic transcription factor PPAR-γ. Lipid droplet accumulation was found to be associated with endoplasmic reticulum (ER) stress activation and autophagic modulation valuated by means of BIP increase, LC3 lipidation and AMP-activated protein kinase (AMPK) phosphorylation, and p62 accumulation. Interestingly, XBP-1, an ER stress-induced lipogenic transcription factor, was found to preferentially localize in the nucleus rather than in the cytoplasm of KO MEFs. These data demonstrate that, upon starvation, c-Flip affects lipid accumulation, ER stress and autophagy, thereby pointing to an important role of c-Flip in the adaptive response and ER stress response programs under both normal and pathological conditions

Ion channel expression in human melanoma samples. in silico identification and experimental validation of molecular targets

Expression of 328 ion channel genes was investigated, by in silico analysis, in 170 human melanoma samples and controls. Ninety-one members of this gene-family (i.e., about 28%) show a significant (p 0.90 and p 90% in most cases). Such five genes (namely, SCNN1A, GJB3, KCNK7, GJB1, KCNN2) are novel potential melanoma markers or molecular targets, never previously related to melanoma. The “druggable genome” analysis was then carried out. Miconazole, an antifungal drug commonly used in clinics, is known to target KCNN2, the best candidate among the five identified genes. Miconazole was then tested in vitro in proliferation assays; it dose-dependently inhibited proliferation up to 90% and potently induced cell-death in A-375 and SKMEL-28 melanoma cells, while it showed no effect in control cells. Moreover, specific silencing of KCNN2 ion channel was achieved by siRNA transfection; under such condition miconazole strongly increases its anti-proliferative effect. In conclusion, the present study identified five ion channels that can potentially serve as sensitive and specific markers in human melanoma specimens and demonstrates that the antifungal drug miconazole, known to target one of the five identified ion channels, exerts strong and specific anti-melanoma effects in vitro

Cancer microenvironment and endoplasmic reticulum stress response

Different stressful conditions such as hypoxia, nutrient deprivation, pH changes, or reduced vascularization, potentially able to act as growth-limiting factors for tumor cells, activate the unfolded protein response (UPR). UPR is therefore involved in tumor growth and adaptation to severe environments and is generally cytoprotective in cancer. The present review describes the molecular mechanisms underlying UPR and able to promote survival and proliferation in cancer. The critical role of UPR activation in tumor growth promotion is discussed in detail for a few paradigmatic tumors such as prostate cancer and melanoma

Toward the implementation of the income stabilization tool: An analysis of factors affecting the probability of farm income losses in Italy

Over the last years, EU agricultural farms suffered an increased sensitivity to market fluctuations in terms of both production flows and incomes, due especially to climate change and market globalisation. In addition to previous instruments as insurances and mutual funds, the new reform of European Common Agricultural Policy 2014-2020 proposes a new instrument, namely the Income Stabilisation Tool (IST), in order to specifically support farmers' severe income drops. This study aims at contributing to the lively debate on risk management linked to the implementation of the IST by estimating the probability of income loss in relation to farm attributes in Italy

A counter insurgency study an analysis of local defenses

Local Defenses are view by many counterinsurgency strategists as an essential element in defeating an insurgency. Providing a population with a local defense organization will strongly support the government's strategy of extending its security and control over the rural areas affected by insurgent organizations. However every insurgency is unique and demands a unique counterinsurgency strategy to be defeated. There always will b e an important commonality: insurgent organizations need popular support to subsist. The final success of the government or the insurgents will be determined by the capacity of either both to win and retain the support among the rural population. The analysis of the four cases presented in this study clearly demonstrates each government's approach to the insurgent problem, including the use of local defenses to protect rural populations from insurgent attacks and influence. The organization of localdefenses during the Malayan Emergency, the El Salvador's Civil War, as well as the Vietnam War and the Terrorist Epoch in Peru proved to be a force multiplier for the government's effort, at least during the time period in which they were effectively implemented. The contribution of this analysis is not that of providing a framework or recipe for strategists to implement this kind of organizations. Rather, the contribution of this study is on a set of variables to be considered when planning the implementation of local defenses as part of a counterinsurgency effort.http://archive.org/details/acounterinsurgen109451424Lieutenant Commander, PeruMajor, United States ArmyApproved for public release; distribution is unlimited

Exploring the Interplay of Risk Attitude and Organic Food Consumption

This exploratory study seeks to advance the research on consumers’ preference for organic food eating by investigating the potential association between organic food consumption (OC) frequency and the individual risk attitude. Moreover, inspired by the literature, we investigate also the association between OC and the perceived OC related health improvement (i.e. the avoidance of health risks), subjective trust toward the certification, social norms and several lifestyle factors. Based on a direct survey of 223 Italian college students, a lottery task was used to characterize the individual risk attitude and a simultaneous equation model was estimated. This study marks a beginning by showing a significant relationship between being risk averse and a high organic food eating, offering a hint for future research avenues in the organic domain

GROWTH, LOCAL STRUCTURAL AND ELECTRONIC PROPERTIES, AND BAND ALIGNMENT AT SRTIO3-BASED ALL-OXIDE HETEROJUNCTIONS

Recent advances in the growth of epitaxial oxide thin films have fostered a steady increase of research on oxide heterojunctions, which are now produced with unprecedented quality. Applications of these systems in the field of electronics, photovoltaics and photocatalysis strongly rely on the capability to master band gap engineering on the atomic scale. Strontium titanate (SrTiO3) is the substrate of choice commonly used in the production of all-oxide heterostructures, as in many cases these systems display a two dimensional electron gas (2DEG) confined at the interface, such as in the LaAlO3/SrTiO3 junction. The band offsets at the interface determine on which of the constituent materials the 2DEG will be confined, and provide additional information on the degree of confinement as well, so the study of the band alignment in oxide-based heterostructures is of crucial importance. Novel properties and functionalities can be achieved upon substitution of LaAlO3 with other oxide materials, an example being the BiFeO3 perovskite. In fact, BiFeO3 is a multiferroic material, and a complete control of the BiFeO3/SrTiO3 heterostructure may allow the tuning of the 2DEG at the interface through the application of an external electric or magnetic field. A 2DEG is also observed in the Al2O3/SrTiO3 junction, which show electron mobilities greater than those previously measured in perovskite-based heterojunctions. Despite in many structures the 2DEG is not observed, different interesting applications can still be obtained, such as in the CuO/SrTiO3 heterostructure, which is a promising material for the production of solar cells and for photoelectrochemical water splitting applications. The main objective of this work is to demonstrate the growth of high quality BiFeO3/SrTiO3, Al2O3/SrTiO3 and CuO/SrTiO3 heterostructures by off-axis sputtering, and to provide a detailed analysis of the interface properties. The band alignment at the interface is thus measured and discussed for each of these materials