Introduction: Eating behavior often becomes unhealthier during the transition from adolescence to young adulthood, but not much is known about the factors that drive this change. We assess the available evidence on this topic through a literature review and pay special attention to the research designs employed in the studies available as well as the modifiability of the factors investigated in previous research. Method: We systematically conducted a scoping review by searching literature published in or after 2000 in three databases that described one or more factors associated with eating behavior or changes in eating behavior during the transition from adolescence to adulthood in the general population. Our search identified eighteen articles meeting these inclusion criteria. The socio-ecological DONE (Determinants of Nutrition and Eating) framework, a recently developed dynamic framework of factors shaping dietary behavior, was used to structure and categorize the factors identified. Results: Most factors identified in the literature were individual-level factors (67%) such as food beliefs, time constraints, and taste preferences; on the other hand, interpersonal-level factors , environmental-level factors  and policy-level factors  have been reported on less extensively. Furthermore, most factors discussed in the literature have been classified in the DONE framework as not easily modifiable. Moreover, previous studies largely used static research designs and focused primarily on one specific population (US freshmen). Discussion: This systematic scoping review identified several gaps in the available literature that hinder insight into the drivers of eating behavior (change) during the transition from adolescence to young adulthood. There is an urgent need for research on broader populations, employing dynamic repeated-measures designs, and taking modifiability of factors into account. The transition from adolescence to young adulthood, typically thought to occur between 18 and 25 years of age . This trCorroborating this assumption, it has been shown that changes in eating behavior  are important contributors to weight gain during the transition from adolescence to young adulthood ,5,6,7. TWhereas substantial evidence shows that emerging adulthood carries an increased risk for unhealthy eating ,5,6,7,8,In order to synthesize the available evidence about the factors shaping eating behavior during the transition from adolescence to young adulthood, we conducted a scoping review of currently available literature. This review accumulates evidence available about the factors driving eating behavior in emerging adulthood and organizes the factors that have been identified in the literature according to a socio-ecological framework of determinants of eating behavior, the multidisciplinary DONE (determinants of nutrition and eating) framework . FurtherA scoping literature search was systematically performed in three databases: Web of Science (WoS), PubMed (PM), and PsycInfo (PI). A scoping literature review ,14 has aThe search was conducted by the first (FMS) and last (TD) authors, with TD searching one database (WoS) and FMS searching two databases (PM and PI). FMS and TD also screened and selected the records that came out of these respective databases, discussing any record that raised questions until agreement was reached. Remaining doubts or disagreements were further discussed and resolved with the second author (BR). Both authors involved in the search and selection process (FMS and TD) have extensive background in performing systematic literature searches, having been previously involved in multiple successful systematic literature reviews ,19,20,21We maintained the following inclusion criteria: (1) The article had to mention at least one factor that was associated with eating behavior. For the purposes of this article, we were interested in actual eating behaviors such as food intake , food choice, and portion size as outcome measures. We excluded articles focusing on outcomes such as attitudes or intentions regarding food or eating, articles focused on weight change rather than eating behavior, as well as articles focusing on problematic eating behaviors as disordered or pathological eating, emotional eating, and night-time eating; (2) Participants in the study had to come from the general population, so studies on specific subgroups  were excluded; (3) In order to ensure that studies truly focused on determinants of eating behavior during the transition period from adolescence to young adulthood, we set the criterion that the time frame of prospective quantitative studies should include the transition period. Quantitative studies employing a sample of college or university students were therefore included only when the sample consisted of freshmen; those incorporating a larger age range  were excluded. Qualitative studies were included if they were aimed specifically at the transitional period (retrospectively), regardless of when the study was conducted. N = 8 for WoS; N = 3 for PM; N = 6 for PI) as well as between the databases (N = 45). After removing these duplicates, articles were fully screened. A total of 94 articles were excluded after full screening. Exclusion of articles happened for seven reasons : (1) the sample did not meet the age-related inclusion criterion that was set (N = 50); (2) the sample represented a specific subgroup rather than the general population (N = 13); (3) the study outcomes were related to weight (change) rather than eating behavior (N = 9); (4) the article described changes in eating behavior but did not investigate any factors associated with such changes (N = 8); (5) the article described an intervention (N = 7); (6) the study outcomes were related to emotional, external, or disordered eating (N = 5); (7) the study outcome was attitudes or intentions about eating, rather than actual eating behavior (N = 2). After full screening of the remaining articles, a set of eighteen unique articles [articles ,38,39,40In order to assess to which extent the studies included in the review are able to capture the dynamic process by which changes in shaping factors affect subsequent changes in eating behavior, the study design of each article included in the review was recorded. This was done using the classification proposed by Renner and colleagues  depictedThe multidisciplinary DONE (Determinants of Nutrition and Eating) framework  was emplAn important advantage of the framework is that the factors’ modifiability, relationship strength and population-level effect have been rated by almost 200 experts . Together, these ratings provide an indication of priority for research for each of the factors included in the framework. For purposes of the current perspective, we will focus on the ratings of modifiability only. Possible modifiability scores range from 1 (low modifiability) to 3 (high modifiability). Average modifiability per stem-category  as judged by the experts involved in the development of the DONE framework, is used as an indicator for the modifiability of the specific factors identified in the current literature search. This rating indicates to what extent experts in the field consider it theoretically possible to modify the average determinant in this stem-category.changes in eating behavior that occur during the transition from adolescence to young adulthood. While longitudinal study designs are better equipped to do this, all but one of the longitudinal studies identified in the literature search  used a simple or adjusted longitudinal design [The eighteen articles identified described results of original research on factors associated with (changes in) eating behavior during the transitional period occurring between adolescence and young adulthood. The fact that only eighteen articles were found that met the inclusion criteria corroborates the idea that there is a paucity of studies on this topic. Of the eighteen studies, twelve (67%) of the studies used a sample of college or university students ,38,39,40l design ,33,39,40l design , would bl design . N = 67; 64%) of the factors identified in the eighteen articles were individual-level factors. Within these individual-level factors, the majority of factors (N = 39) were psychological factors, which thus clearly and consistently emerged as drivers of eating behavior during emerging adulthood. The specific leaf-categories from which psychological factors were identified were ‘mood and emotions’ ; ‘self-regulation’ ; ‘health cognitions’ ; ‘food knowledge, skills and abilities’ ; ‘food beliefs’ ; ‘food habits’ ; and ‘weight control cognitions and behaviors’ . Other individual-level factors were situational factors , biological factors  and demographic  These factors generally had relatively low modifiability scores (average modifiability across the leaf-categories = 1.83 out of a maximum of 3.00), with modifiability judged slightly higher for factors from the ‘psychological’ and ‘situational’ stem-categories than for factors from the ‘biological’ and ‘demographic’ stem-categories. One hundred and five distinct factors were described in the articles (see N = 18 factors were found (17%), which were sorted into the specific leaf-categories of ‘family food culture’ ; ‘household socio-economic status’ ; ‘social influence ; ‘social support’ ; ‘parental behaviors’ ; and ‘cultural cognitions’ . Modifiability of these factors was relatively low as well (average modifiability across the leaf-categories = 1.82 out of 3.00). At the environmental level, 18 factors were found (17%). Most of these environmental factors were related either to the product  or to the micro-environment . At the meso- and macro-environmental level, on the other hand, only N = 3 factors were found . Importantly, modifiability of factors at the environmental level was judged by the experts to be higher (average modifiability across the leaf-categories represented in the review = 2.15 out of 3.00). Finally, only N = 2 factors were identified at the policy level (2%), the most distant socio-ecological level in the DONE framework . Modifiability of policy-level factors was judged to be moderate (modifiability of the leaf-category represented in the review = 2.11 out of 3.00). Fewer upstream factors were identified. At the interpersonal level, To summarize, our literature review identified eighteen articles describing factors shaping eating behavior change in emerging adulthood. Synthesis of these articles indicated that all but one of the studies described in these articles employed static research designs . Only ochange; secondly, the lack of focus on the modifiability of the factors shaping eating behavior change during emerging adulthood; and thirdly, the strong focus on college freshmen as study populations.A scoping review of the available literature on the factors shaping eating behavior (change) during the transition from adolescence to young adulthood confirmed that there is a paucity of research on this topic; only eighteen articles were identified that reported on shaping factors of eating behavior in emerging adulthood. Synthesis of these articles indicated that most studies employed static research designs, mainly studied college students, and primarily investigated individual-level factors which had rather low modifiability—more upstream factors (which tend to have higher modifiability scores) were identified much less often. From this research synthesis, we thus identify several important limitations of the current body of research. In our perspective, three main issues are of relevance: firstly, the lack of studies using a design that can properly establish which factors truly affect eating behavior Importantly, the nature and design of the studies that have been conducted to date are not ideally suited to determine which factors are systematically and strongly associated with changes in eating behavior throughout the transition. The scoping review revealed that previous quantitative studies have overwhelmingly employed static designs  which factors are strongly and systematically associated with changes in eating behavior; and (b) which of these factors are modifiable (presuming that changing these factors will subsequently change the behavior). Therefore, two crucial aspects for new research in this area are the use of dynamic research designs capable of capturing changes in both shaping factor and behavioral outcome, and a focus on modifiability of the shaping factors under investigation. Scholars should focus not only on individual-level factors, but pay substantial attention as well to more upstream factors, as there are indications  that fa


Pluripotency is orchestrated by distinct players and chaperones and their partners have emerged as pivotal molecules in proteostasis control to maintain stemness. The proteostasis network consists of diverse interconnected pathways that function dynamically according to the needs of the cell to quality control and maintain protein homeostasis. The proteostasis machinery of pluripotent stem cells (PSCs) is finely adjusted in response to distinct stimuli during cell fate commitment to determine successful organism development. Growing evidence has shown different classes of chaperones regulating crucial cellular processes in PSCs. Histones chaperones promote proper nucleosome assembly and modulate the epigenetic regulation of factors involved in PSCs’ rapid turnover from pluripotency to differentiation. The life cycle of pluripotency proteins from synthesis and folding, transport and degradation is finely regulated by chaperones and co-factors either to maintain the stemness status or to cell fate commitment. Here, we summarize current knowledge of the chaperone network that govern stemness and present the versatile role of chaperones in stem cells resilience. Elucidation of the intricate regulation of pluripotency, dissecting in detail molecular determinants and drivers, is fundamental to understanding the properties of stem cells in order to provide a reliable foundation for biomedical research and regenerative medicine. Embryonin vitro  brought in vitro . OCT4, N network .Although many studies have been conducted to understand the exact mechanisms by which the TFs and additional factors regulate pluripotency, much remains to be elucidated. Studies are still being conducted to understand TFs individual and integrated functioning. By unmasking pluripotency mechanisms, it will be possible to use PSCs more safely and harness their therapeutic potential, also serving as a model to understand early development, important cellular processes and diseases. Besides transcriptional regulation, other mechanisms are being recently discussed as relevant to understanding the maintenance of pluripotency in stem cells, such as chromatin conformation and proteome quality control assisted by molecular chaperones. In this review we discuss aspects of PSCs maintenance, such as TFs regulation and chromatin conformation in PSCs, as well as the relationship of chaperones, co-chaperones and ubiquitin-proteasome system (UPS) with the control of TF levels and pluripotency in PSCs. A comprehensive and integrated understanding of the events – from transcription, translation to post-translational processes – that govern pluripotency is needed to answer questions that remain unanswered in the field of PSCs.Pluripotent stem cells are regulated by a series of interconnected cellular processes that pass through the transcription, translation, and final destination of proteins through different post-translational modifications. The state of chromatin conformation is important for the exposure or concealment of regulatory regions in DNA. Regulatory regions are possible targets of several TFs, that will associate in a specific way in the DNA molecule and regulate the transcription of several genes.Several studies described the essentiality of the TFs OCT4, SOX2, and NANOG to pluripotency. These TFs form an interconnected self-regulating core, cooperatively associating with their own promoters and co-occupying more than 300 targets in an integrated manner, finely regulating their own and also their targets expression, repressing differentiation genes and activating pluripotency . Other TAlthough pluripotency is controlled by the expression of a network of TFs, the levels of this expression must be highly regulated in order to maintain the pluripotent state. PSCs balance between self-renewal and differentiation potential. Some researches , is used in cell culture . Brieflyin vivo , and is in vivo may contribute to the understanding of their existence as part of the development of organisms, or as artifacts of cell culture.Interestingly, the existence of different statuses of pluripotency has been reported, naïve and primed, with great variety in transcriptional and epigenetic profile  network (PN) ensures the balance of the proteome by coordinating protein synthesis, folding and conformational maintenance; and protein degradation. PN is achieved by an orchestrated system of proteins, including molecular chaperones and their regulators, which help proteins to reach its functionally active conformation, without being part of its final structure. In addition, the UPS exerts a post-transcriptional control on the levels of proteins, such as TFs, which is important to pluripotency maintenance .Considering the fine mechanics of chromatin conformation control, the importance of PN for the maintenance of cellular functions, both in health and in diseases, the increased expression of PN elements in PSCs, as well as an increased activity of PN in these cells, many studies have been conducted to understand the control of pluripotency from the perspective of these events. Regardless of the mediation of chaperones, an important fraction of polypeptides usually exhibit errors in folding or refolding ; consequThe protein levels of pluripotency TFs must be finely regulated for the maintenance of PSCs specific properties. It has been shown that both downregulation and upregulation beyond the required levels of some TFs, such as OCT4 , leads tInvestigating the total profile of ubiquitinated proteins in mESCs and iPSCs, Upon the downregulation of PSMD14, a 26S proteasome non-ATPase subunit, the loss of OCT4 expression is observed, which is linked to an apparent dysfunction of the deubiquitinating enzymatic activity of PSMD14 . FurtherThe proteasome activity is enhanced in hESCs and iPSCs, with a loss of activity being observed as the cells differentiate, with a concomitant increase in differentiation factors, such as PAX6, as well as a decrease in the expression of pluripotency TFs . FurtherThe COP9 signalosome (CSN) is an important protein complex that prevents protein degradation, and was previously implicated in pluripotency maintenance . ExperimMore specifically, it has been previously demonstrated that OCT4 can be post-translationally modified by ubiquitin in both mESCs and hESCs. WWP2 protein was identified as the first to post-translationally modify OCT4, functioning as an E3 ligase . WWP2 prInterestingly, evidence shows that WWP2 also plays a role in the regulation of SOX2. WWP2 HECT domain recognizes methylation on lysine 119 (K119me) of SOX2, modification that stimulates SOX2 ubiquitination and subsequent degradation via proteasome pathway . TreatmeNANOG was previously described as a target for polyubiquitylation by the F-box protein family member FBXW8, and consequent degradation by proteasome . When phThe role of deubiquitinases (DUBs) started to be investigated in the context of proteostasis maintenance in PSCs. In brief, DUBs remove ubiquitin from protein substrates in order to maintain their targets stability. A recent study in mESCs, scanning for potential deubiquitinases regulators of NANOG, found that deubiquitinase ubiquitin-specific protease 21 (USP21) stabilizes NANOG protein levels removing K48-linked polyubiquitylation . USP21 dA summary of the proteasome-related proteins presented in this section and their function in PSCs can be found in As mentioned before, several factors affect the expression of pluripotency genes, and epigenetic modulations of transcription have been broadly studied to gain insight into the mechanisms which rule the rapid proliferation and turnover of pluripotent cells into differentiation. In the context of chaperone proteins, histone chaperones have recently been identified as important factors in regulating pluripotency. Here, we discuss the importance of histone chaperones in the modulation of transcription in PSCs, mainly ESCs, and their role in maintaining pluripotency through epigenetic modifications.Chromatin in eukaryotes is organized in complexes called nucleosomes composed of 147 pairs of bases of DNA associated with a core of small basic proteins, the histones, which form an octamer of two copies of each protein H2A, H2B, H3, and H4, binding a linker histone H1 . HistoneHistone chaperones are molecules that associate with histones and present an important role in histones dynamics. These chaperones are responsible for the transferring of histones to the DNA, and they can modulate histones modifications as acetylation and methylation or remodeling nucleosomes during transcription, among other important functions. Histone chaperones can work single, as the chaperone anti-silencing function 1 (ASF1), or form complexes as chromatin assembly factor 1 (CAF1) and the facilitates chromatin transcription (FACT), presenting relevant roles in post-translational histone modifications .Embryonic stem cells present rapid changes in transcription associated with transition from pluripotency to a more differentiated state, therefore their chromatin is characterized by an open state with a less condensed structure and predominance of variant histone modified post-translationally  involved in transcription activation . One of HIRA is also required in mESC for the formation of H3.3 complex with polycomb repressive complex 2 (PRC2), which control gene transcription during lineage commitment in these cells through trimethylation of lysine 27 on histone H3 (H3K27me3) . This coRunt-related transcription factor 1 (RUNX1) is essential for hematopoietic cells transition and HIRA can modulate RUNX1 targets participating of mESCs differentiation process to hemogenic endothelium through its interaction with RUNX1 and deposition of H3.3 variant . AdditioTranscriptional regulator ATRX (ATRX) is a member of SNF2 family of chromatin remodeling factors and presents histone chaperone activity, forming a chromatin-remodeling complex with the death domain-associated protein DAXX. ATRX and DAXX associate with H3.3 in a HIRA-independent manner modulating H3.3 deposition at telomeres and repression of telomeric RNA in mESCs . DAXX fuASF1a and ASF1b are present; ASF1a depletion is embryonic lethal while ASF1b was correlated with differentiation potential during early embryonic stages and gonad development  complex is formed by 3 subunits P150, P60, and P48/RbAp48a and is related to processes involved in DNA synthesis and repair , and alsCheloufi and colleagues recently demonstrated the participation of CAF-1 in the reprogramming of mouse fibroblasts into iPSCs, since the negative modulation of CAF-1 expression produces a decrease in somatic cells heterochromatin domains, increasing the binding of SOX2 to pluripotency specific targets and somatic cellular plasticity . InteresThe histone chaperone FACT is a complex composed by two subunits, SSRP1 and SPT16, and participates in transcription elongation . In mESCSeveral other histone chaperones have presented important functions on ESCs biology and somatic cells reprograming. The mammalian high-mobility group AT-hook 2 (HMGA2) are highly expressed in ESCs and not translated in somatic normal cells, working as a replication fork chaperone stabilizing its integrity, in a DNA repair mechanism. During the proliferation of highly replicating cells as ESCs or cancer cells, HMGA2 expression is irrelevant, however, when forks are arrested; HMGA2 binds with high affinity to branched DNA, stabilizing stalled forks and preventing DNA mutations .Another histone chaperone, nucleoplasmin-2 (NPM2), was recently associated with human dermal fibroblasts reprogramming. NPM2 associated with its histone variants TH2A and TH2 can improve the reprogramming modulated by OCT4, SOX2, KLF4, and c-MYC, generating iPSCs in a more naïve state compared to the classical TFs alone . NPM2 phSPT6 is a histone chaperone and works as a transcription elongation factor in mESCs, modulating negatively H3K27 acetylation and methylation through its interaction with PRC2 core subunits, which maintains pluripotency and avoids differentiation mediated by the accumulation of H3K27me3 deposition at ESC critical super-enhancers . SET is Finally, it is well known that PSCs present rapid turnovers to maintain an undifferentiated state or for entry into cellular commitment. Epigenetic mechanisms of gene activation that depends on histone variants allow these rapid turnovers observed in stem cells, especially PSCs, and histone chaperones are determinant for the refined work of histones modifications. Briefly, here we described how histones chaperones present relevant roles on development, pluripotency maintenance and somatic cell reprograming, as their main function is to modulate these essential epigenetic changes related to histones modifications and deposition, which have great participation on the activation and repression of TFs. Elements of the PNs are enhanced in PSCs, such as chaperones and co-chaperones. Chaperones are classified in heat-shock proteins (HSPs) such as small heat shock proteins (sHSPs), HSP60, HSP70, HSP90, and HSP100. HSP70 and HSP90 are the main families of chaperones induced in response to cellular stress and they act along TFs known as heat shock factors (HSF), which regulate several genes encoding chaperones .de novo folding to establish functional structures (The heat shock factor 1 (HSF1) creates a dormant complex with HSP90 and HSP70 under balanced conditions, however, in the presence of misfolded proteins and cytotoxic protein aggregation, chaperones detach from HSF1, which then induces members of the PN to decrease production of new clients . When thructures . HSPs arructures .Both protein quality control and the maintenance of the PN are indispensable for cellular biology and for the health of whole organism. Important additional pathways such as the cytosolic stress response and the unfolded protein response (UPR) are part of this complex that contributes to circumvent the accumulation of misfolded molecules . HoweverHeat shock protein 90 (HSP90) is a highly conserved stress response protein in eukaryotic cells, being the most abundant protein in unstressed cells, modulates the activity, turnover and trafficking of several proteins and participates of signal transduction. HSP90 forms heterocomplexes with different proteins through the formation of a complex composed by HSP90 and another four chaperones – HSP70, HSP Organizing Protein (HOP), HSP40, and p23 – binding to several clients as steroid receptors. The assembly requires HSP90 and HSP70 bound by HOP and incorporates HSP40 and p23 during the process, assisting the folding and refolding of naïve proteins for properly active conformation . This chSeveral studies support the importance of HSP90-HOP-HSP70 complex on PSCs biology. Recent data from our group determined that HOP, HSP70, and HSP90 are found as cargo of extracellular vesicles (implicated in intercellular communication) of mESCs, suggesting their participation in processes related to development, since the microenvironment is important for pluripotency maintenance through the modulation of several signaling pathways ,b.Constitutive HSP90 is required for mouse embryos development and maintenance of pluripotency via HOP-STAT3 interaction ; moreoveHSP90 has been described in literature as an essential molecule for pluripotency maintenance. The inhibition of HSP90 expression decreases OCT4, NANOG and pSTAT3 levels and leads to mESCs differentiation, preferentially to the mesoderm layer. HSP90 associates with OCT4 and NANOG to protect them from ubiquitin proteasome degradation  and is aHSP90 partner HOP is essential for mouse development since its knockout (KO) is embryonic lethal and HOP KO mouse do not reach E10.5 development stage . HOP wasHSP70 is constitutively expressed and its expression can be inducted during stress. In hESCs, constitutive HSP70 expression is slightly increased compared to somatic cells and is located into the cytoplasm and nucleus independent of differentiation status. However, in hESCs HSP70 can also be found on cell surface , which iThe HSPS from the family HSP70 HSPA1A (HSP70A1), HSPA1B (HSP70B1), and HSPA9A  are highly expressed in mESCs compared to differentiated cells and are associated with increased resistance of these cells against genotoxic stress . AdditioRecently, it was demonstrated that upregulation of HSP70 in iPSCs caused by stress as hypoxia promotes survival through the inhibition of apoptosis pathways, however, it also inhibits STAT3 phosphorylation leading to differentiation  and incrDifferentiation of mESCs into neurogenic EBs (NEBs) leads to a decrease in the expression of mortalin, a chaperone from the family of HSP70 (HSP70A9) . AnotherHSP60 is a target gene of c-MYC  and other TFs, epigenetic regulators and specific signaling pathways are key elements to orchestrate pluripotency. Although these well-established TFs are of undeniable importance for pluripotency maintenance, the search for additional TFs that are able to interact with core factors to regulate processes related to pluripotency should be enhanced, and the role of an expanded core has just begun to be dissected. Moving forward to the debate on the importance of TFs for pluripotency and stemness regulation, this will be a fruitful area for further research.As presented here, emerging evidence points out the network plasticity of chaperones controlling the activities of key players involved in pluripotency maintenance. Chaperones play roles from shaping chromatin dynamic to controlling transcriptional regulation of pluripotency genes, to the assistance of proper folding of these genes when translated into proteins in different PSCs models , 2. The This review brings together classical and recent research on the control of pluripotency, going through broad important cellular processes related to this unique and promising feature. The diversity of these processes embraces different levels of cellular regulation and shows how complex is the understanding of the pluripotency context.CF and RI conceived the presented idea and proof outline, did the literature review, organized the tables, and wrote distinct topics. MP did the literature review and wrote a topic. MM-E did the literature review, wrote a topic, and prepared the illustration art-, 2. ML cThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Blood donation screening for West Nile virus (WNV) was mandatory in the Lazio region in 2017 and 2018 (June-November) according to the national surveillance plan. In these years, all five donations reactive in WNV nucleic acid amplification tests harboured instead Usutu virus (USUV). Clade ‘Europe 2’ was identified in four blood donations and a 2018 mosquito pool. The cocirculation of WNV and USUV in Lazio warrants increased laboratory support and awareness of possible virus misidentification. In the summer and autumn seasons of 2017 and 2018, following serological identification of West Nile virus (WNV) infection in horses, mandatory WNV screening with nucleic acid amplification test (NAT) was established for blood donations in some provinces of the Lazio Region in central Italy: Viterbo (starting from 18 August 2017), Latina (12 September 2018) and Rome (10 October 2018). During this period, five blood donations were reactive in WNV NAT screening. In none of these samples could WNV positivity be established by further tests conducted at the Regional Reference Laboratory. In the present study, we describe serological, detailed molecular and phylogenetic analyses undertaken to better characterise the WNV positivity observed during the screening. For the Lazio Region, centralised blood screening for WNV is performed at the Biological Qualification Center (CQB) for NAT blood screening at the Sandro Pertini Hospital in Rome, using NAT . Blood donations reactive in WNV NAT screening and confirmed twice by the same method at the CQB, are considered not suitable for the transfusion service and are discarded. Plasma aliquots of these donations are sent to the Regional Reference Laboratory for Arbovirosis  for confirmatory testing and further diagnostic investigation. Donors who provided WNV NAT-positive donations are recalled 2–3 weeks after donation, to complete serological investigation.All five donors reactive in repeated WNV NAT were asymptomatic and fulfilled the eligibility criteria concerning WNV risk. According to Ministerial Decree of 2 November 2015, in order to guarantee self-sufficiency in blood components during the summer period, it is recommended to use the WNV NAT test as an alternative to the provision of 28 days suspension of donors who have spent at least one night in areas at risk of WNV infection [Plasma samples were concentrated by ultracentrifugation and tested with a nested pan-flavivirus-RT-PCR targeting the NS5 gene  and not WNV. These results were further confirmed with a USUV-specific real-time RT-PCR, modified from Nikolay et al. , and by The serological investigation revealed that four donors did not have WNV- or USUV-specific antibodies at donation, while donor #1 had both IgG and IgM specific for WNV. In follow-up samples, obtained 2–3 weeks apart, USUV seroconversion was observed for donors #2 and #5, while the USUV-specific antibody titre exceeded the WNV-specific antibody titre in donor #1 .C. pipiens were divided in pools of at most 100 specimens and analysed by RT-PCR for virus detection. According to the national integrated surveillance plan for WNV and USUV, the Istituto Zooprofilattico Sperimentale delle Regioni Lazio e Toscana performed the entomological investigation: adult mosquitoes were sampled at sites with known virus circulation (areas where horses positive for anti-WNV antibodies or blood donors reactive in WNV NAT had been identified) using traps of the model Italian Mosquito Trap , BG Sentinel  and Gravid . The sampling protocol was defined depending on the number of WNV cases in the area and on the number of caught mosquitoes. After mosquito sorting and identification, those of the species C. pipiens were caught, divided into 38 pools and tested by RT-PCR for the presence of WNV and USUV RNA. Among the 38 tested pools, 14 were positive for USUV (minimum infectious rate: 1.3); they originated from two municipalities in the Latina province. Sequencing indicated that these viruses belonged to the ‘Europe 2’ clade and were very similar to those obtained from blood donors #2 to #5 . These results were confirmed by the National Reference Centre for Foreign Animal Diseases (CESME), Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise.We also performed molecular analysis on four USUV-positive blood donations identified in 2017 by the Regional Reference Laboratory of the Lombardy Region  in northern Italy, where endemic cocirculation of different USUV strains was reported . Their mUSUV is a mosquito-borne flavivirus, member of the Japanese encephalitis virus group, with six distinct clades so far recognised . Its natTurdus merula); however, a retrospective study showed that it has been present in Italy at least since 1996 [USUV was isolated in South Africa in 1959 and was first reported in Europe in 2001, causing mass mortality of Eurasian blackbird  . In 2017Partial molecular characterisation indicated close genetic relationships between human and mosquito USUV strains detected in Lazio region; all belonged to the ‘Europe 2’ clade. The only USUV strain of clade ‘Europe 3’, detected in donor #1, was presumably acquired in Switzerland, according to the travel history of the donor; in addition, this donor’s place of residence, Argentina, accounted for the serological pattern observed at donation, that was consistent with previous exposure to other related flaviviruses.The presence of WNV lineage 2 detected by molecular assay in the brain of a symptomatic horse in the province of Latina is evidence of the cocirculation of WNV and USUV in this area. Data from blood donors in Lombardy confirmed the cocirculation, in northern Italy, of different USUV strains (belonging to clades ‘Europe 2’ and ‘Europe 4’). This finding is in line with previous reports  and indiThis is the first report showing circulation of USUV virus in the Lazio region, both in mosquito vectors and in human hosts, while so far, USUV has not been detected in other animal hosts (neither equids nor birds).This study corroborates the hypothesis that USUV can cause clinically asymptomatic infection in humans ; this, iPublic health authorities, blood transfusion services and clinicians should be aware of the expanding risk of USUV infection in humans, especially during the summer, and advanced laboratory support is required to address the problem of possible virus misidentification by screening tests.


Though enriched in neurons, CN can become strongly induced in subsets of activated astrocytes under different pathological conditions where it interacts extensively with the nuclear factor of activated T cells (NFATs). Recent work has shown that regions of small vessel damage are associated with the upregulation of a proteolized, highly active form of CN in nearby astrocytes, suggesting a link between the CN/NFAT pathway and chronic cerebrovascular disease. In this Mini Review article, we discuss CN/NFAT signaling properties in the context of vascular disease and use previous cell type-specific intervention studies in Alzheimer’s disease and traumatic brain injury models as a framework to understand how astrocytic CN/NFATs may couple vascular pathology to neurodegeneration and cognitive loss.Calcineurin (CN) is a Ca Cerebrovascular pathology is one of the leading causes of cognitive loss and mortality. While stroke is usually the most devastating form of cerebrovascular disease, other forms of vascular damage and dysfunction including microinfarcts, microhemorrhages, cerebral amyloid angiopathy and cerebral hypoperfusion are more insidious and can lead to chronic and progressive cognitive loss, especially in aged individuals. These vascular contributions to cognitive impairment and dementia (VCID) are the second leading cause of dementia, behind Alzheimer’s disease, and frequently co-exist with other neurodegenerative conditions  and its dysregulation in astrocytes as a pathological mechanism and potential target for neurodegeneration and cognitive loss due to cerebrovascular damage.Brain ischemia results when stroke or other forms of VCID block the blood supply to parts of the brain, resulting in depletion of oxygen and glucose. This depletion rapidly exhausts the energy production of neural cells and their ability to maintain the normal balance of ions across cellular membranes, thus causing excitotoxicity and Cats Choi, . Ca2+ ov2+/calmodulin. CN consists of a catalytic subunit (PPP3CA) and a Ca2+ binding regulatory subunit (PPP3R1). When cellular Ca2+ levels are low, the phosphatase activity of CN is held in check by an autoinhibitory domain located near the C terminus of the catalytic subunit. The interaction of Ca2+ with the CN regulatory subunit and calmodulin leads to a physical interaction between the CN catalytic subunit and Ca2+/calmodulin, which, in turn, displaces the AID and frees the catalytic core from inhibition. When cellular Ca2+ levels fall, calmodulin is released from the catalytic subunit and AID-mediated inhibition of phosphatase activity is reinstated , thus perpetuating de-phosphorylation of the myriad of CN targets  vectors equipped with the human GFAP promoter Gfa2 , healthier neurite morphology, reduced synaptic hyperexcitability, and normalized NMDA-to-AMPA receptor activity ratios (Sompol et al., Blockade of CN interactions with NFAT transcription factors, using the peptide VIVIT, has been associated with many beneficial effects in cell culture and intact animal models of neurodegeneration. VIVIT mimics the CN-binding PxIxIT motif found in the regulatory region of NFATs 1–4 (Aramburu et al., Interestingly, many of the same telltale signs of glutamate toxicity, including a loss of GLT1 and neuronal hyperactivity, have been noted in experimental models of ischemia and stroke (Maragakis and Rothstein, Cerebrovascular pathology is one of the leading causes of dementia and a frequently identified comorbid factor in many neurologic diseases, such as Alzheimer’s disease. Numerous studies have reported a role for CN hyperactivity in the pathophysiologic sequelae coupling vascular disruption and damage to neuronal death and cognitive loss. Mounting evidence suggests that CN/NFAT signaling may play a particularly important role in neural changes that arise with astrocyte activation in many different neurodegenerative diseases, including cerebrovascular disease. However, no studies to date have tested the specific involvement of astrocytic CN/NFAT signaling in either global ischemia models, models characterized by localized damage to microvessels, or in models that develop chronic vascular inflammation and microhemhorrages. Based on the observations discussed above, we hypothesize that acutely and chronically developing vascular damage will lead to the activation of astrocytes and hyperactivation of CN/NFAT signaling Figure . In thisSK and CN researched and wrote this manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The consumption of an olive oil rich diet has been associated with the diminished incidence of cardiovascular disease and cancer. Several studies have attributed these beneficial effects to oleic acid (C18 n-9), the predominant fatty acid principal component of olive oil. Oleic acid is not an essential fatty acid since it can be endogenously synthesized in humans. Stearoyl-CoA desaturase 1 (SCD1) is the enzyme responsible for oleic acid production and, more generally, for the synthesis of monounsaturated fatty acids (MUFA). The saturated to monounsaturated fatty acid ratio affects the regulation of cell growth and differentiation, and alteration in this ratio has been implicated in a variety of diseases, such as liver dysfunction and intestinal inflammation. In this review, we discuss our current understanding of the impact of gene-nutrient interactions in liver and gut diseases, by taking advantage of the role of SCD1 and its product oleic acid in the modulation of different hepatic and intestinal metabolic pathways. Indeed,According to conventional dietary recommendations, a healthy diet should include a daily consumption of fats: an inadequate quantity of fat might increase the risk of conditions such as depression, psoriasis, Alzheimer’s disease and schizophrenia ,5. To daEVOO represents the main source of fat in the Mediterranean diet . It is cThe SFO contains a large amount of PUFA (linoleic acid 60% to 70%), but MUFA (oleic acid), SFA (stearic acid) and tocopherol are present as well in adequate quantities ,16. ThisDifferent from the previous two fatty acids, fish oil is a good source of the B12 and D vitamins and omega-3 FAs (EPA and DHA) ,21. The Palm oil is constituted of 51% SFA , 38% MUFA, 11% PUFA , carotenIt is becoming evidently clear that the various dietary oils differ not only in the FA profile, but in the content of non-saponifiable species as well. Studies on olive oil have regularly been more focused on these non-saponifiable components. On the contrary, the direct role of oleic acid has been neglected. Different from other FA, oleic acid is not an essential FA, since it can be de novo synthesized by stearoyl-CoA desaturase 1 (SCD1). Exploring SCD1, the master regulator of oleic acid synthesis, may therefore offer important insights on the role of the oleic acid largely present in olive oil. In this light, in vivo and in vitro studies in which the activity of SCD1 has been shut down represent a valid opportunity to evaluate the direct contribution of oleic acid supplementation with the diet (i.e. EVOO). To date, several clinical trials are currently conducted focusing on SCD1  aiming tLipids ingested with the diet are primarily digested and absorbed in the small intestine, followed by delivery to their sites of action in the body by crossing the liver. Thus, the gastrointestinal tract and the liver are at the nexus between a vast source of nutrients and the rest of the body and are connected both anatomically and functionally. The gut-liver axis is characterized by bidirectional traffic: nutrients and factors derived from gut lumen reach the liver through the portal circulation, while bile acids are released in the small intestine from the biliary duct.In this review, we will particularly focus on the role of the major component of olive oil—the oleic acid. We will exploit the contribution of SCD1 and its product oleic acid in health and diseases, with particular attention to the gut-liver axis. Although it might appear controversial in some cases, it is evidently clear that oleic acid plays an essential part in the development and the homeostasis of our organism, and possible disruption of its pathway may result in disease onset and progression.In order to characterize the role of oleic acid in health and disease, numerous studies and clinical trials have been conducted . SeveralFurthermore, oleic acid displays the ability to prevent SFA-induced inflammation. In high fat diet fed mice, oleic acid administration ameliorated insulin sensitivity, reduced pro-inflammatory cytokines levels as Interleukin-1β, Interleukin-6 and Tumor Necrosis Factor-α and up-regulated the anti-inflammatory Inteleukin-10 and adiponectin levels . ThroughThe peculiar beneficial role of oleic acid has been observed in several diseases including coronary heart disease, rheumatoid arthritis, and cancer. Oleic acid consumption prevents the risk of developing rheumatoid arthritis by increasing leukotriene A3 levels, a potent inhibitor of pro-inflammatory LTB4 . In coloInterestingly, in animal models, oleic acid is able to induce lung injury miming acute respiratory distress syndrome (ARDS). Oleic acid administration induces direct toxicity to the endothelial cells characterized by endothelial necrosis, epithelial injury and neutrophil infiltration ,58. ReceFinally, oleic acid displays a pivotal role in the development of the brain, the organ with the highest lipid content of the body second to white adipose tissue. In this organ, lipids are essential for the correct homeostasis and alterations in lipid metabolism are linked to neurological diseases . Oleic aSCD is an enzyme anchored to the endoplasmic reticulum, where it catalyzes the biosynthesis of MUFA from SFA, either derived from the diet or synthesized de novo. SCD is Δ9-fatty acyl CoA desaturases that catalyzes the formation of a double bond in the cis-delta-9 position of saturated fatty acyl CoA. The preferred substrates of SCD are palmitic (C16:0) and stearic (C18:0) acids, which are converted into palmitoleic (C16:1 n-7) and oleic (C18:1 n-9) acids, respectively  Figure . These pSCD is a highly conserved enzyme with multiple isoforms sharing similarities in protein sequences . To dateThe existence of multiple SCD1 isoforms, sharing a high sequence homology and catalyzing the same biochemical reaction, poses difficulties in order to distinguish the role of each single isoform and its metabolic contribution. To this end, substantial insights into SCD1 functions have been gained by the use of specific mouse models, in which the expression of SCD1 is downregulated or its activity is inhibited. Several genetically engineered whole body and tissue specific SCD1 knockout models have been utilized  as well Although many efforts have been made utilizing total body SCD1KO mice, the confirmation that these mice display altered skin permeability enhancing energy expenditure and protection from high fat diet-induced obesity , has ledThe central contribution of FA to cellular inflammation has been extensively demonstrated . RecentlInflammation caused by abnormal levels of lipids remaining unresolved for an extended period of time may lead to cellular stress and dysfunction, resulting in lipotoxicity, one of the major causes of pathologies including obesity, insulin resistance and cardiovascular disease ,94. The In addition, SCD1 global depletion increases insulin sensitivity and glucose utilization . This miOverall, these metabolic diseases are characterized by a metabolic shift toward biosynthetic reactions aimed to supply new lipid macromolecules. Interestingly, enhanced FA biogenesis is a hallmark of cancer as well. Whereas in obesity and diabetes, the elevated lipid biosynthetic activity result in increased production lipids for energy storage, most of the newly synthesized lipid products are utilized for membrane biogenesis in cancer . The metRecent analyses have revealed a fundamental contribution of SCD1 to the metabolic and signaling pathways closely related to cell replication, survival and tumorigenesis . The molDue to the crucial role of lipid biosynthesis in tumor cells, SCD1 deletion could modulate several phenotypic features of cancer. Specifically, SCD1 enhances cancer cell mitogenesis and survival, increases tumorigenic capacity and tumor cell invasiveness ,111. EveThe liver plays a central role as a metabolic hub, where multiple biochemical processes converge to render food nutrients available to the rest of the body. However, an unbalanced diet—rich in lipid and carbohydrates—together with a sedentary lifestyle, may severely compromise the hepatic health status. Indeed, it has been shown that a high energy intake with a concomitant low energy expenditure resulted in hepatic lipid accumulation, one of the key features of non-alcoholic fatty liver disease (NAFLD). NAFLD is the most common disease of the Western Countries, characterized by a spectrum of conditions that vary from hepatic steatosis to severe form of non-alcoholic steatohepatitis (NASH), which may eventually progress toward cirrhosis and end-stage liver diseases . DespiteLipids are essential for several processes that involve cellular and tissue homeostasis. Expression of lipogenic genes, including SCD1, is essential for hepatic maturation and function . Indeed,Genetically engineered whole body and liver specific SCD1 knockout animal models as well as mice—in which SCD1 activity is suppressed—has provided substantial insights into hepatic SCD1 functions ,123,124.Whole body SCD1 ablation resulted in global modifications of gene expression and metabolic processes that cause loss of body fat and increased insulin sensitivity. Specifically, the transcription of several genes involved in de novo lipogenesis were downregulated in the liver of SCD1KO mice, accompanied by an upregulation of genes involved in lipid β-oxidation . When chThe observations obtained from global SCD1 ablation led to the generation of liver specific SCD1KO (LSCD1KO) mice using Cre-Lox system, in order to investigate whether the hepatic SCD1 might be responsible for the observed phenotypes. Differently from whole body SCD1KO, mice lacking SCD1 in the liver were not protected from both genetic and diet-induce obesity, although they displayed lowered content of hepatic MUFA and triglycerides . The losAlthough loss of hepatic SCD1 expression confers beneficial metabolic effect in terms of protection from fat accumulation, several studies have pointed out that SCD1 and its product oleate are necessary to preserve liver function in a multiplicity of stressful circumstances. Mice fed with a Methionine and Choline Deficient (MCD) diet—a model of steatohepatitis—displayed diminished expression of SCD1 in the liver ,132. WheAlthough the contribution of SCD1 to NAFLD is still not fully depicted, recent studies have highlighted the important association between SCD1 expression and hepatocarcinoma (HCC) progression. The expression of SCD1, as well as genes involved in FA metabolism, is upregulated in HCC, and is strictly related to a poor prognosis ,136,137.Overall, these studies have suggested that SCD1 expression and the activity or oleic acid supplemented with the diet or originated from other tissues subsequently delivered to the liver are fundamental for the proper development and function of the liver. Although the analysis regarding SCD1 in HCC seems to offer a different scenario, where SCD1 activity contributes to tumor progression, they depict an established situation without taking into account the contribution of SCD1 to tumor onset and the possible beneficial effect of oleic acid supplementation. Further studies focused on the part played by SCD1 in the first steps of liver tumor formation as well as on the role in HCC development of oleic acid synthetized by other tissues are required.Different from the liver, the role of intestinal SCD1 has been less characterized. The few studies conducted depict an incoherent scenario, where SCD1 may both confer protection or worsen intestinal diseases . Here, wThe analysis of cancerous specimens collected from patients with stage II colon cancer (CRC) revealed that an elevated expression of SCD1 together with other three genes involved in lipid metabolism  delineate a high-risk group of patients, characterized by a worse clinical outcome . Indeed,-/-) in the intestinal epithelium presented an increased expression of inflammatory markers and crypt proliferative genes compared to the control group [-/- mice crossed with LDLR null mice [-/- mice are backcrossed with ApcMin/+ mice, a genetic model of CRC highly susceptible to spontaneous intestinal adenoma formation, they developed and larger and more tumors compared to the mice expressing SCD1 in the gut. However, the consumption of an oleic acid-enriched diet ameliorated the phenotype, reducing intestinal inflammation and tumor development [However, the comparison between adjacent normal tissue and cancerous tissue of a cohort of CRC patients revealed that SCD1 expression and activity are decreased in cancerous specimens. Indeed, cancerous tissue displayed higher content of SFA and a lower amount of MUFA compared to normal one . Moreoveol group . These rull mice . Howeverelopment . OverallThe close relationship between the liver and the gut under the light of SCD1 have been fairly characterized. Gut microbiota may variously affect the function of SCD1 in mice liver. Indeed, hepatic SCD1 levels are substantially reduced in germ free and microbiota ablated mice, thus indicating that SCD1 is partly regulated via gut microbial metabolites . MoreoveIntensive studies have demonstrated the fundamental contribution of oleic acid to health status maintenance. At the same time, the rate limiting enzyme for its production, SCD1, has emerged as a key regulator in serious diseases associated with inflammation and stress. Despite the large number of data, the role of SCD1 in these disorders is complex and, in many cases, conflicting results have been obtained, most likely due to variation of the experimental settings as well as of the strategies for the suppression of SCD1 activity . Therefo


Research conducted in the last two decades suggests that neuromuscular electrical stimulation of the lower limb muscles (NMES) may be a „bridge” to conventional exercise or an alternative for patients with advanced chronic heart failure (CHF), non-compliant or non-responsive to physical training. Through stimulating the work of the skeletal muscles, NMES increases the functional capacity, muscle mass and endurance in patients with CHF. A beneficial effect of NMES on functional capacity, vascular endothelial function, quality of life and aerobic enzymes activity has been shown. A significant benefit of this novel therapy in heart failure is the fact that the procedure can be home-based, after prior guidance of the patient. The major causes of HF are coronary artery disease, hypertension, cardiomyopathy, and valvular heart disease [1].Heart failure (HF) is defined as a complex clinical syndrome that can result from any structural or functional cardiac disorder that impairs the ability of the ventricle to fill with or eject blood [2]. The mechanisms responsible for the development and progression of HF are the subject of rigorous investigation.Following these pathologies, a process known as “cardiac remodeling” may occur. This pathophysiological remodeling of the heart involves changes in the structure and function of cardiac myocytes in the affected myocardium either directly or indirectly. These changes can lead to substantial alterations in the shape and volume of the heart and progressive ventricular dysfunction and clinically evident HF [1]. Over the past 20 years, significant development was noted in the treatment of HF by the introduction of multiple drugs  and cardiovascular procedures (cardiac resynchronization therapy). Nevertheless, despite the use of these modern methods of treatment, the latest European data show all-cause mortality at one year in hospitalized or outpatients with HF to be 17% and 7% and a readmission rate of 44% and 32%, higher in those with reduced left ventricle ejection fraction (LVEF) [3].The substantial scientific research conducted on HF is due to its high mortality and prevalence, affecting approximately 1-2% of the adult population and rising to over 10% in patients aged over 70 years [4].Given the poor prognosis of HF patients despite the use of multiple strategy-based treatments, scientists have been encouraged to search for new pathophysiological pathways that could be therapeutically targeted to the patient’s advantage including oxidative stress [Oxidative stress represents an imbalance between the production of reactive oxygen species (ROS) and endogenous antioxidant defense mechanisms. These molecules are produced in the cells and play distinctive roles in both physiological signaling processes as well as pathological pathways when their production becomes impaired.5].The cells constantly generate ROS, of which the principal molecule is the superoxide anion radical. Superoxide anions are synthesized as an unavoidable by-product of mitochondrial electron transport and enzymatically by xanthine oxidase. Other potential sources include prostanoid metabolism, catecholamine autoxidation, NAD (P) H oxidase activity, and the NO synthases . There are multiple proposed mechanisms through which oxidative stress might impair cardiac function and the first is by damaging cellular proteins and membranes, thus inducing cellular dysfunction or death through apoptosis and necrosis.Over the past several decades, oxidative stress has been involved in experimental and clinical studies involving HF patients. Markers of oxidative stress are elevated in CHF patients and have been associated with myocardial dysfunction, overall severity and poor prognosis of HF [9]. Because ROS also modulate the activity of various intracellular signaling pathways and molecules, they can induce specific changes in proteins involved in myocardial excitation-contraction coupling [4]. ROS are involved in fibroblast proliferation and collagen synthesis, and also in matrix metalloproteinases (MMP) activation and increased expression [10]. Moreover, ROS activate a broad variety of hypertrophy signaling kinases and transcription factors and mediate apoptosis.The superoxide anion is a potent inactivator of the signaling molecule NO that is involved in vascular endothelial dysfunction, with consequent loss of other physiological effects of NO [11].The importance of oxidative stress is increasing due to the involvement in the pathophysiological mechanism of cardiac remodeling responsible for the development and progression of HF [12].The results of the study conducted by Kameda et al. showed that that myocardial oxidative stress is an important regulator of MMP activity and that this contributes to ventricular remodeling and left ventricular dilatation and dysfunction in patients with ischemic heart disease [13].Kingery et al. concluded in a recent study that leukocyte iNOS is required for local and systemic inflammatory activation and cardiac remodeling in ischemic HF and that activated macrophages in HF patients may directly induce myocyte contractile dysfunction and oxidant stress [14]. This can be explained, at least in part, by the difficulty of directly targeting ROS and thus minimizing the harmful effects on other physiological signaling pathways.Randomized clinical trials have been discouraging until now despite substantial pre-clinical data showing positive effects of reduced ROS activity by using antioxidant therapy in cardiovascular diseases. Vitamin E supplementation showed initial promise in observational studies but did not demonstrate any clinical benefits in the prevention or treatment of HF patients [15][16].Recent research has shown that ROS signaling pathways are a complex system, and in many cases, essential for normal cardiovascular physiology, and that perturbation of these pathways resulting in dysregulated ROS is implicated in most cardiovascular diseases [17]. The pathophysiological process of HF will eventually lead to skeletal muscle weakness and atrophy, and when the symptoms will affect daily activities, to a sedentary lifestyle and social isolation with an impact on the prognosis of the patient. CHF-related skeletal muscle dysfunction is the result of an ongoing imbalance in the activation of anabolic and catabolic pathways [18] and has been shown to have significant prognostic importance [19].The main symptom of HF is the progressive decrease in functional capacity associated with dyspnea with prognostic implications independent of LVEF [1]. Unfortunately, this recommendation is less enacted, recent studies showing that less than 20% of patients with HF participate in such a program [20].Targeted patient management in HF consists of an integrated system which includes education programs, pharmacological, interventional therapy and cardiac rehabilitation programs. The European Society of Cardiology strongly recommends sustained and individualized physical activity in a standardized cardiac rehabilitation program [21], Piepoli et al. have shown in a total of 801 analyzed patients (395 received exercise training and 406 were controls) that exercise training leads to a reduction of 23% in the relative risk for death or hospitalization in HF patients. In the HF-ACTION large randomized control trial, after adjusting key prognostic factors, the investigators showed a reduction in cardiovascular mortality and hospitalization rate [22].In the meta-analysis carried out in 2004 .Numerous studies, confirmed by meta-analyses, indicate that regular exercise training reduces cardiovascular mortality and events, coronary heart disease, heart failure and atherosclerosis .Cardiac rehabilitation through physical exercise is known to improve exercise tolerance, quality of life and decrease the rehospitalization rates of patients with HF [33-37].It has also been proved that physical exercise has beneficial effects on endothelial dysfunction, neurohormonal activation, oxidative stress, inflammatory activation and depressive symptoms .Anti-inflammatory effects have been reported in several interventional studies. Exercise therapy has been shown to reduce inflammatory markers, particularly CRP, TNF-41][42] or in aged adults [43]. Also, in the HF population, the same discrepancies remain, as some randomized trials did not show a significant decrease in inflammatory biomarkers in patients who received optimal medical therapy [22][44].It is worth highlighting that a minority of interventional and randomized controlled trials did not detect a significant effect of regular exercise on systemic inflammatory biomarkers in adults [45]. The differences can be attributed to the smaller sample size used in the examined clinical trials.A meta-analysis found only five randomized controlled trials that examined the effects of regular aerobic exercise (of at least 4-week duration) in adults and concluded that aerobic exercise did not reduce C-reactive protein (CRP) levels [46-48]. However, Brooks et al. [49] reported that 16 weeks of resistance training reduced CRP and increased adiponectin levels in older diabetic patients.The effect of resistance training on inflammation was the topic of research in several studies and the reported results were mostly negative [50], liver [51], plasma [52], arteries [53], and skeletal muscles [54][55]. In elderly people, regular exercise reduced serum/plasma levels of myeloperoxidase, a marker of inflammation and oxidative stress [56], and thiobarbituric-reactive acid substances, a marker of lipid peroxidation [57].Regular exercise improves age-associated oxidative stress in the heart . Prior reviews suggested that NMES produces similar improvement in a 6-min walk distance (6MWD) test, a simple test used to detect functional capacity, when compared to conventional aerobic exercise training used by cycle ergometer. Also, previous studies have shown that resistance training improved the distance in a 6-min walk test in patients with HF [61][62].NMES has been consistently shown to elicit positive effects on functional capacity and skeletal muscle adaptations in patients with HF and unable to participate in traditional aerobic and/or resistance training programs at an appropriate stimulus . Several studies have shown higher improvements in peak VO2 following NMES in HF patients with low functional capacity compared to those with average exercise capacity [20]. The benefits of NMES therapy in HF patients also occurred in an older age group (age 75 ± 4 years) [63].Peak oxygen uptake  and CRP (-62% in average), showing anti-inflammatory effects.Dobcek et al. [70] showed that circulating cytokine TNF-α and the IL-10/TNF-a ratio were significantly modified after the NMES program in favor of the anti-inflammatory action, whereas their control group remained unaffected. In this study, there was a significant reduction in the circulating cellular adhesion molecules (soluble ICAM-1 and soluble VCAM-1) in the NMES group indicating an NMES-induced beneficial modulation of endothelial–monocyte cellular interactions in chronic heart failure.Karavidas et al. [67] found that the levels of citrate synthase (CS) and glyceraldehydes phosphate dehydrogenase (GAPDH), enzymes that indicate the citric acid cycle and glycolysis activity, displayed significant changes through the NMES program. CS activity increased whereas GAPDH activity decreased. In the control groups, neither CS nor GAPDH were significantly changed using sham stimulation. During follow-up, the groups differed significantly in terms of CS and GAPDH.Nuhr et al. [65] are in agreement with previous research which has shown that NMES combined with muscle training might induce an increase in the oxidative capacity of human muscle (in the activity of succinate dehydrogenase) even greater than that induced by muscle training alone.The findings in the study of Vaquero et al. [It appears that, in fact, NMES has beneficial effects on inflammation and oxidative stress, both of these mechanisms with implications in the pathophysiological mechanism of HF progression.To improve the poor prognosis of patients with HF we need to develop therapeutic strategies based on a novel understanding of the pathophysiology of HF. The NMES therapy showed a beneficial effect in a specific type of HF population with advanced disease, an area where the physician and the patient need the most help. The approach of regulating oxidative stress throughout NMES therapy may contribute to establishing effective treatment strategies for HF.The authors declare that there is no conflict of interest.


To assess the efficacy and short- and mid-term results of endovascular surgery and hybrid surgical procedures in treatment of complex aortic dissection. Clinical data of 90 patients with complex aortic dissection admitted from June 2012 to June 2018 were retrospectively analyzed. Among the patients, 60 cases were male and 30 cases were female, and their ages were ranged from 32 to 79, with an average age of 55 years old; different endovascular techniques and/or hybrid procedures were performed in these patients. Technical success rate was 100% for the entire group of patients. Type I endoleak occurred in 8 patients immediately after stent-graft placement, which in 2 cases disappeared after a proximal Cuff placement, and the other cases received no special treatment. Follow-up was conducted from 1 month to 72 months, with an average of 36.3 months, and no stent-graft migration or organ ischemia was noted. In the follow-up patients, no type I endoleak occurred but type II endoleak was found in 2 cases, which were cured without treatment; no patient had paraplegia. Endovascular surgery and hybrid procedures have demonstrable mid- and long-term efficacy in treatment of complex aortic diseases. However, this conclusion still requires multicenter, large-sample studies to further confirm. Endovascular repair is the first choice for treatment of aortic dissection (AD). Hybrid procedures are the combined application techniques of endovascular repair and various surgical bypass grafts. The treatment of aortic disease by hybrid surgery causes the original complex, high-risk surgery feasible, minimally invasive, and low-risk. In our hospital from June 2012 to June 2018, a total of 283 cases of aortic dissection were treated. 90 cases of complicated aortic dissection were treated with chimney technique, fenestration, and hybrid surgery and achieved good mid- and long-term effects. It is reported below.Inclusion criteria were as follows: (1) complying with diagnostic criteria for Stanford type A or stable Stanford type B aortic dissection [follows:  complyinssection  and confExclusion criteria were as follows: (1) mental/psychiatric disorders; (2) serious abnormalities in vital organs such as lung, liver and kidney, or serious diseases of the hematopoietic system and immune system; (3) inability to cooperate or withdraw for various reasons; (4) pregnancy or lactating women;There were 90 cases of aortic dissection in this study, 60 males and 30 females, aged 32 to 79, mean 55 years old. The course of hypertension ranged from 8 to 18 months. The general information of patients is shown in The results of aortic CTA examination  were useThe left subclavian artery (LSA) chimney technique was used in 53 cases, the left common carotid artery chimney technique was used in 2 cases, and the double chimney technique of left common carotid artery and left subclavian artery was used in 1 case. There were 1 case of fenestration using the truncus brachiocephalicus, left common carotid artery and left subclavian artery, 1 case of restrictive bare stent, and 5 cases of vascular artificial blood vessel reconstruction technique. The iliac artery percutaneous transluminal angioplasty (PTA) was performed in 2 cases of aortic dissection patients with severe stenosis of the access vessel. Nine cases were implanted with stent or catheter thrombolysis due to ischemia of superior mesenteric artery (SMA), and 4 cases were treated by hybridization technique. One case of aortic dissection combined with ischemic necrosis of lower extremity arteries was treated with middle and upper thigh amputation in the first stage after TEVAR. Five cases of aortic dissection were narrowed in the distal cavity and implanted with a limiting metal bare stent before implantation of the stent artificial blood vessel; 18 cases of aortic dissection and superior mesenteric ischemia were implanted with the stent. After the operation, the patient was cured by the implantation of the stent in the superior mesenteric artery or thrombolysis with Unifuse catheter. Two cases of Stanford type A aortic dissection were treated with cephalic tract fenestration, left common carotid artery, and left subclavian artery fenestration. Two patients with thoracic and abdominal aortic pseudoaneurysm were treated with superior mesenteric artery sulcus and abdominal cavity occlusion. Eight cases of type I endoleak occurred immediately during operation, and two cases disappeared after releasing a Cuff at the proximal end. Other patients did not receive special treatment. The following six types of endograft were used for endovascular stenting: Hercules (Shanghai Wei Chuang Xinmai)(n=41), Ankura (Shenzhen Xianjian)(n=38), Valiant (Medtronic)(n=17), Relay (n=15), Zenith TX2 (Cook)(n=22), and Fluency (Bard)(n=26). Patient's surgical results and follow-up are shown in Follow-up was performed from 3 months to 72 months, with an average of 36.3 months. There were no graft displacement and organ ischemia. There was no type I endoleak in the follow-up patients . Stents In the aortic dissection TEVAR, the proximal part of the stent graft should be at least 1.5~2.0 cm beyond the proximal end of the dissection, which can effectively isolate the true and false lumen of the dissection; try to prevent the opening of the left subclavian artery from being partially or completely covered. If the left vertebral artery is the dominant artery, chimney technique or arterial vascular bypass can be used to remedy . Hybrid For patients with segmental stenosis of the radial artery, covered stent grafts can be performed after the radial artery PTA. Two patients in this group were cured by this method. For the patients with stenosis of the external iliac artery and the iliac artery, the retroperitoneal or transabdominal approach of the common iliac artery or abdominal aorta artificial vascular reconstruction approach is performed firstly and then endovascular treatment can be performed. In this group, 2 patients with aortic dissection were recovered by TEVAR surgery after common right iliac artery and abdominal aortic artificial vessel approach reconstruction. The follow-up effect was good.Patients with aortic dissection often suffer from large pressure in the false lumen due to large rupture and narrow true lumen, which may lead to superior mesenteric artery stenosis or thrombosis, resulting in intestinal ischemia, or lower limb ischemic necrosis due to abdominal aorta or true iliac artery occlusion –21. In mThe most common complications of endovascular repair and hybrid surgery are endoleak, stent displacement, aortic injury, distal arterial embolism, spinal cord ischemia, and graft syndrome –31. In tIn summary, for patients whose left vertebral artery is the dominant artery and the proximal anchoring zone is insufficient, the left subclavian artery or the left common carotid chimney technology is a good solution to the brain blood supply –34. Hybr


Viruses have been infecting their host cells since the dawn of life, and this extremely long-term coevolution gave rise to some surprising consequences for the entire tree of life. It is hypothesised that viruses might have contributed to the formation of the first cellular life form, or that even the eukaryotic cell nucleus originates from an infection by a coated virus. The continuous struggle between viruses and their hosts to maintain at least a constant fitness level led to the development of an unceasing arms race, where weapons are often shuttled between the participants. In this literature review we try to give a short insight into some general consequences or traits of virus–host coevolution, and after this we zoom in to the viral clades of adenoviruses, herpesviruses, nucleo-cytoplasmic large DNA viruses, polyomaviruses and, finally, circoviruses. Viruses are obligatory cellular parasites, developing with their hosts since the dawn of life. Coevolution, when the virus and the host reciprocally affect each other’s evolution, is often detected. According to the Red Queen Hypothesis, both the parasite and the host are perpetually struggling to maintain a constant fitness level  the primordial virus world or the virus first hypothesis claims that the ancestors of viruses existed already in the pre-cellular world; (ii) the escaped genes theory describes viruses as mobile genetic elements, which became independent of their host cells; (iii) whereas according to the cellular regression theory, viruses are regressed intracellular parasites Holmes .According to the primordial virus world hypothesis, in the primordial soup multiple pre-cellular and pre-viral Ur-organisms competed with each other. The last universal common ancestor—the first cellular life form—emerged from these primordial replicators, and at least a fraction of current viruses might originate from the remaining ones  are viral genes or sometimes complete genomes inserted into host genomes. As genome integration is a compulsory step in retroviral replication, most of these elements are of retroviral origin, but further virus families were detected as well: hepadnaviruses, adeno-associated viruses, herpesviruses and others  or organ  against these genetic parasites to protect themselves and their genomic integrity.Most strikingly, antiviral defence mechanisms often have viral origins too  A, F and G  fibre seems to be of atadenoviral origin , while the family Alloherpesviridae contains the HVs of amphibians and fish (Anamnia), and the family Malacoherpesviridae contains the HVs of molluscs. The three families were clustered under the novel order Herpesvirales  implying the common origin of HVs and tailed bacteriophages  are large (150–200 nm) icosahedral, enveloped viruses having a so-called tegument layer between the capsid and the envelope. HVs possess a linear, non-segmented, double-stranded DNA genome (135–295 kbp)  of Herpesviridae emerged probably before the mammalian radiation, 60 to 80 million years ago, while the diversification time point of the three subfamilies had been around 200 million years ago  existed around 400 million years ago based on similar phylogenetic branching patterns of HVs and host lineages  has been isolated from cattle worldwide and this species was thought to be the original host of the virus. However, BoHV-4 has been reported from wild buffalo and other ruminants as well   (Aswad and Katzourakis Branchiostoma floridae (Florida lancelet or amphioxus) contains several genes showing homology to herpesviral genes, implying the existence of a HV associated with this invertebrate chordate. The terminase and polymerase gene sequences from the putative amphioxus HV show higher similarity to those of the mollusc HVs than to any vertebrate HVs ] to collect the above-mentioned virus families  are a group of DNA viruses including the families Mimiviridae and pandoraviruses. Branch #2 gathered the families Asco-, Irido-, Marseille- and Pithoviridae. This branch contained the widest variation of genome sizes (100 kb to 1500 kb). Branch #3 included only non-giants in two distinct clades: Asfarviridae (African swine fever virus and its protist-infecting relatives) and Poxviridae. According to the present knowledge, a major host switch took place two times along this branch. Once during the early history of poxviruses, which infect both arthropods and vertebrates, two animal phyla that radiated from the common ancestor more than 500 million years ago  viruses are the most prominent gene gainers, whereas NCLDVs infecting animal hosts have undergone considerable gene losses, a sort of ‘genome contraction’ during their evolution from ancestral protist viruses. It was hypothesised that in animals, the selective pressure for virus genome size is stronger than in protists, yet its mechanism needs to be analysed  according to the ICTV. The taxonomical classification of PyVs is based on the genetic distance of the large tumour antigen. Still, some PyVs cannot be categorised into any accepted genus, so these require additional genera in the future  are non-enveloped icosahedral DNA tumour viruses with a double-stranded, cca. 5000-bp-long circular DNA genome. Approximately 80 accepted PyV species form the family The organisation of the circular PyV genome is highly conserved: the 5–7 genes are located on both strands, and distributed into early and late regions. There is a non-coding control region as well. The early coding region contains regulatory proteins: the small and the large tumour antigen. The late coding region contains the genes of the structural proteins, like the major (VP1) and two minor capsid proteins. The approximately 500-bp-long non-coding control region contains regulatory elements and transcription promoters, the origin of DNA replication  is oncogenic like several other mammalian PyVs.PyVs are known to infect mammals and birds, but using viral metagenomics, PyVs were identified and characterised also in fish and arthropods  infects birds of diverse families  are vertebrate-infecting, single-stranded DNA (ssDNA) viruses with one of the smallest (~2 kb) virus genomes (Parrish rep and cap) encoding the replication-associated (Rep) and the capsid (Cap) proteins, respectively  and broadened the perspective for the coevolutionary studies. (3) Concerning NCLDVs, we have discussed that many earlier unrelated virus families with large/giant capsid and genome sizes and even more divergent host range (from protists to vertebrates) can be phylogenetically linked based on a subset of core genes. This phylogenetic reconstruction demonstrated a ‘turbulent evolution’, which was dominated by gene gain and gene loss via horizontal gene transfer both from other viruses and from host genomes. (4) In connection with polyomaviruses (PyV) the recombination among PyV lineages generating novel virus species was also mentioned, and the importance of the intra-host molecular evolution was discussed in more detail. It was demonstrated that point mutations and rearrangements in non-coding control regions both can critically alter the pathogenic potential, immunogenicity and target organs of related PyVs of a single host species. (5) In the case of circoviruses (or more broadly CRESS DNA viruses), higher mutation rates and rolling circle type replication of their ssDNA, alongside with recombination events and integration into host genomes as well as presumed cross-species transmissions of distant hosts were simultaneously forming the coevolutionary processes.As seen from the example of this latest mentioned dispute concerning the appearance of circoviruses, there are a number of unsettled questions in connection with the coevolutionary processes observed in DNA viruses and their hosts. In the paragraphs above, we have listed these major processes. (1) Apparent speciation and coevolution of adenovirus lineages (genera) with the different vertebrate families. Here, host switches to new taxa were marked both with shift in the genome content  and more severe pathology in the non-coevolved hosts. (2) In connection with herpesviruses (HVs) a longer coevolutionary past was supposed, largely synchronous with the more divergent (vertebrate and invertebrate) host lineages. Integrated HV genomes in host chromosomes were demonstrated to provide new data about the genome content of ancient HVs and a new HVs lineage. Moreover, the discovery of a HV core gene homologue in tailed bacteriophages established the phylogenetic relatedness between two seemingly unrelated virus orders  hosts (Welch et al. Viruses have always been around. Their parasitic lifestyle had an enormous driving effect on the evolution of all organisms, and thus life would not be the same without them. Future research will hopefully shed light on the exact origin and coevolutionary history of all viruses.


The UN Sustainable Development Goals (SDGs) call for under-5 mortality to be no greater than 25 deaths per 1000 livebirths. Since 1990, there has been tremendous progress towards this goal, with a reduction from 142 deaths per 1000 livebirths to 65 deaths per 1000 livebirths for the poorest quintile in low-income and middle-income countries (LMICs).6 In our work,6 we found protection efficacy against severe rotavirus diarrhoea in Bangladesh was 74%, and that rotavirus vaccine failure was associated with a chronic inflammatory intestinal disease called environmental enteric dysfunction (EED). EED was present in 80% of infants aged 12 weeks in an urban slum of Dhaka, Bangladesh, as measured by faecal biomarkers of inflammation.7 EED was associated with oral vaccine failure but not failure of systemically administered vaccines, such as the diphtheria, tetanus, and pertussis vaccine. We measured rotavirus vaccine failure as the occurrence of rotavirus-caused diarrhoea in a vaccinated child. Biomarkers of EED and maternal health accounted for 24% of rotavirus vaccine failures.7A further problem with control of diarrhoeal diseases is that the oral rotavirus vaccine is less effective in LMICs.8 It is probable that rotavirus vaccine failure is in part due to a failure of vaccine virus replication resulting from an active gut antiviral immune response.8One cause of EED is enterovirus infection. We found that enterovirus in stool on the day of vaccination was associated with diminished rotavirus IgA vaccine response.Despite the issue of a less effective response in LMICs, the WHO Strategic Advisory Group of Experts on Immunization has recommended rotavirus vaccination as part of the Expanded Program on Immunization for all regions of the world. This recommendation is in large part because of the benefit in reduction in child mortality and morbidity from even a partly effective vaccine.The Lancet Infectious Diseases, Andrew Clark and colleagues9 examined all randomised controlled trials of rotavirus vaccination published before April 4, 2018. They classified the trials’ settings into low-mortality, medium-mortality, and high-mortality strata, and estimated stratum-specific vaccine efficacy over time from a Bayesian hierarchical Poisson meta-regression model, which accounted for observed study-level characteristics and unmeasured heterogeneity among the studies. Clark and colleagues9 also converted the cumulative vaccine efficacy to instantaneous vaccine efficacy to account for the potential temporal changes of rotavirus failure risk and tested for waning efficacy over the first year of life. The result was a validation of low rotavirus vaccine efficacy in low-income and high-mortality settings for the prevention of severe rotavirus diarrhoea, with only 44% effectiveness  after 12 months compared with 94% (87–98) effective in high-income and low-mortality settings. Vaccine efficacy waned more rapidly in high-mortality settings leading to the important observation that improvements in efficacy need to address both instantaneous and prolonged immunity.In The research questions arising from this Article are centred on how to improve vaccine efficacy. Rotavirus vaccination expansion throughout high-mortality countries should help to meet the SDG goals, and even small improvements in vaccine efficacy will have a measurable effect on infant mortality due to diarrhoeal diseases. Interference from EED and enterovirus suggests that the gut metabolome and microbiome could have a role, and any metabolites found to be associated with vaccine success might hold potential as vaccine adjuvants. Avoidance of interfering enterovirus infection and EED by administering the first dose of the vaccine at birth is another potentially beneficial strategy.10 A heat-inactivated parenteral vaccine can induce mucosal immunity in animal models,13 and a parenteral subunit vaccine has been shown to be safe and efficacious in a clinical trial with infants in South Africa,14 spurring a phase 1 and 2 trial that is underway. Indeed, the ability to effectively induce mucosal immunity using parenterally administered vaccines with select adjuvants might render the oral rotavirus vaccine unnecessary in the not-too-distant future.15A promising alternative strategy is to move from a live virus oral vaccination to an inactivated vaccine administered intramuscularly or intradermally, either alone or in combination with oral vaccine, as borne out by experience with the poliovirus vaccination.


Amyotrophic lateral sclerosis (ALS) is one of the most threatening neurodegenerative disease since it causes muscular paralysis for the loss of Motor Neurons in the spinal cord, brainstem and motor cortex. Up until now, no effective pharmacological treatment is available. Two forms of ALS have been described so far: 90% of the cases presents the sporadic form (sALS) whereas the remaining 10% of the cases displays the familiar form (fALS). Approximately 20% of fALS is associated with inherited mutations in the Cu, Zn-superoxide dismutase 1 (SOD1) gene. In the last decade, ionic homeostasis dysregulation has been proposed as the main trigger of the pathological cascade that brings to motor-neurons loss. In the light of these premises, the present review will analyze the involvement in ALS pathophysiology of the most well studied metal ions, i.e., calcium, sodium, iron, copper and zinc, with particular focus to the role of ionic channels and transporters able to contribute in the regulation of ionic homeostasis, in order to propose new putative molecular targets for future therapeutic strategies to ameliorate the progression of this devastating neurodegenerative disease. Amyotrophic lateral sclerosis (ALS) is a progressive and devastating neurological disease characterized by the loss of Motor Neurons (MNs) in spinal cord, motor cortex and brainstem  that includes the majority of ALS cases and, the familiar form (fALS) that regards approximately 5–10% of cases  such as calbindin and parvalbumin , due to the quite weak cytosolic Ca2+ buffering property of these transporters, resulting in an excess of mitochondrial Ca2+ and ROS production. Moreover, the entry of Ca2+ into the mitochondria contributes to establish the chronic depolarization of the mitochondrial membrane, which consequently determines the release of pro-apoptotic proteins and the activation of enzymes able to activate other cell death pathways  in ALS pathogenesis because animals affected by ALS show a strong reduction in its expression and activity at muscular and neuronal levels, whereas strategies able to delay ALS progression worked also through NCX3 activation and overexpression  mutation, causes strong ROS generation. In these patients, riluzole, a drug able to increase survival rate, has shown to prevent MNs deterioration and moderately reduce excitotoxicity and cell loss by blocking tetrodotoxin-sensitive sodium channels which are associated with damaged neurons. Deregulation of intracellular calcium homeostasis has also been described in the terminal motor axons of subjects affected by ALS, as well as in the spinal MNs of ALS animals, both in the sporadic and familial forms  conductance and reduced potassium (K+) currents, have been described and related to the evolution of ALS signs and symptoms such as neurodegeneration and fasciculation. In fact, membrane hyperexcitability observed in ALS due to Na+ and K+ conductances abnormalities, leads to muscle cramps and fasciculations, and promotes a neurodegenerative cascade mediated by Ca2+-dipendent processes. In addition, modulation of axonal Na+ channel function in ALS resulted in amelioration of symptoms and stabilization of axonal excitability parameters. Axonal ion channel dysfunction evolves with disease progression and correlates with survival, thus representing a potential therapeutic biomarker in ALS , and, specifically, it is not clear whether any difference occurs with the processes identified in subjects affected by the sporadic form of ALS (sALS). Furthermore, several studies documented a deregulation of voltage-dependent Na+ currents (Nav) and increased persistent sodium current (PICNa) in primary neuronal cultures obtained from ALS mice. At a molecular level, voltage-gated Na+ channels are composed by a single 260 kDa α subunit and one or more β subunits of 30–40 kDa. The α subunits constitute the actual ion channel and include sensors for voltage dependence. These subunits represent the working cores of the excitation process, whereas β subunits control the kinetics of channel activation and inactivation in dependence from the voltage and the localization of the channel on the plasmamembrane  in the brainstem and cortex of ALS rats and in cultured ALS cortical astrocytes occurs. Since these channels are required for the maintenance of a functional BBB astrocytic lining these results strongly suggest a role for the control of water and K+ homeostasis in the disturbance of motor neuron survival during ALS  integrity, may alter the neuronal microenvironment, and may cause motoneuronal dysfunction and death  and the Cu efflux pump (ATP7A) increases and decreases, respectively, in the spinal cord of mutant animals. Furthermore, also the expression level of metallothioneins (MT), a class of proteins with a very high affinity for copper, is augmented in the spinal cord of G93A SOD1 mice  can induce oxidative stress directly catalyzing the formation of strong oxidant such as lipid hydroperoxides from hydrogen peroxide and hydroperoxides via a Fenton-like reaction ], a molecule with low toxicity, able to deliver copper in the CNS within minutes, prevents early mortality of G93A SOD1 mice over-expressing CCS protein  administration to G93A symptomatic SOD1 mice delays onset of paralysis and extends lifespan  although intracellular concentration of copper is extremely higher compared to wild type animals  of cerebral cortex, hippocampus and amygdala  and zinc binding proteins such as metallothioneins (MT)  and the Zrt-, Irt-related proteins or ZIP (ZIP 1-14), which move zinc ions from cytosol to extracellular space or lumens of intracellular compartments and from extracellular space and lumens to cytosol, respectively  alterations of proteins involved in both iron influx and sensing of intracellular iron concentrations; (2) blockage of anterograde axonal transport with a consequent iron accumulation in ventral motor neurons; and (3) increased mitochondrial iron load in neurons and glia leading to neurodegeneration copperII [CuII(atsm)], a metal complex which provides Cu to the mutant protein and decreases the abundance of Cu-deficient SOD1 in treated mice at the spinal cord level  have been successfully tested in SOD1 G93A animals where they were able to delay the onset of the disease and to increase mice life span. These compounds have been approved in late fifties of last century for the treatment of Wilson disease, a pathology caused by an autosomal recessive mutation in the gene that encodes the transmembrane protein ATPase (ATP7B), a copper-dependent P-type ATPase. Clinical features of Wilson disease are the accumulation of copper ions in different body organs like in the liver and in the brain (Brewer et al., Copper intracellular levels could be also lowered by increasing MT expression. Indeed, these small cysteine-rich proteins have a very high affinity for heavy metals and particularly for copper ions Hamer, . InteresThe protein aggregation is another important neuropathological hallmark of ALS and precisely in order to counteract this aspect of the disease, Evans et al.  tested tCollectively, in the present review we summarized the recent data concerning the crucial role of several ions and redox metals in ALS pathogenesis Table . In partGP, RS, NB, VV, LA, SaA, and LC manuscript orgranization. OC, AV, PC, NG, SeA, PB, and CF study of the state of art.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.