Alzheimer’s disease (AD) is ultimately linked to the amyloid precursor protein (APP). However, current research reveals an important synaptic function of APP and APP-like proteins (APLP1 and 2). In this context various neurotrophic and neuroprotective functions have been reported for the APP proteolytic fragments sAPPα, sAPPβ and the monomeric amyloid-beta peptide (Aβ). APP is a metalloprotein and binds copper and zinc ions. Synaptic activity correlates with a release of these ions into the synaptic cleft and dysregulation of their homeostasis is linked to different neurodegenerative diseases. Metal binding to APP or its fragments affects its structure and its proteolytic cleavage and therefore its physiological function at the synapse. Here, we summarize the current data supporting this hypothesis and provide a model of how these different mechanisms might be intertwined with each other. Alzheimer’s disease (AD) is a fatal neurodegenerative disorder and a severe burden of our aging societies  and several unstructured regions  in a slightly distorted square pyramidal geometry (a type 2 non-blue site) to three protein ligands  and two water molecules  almost as high as to CuBD. Copper-binding to the GFLD again correlates with the reduction of a neighboring disulfide bridge (Cys98-Cys105) pointing to redox-activity also of this site. Furthermore, the match of geometry and functionality is suggestive for a complementation of the two copper-binding sites by either a conformational change within the E1 domain into a “closed” conformation  and with low affinity to zinc  harbors two or three different metal binding sites, here designated as M1–3  spread on three helices  to regulate homo- and hetero-dimerization with APP and APLP2  become progressively more acidic down to pH 6 in the trans-Golgi and <5 in lysosomes  in the amyloid plaques , ceruloplasmin, tyrosinase and dopamine β hydroxylase , or WD. Loss of ATP7A function causes growth failure, brittle hair, hypopigmentation, arterial tortuosity and neuronal loss most prominent in the hippocampus and cerebellum  discussed involve S-nitrosylation, oxidation and allosteric modulation, increased anchorage of the neurotransmitter receptors to the membrane, and modulation of neurotransmitter receptor function  receptors), Glycine and other surface receptors, such as TrkB. Furthermore, zinc can bind to and regulate ProSAPs/Shanks scaffolding proteins of the postsynaptic density (PSD), involved in synaptic signaling. Indeed, in some cerebral areas nearly 50% of the glutamatergic synapses are actually “glu-zinc-ergic” (Watt et al., In contrast to copper ions there is a substantial amount of zinc loosely bound to biomolecules, designated as reactive or chelatable zinc, which is implicated in neuronal signaling. Reactive zinc is largely distributed within presynaptic vesicles in some axon terminals throughout the telencephalon and co-localizes with a subset of glutamatergic neurons (Frederickson et al., Synaptic function of APP is discussed in detail in other chapters of this special issue. Briefly, loss of APP function leads to a reduced number of dendritic spines (Watt et al., in vitro interaction of APP, and mutations, abolishing copper-binding to the GFLD, reduce APP synaptogenic activity in a cellular assay system (Baumkötter et al., As pointed out before, copper binds APP at different sites within the E1 and E2 domain, causing structural changes and altered dimerization properties and heparin binding characteristics (Baumkötter et al., in vivo conditions at the synapse. Consistently, addition of 50 μM zinc to cells expressing APLP1 caused a lateral concentration at cell-cell contact sites (Mayer et al., Different reports suggest binding of APP, APLP1 and to a minor extent also APLP2 to zinc with affinities in the low micromolar range (Bush et al., Despite some major gaps in our understanding of APP/APLPs synaptic function, the current data as presented in this review article strongly suggest that activity-dependent changes in zinc and copper concentrations in the synaptic cleft can be sensed by the APP/APLP family. In turn, they seem to modulate neurotransmission by different pathways including neurotrophic activity of sAPPα or trans-cellular dimerization/signaling. However, one major gap in our current understanding, especially in respect to the function of copper, is the limitation of available sensitive sensors, allowing determination of local transient changes in copper concentration. In this regard, live-cell optical imaging with fluorescent sensors offers a potentially powerful approach for interrogating aspects of labile copper accumulation, speciation, trafficking, and redox function in living systems at the molecular level. Such reagents have greatly facilitated the study of calcium and zinc in cell biology, but analogs tools for cellular copper remain underdeveloped (Zeng et al., KW, AA, CUP and SK wrote this review article.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.


Lin et al. reported that young, asymptomatic APOE4 targeted-replacement mice treated with rapamycin had restored CBF and BBB integrity to the level of wild-type controls [Alzheimer's disease (AD) is the most common form of dementia and the 6th leading cause of death in the US. Inheritance of the Apolipoprotein ε4 (APOE4) allele is the strongest genetic risk factor for late-onset AD - APOE4 carriers accumulate beta-amyloid (Aβ) and neurofibrillary tau tangles earlier and with more extensive pathology compared to non-carriers. However, decades before the aggregation of Aβ and tau, cognitively normal APOE4 carriers have developed neurovascular deficits, including reduced cerebral blood flow (CBF) and impaired bloodbrain barrier (BBB) integrity . The abocontrols . The preRapamycin is a drug approved by US Food and Drug Administration (FDA). It has been widely used in clinical settings and was originally used as an immunosuppressive agent to prevent rejection of organs in transplant patients. By 1991, Heitman and co-workers discovered a kinase, Target of Rapamyin (TOR) . In mammLin et al. demonstrated that mTOR inhibition reduced proinflammatory pathways in brain vasculature that otherwise impair BBB integrity in the APOE4 transgenic mice [Lin et al. previously reported that inhibition of mTOR activates endothelial nitric oxide synthase and causes the release of nitric oxide, a vasodilator, which in turn increases CBF [Lin et al. found that rapamycin restored their CBF and vascular density, which were associated with reduced accumulation of Aβ and cerebral amyloid angiopathy, and improved memory [nic mice . Loss ofnic mice . Restorinic mice . In addiases CBF . In miced memory . CollectLin et al. further demonstrated that rapamycin restores and preserves neurovascular functions in the APOE4 transgenic mice using MRI. Their results may provide the basis for future AD prevention trials in human APOE4 carriers.In conclusion, mTOR inhibition has been shown to increase lifespan and healthspan in various species. Rapamycin  has been approved by the FDA since 1999 for various uses in humans, and these compounds have been given to cancer patients for relatively long periods of time with little change in the quality of life.


Irradiated ice-covered ocean worlds with rocky mafic mantles may provide the conditions needed to drive the emergence and maintenance of life. Alkaline hydrothermal springs—relieving the geophysical, thermal, and chemical disequilibria between oceans and tidally stressed crusts—could generate inorganic barriers to the otherwise uncontrolled and kinetically disfavored oxidation of hydrothermal hydrogen and methane. Ionic gradients imposed across these inorganic barriers, comprising iron oxyhydroxides and sulfides, could drive the hydrogenation of carbon dioxide and the oxidation of methane through thermodynamically favorable metabolic pathways leading to early life-forms. In such chemostatic environments, fuels may eventually outweigh oxidants. Ice-covered oceans are primarily heated from below, creating convection that could transport putative microbial cells and cellular cooperatives upward to congregate beneath an ice shell, potentially giving rise to a highly focused shallow biosphere. It is here where electron acceptors, ultimately derived from the irradiated surface, could be delivered to such life-forms through exchange with the icy surface. Such zones would act as “electron disposal units” for the biosphere, and occupants might be transferred toward the surface by buoyant diapirs and even entrained into plumes. Key Words: Biofilms—Europa—Extraterrestrial life—Hydrothermal systems. Astrobiology 17, 1265–1273. Such hydrogenations yield an ever-renewed stock of highly specified organic molecules—the so-called CHNOPS with a typical bonding motif -C–C(H2)–N(H)–C–O  reactions as salty ocean waters are reduced to hydrogen and formate on gravitation into the primitive ultramafic crust  that are potentially more amenable as acceptors along metabolic pathways than just CO2 or carbonate  is a lower potential acceptor but has the advantage of also providing further carbon through a variant of the acetyl coenzyme-A pathway  from hydrothermal activity, and an ocean with dissolved oxidants providing a geochemical gradient, the first life-forms could potentially follow some of the same metabolic pathways known to be at, or near, the root of the earliest life on Earth, that is, acetogenesis and methanotrophy , leading to effective delivery through the ocean to the ice-water interface  along the ice-water interface to reach sites of down-welling convection in the ice that might bring radiolytically produced oxidants to the ocean  at the same time (Vargas et al.,et al.,et al.,et al.,et al.,et al.et al.,et al.,et al.,et al.,The dynamic communities comprising any sub-ice-shell biosphere could constitute a habitable zone, manifested as microbial mats concentrated around oxidant-rich ice-shell sites at the down-welling regions of thermally unstable regions at the ice-water interface (Goodman et al.,et al.,et al.,et al.,et al.,et al.,Future missions to search for life on Europa (Hand


Recently, Birte Hellwig and colleagues from the Department of Statistics, TU Dortmund University have published a study to predict late metastasis in breast cancer . In breIn the present study Hellwig et al. (2016) used a In recent years numerous studies have been performed to predict prognosis of cancer (Hammad, 2013; Hammad


Neutrophils are well known for their role in infection and inflammatory disease and are first responders at sites of infection or injury. Platelets have an established role in hemostasis and thrombosis and are first responders at sites of vascular damage. However, neutrophils are increasingly recognized for their role in thrombosis, while the immunemodulatory properties of platelets are being increasingly studied. Platelets and neutrophils interact during infection, inflammation and thrombosis and modulate each other’s functions. This review will discuss the consequences of platelet–neutrophil interactions in infection, thrombosis, atherosclerosis and tissue injury and repair. Platelets are anuclear cell fragments that circulate in healthy humans at about 150,000–400,000 per microliter blood. Platelets are well known for their role in thrombosis and hemostasis. Inherited or acquired defects in platelet count and/or function can be associated with bleeding complications. Conversely, platelets are key players in thrombotic disease and drugs inhibiting platelet function are vital in treatment and prevention of arterial thrombotic diseases such as myocardial infarction and stroke. Recent studies show that platelets have many functions beyond physiological or pathological thrombus formation. They play a role in inflammation, are effectors of injury in a variety of pulmonary disorders and syndromes, facilitate tissue repair and act in the growth and development of metastases of various cancers  have been shown to crawl along and adhere to the surface of (activated) endothelial cells, eventually forming an almost continuous layer on the vascular endothelium (von Bruhl et al. At least three distinct mechanisms may be involved in platelet-dependent, neutrophil-mediated induction of thrombosis. First, neutrophils have been demonstrated to transfer tissue factor, the physiological initiator of coagulation, to platelets during thrombus formation in vitro (Giesen et al. The ‘blood-borne’ TF concept challenges the dogma that tissue factor is normally not located on cells in contact with blood and the source and quantity of blood borne TF is subject to ongoing debate (Butenas et al. A second mechanism by which the platelet–neutrophil axis contributes to initiation and propagation of thrombosis is through generation of NETs (Fuchs et al. A third mechanism linking the platelet–neutrophil axis to thrombosis relates to neutrophil constituents modulating hemostasis. For instance, neutrophil cathepsin G and elastase inactivate natural anticoagulant systems including tissue factor pathway inhibitor, thrombomodulin and antithrombin (von Bruhl et al. Platelets have long been implicated in acute coronary events. Their key role is evidenced by the success of platelet inhibitory drug in treatment and prevention of acute arterial thrombotic events (Jamasbi et al. Sterile tissue injury as for example encountered in injury by toxins (Miyakawa et al. For example, acetaminophen-induced liver injury results in an influx of platelets and neutrophils in the liver microcirculation (Miyakawa et al. Platelets and neutrophils have been implicated as drivers of ischemia/reperfusion injury in liver (Yadav et al. Platelets trigger the release of NETs in a model of experimental transfusion-associated acute lung injury and thereby aggravate disease (Caudrillier et al. Platelet–neutrophil interactions have been shown to facilitate repair in a model of heat-induced liver injury (Slaba et al. Platelets are well known to be required for liver regeneration following partial hepatectomy and it has been suggested that growth factors stored within platelet granules drive platelet-mediated liver regeneration (Lesurtel et al. Finally, NETs have also been demonstrated to contribute to organ injury in sepsis (Czaikoski et al. Experimental and clinical evidence for a role of platelet–neutrophil crosstalk in a variety of clinical conditions including inflammation, thrombosis, atherosclerosis and tissue injury is rapidly emerging. Agents blocking PSGL-1, P-selectin, or MAC-1 that have pleiotropic effects that include inhibition of platelet–neutrophil interactions have been trialed in humans (Mertens et al. Inhibitors of PAD4 may have merit given the key role of PAD4 in NET formation and the intact NET-independent anti-inflammatory properties of PAD4-deficient neutrophils (Li et al. A clear advantage of targeting NETs in thrombotic syndromes is the lack of effect of the intervention on physiological hemostasis. As bleeding is an important side effect of current therapies for venous and arterial thrombosis and disseminated intravascular coagulation, the development of agents with a better risk/benefit ratio in terms of thrombotic potency versus bleeding risk would be a significant improvement.Although platelets are traditionally seen as key players in thrombosis and hemostasis and neutrophils as prime inflammatory cells, recent data demonstrate a complex interplay between these cells with important immune functions for platelets and a clear role of neutrophils in thrombotic diseases. The prime mode of interaction between platelets and neutrophils and the downstream effects of this interaction are summarized in Fig.


HTT is ubiquitously expressed and acts systemically, meaning blood, which is readily available and contains cells that are dysfunctional in HD, could act as a surrogate for brain tissue. We conducted an RNA-Seq transcriptomic analysis using whole blood from two HD cohorts, and performed gene set enrichment analysis using public databases and weighted correlation network analysis modules from HD and control brain datasets. We identified dysregulated gene sets in blood that replicated in the independent cohorts, correlated with disease severity, corresponded to the most significantly dysregulated modules in the HD caudate, the most prominently affected brain region, and significantly overlapped with the transcriptional signature of HD myeloid cells. High-throughput sequencing technologies and use of gene sets likely surmounted the limitations of previously inconsistent HD blood expression studies. Our results suggest transcription is disrupted in peripheral cells in HD through mechanisms that parallel those in brain. Immune upregulation in HD overlapped with Alzheimer’s disease, suggesting a common pathogenic mechanism involving macrophage phagocytosis and microglial synaptic pruning, and raises the potential for shared therapeutic approaches.There is widespread transcriptional dysregulation in Huntington’s disease (HD) brain, but analysis is inevitably limited by advanced disease and postmortem changes. However, mutant  HTT gene and is characterised by motor, cognitive and psychiatric features. Onset occurs around 45 years on average and inversely correlates with CAG repeat lengthHuntington’s disease (HD), the most common monogenic neurodegenerative disorder in the developed worldHTT is ubiquitously expressed589101415519HD research has traditionally focused on the brain due to the presence of characteristic mutant huntingtin protein aggregateset al.2324et al.Transcriptional dysregulation is a central feature of HD pathogenesisIn the current study we present a transcriptomic analysis of whole blood in human HD using RNA sequencing (RNA-Seq). We studied differential expression of individual gene transcripts and enrichment of differential expression in gene sets in two independent cohorts from Track-HDPremanifest gene carriers had a mean total motor score (TMS) of 2 and total functional capacity (TFC) of 13  between premanifest and manifest HD in either the Track-HDHTT gene carriers were combined to increase the analytical power in a comparison of HD and controls. Once again there were no individually significant transcripts in independent or combined cohorts, but the differential expression analysis in the combined cohort is given in Attempting to identify both HD specific and stage-specific changes in gene expression (mRNA) level, we compared premanifest, manifest and control subjects, whilst controlling for age and gender. C) of 13 , indicatWe next asked whether networks of genes with similar functional annotation were dysregulated in HD relative to controls. Pathway annotations were collated from publicly available gene ontology databases to form a set of generic pathways using the same method as the recent HD genome-wide association study (GWAS) of modifiers of age at onsetGene set enrichment analysis (GSEA), with a false discovery rate  threshold of q < 0.05 to correct for multiple testing, identified 53 upregulated  and 14 det al.et al.Through RNA-Seq, Miller, et al.et al.A limitation of using curated pathways from databases is the incomplete or incorrect annotation. One way to overcome this is to use gene co-expression, because genes that are co-expressed often have related functions. WGCNA identifies clusters (modules) of genes with highly correlated expression, constructing original, unbiased gene co-expression networks based on observed dataGSEA for brain co-expression modules was applied to our combined Track-HD and Leiden blood expression dataset. Immune- and inflammatory-related brain modules were upregulated in HD blood, and notable downregulated modules included synaptic function, proteasomal degradation, mitochondrial function and transcription. The 10 most significantly up and downregulated modules in the combined dataset that were also nominally significant (p < 0.05) in both independent cohorts are given in –4) correlation between dysregulation p-value in the direction of interest (positive) in HD blood and degree of module membership (kME)The module membership (kME) of a gene is measured by the correlation of its expression with the eigengene, which is representative of all gene expression profiles in the module et al.et al.−16) excess , suggesting some concordance in signal at the individual gene level. Furthermore, a significant excess of generic pathways was found to be significantly (p < 0.05) dysregulated in both datasets, most markedly in the positive (p < 0.001) direction, but also negative (p = 0.028), thus showing an overlap in biological signal. Pathways significantly upregulated in both datasets are mainly related to immune response  in the 112 gene positive Track-HD subjects . After cWe then tested whether generic pathways that were significantly enriched for upregulated  or downr−7) and 66 , which were also dysregulated in the HD caudateSimilarly, we tested whether modules dysregulated in HD blood relative to controls  also coret al.−3). Using the same method as for concordance with Labadorf, et al.et al.−7). Thus, we conclude that analysis of TMS in the Track-HD cohort broadly supported the associations reported in Mastrokolias et al.Mastrokolias et al.−4).In Alzheimer’s disease, an early inflammatory response involving microglia contributes to pathogenesis3637et al.yellow, being particularly highly enriched (combined Track-HD and Leiden p < 1 × 10−16). Notably, this module has immune and microglia-specific functionset al.Zhang, HD research has focused on the brain as the most conspicuous clinical features can be clearly linked to progressive degeneration of specific brain regions45We conducted RNA-Seq of whole blood in two independent cohorts of HD patients. Using gene set enrichment analysis (GSEA) with publicly-available pathway databases and WGCNA modules from HD and control brain datasets, we identified dysregulated genes and gene sets in blood that replicated in both independent cohorts and correlated with clinical motor signs (TMS). These correspond to the most significantly dysregulated modules in caudate nucleus, the most prominently affected region in HD brain. This suggests mutant huntingtin drives a common pathogenic signature in both blood and brain.et al.944RNA-Seq more comprehensively and accurately quantifies mRNA than hybridisation-based microarrays or tag-based methodsDespite these limitations, gene set enrichment analysis identified significantly overlapping dysregulated pathways in the Track-HD and Leiden HD blood datasets, even though they differed in age and disease severity. Thus, through grouping transcripts into biologically relevant pathways and co-expressed transcripts, we could highlight areas of dysfunctional biology in HD. The observed upregulation of immune-related pathways is consistent with that previously identified in transcriptional and functional studies57910PGC-1α, a member of the dysregulated ATP metabolic process pathway  and 66 (CNneg1), were also significantly dysregulated in the same direction in both independent blood datasets. Compared with other brain regions, the caudate has the largest number of expression changes and the highest correlation with HDet al.et al.Our demonstration of a transcriptional signature common to both HD blood and brain supports the use of blood cells to study aspects of HD biology. HD model systems, such as mice, only recapitulate aspects of disease and must be compared to the relevant data in human tissue5519et al.et al.et al.In AD, amyloid plaques are surrounded by chronically activated microglia36All experiments we performed in accordance with the Declaration of Helsinki and approved by the University College London (UCL)/UCL Hospitals Joint Research Ethics Committee and the LUMC IRB. Peripheral blood samples were donated by genetically-diagnosed HD patients and controls, and all subjects provided informed written consent.Manifest subjects demonstrated motor abnormalities that were unequivocal signs of HD, as evidenced by total motor scores (TMS) over 5 on the Unified Huntington’s Disease Rating Scale (UHDRS). Premanifest gene carriers had a burden of pathology score (age x [CAG – 36.5))The Track-HD cohort consisted of 54 premanifest gene carriers, 63 manifest HD subjects and 23 controls. These were a representative sample from the Track-HD study , preseleThe Leiden cohortWhole blood was collected in two PAXGene Blood RNA tubes  per subject, and immediately placed upright at room temperature. They were checked at 5 hours for incomplete mixing or separation, and any showing separation were remixed with a further 10 inversions. Tubes were stored overnight at −20 °C and transferred to −80 °C the following morning. They were sent on dry ice to Biorep within 30 days.TM method . Quality control measures were made on globin-reduced samples on the Bioanalyzer RNA 6000 Nano kit .Total RNA extraction was performed using the PAXGene Blood RNA kit , following the supplier’s instructions. Each solution in the kit was divided into aliquots to process batches of 12 samples. Replicate tubes for each subject were processed on different days. RNA was stored at −80 °C before proceeding with the quality measurements and further use. RNA was collected by centrifugation, washing with 70% ethanol, and resuspended in buffer. Quality measurements of total RNA were made using spectrophotometric analysis (Nanodrop), 260/280 ratio denaturing agarose gel, and the RNA 6000 Nano kit for the Agilent Bioanalyzer . Samples were globin reduced using the GLOBINclearTM Poly-A mRNA method (Illumina). In short, poly-A mRNA transcripts were captured from total RNA using poly-T beads and cDNA generated using random hexamer primingIndexed cDNA sequencing libraries were prepared using the TruSeqSequencing failed for six Track-HD samples, including four premanifest, one manifest and one control subject. Quality control analysis was performed using the RNA-SeQC packagehttp://www.ensembl.org/info/data/ftp/index.html, obtained in gtf format, genome build GRCh38.3, gene build updated in June 2015). To remove residual batch effects the R package svaseq was usedRNA-Seq data were aligned to the human reference genome hg19 using TopHat2Enrichment of differential expression among gene sets corresponding to biological hypotheses (pathways) was tested using the Gene Set Enrichment Analysis (GSEA) methodet al.The set of 124 HD brain expression modules derived by Neueder and Bateset al.A set of 117 co-expression modules derived from the Gibbs, We generated a set of 213 co-expression modules from Braineacet al.The set of 111 co-expression modules from Zhang, Weaknesses of relying on public databases to provide pathways for analysis include their restriction to prior biological knowledge and the poor annotation of many genes. To overcome this annotation gap, we also tested the following sets of gene co-expression modules for enrichment of dysregulation:et al.et al.Labadorf, et al. datasets.We used a similar method to test for concordance of fold change in genes between the Track-HD and Mastrokolias All data is deposited at the European Genome-phenome Archive (EGA) and accessible through the authors or the NeurOmics consortium.How to cite this article: Hensman Moss, D. J. et al. Huntington’s disease blood and brain show a common gene expression pattern and share an immune signature with Alzheimer’s disease. Sci. Rep.7, 44849; doi: 10.1038/srep44849 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


The Mediator complex was originally discovered in yeast, but it is conserved in all eukaryotes. Its best-known function is to regulate RNA polymerase II-dependent gene transcription. Although the mechanisms by which the Mediator complex regulates transcription are often complicated by the context-dependent regulation, this transcription cofactor complex plays a pivotal role in numerous biological pathways. Biochemical, molecular, and physiological studies using cancer cell lines or model organisms have established the current paradigm of the Mediator functions. However, the physiological roles of the mammalian Mediator complex remain poorly defined, but have attracted a great interest in recent years. In this short review, we will summarize some of the reported functions of selective Mediator subunits in the regulation of metabolism. These intriguing findings suggest that the Mediator complex may be an important player in nutrient sensing and energy balance in mammals. Gene transcription in eukaryotic cells is orchestrated through extremely complicated processes and multiple steps, including initiation, elongation, and termination, with the initiation being the most studied regulation step of gene expression. Disturbance of transcription initiation often results in serious diseases, such as cancer, in humans  has sharply increased in the past decades. Dysregulation of genes in the pathways controlling glucose and/or lipid metabolism is common in states of insulin resistance and T2D, especially when patients are also diagnosed with non-alcoholic fatty liver disease (NAFLD) or cardiovascular disease  and PGC1 families of transcription cofactors, MED1 contains two LXXLL motifs  that provide the binding surfaces for various nuclear receptors, and either one of the LXXLL motifs is sufficient for protein–protein interactions -selective genes  and HMG-CoA synthase (Hmgcr) (Amemiya-Kudo et al. The functions of MED15 and other mediator subunits in worms have been recently reviewed (Grants et al. RhoA/MAL or Ras/ELK1, by directly interacting with them in response to upstream signals (Stevens et al. MED23 has been well studied in mammalian model systems. The importance of MED23 in viability has been shown with the embryonic lethality when MED23 was knocked out in mice (Balamotis et al. It is known that HNF4α regulates a various set of genes that are not only involved in early development but also in liver and pancreatic cell differentiation, glucose metabolism, and lipid homeostasis (Odom et al. i.e., CDK19, MED12L, and MED13L (Daniels et al. Originally considered as a part of a transcriptional repressor in the large Mediator complex, the kinase module can repress or activate gene expression through kinase-dependent or kinase-independent mechanisms (Malik and Roeder The CDK8–CycC dimer negatively regulates de novo lipogenesis by reducing nuclear SREBP-1a or SREBP-1c protein stability (Zhao et al. In addition to CDK8 function in the negative regulation of de novo lipogenesis through its kinase activity, it has been shown that CDK8 has a positive role in response to serum, likely at the elongation step instead of the initiation step where CDK8 is a negative regulator (Donner et al. Through MED13, the MED12–MED13 dimer links the small mediator as well as the CDK8–CycC dimer (Tsai et al. To date, not much information is available on the functions of MED12L and MED13L. Expressed in both heart and brain, MED13L is associated with early development of both heart and brain since its missense mutations and gene interruption are found in patients with congenital heart defect, learning disabilities, and facial anomalies (Muncke et al. Since the discovery of the Mediator complex as a transcription cofactor of eukaryotes, its subunits have been associated with various biological processes and several diseases ranging from developmental defects to cancer in animal models and humans. The metabolic functions of the Mediator complex have become increasingly significant. In most cases, the Mediator complex functions as a bridge to connect and integrate specific transcription factors to the basal transcription machinery, resulting in expression changes of a selective set of genes. In addition to the presence of many subunits, tissue-specific expression and possible diverse assembly states at different physiological conditions further complicate the biological functions of the Mediator complex. Future studies on the role of the Mediator complex in metabolism may include identification of how the metabolic or nutrient signals may regulate the assembly states and subunit compositions and investigation of tissue-specific functions of each subunit in regulating nutrient and energy metabolism in vivo.


Erythrocytes regulate vascular function through the modulation of oxygen delivery and the scavenging and generation of nitric oxide (NO). First, hemoglobin inside the red blood cell binds oxygen in the lungs and delivers it to tissues throughout the body in an allosterically regulated process, modulated by oxygen, carbon dioxide and proton concentrations. The vasculature responds to low oxygen tensions through vasodilation, further recruiting blood flow and oxygen carrying erythrocytes. Research has shown multiple mechanisms are at play in this classical hypoxic vasodilatory response, with a potential role of red cell derived vasodilatory molecules, such as nitrite derived nitric oxide and red blood cell ATP, considered in the last 20 years. According to these hypotheses, red blood cells release vasodilatory molecules under low oxygen pressures. Candidate molecules released by erythrocytes and responsible for hypoxic vasodilation are nitric oxide, adenosine triphosphate and S-nitrosothiols. Our research group has characterized the biochemistry and physiological effects of the electron and proton transfer reactions from hemoglobin and other ferrous heme globins with nitrite to form NO. In addition to NO generation from nitrite during deoxygenation, hemoglobin has a high affinity for NO. Scavenging of NO by hemoglobin can cause vasoconstriction, which is greatly enhanced by cell free hemoglobin outside of the red cell. Therefore, compartmentalization of hemoglobin inside red blood cells and localization of red blood cells in the blood stream are important for healthy vascular function. Conditions where erythrocyte lysis leads to cell free hemoglobin or where erythrocytes adhere to the endothelium can result in hypertension and vaso constriction. These studies support a model where hemoglobin serves as an oxido-reductase, inhibiting NO and promoting higher vessel tone when oxygenated and reducing nitrite to form NO and vasodilate when deoxygenated. How erythrocytes modulate vascular tone has been widely studied over the last two decades. The vasodilation of the vasculature under hypoxic conditions has inspired much research ranging from the effect of oxygen partial pressure on smooth muscle cell contractility and endothelial nitric oxide synthase (eNOS) activity to nitrite reduction by hemoglobin (Hb) inside erythrocytes and subsequent production of nitric oxide Figure . Here we2 molecule. As hemoglobin binds oxygen, conformational changes occur to the protein and the binding affinity of the protein for molecular oxygen increases. Thus, binding is cooperative so that binding subsequent molecules of oxygen becomes easier after initial binding. Cooperative binding can be understood by a two-state model: The high affinity R-state and the low affinity T-state   by eNOS, which requires oxygen as a substrate along with L-arginine    extremely fast (6–8 × 10However, as demonstrated above in the work by Chu et al. endothelial NO does play a physiological role in vascular function (Chu et al., One can explain this paradox of NO bioavailability and Hb concentration in the blood by considering Hb encapsulation by RBCs. Experiments where RBCs are mixed with NO in suspension or in a stopped flow apparatus show that RBCs scavenge NO 1,000 times slower than cell-free Hb (Liu et al., In addition to these changes under static conditions, a cell free zone created by the fluid dynamics of blood flow further reduces NO scavenging. The velocity of blood near the vessel wall is slower than in the center of the vessel due to friction. This creates a pressure gradient that pushes RBCs toward the center of the vessel away from the NO producing endothelium. Bugliarello and Sevilla, Cokelet and Liao et al. experimentally showed the presence of a cell free zone and computational modeling by Butler et al. supported its ability to reduce NO scavenging (Bugliarello and Sevilla, In disease, RBC membrane abnormalities promote thrombosis Ataga, . Setty eIn addition to adhesion, polymerization of hemoglobin in sickle cell disease leads to rigid, inflexible red blood cells, which effect blood rheology. Moreover, oxygenated sickle RBC also displays changes in cell mechanics and therefore blood rheology, attributed to altered hemoglobin concentration in the cell and changes to the cell membrane (Nash et al., Work by Bor-Kucukatay et al. suggest vascular function, specifically nitric oxide production could play a role in RBC deformability and rheological alterations to RBCs during hypertension (Bor-Kucukatay et al., As reviewed above, encapsulation of Hb inside RBCs greatly reduces its ability to scavenge NO produced by the endothelium. However, in multiple diseases RBC hemolysis occurs introducing cell-free Hb into the vasculature (Reiter et al., In vitro findings suggest cell-free hemoglobin has the potential to release hemin and take part in oxidative reactions resulting in oxidative stress and inflammation (Miller et al., Cell-free hemoglobin reactions in the vasculature and surrounding tissues through extravasation may add to disease conditions (Schaer et al., In addition to releasing Hb, hemolysis also releases arginase, an enzyme that converts L-arginine to ornithine, into the blood stream. L-arginine is the substrate for nitric oxide synthesis by eNOS and therefore, the release of arginase further diminishes NO production and vascular function (Morris et al., Red blood cell breakdown not only decreases NO bioavailability through NO scavenging by cell-free Hb, but also by production of red blood cell microparticles (Donadee et al., RBC hemolysis contributes to the vascular pathology of diseases and disorders such as thalassemia, hereditary spherocytosis, Glucose-6-phosphate dehydrogenase deficiency, paroxysmal nocturnal, hemoglobinuria, and autoimmune hemolytic anemia (Johnson et al., In contrast to the role played by RBCs in diminishing NO. Research shows deoxy-RBCs promote vasodilation in the presence of nitrite (Cosby et al., 2 caused vessel dilation and an increased concentration of ATP in the effluent (Dietrich et al., ATP activates purinoceptors on endothelial cells leading to the production of NO and alteration of vascular tone (Ralevic and Burnstock, Addition of nitrite to deoxyRBCs showed an enhanced effect on vasodilation when infused (Cosby et al., in vivo SNO-Hb must form during the oxygenation/deoxygenation cycle of RBCs. This leads to the question of how SNO-Hb forms. Huang et al. and Xu et al. ruled out allosterically-controlled transfer of NO from HbNO to the beta-93 cysteine of Hb (Xu et al., The second proposed mechanism for the effect deoxyRBC and nitrite on vascular function is the formation of SNO-Hb in the presence of nitrite and subsequent release of NO from deoxygenated SNO-Hb. Research has demonstrated the hypoxic release of NO from SNO-Hb and the ability of the released NO to relax vessels (Stamler et al., In vitro studies have rejected a significant role in the reduction of nitrite by other RBC molecules such as xanthine oxidoreductase and carbonic anhydrase supporting the hypothesis that hemoglobin plays a dominant role (Liu et al., The last proposed mechanism of RBC vasodilation in the presence of nitrite is the reduction of nitrite to NO by Hb. Brooks first studied the nitrite Hb interaction in 1937 and this work was later extended by Doyle in 1981 Brooks, ,b, to fo2O3 formation (Basu et al., + for the RBC membrane, which in turn could also lead to NO escape through nitrosylation of AE1 (Salgado et al., However, once NO is formed inside a RBC, it must somehow escape rapid scavenging by the intraerythrocytic Hb. Proposed mechanisms of escape of NO bioactivity from the RBC have included S-nitrosothiol formation and the role of the RBC membrane. Research by Basu et al and Roche et al suggested S-nitrosothiol formation could occur through a metHb-nitrite mediated NOther heme-globin nitrite reducers may play a role in modulating vascular function. Deoxymyoglobin reduces nitrite faster than Hb (Shiva et al., The production of NO by deoxyRBCs in the presence of nitrite and other NO donors provide a potential therapeutic role for plasma nitrite in reducing vasoconstriction, RBC adhesion, and potentially improving RBC deformability (Space et al., Red blood cells play an important role in vascular function. Through the compartmentalization of Hb, RBCs deliver oxygen, minimize NO scavenging, sense oxygen tension, and deliver NO bioactivity in hypoxia. In healthy conditions, RBCs promote hemostasis through the well-regulated delivery of oxygen and the balance of NO scavenging and production. As data has shown, the RBC, once only viewed as a sink for NO, has the ability to produce NO bioactivity under hypoxia. The important role of the RBC in vascular function becomes more evident in diseases that alter or compromise the RBC membrane leading to conditions of oxidative stress, hypertension, thrombosis, and vaso-occlusion.CH, MG, and DK-S together conceptualized the article and its content. CH wrote the initial draft of the manuscript and further edited the manuscript. MG and DK-S heavily edited the 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.


Alternatively, resigned individuals may simply be numb to the negative effects of exclusion regardless of their momentary expectations (Bernstein and Claypool, Future research should examine if and how expectations matter for chronically excluded individuals. Williams  refers tExclusion paradigms often blindside participants with exclusion (Williams and Wesselmann, whether participants will choose to respond pro- or anti-socially to exclusion, as well as the degree of their response, which is a current paradox in the literature (Wesselmann et al., get-acquainted paradigms (Wesselmann et al., life alone paradigm (Twenge et al., in vivo paradigms.Participants' expectations of inclusion also likely influence Situational factors that may influence expectations of relational evaluation could be the psychological closeness of the sources of inclusion (e.g., a romantic partner; Arriaga et al., role-based exclusion, Nezlek et al., Individual factors may also influence one's expected relational evaluation levels. For example, narcissistic individuals may expect high relational evaluation and thus respond with more aggression than non-narcissists when their expectations are violated (Bushman and Baumeister, Researchers should investigate the specific role that relational evaluation plays in the consequences of exclusion, and how situation-level and individual-level characteristics influence this construct. EW, JW, and MB each contributed to the theoretical arguments in this article.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.


Myocardial infarction afflicts close to three quarters of a million Americans annually, resulting in reduced heart function, arrhythmia, and frequently death. Cardiomyocyte death reduces the heart’s pump capacity while the deposition of a non-conductive scar incurs the risk of arrhythmia. Direct cardiac reprogramming emerged as a novel technology to simultaneously reduce scar tissue and generate new cardiomyocytes to restore cardiac function. This technology converts endogenous cardiac fibroblasts directly into induced cardiomyocyte-like cells using a variety of cocktails including transcription factors, microRNAs, and small molecules. Although promising, direct cardiac reprogramming is still in its fledging phase, and numerous barriers have to be overcome prior to its clinical application. This review discusses current findings to optimize reprogramming efficiency, including reprogramming factor cocktails and stoichiometry, epigenetic barriers to cell fate reprogramming, incomplete conversion and residual fibroblast identity, requisite growth factors, and environmental cues. Finally, we address the current challenges and future directions for the field. As the leading cause of death in the United States, heart disease accounts for one out of every four mortalities —that are necessary and sufficient to convert fibroblasts to induced cardiomyocyte-like cells (iCMs) both in vitro  inhibition directs cells toward a cardiac lineage fate (Karamboulas et al., Widespread epigenetic repatterning occurs during direct cardiac reprogramming. Zhao et al. performed chromatin immunoprecipitation sequencing of H3K4me2, which marks the promoters and enhancers of transcriptionally active genes, and demonstrated that one week after GHMT cocktail reprogramming 47% of the H3K4me2 peaks had shifted to align with those of primary cardiomyocytes (Zhao et al., Given that extensive repatterning of histone modifications occurs during the reprogramming process, modulation of the deposition and removal of histone modifications may promote greater reprogramming efficiency. Hirai et al. used a small molecule inhibitor of Ezh2, the catalytic component of the PRC2 complex catalyzing H3K27me2/3, to demonstrate that Ezh2 inhibition early in reprogramming increases reprogramming efficiency (Hirai and Kikyo, To identify key epigenetic barriers to the reprogramming process, we conducted a comprehensive loss-of-function screen of epigenetic modifying factors and identified Bmi1 as a critical epigenetic inhibitor of reprogramming (Zhou et al., Ebf1 is a key target of Mll1, and the reduction in Ebf1 expression caused by Mll1 inhibition mediates the observed increase in reprogramming efficiency (Liu et al., In a similar approach, Liu et al. conducted a gain-of-function screen of cardiac development epigenetic modifiers and transcription factors and identified the H3K4 methyltransferase Mll1 as a barrier to direct cardiac reprogramming (Liu et al., Direct cardiac reprogramming is accomplished through the activation of cardiac gene expression but must also be accompanied by complete silencing of the original fibroblast signature. Characterization of epigenetic repatterning during reprogramming by Liu et al. demonstrates a gradual repression of fibroblast loci (Liu et al., Maintenance of residual fibroblast gene expression presents a roadblock to the successful reprogramming of functionally mature cardiomyocytes. MicroRNAs miR-1 and miR-133 enhance direct cardiac reprogramming by suppressing fibrotic gene expression (Muraoka et al., in vitro.In another approach to erasing fibroblast identity, studies have used inhibition of pro-fibrotic signaling pathways to enhance reprogramming efficiency (Ifkovits et al., in vivo (Mohamed et al. Mohamed et al. demonstrate that repression of residual fibroblast signature using TGFβ and Wnt inhibition also enhances reprogramming and cardiac function Recent studies have examined the effect of intracellular signaling pathways on direct cardiac reprogramming. Zhou et al. modulated intracellular signaling pathways by screening a library of 192 protein kinases to assess the effect on GHMT transcription factor reprogramming (Zhou et al., Abad et al. screened seven small molecule compounds with a demonstrated role in iPSC reprogramming and found that the Notch inhibitor DAPT enhances GHMT transcription factor cocktail reprogramming (Abad et al., in vitro and in vivo (Mohamed et al., in vitro (Mohamed et al., in vivo (Mohamed et al., Mohamed et al. screened a library of 5,500 small molecule compounds in an unbiased, high throughput approach to determine cell signaling pathways that modulate reprogramming and identified the WNT and TGFβ signaling pathways as barriers to reprogramming (Mohamed et al., in vivo over in vitro (Qian et al., In vivo reprogrammed iCMs are more similar to endogenous cardiomyocytes than in vitro reprogrammed iCMs. This suggests that unidentified extrinsic factors in the in vivo microenvironment such as topographic cues, mechanical forces, growth factors, cytokines, or paracrine signaling play an important role in promoting iCM maturation.Multiple studies have noted greater conversion efficiency or more complete functional maturation of iCMs reprogrammed in vitro reprogramming generated incompletely converted, immature iCMs (Yamakawa et al., in vitro generation of functionally mature iCMs that contract spontaneously and exhibit calcium oscillations (Yamakawa et al., Lack of requisite growth factors constitutes a barrier to reprogramming. Yamakawa et al. noted that under serum-based culture conditions, in vitro substrate (Chopra et al., In addition to growth factors, environmental cues are important in developing the functional maturity of iCMs. Cultured cardiomyocytes respond differently to the stiffness of the in vitro substrate stiffness or mechanical stretch, reprogramming does respond to other topographical cues. Morez et al. demonstrated that the forward programming of cardiac progenitor cells using the cardiac lineage transcription factor cocktail Myocardin, Tbx5, and Mef2c is enhanced by topographical cues which modulate histone acetylation (Morez et al., + adult progenitor cells were reprogrammed on flat or microgrooved collagen I coated polydimethylsiloxane substrates. Reprogramming efficiency and sarcomere assembly are enhanced on microgrooved substrates compared to flat substrates (Morez et al., While direct cardiac reprogramming is unaffected by in vitro cell culture substrates directly alters reprogramming efficiency and maturity.The identity and composition of the extracellular matrix also impacts cardiomyocyte phenotype and reprogramming. Substrate adhesive ligands alter cardiomyocyte sarcomere organization and maturation through integrin signaling. Sarcomeres are well organized and polarized when cardiomyocytes are cultured on fibronectin coated polyacrylamide substrates but not on collagen I coated polyacrylamide (Chopra et al., in vivo, Li et al. cultured reprogramming iCMs in a 3D fibrin hydrogel and demonstrated that 3D culture enhances direct cardiac reprogramming (Li et al., in vivo compared to in vitro (Li et al., To more accurately mimic environmental stimuli in vivo conditions also has the potential to improve reprogramming. Promoting angiogenesis through preconditioning with VEGF increases in vivo reprogramming efficiency and improves therapeutic restoration of cardiac function (Mathison et al., Myh7 positive cardiomyocytes in the infarct zone of GMT treated hearts and increases ejection fraction by four-fold (Mathison et al., in vivo reprogramming efficiency (Qian et al., Manipulation of Additional research is required to translate direct cardiac reprogramming into a clinical therapy. Necessary steps include continued basic research, research in large animal models, improvement in human reprogramming, and bioengineering of delivery mechanisms.A better understanding is needed of the mechanism of late stage reprogramming events and iCM maturation. Research in this area is currently hindered by inefficiency in the reprogramming process. Asynchronous, heterogeneous cell populations produce low rates of fully reprogrammed cells making it difficult to acquire sufficient cell numbers to study late stage reprogramming events. Early stage studies are aided by the comparative synchrony of cells early in the reprogramming process that provides a large sample population.in vivo monitoring systems, and genetic manipulation, making the mouse a particularly productive model. However, the mouse exhibits significant cardiovascular differences compared to humans. In addition to obvious differences such as small size and short lifespan, mice differ from humans in a range of anatomical, physiological, energetic, electrophysical, and mechanical properties that include heart rate, coronary artery structure, and contraction/relaxation kinetics. Large animal models such as the dog, sheep, or pig have greater physiologic resemblance to humans with similar body size, heart size, and heart rate. In fact, physiological similarities between pigs and humans are close enough to make the pig an ideal xenotransplant donor. Recent progress in genetic manipulation of pigs will contribute to the use of the pig as a cardiovascular disease model.Research in large animal models is also required to move direct cardiac reprogramming toward clinical application. The efficiency of housing, breeding, and handling rodents has made them the most widely used animal models in biomedical research. Additionally, a wealth of tools has been developed specifically for murine research, including imaging techniques, Although significant progress has been achieved in uncovering the molecular barriers to direct reprogramming in mice, research in reprogramming human cells lags far behind. Reprogramming human fibroblasts requires the addition of extra factors but yields far lower conversion efficiency. Spontaneously beating cells are rare, indicating that more work is required to translate findings from the mouse to human and uncover undiscovered molecular barriers in human reprogramming.in vivo research uses direct injection of retroviral vectors into the infarct zone. While conceivably, direct cardiac injection could be achieved in some myocardial infarct patients during coronary bypass surgery, a non-invasive delivery method is preferable. Additionally, retroviral vectors integrate into the host cell genome, incurring the risk of gene disruption and cellular transformation. The ideal delivery vector would be non-integrating with high transfection efficiency, specificity for the target cell type, and adequate capacity to accommodate multiple reprogramming factors. Adenoviruses are promising viral vectors for the delivery of reprogramming factors. The most commonly employed vector in clinical trials, adenoviruses are non-integrating, with large capacity and high transduction efficiency. Additionally, recent research using small molecules to achieve reprogramming and developments in nanoparticle delivery systems offer potential alternatives to viral vector reprogramming factor delivery.Finally, a safe and efficient delivery system is required for the translation of direct cardiac reprogramming to clinical use. Current


The International Conference on Bioinformatics (InCoB) has been publishing peer-reviewed conference papers in BMC Bioinformatics since 2006. Of the 44 articles accepted for publication in supplement issues of BMC Bioinformatics, BMC Genomics, BMC Medical Genomics and BMC Systems Biology, 24 articles with a bioinformatics or systems biology focus are reviewed in this editorial. InCoB2017 is scheduled to be held in Shenzen, China, September 20–22, 2017. Abbas and Bahig  investigMycobacetrium tuberculosis could explain how azathioprine could suppress the pathogenicity of this organism in transplant patients. Le and Ou [Molecular recognition factors (MoRFs) located within longer intrinsically disordered proteins perform important cellular functions such as signalling and regulation, but are very difficult to accurately detect. Sharma et al.  have adde and Ou  have devGutierrez and Nakai  have buiCastiglione et al.  have devOne of the fundamental assumptions of structural bioinformatics is that hydrophobic residues prefer to be buried in protein structures. By analysing the standard Barton502 dataset using support vector regression on informative physicochemical properties, Liou et al.  show thaSmall organic molecules as drug lead compounds is an important area of structural bioinformatics research, where knowledge of the target’s 3D structure enables rational ligand design, for therapeutic applications. Using a series of derivatives from the influenza drug zanamivir, Dholakia et al.  have genGiven the spiralling cost of drug development, a growing number of approved drug molecules are being considered for medical conditions different from their original purpose, known as repurposing or repositioning. As the number of known drug-target interactions far exceeds the potential interactions of these drugs with other potential targets, predicting the repurposing of a drug appears to be extremely challenging. Ezzat et al.  have addComputer-based morphological analysis present a rapid and efficient way to analyse biological samples. Skull analysis can be used for clinical applications as well as for species classification. Based on human orthodontic parameters used in clinical analysis, Mosleh et al.  have devIn pathology applications, cell-based microscopic analysis suffer from out-of-focus images due to depth effects. Intarapanich et al.  have devThe quality of the articles reviewed in this introduction are comparable with earlier InCoB conferences while the subject coverage has extended substantially into bioimaging. Next year's conference, InCoB2017 will be held in Shenzhen from Sept. 20–22, 2017 , with pa


Bladder-related pain is one of the most common forms of visceral pain, and visceral pain is among the most common complaints for which patients seek physician consultation. Despite extensive studies of visceral innervation and treatment of visceral pain, opioids remain a mainstay for management of bladder pain. Side effects associated with opioid therapy can profoundly diminish quality of life, and improved options for treatment of bladder pain remain a high priority. Endocannabinoids, primarily anandamide (AEA) and 2-arachidonoylglycerol (2-AG), are endogenously-produced fatty acid ethanolamides with that induce analgesia. Animal experiments have demonstrated that inhibition of enzymes that degrade AEA or 2-AG have the potential to prevent development of visceral and somatic pain. Although experimental results in animal models have been promising, clinical application of this approach has proven difficult. In addition to fatty acid amide hydrolase  and monacylglycerol lipase , cyclooxygenase (COX) acts to metabolize endocannabinoids. Another potential limitation of this strategy is that AEA activates pro-nociceptive transient receptor potential vanilloid 1 (TRPV1) channels. Dual inhibitors of FAAH and TRPV1 or FAAH and COX have been synthesized and are currently undergoing preclinical testing for efficacy in providing analgesia. Local inhibition of FAAH or MAGL within the bladder may be viable options to reduce pain associated with cystitis with fewer systemic side effects, but this has not been explored. Further investigation is required before manipulation of the endocannabinoid system can be proven as an efficacious alternative for management of bladder pain. Intravenous treatment of rats with URB937, a FAAH inhibitor with minimal penetration of the CNS decreased activity of bladder-specific afferent nerve in response to controlled filling of the bladder (Aizawa et al., AEA has been shown to be an agonist of the TRPV1 pro-nociceptive channel (Tognetto et al., Clinical trials of FAAH inhibitors failed to alleviate pain associated with osteoarthritis (Huggins et al., Although 2-AG is far more abundant than AEA, less work has been done investigating the therapeutic potential of inhibition of MAGL to ameliorate visceral pain. In a mouse chronic neurogenic pain model, MAGL inhibition alleviated neuropathic pain (Kinsey et al., A major obstacle to management of pain by inhibiting MAGL is the observation that increased systemic 2-AG actually results in enhanced response to painful stimuli due to desensitization of CB1 (Schlosburg et al., Endocannabinoids have been demonstrated to suppress inflammation in bladder (Wang et al., Mechanisms of suppression of inflammation by endocannabinoids are not completely understood, but AEA has been shown to inhibit mitogen-induced T- and B-cell lymphocyte proliferation by increasing apoptosis (Schwarz et al., in vitro damage to hippocampal slices exposed to β-amyloid by a process mediated by CB1 binding that resulted in diminished ERK 1/2 phosphorylation, decreased NFκB activation, and reduced COX-2 expression (Chen et al., Binding of 2-AG to CB1 inhibits cyclooxygenase-2 in nerves resulting in suppression of MAPK/NFκB signaling (Zhang and Chen, Inflammation plays a key role in release of substances that modulate nociception. Thus, it is highly likely that the analgesic effects of endocannabinoids may in part be due to their anti-inflammatory effects.Evaluation of bladder pain in rodent models can be difficult, and the presence of bladder pain is most often inferred by evaluating referred mechanical sensitivity of the hind paws or abdominal wall or activity of bladder afferent nerves (Sadler et al., in vivo with cyclophosphamide to induce inflammatory cystitis (Walczak et al., ex vivo in the presence or absence of intravesical cannabinoid agonists, and it was determined that intravesical cannabinoids suppressed afferent nerve fiber firing in inflamed bladders via CB1 activation (Walczak et al., Bladders and associated afferent nerves were isolated from mice treated Manipulation of the endocannabinoid system has emerged as an appealing alternative to opioids for management of severe bladder pain. However, the potential for undesirable side effects or lack of efficacy remain significant obstacles to advancement of this therapy. Emergence of dual inhibitors of endocannabinoid degradation and either activation of TRPV1 or COX may address many of the limitations of this approach. Information on the activity of endocannabinoids synthesized in peripheral tissues remains extremely limited. It remains unclear whether or not strategies to address organ-specific pain by manipulation of endocannabinoids is a viable option, but this is an intriguing alternative that has the potential to provide effective analgesia with minimal systemic side effects.All studies were approved by the University of Wisconsin Animal Care and Use Committee prior to performance. Procedures were consistent with guidelines provided by the Guide for the Care and Use of Laboratory Animals, 8th Edition, The National Academies Press, Washington, DC, USA.DB and ZW participated in the concept experimental design, analysis and writing. ZW was directly involved in performing experiments.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.


However, remyelination often fails in chronic neurological diseases, such as progressive multiple sclerosis. The lack of currently approved pro-remyelination therapies highlights the need to elucidate the cellular and molecular mechanisms underpinning this regenerative process. Whereas some T lymphocyte subsets such as Th1 and Th17 are implicated in inducing myelin injury, a recent study by Dombrowski et al. reveals a novel role for regulatory T cells (T Regeneration can occur efficiently in the central nervous system (CNS) in the form of remyelination, whereby new myelin is ensheathed around axons to reinstate trophic/ metabolic support and insulation for electrical impulse conduction. This process requires oligodendrocyte progenitor cells (OPCs) to migrate to areas of injury, proliferate, and differentiate into myelin-producing oligodendrocytes. Remyelination is limited or fails altogether in various neurological disorders, most prominently in progressive multiple sclerosis (MS), leading to axon dysfunction or loss. Although it is recognized that this failure largely reflects impaired oligodendrocyte differentiation , demonstrating that these cells directly stimulate remyelination independent of immunomodulation.While it is now recognized that the innate immune system (i.e. macrophages) supports remyelination  were detected in lesions at the time of oligodendrocyte differentiation and remyelination initiation (Dombrowski et al., regs. The effects of Treg depletion were mirrored in a distinct demyelination model (Dombrowski et al., regs in remyelination is not dependent on the mode of demyelination nor is it restricted to the spinal cord.Using a focal model of toxin-induced demyelination in the mouse spinal cord, where the temporal distinction between myelin damage and remyelination allows investigations of the regenerative process in isolation, Treg depletion in both models did not alter numbers of total oligodendrocyte lineage cells nor proliferating OPCs, suggesting effects of Tregs on the differentiation of OPCs into mature myelinating oligodendrocytes. Indeed, exposing brain explants to Tregs or their conditioned media enhanced oligodendrocyte differentiation, myelination, and remyelination, in comparison to non-polarized CD4+ T cells (Dombrowski et al., regs on OPCs was confirmed in vitro, where Treg conditioned media enhanced the differentiation of isolated OPCs and accelerated myelination in OPC-neuronal co-cultures (Dombrowski et al., reg conditioned media was carried out to identify pro-remyelination factors, and identified high expression of the growth regulator CCN3 (Dombrowski et al., reg conditioned media, as use of a blocking antibody or specific depletion from the conditioned media abolished the pro-differentiation and pro-myelination effects (Dombrowski et al., reg-derived CCN3 was sufficient to support these responses (Dombrowski et al., Tregs are direct drivers of oligodendrocyte differentiation and remyelination, thereby revealing a novel regenerative function for Tregs beyond immunomodulation. Considering this study, one must now acknowledge that the adaptive immune system is not solely involved in damage induction and modulation of inflammation, but is also a critical component of the regenerative process that follows. A recent study demonstrated that MS patient-derived CD4+ T cells injected into demyelinated mouse CNS show high variability in their ability to support remyelination (El Behi et al., regs to stimulate remyelination. Overall, these studies support that further investigations into the pro-remyelination function of Tregs and CCN3 in MS are warranted for development of novel regenerative therapies.Altogether, these data demonstrate that T


Machine learning techniques have been increasingly applied in the medical imaging field for developing computer-aided diagnosis and prognosis models. Multimodal medical imaging can provide us with separate yet complementary structure and function information of a patient study and hence has transformed the way we study living bodies. Therefore, using machine learning techniques to deal with multimodal medical images is much more challenging due to the diversity of biophysical-biochemical mechanisms. In these years, researchers mainly adapt modern machine learning and pattern recognition techniques such as supervised, unsupervised, semisupervised, and deep learning to solve multimodal medical imaging related problems.To record the ideas of talents and gather more contributions to these fields, this special issue was launched and supported by this journal. This special issue focuses on the new imaging modalities/methodologies and new machine learning algorithms/applications for the further development in the multimodal medical imaging field, which will provide opportunities for academics and industrial professionals to discuss the latest issues and progresses in the area of multimodal medical imaging. The papers contained in this special issue address the development and application of medical image segmentation, registration, fusion, classification, image restoration, image retrieval, and computer-aided diagnosis.In “Estimation of Response Functions Based on Variational Bayes Algorithm in Dynamic Images Sequences,” B. Shan proposes a nonparametric Bayesian model to estimate the response functions in dynamic medical imaging, in which the nonparametric Bayesian priors are designed to favor desirable properties of the functions and used to improve the estimation of response functions.In “Two-Layer Tight Frame Sparsifying Model for Compressed Sensing Magnetic Resonance Imaging,” S. Wang et al. propose a two-layer tight frame sparsifying model for compressed sensing magnetic resonance imaging (MRI) by sparsifying the image with a product of a fixed tight frame and an adaptively learned tight frame, which is solved by a three-level Bregman numerical algorithm and enables accurate MRI reconstruction from highly undersampled data with efficiency.In “Many is Better than One: An Integration of Multiple Simple Strategies for Accurate Lung Segmentation in CT Images,” Z. Shi et al. present a novel computerized tomography (CT) lung image segmentation method by integrating multiple strategies, including the guided filter to smooth the image, the optimized threshold to get binary image, region-growing strategy to extract thorax regions, and random walk algorithm to segment lung regions and to get the state-of-the-art segmentation accuracy.In “Pulmonary Nodule Detection Model Based on SVM and CT Image Feature-Level Fusion with Rough Sets,” T. Zhou et al. present a pulmonary nodules detection algorithm based on support vector machine (SVM) and CT image feature-level fusion with rough sets to improve the detection accuracy of pulmonary nodules in CT image. Both the unreasonable feature structure and the nontightness of feature representation are taken into consideration in this pulmonary nodules detection algorithm.In “Multigrid Nonlocal Gaussian Mixture Model for Segmentation of Brain Tissues in Magnetic Resonance Images,” Y. Chen et al. propose a novel segmentation method based on the regional and nonlocal information to overcome the impact of image intensity inhomogeneities and noise in human brain magnetic resonance images.In “DTI Image Registration under Probabilistic Fiber Bundles Tractography Learning,” Z. Guo et al. propose a diffusion tensor imaging (DTI) image registration method under probabilistic fiber bundles tractography learning, where the residual error model is modified with finite sample set and the calculated deformation field is then registered on the DTI images.In “Automated Segmentation of Coronary Arteries based on Statistical Region Growing and Heuristic Decision Method,” Y. Tian et al. propose a fully automated coronary artery segmentation from cardiac data volume based on a statistics region growing together with a heuristic decision to further help cardiovascular radiologists detect and quantify stenosis.In “Rapid Retrieval of Lung Nodule CT Images Based on Hashing and Pruning Methods,” L. Pan et al. propose a new retrieval framework based on a hashing method for lung nodule CT images, which can translate high-dimensional image features into a compact hash code to greatly reduce the retrieval time and memory space. Moreover, a pruning-based decision rule is utilized in this algorithm to improve its retrieval precision.In “The Classification of Tongue Colors with Standardized Acquisition and ICC Profile Correction in Traditional Chinese Medicine,” Z. Qi et al. design a tongue color classification approach using a standardized tongue image acquisition process, color correction, and several machine learning techniques for tongue inspection-based diagnosis in traditional Chinese medicine.In “Diagnostic Method of Diabetes Based on Support Vector Machine and Tongue Images,” J. Zhang et al. develop a SVM-based diagnostic method for diabetes using standardized tongue images. This work shows the potential of applying digitalized tongue images, which are usually used in traditional Chinese medicine, to the diagnosis of diabetes.In “A Computer-Aided Analysis Method of SPECT Brain Images for Quantitative Treatment Monitoring: Performance Evaluations and Clinical Applications,” X. Zheng et al. introduce and validate a computer-aided analysis method to achieve the quantitative treatment monitoring based on single-photon emission computed tomography (SPECT) images, which can provide a convenient solution to generate a parametric image and derive the quantitative indexes from the longitudinal SPECT brain images for treatment monitoring.The papers in this special issue provide a useful message of machine learning techniques in dealing with multimodal medical images. This unique and informative collection of papers highlights the direction of related studies. This special issue illustrates the important role that machine learning techniques play in the multimodal medical imaging fields.


Escherichia coli Bacteriophages Differ in Their Efficacy and Ability to Stimulate Cytokine Release In vitro,” Dufour et al. suggest that the level of cytokine response generated in our paper is due to remaining bacterial debris rather than true differences between individual phages , which to our understanding is achieved through lipopolysaccharide specific binding to a phage derived protein. However, as Dufour et al. rightly point out, “bacterial lysates may also contain several pathogen-associated molecular patterns (PAMPs) able to elicit a pro-inflammatory response, such as flagellin (sensed by TLR-5), unmethylated CpG Oligodeoxynucleotide DNA (TLR-9), lipoteichoic acid from Gram-positive bacteria (TLR-2) and triacyl lipopeptides (TLR-1 with TLR-2) . The additional purification step (the EndoTrap Blue system) binds endotoxins based on the conserved core of lipopolysaccharide (With hindsight and a thorough data audit, we would certainly concur that at least some component of the observed cytokine responses are due to remaining contaminants and that additional purification steps such as chromatography will further reduce possible contaminants. However, such additional steps could come at a cost, reduce overall phage concentration and potentially require additional concentration steps (Boratyński et al., However, while it is likely that some component of the cytokine response generated is due to remaining contaminants, additional variation could also be introduced through factors which are uncontrollable, such as genetic variation in Toll receptors between different donors (Netea et al., Despite the suggestion to the contrary provided by the Dufour et al. commentary, the reduction and quantification of endotoxin in phage preparations remains an active area of research (Branston et al., MKM, YH, MN, CC, ES-E, and AN: Writing of the reply and approved it for publication.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 author reports that Newcastle disease virus (NDV)-infected umbilical cord-derived mesenchymal stem cells (MSCs) may provide a novel effective therapeutic approach for targeting glioma stem cells (GSCs) and glioblastoma multiforme (GBM), and for sensitizing these tumors to γ-radiation [I read with great interest the article by Kazimirsky et al.  entitledInternational Journal of Cancer [Stem Cell Research & Therapy.A couple of additional points can be made on this topic. The findings of Kazimirsky et al.  regardinf Cancer  and an af Cancer –6. In adf Cancer  stated tf Cancer . In partf Cancer . From thContinuing with this important issue, in an article with 70 references none of the previously published work is discussed or cited by Kazimirsky et al. . As we k


Traditionally, the role of “emotion” has received little attention in research studies of decision-making  that can be used to simulate dynamic real-life decision-making behavior as well as test the SMH  in each trial and large losses  in some trials. The selection of decks C and D results in a small gain (US$50) in each trial and small losses  in some trials. On average, selections from decks A and B over 10 trials will cause decision-makers lose US$250, and as such, these are defined as disadvantageous decks. Conversely, selections from decks C and D over 10 trials will cause decision-makers gain US$250, so these are defined as advantageous decks. The advantageous decks  provide small immediate gains in each trial, but the long-term outcome is positive; by contrast, the disadvantageous decks  provide large immediate gains in each trial, but the long-term outcome is negative . Consequently, findings related to the PDB phenomenon and gain/loss frequency have clearly echoed the main points reported in previous literature concerning behavioral decision-making  Must et al. review IGT and depression-related issues; (2) Brevers et al. review studies on IGT and gambling disorders; (3) Linnet provide a review of IGT in the context of dopamine and gambling disorders; (4) Cassotti et al. review IGT in relation to developmental studies; (5) Turnbull et al. consider IGT performance as the processing of emotion-based learning; (6) Overman and Pierce examine the effects of real plus virtual cards and additional trials; and (7) van den Bos et al. provide a global overview of rodent version of the IGT.Category II: Clinical examinations: (1) Sallum et al. discuss the IGT and attention deficit hyperactivity disorder; (2) Xiao et al. combine the IGT and functional magnetic resonance imaging (fMRI) in order to investigate adolescent smoking behavior; (3) Singh describe the connection between sleep deprivation and IGT performance; and (4) de Oliveira Cardoso et al. provide a behavior-image study that investigates the correlation between frontal and cerebellar lesions and IGT performance.Category III: Model construction: (1) Worthy et al. compare predictability between win-stay/lose-shift and Value-Plus-Preservation (VPP) models in the IGT; (2) Steingroever et al. validate the predictive power of the Prospect Valence Learning–Delta model; (3) Dai et al. provide an improved cognitive model for predicting IGT choice behavior; (4) Lin et al. refine a simplified model for estimating IGT performance; and (5) Ahn et al. compare three advanced IGT-related computational models.Category IV: Theoretical integration: (1) Okdie et al. provide a statement on construal level theory for IGT-related performance; (2) Bull et al. consider sensitivity toward reward and punishment in healthy IGT participants; (3) Singh suggest a potential role for reward and punishment during the IGT; and (4) Singh consider the influence of sex-differences, handedness, and lateralization on IGT performance.Category V: Brain imaging technology: (1) He et al. combine IGT and fMRI to investigate decisions involving unhealthy food; (2) Mapelli et al. utilize the IGT and event-related potentials (ERPs) to depict the behavioral performance and brain activation of patients with Parkinson's disease; (3) Tamburin et al. combine the IGT and ERPs to detect choice behavior and brain activation in patients with chronic lower-back pain; and (4) Fernie and Tunney describe a study on the correlation between SCRs and knowledge effects in the IGT.The articles selected for inclusion in this special issue provide good coverage of neuroimaging modalities  used in previous IGT experiments. However, there might still be some room for a data-driven data analysis method  What types of brain lesions  does the IGT truly measure? (2) Can SCRs be combined with the IGT to form a critical index of somatic markers? (3) Does the IGT measure ability for implicit or explicit learning? (4) Does EV or gain/loss frequency primarily guide decision-making behavior in the IGT? (5) Is it possible to devise a more sensitive data analysis method that can allocate more specific brain responses to the precise behaviors of IGT performance, such as the events of win, loss, and the switching of card decks? We recommend that future studies of IGT consider these questions seriously and provide in-depth investigations and discussions.Y-CC, C-HL, and J-TH discussed the main structure of this article. Y-CC and C-HL drafted the preliminary title, literature review, and chapter categorization, as well as the initial draft. J-RD and C-HL provided additional viewpoints for future development in the use of brain imaging for studying the IGT. J-TH and J-RD provided final refinements to this article.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 term “nutraceutical”, derived from the words “nutrition” and “pharmaceutical” was coined in 1989 to describe substances that could be used as foods that possess health benefits. The range of nutraceutical products is widely diverse and may be divided into three main categories: (i) natural products, comprising, among others, herbs and spices; (ii) dietary supplements; and (iii) functional foods. Based on food sources, nutraceuticals can be classified as: (i) dietary fibres of plant origin; (ii) probiotics, which are live microbial feed supplements; (iii) prebiotics, dietary ingredients that selectively alter the composition or metabolism of gut microbiota; (iv) polyunsaturated fatty acids, omega-3 or omega-6 fatty-acids; (v) antioxidant vitamins, including vitamin C, vitamin E and carotenoids; (vi) biologically active phytochemicals like polyphenols; and (vii) spices, whose main components are terpenes and other constituents of essential oils.In the recent years, the field of nutraceuticals has progressed from being a mere conceptual area within biomedical research to a value-added industry with a promising future ahead. Indeed, lately, both the academic community and the general population have become increasingly interested in the health benefits and potential risks pertaining to beverages and food products beyond their basic nutritional assets. In addition, continuing scientific research has provided evidence regarding the biologically active compounds and underlying physiological mechanisms, highlighting the importance of nutraceuticals as complements to the conventional therapies and medications allowing dose reduction and decreasing the occurrence of adverse effects. Despite the undeniable progress in the field of nutraceuticals, there are still several issues that remain to be addressed. These include clinical evidence supporting in vitro claims, regulatory aspects and assurance of nutraceuticals’ identity, quality and safety.International Journal of Molecular Sciences comprises one review article and 16 original research papers that provide important insights into the state of the nutraceuticals field. Nosrati et al. [Fucus spiralis in a human cell in vitro model (MCF-7 cells). Han et al. [The special issue “Nutraceuticals in Human Health and Disease” in the i et al.  use mixei et al.  charactei et al.  investign et al.  report tFicus umbellata constituents that appears to be due to apoptosis induction through the ROS-dependent mitochondrial pathway. The work conducted by Dai et al. [Sophora flavescens, inhibits influenza A virus replication and possesses anti-inflammatory activities and that the underlying mechanism may be linked to its ability to inhibit IAV-induced activations of the TLR4, p38 MAPK, and NF-κB pathways.The studies conducted by Forney et al.  provide i et al.  reveals Acanthopanax senticosus in repressing radiation-induced protein expression changes on prefrontal cortex, and suggest that this compound may be a promising alternative medicine for the treatment of radiation injury in the brain yet further testing are needed to understand whether the bioactive ingredients could be effective through the blood–brain barrier. Klimaszewska-Wiśniewska et al. [Studies developed by Liu and Dey  suggest a et al.  provide Collectively, these studies support a role for nutraceuticals in the prevention and amelioration of several diseases. Their innovation in the field and their successful future clinical application will be governed by evidence regarding safety, purity and efficacy. The use of these compounds in clinical practice is emerging, yet important pharmaceutical and clinical issues remain to be answered.


To lessen the “wear and tear” of existence, cells have evolved mechanisms that continuously sense DNA lesions, repair DNA damage and restore the compromised genome back to its native form. Besides genome maintenance pathways, multicellular organisms may also employ adaptive and innate immune mechanisms to guard themselves against bacteria or viruses. Recent evidence points to reciprocal interactions between DNA repair, DNA damage responses and aspects of immunity; both self-maintenance and defense responses share a battery of common players and signaling pathways aimed at safeguarding our bodily functions over time. In the short-term, this functional interplay would allow injured cells to restore damaged DNA templates or communicate their compromised state to the microenvironment. In the long-term, however, it may result in the (premature) onset of age-related degeneration, including cancer. Here, we discuss the beneficial and unrewarding outcomes of DNA damage-driven inflammation in the context of tissue-specific pathology and disease progression. To withstand the hazards of existence, multicellular organisms need to preserve their bodily functions for long periods of time and protect themselves against pathogens. Taking the cell as a point of reference, the maintenance is directed inwards to counteract macromolecular damage. This often involves restoring injured nucleic acids back to their native form  and interleukin-8 (IL-8) , we recently showed that persistent DDR triggers a chronic auto-inflammatory response leading to severe fat depletion in mice (Karakasilioti et al., Ercc1F/− fat depots showed hallmarks of persistent DDR together with the marked up-regulation of pro-inflammatory factors, the infiltration of activated macrophages as well as the release of DAMPs known to initiate and perpetuate immune responses (Karakasilioti et al., Ercc1F/− fat depots in vivo and in adipocytes ex vivo showed that persistent DNA damage signaling triggers the induction of IL-6, IL-8, and TNFα by promoting transcriptionally active histone marks and the dissociation of nuclear receptor co-repressor complexes from promoters; the response required ATM and it was instigated in a DNA lesion- and cell type-specific manner. In support of these findings, NF-κB is stochastically activated in tissues of naturally-aged and Ercc1−/Δ mice (unlike Ercc1−/− mice, the Ercc1−/Δ animals maintain about 10% of the wild-type ERCC1 protein levels and develop progressive, degenerative changes that markedly resemble those seen in natural aging (Tilstra et al., Ercc1−/Δ mice. Moreover, inhibition of IKK/NF-κB activity reduced cellular senescence and oxidative damage in DNA and proteins (Tilstra et al., Zmpste24−/− and LmnaG609G/G609G progeroid animals. As in Ercc1−/Δ animals, genetic and pharmacological inhibition of NF-κB signaling can ameliorate the age-associated features and extend the lifespan of these animal models (Osorio et al., Atm−/− animals present with infiltration of neutrophils and lymphocytes in the lungs and increased mRNA levels of pro-inflammatory e.g., IL-6, TNF cytokines (Eickmeier et al., Cellular senescence is a term used to describe cells that cease to divide in culture and has been one of the first paradigms to link DNA damage and immunity to disease (Campisi and d'Adda di Fagagna, per se but persistent DDR that triggers the repertoire of innate immune responses (Fumagalli and d'Adda di Fagagna, ssDNA intermediates generated during e.g., transcription or DNA replication may also activate DDR and trigger a pro-inflammatory response (Abe et al., DNA damage-driven inflammation can be both beneficial and detrimental for organismal survival Figure . To undeFuture strategies aimed at identifying new players or delineate key pathways may shed light on the biochemical crosstalk DNA repair and immune factors allowing us to gain insights onto how both systems contribute to disease origin and progression at old age. In this regard, the use of e.g., tissue-specific or tagged knockin animals and high-throughput proteomics and genomics approaches will likely prove valuable toward the development of rationalized interventions (Tilstra et al., AI and EG researched the literature, prepared a schematic first draft, and Figure 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.


Coffee is the most consumed beverage worldwide. Epidemiological studies with prospective cohorts showed that coffee intake is associated with reduced cardiovascular and all-cause mortality independently of caffeine content. Cohort and case-control studies reported an inverse association between coffee consumption and the degree of liver fibrosis as well as the development of liver cancer. Furthermore, the beneficial effects of coffee have been recently confirmed by large meta-analyses. In the last two decades, various in vitro and in vivo studies evaluated the molecular determinants for the hepatoprotective effects of coffee. In the present article, we aimed to critically review experimental evidence regarding the active components and the molecular bases underlying the beneficial role of coffee against chronic liver diseases. Almost all studies highlighted the beneficial effects of this beverage against liver fibrosis with the most solid results indicating a pivot role for both caffeine and chlorogenic acids. In particular, in experimental models of fibrosis, caffeine was shown to inhibit hepatic stellate cell activation by blocking adenosine receptors, and emerging evidence indicated that caffeine may also favorably impact angiogenesis and hepatic hemodynamics. On the other side, chlorogenic acids, potent phenolic antioxidants, suppress liver fibrogenesis and carcinogenesis by reducing oxidative stress and counteract steatogenesis through the modulation of glucose and lipid homeostasis in the liver. Overall, these molecular insights may have translational significance and suggest that coffee components need clinical evaluation. Coffea, a member of the Rubiaceae family, which includes hundreds of different species. Commercial production mainly exploits the seeds of Coffea arabica (arabica coffees), which represent about 70% of the world market, while Coffea canephora (commonly known as robusta coffees), which has a more bitter taste than arabica, is used principally for instant and espresso coffees (www.ico.org).Coffee is the most consumed beverage worldwide with an impressive impact on the economy of producing countries. Coffee is prepared from the seeds of the coffee plant, genus The association between coffee consumption and all-cause and cause-specific mortality, especially cardiovascular mortality, has been investigated in independent studies evaluating prospective cohorts, with most solid studies finding an inverse association with all-cause and cardiovascular mortality for both caffeinated and decaffeinated blends ,2. RecenIn this review, we aimed to summarize experimental evidence regarding molecular mechanisms underlying the hepatoprotective effects of coffee by reviewing in vitro and in vivo studies. A search of the electronic databases Medline and Embase was performed to retrieve relevant articles published up to May 2016. A combination of the keywords “coffee”, “caffeine”, “coffee polyphenols“, “chlorogenic acid”, “cafestol”, “kahweol”, “melanoidins”, “liver steatosis”, “liver damage”, “liver injury”, “liver inflammation”, “liver fibrosis”, “liver cirrhosis”, “liver cancer” was searched. All published studies in the English language were considered. Additional papers were retrieved by references of epidemiological studies on coffee and liver diseases. After careful evaluation, the following studies were excluded from the review: (i) studies reporting in vivo effects on acute models of liver damage or failure ; (ii) studies reporting in vitro effects on metabolic processes without assessing effects on cell steatosis or apoptosis (pure metabolic studies).Convincing results from a meta-analysis of 28 prospective studies have indicated that both caffeinated and decaffeinated coffee consumption is associated with lower risk of diabetes in a dose-response manner . ConsistPparγ), cluster of differentiation (Cd36), fatty acid binding protein 4 (Fabp4) and Mgat1 genes; more interestingly, CGA treatment also attenuated inflammation in the liver and white adipose tissue accompanied by a decrease in the mRNA levels of macrophage marker genes including EGF-like module-containing mucin-like hormone receptor-like 1 (F4/80), cluster of differentiation (Cd68), integrin Cd11b, integrin Cd11c and tumor necrosis factor alpha (Tnfα), monocyte chemotactic protein 1 (Mcp-1) and C-c chemokine receptor type 2(Ccr2) encoding inflammatory proteins.To our best knowledge, the first experimental study showing a protective effect of a coffee component against liver steatosis was from Rodriguez de Sotillo and Hadley , who shoDue to the tight connection of NAFLD with cardiovascular mortality, it is important to highlight the results from Panchal et al. , who evaRecently, further studies suggested the impact of coffee components not only on metabolic pathways in the liver but also on endoplasmic reticulum stress and autophagy, which are pivot processes in NAFLD . Our groThe association of coffee consumption with fibrosis and cirrhosis development has been investigated in several studies . In a pr4) or thioacetamide and attributed this effect to its nonselective adenosine receptor antagonist activity [Coffee has been consumed for several centuries worldwide for its flavor and taste but also because of the psychoactive effects of caffeine, an alkaloid which is contained in different quantities according to coffee species, techniques of roasting and brewing ,23. To tactivity  Table 24 or thioactivity  showed tactivity  and to ractivity .Arauz et al. and Furtado et al. ,36 indepThe anti-fibrotic effects of caffeine have been also evaluated in in vitro cultures of human and rodent hepatic stellate cells. Shim et al. assessed the effect of caffeine on human hepatic stellate cells (HSC) proliferation and migration and found that caffeine attenuates the progression of liver fibrosis by inhibiting HSC adhesion and activation . Wang etBeside morphological evidence for the anti-fibrotic activity of caffeine coming from the mentioned studies, recently it was elegantly shown by Hsu et al. that caffeine may also favorably impact hemodynamic changes in a rat model of portal hypertension . In this4-induced cirrhosis; the authors found that CGA reduces liver fibrosis and the expression of collagen I and collagen III. Consistently, rats treated with CGA displayed reduced levels of VEGF, TGF-beta and alpha-smooth muscle actin, thus indicating that CGA is able to counteract liver fibrogenesis in rats [Although the anti-fibrotic effects of caffeine are well characterized, few experimental studies have been conducted so far to investigate the effects of polyphenols, such as CGA, in in vitro and in vivo models of liver fibrosis. Shi et al. first explored the effect of the oral administration of CGA in rats with CCl in rats . Success in rats . These f in rats .The epidemiological association between coffee intake and liver cancer prevalence or incidence has been extensively studied throughout the two last decades  Table 3Table 3. At the experimental level, different studies have been conducted so far to identify the molecular determinants of such effects. To our best knowledge, the first experiment that showed the beneficial effects of a coffee component on liver carcinogenesis was conducted by Mori H et al. , who rep4. One week after DEN injection, the groups started receiving coffee or 0.1% caffeine ad libitum for 24 weeks. The groups receiving coffee or caffeine alone not only had a reduction in collagen content but also displayed a significant reduction in the size and area of pre-neoplastic lesions and in the mean number of neoplastic lesions [Katayama et al. assessed the effects of coffee in a liver cancer–prone Long Evans Cinnamon rat and showed that coffee administration for 25 weeks delayed the occurrence of hepatitis, significantly improved survival, reduced the expression of inflammatory cytokines, and reduced the incidence of small pre-neoplastic liver foci in Long-Evans Cinnamon (LEC) rats . Similar lesions .As concerns the molecular targets involved in the chemopreventive effects of coffee, evidence points out the importance of Nrf-2. Cavin et al.  showed aSeveral epidemiological studies have associated the consumption of coffee with a decreased risk of developing chronic liver diseases. Consistently, experimental data suggest that specific coffee components, in particular caffeine and CCA, favorably impact liver steatogenesis, fibrogenesis and carcinogenesis by acting on different molecular and cellular targets . Therefo