Biomarkers encompass a wide range of different measurable indicators, representing a tangible link to physiological changes occurring within the body. Accessibility, sensitivity, and specificity are significant factors in biomarker suitability. New biomarkers continue to be discovered, and questions over appropriate selection and assessment of their usefulness remain. If traditional markers of inflammation are not sufficiently robust in their specificity, then perhaps alternative means of detection may provide more information. Epigenetic drift (epigenetic modifications as they occur as a direct function with age), and its ancillary elements, including platelets, secreted microvesicles (MVs), and microRNA (miRNA), may hold enormous predictive potential. The majority of epigenetic drift observed in blood is independent of variations in blood cell composition, addressing concerns affecting traditional blood-based biomarker efficacy. MVs are found in plasma and other biological fluids in healthy individuals. Altered MV/miRNA profiles may also be found in individuals with various diseases. Platelets are also highly reflective of physiological and lifestyle changes, making them extremely sensitive biomarkers of human health. Platelets release increased levels of MVs in response to various stimuli and under a plethora of disease states, which demonstrate a functional effect on other cell types. Biomarkers encompass a wide range of different measurable indicators, including protein, polysaccharides, and nucleic acids, and are distinct, both quantitatively and qualitatively, from common circulating biomolecules, such as albumin. They represent a tangible link to physiological changes occurring within the body and ideally enable diagnostic and/or prognostic evaluations to be determined with respect to disease. Accessibility, sensitivity, and specificity are significant factors in biomarker suitability. The initiation and progression of atherosclerosis are traditionally measured by internal intima-medial thickness (IMT) . HoweverCardiovascular disease (CVD) is an umbrella term for a class of diseases involving the cardiovascular system and consequently includes heart failure, arrhythmia, angina, hypertension, high cholesterol, and stroke amongst others. Vascular inflammation is a key contributor to CVD etiology, with inflammation and CVD risk factors closely interrelated. CVD risk factors, such as hypertension, and hypercholesterolemia are associated with higher circulating concentrations of inflammatory biomarkers . Types oDysregulation of endothelial homeostasis is characterised by reduced availability of nitric oxide and is an acknowledged marker of early atherogenesis  and a muBiomarkers of associated inflammatory responses yield an alternative/indirect means of detection. Oxidative stress, for example, is widely associated with atherosclerosis, myocardial infarction, and so forth and is known to play a critical role in the genesis and continuance of inflammation through interruption of normal cell signaling mechanisms . As suchA recently published longitudinal study conducted over a twenty-year period investigated the relationship between subclinical atherosclerosis and a host of biomarkers including cytokines/adipokines, thrombosis, and adhesion molecules. IMT measurement was used as control and conducted over four time-points during the investigative period. The test group consisted of 886 individuals with type 1 diabetes. Logistic regression models carried out by the team suggest that individual biomarkers were not predictive of/associated with subclinical atherosclerosis. However, composite scores of acute-phase reactants, cytokines/adipokines, and thrombolytic factors were nevertheless associated with higher levels of atherosclerosis at Epidemiology of Diabetes Interventions and Complications (EDIC) year 12 [While biomarkers of other diseases may be derived from surgical biopsy, in terms of detecting vasculature-derived disease, blood represents perhaps the simplest and most logical of sources from which to obtain biological markers. Analysis of blood and bodily fluids in general has emerged as a form of “liquid biopsy.” This has primarily been used in the field of cancer diagnosis but equally holds promise for diagnosis of other conditions.α), for example, is widely used as a marker of vascular inflammation [α or CD40, may fail to capture the global inflammatory stimulus that the blood conveys to the endothelium. As such, they believe that the complex composition of blood results in a tangible gap between biomarker and clinical outcome. A similar sentiment is shared in another recent publication, where the presence of interferons due to the presence of disease inhibited biomarker interpretation [One of the major difficulties in identifying suitable biomarkers is determining those which independently signal a change within the system, without becoming overwhelmed by the myriad of processes surrounding them. Sensitivity and specificity are paramount . Numerouammation , 11. Cunammation  acknowleretation . This coretation . It has retation . Neverthretation . If trad vice versa. Involvement of hereditary factors may represent a more significant agent in both vascular remodelling and disease development. Indeed, with regard to coronary artery disease (CAD), genome-wide association studies have led to the identification of a large number of single nucleotide polymorphisms (SNPs) associated with increased risk of developing the disease [Data from studies carried out by Brindle et al.  has show disease . HoweverBiomarkers discussed thus far are predominantly diagnostic in nature. However, the emerging field of epigenetics may hold enormous predictive potential. Owing to the existence of numerous conflicting interpretations of what the term meant, epigenetics was finally defined in 2008 as “a stably heritable phenotype resulting from changes in a chromosome without alterations in the DNA sequence” . DNA metFollowing methylation and/or histone N-terminal modification , the epigenome is altered. Effect on gene transcription is nuanced and may vary depending on context. Regulation of gene expression was originally believed to occur through alteration of transcriptional start sites (TSSs). Methylated regions have been linked with particular chromatin features associated with transcriptional repression, especially H3K9me3 and methyl-CpG-binding protein 2 (MECP2). Methylation in other regions of the genome is increasingly accepted as having functional relevance and presents fundamentally more dynamic patterns of methylation than those discovered in TSSs . There, In contrast to silencing, transcriptional stimulation may instead occur as a result of methylation. This has been observed largely in regions located away from TSSs, a phenomenon termed the “DNA methylation paradox” . FurtherEpigenetic drift is the term given to epigenetic modifications as they occur as a direct function with age. While age is a known risk factor for many diseases, age-related methylation has been found to occur differentially at specific sites along the genome. Hence, investigation of epigenetic drift may result in the discovery of key underlying biomarkers of numerous age-related pathologies . Researcα, PPAR-δ, TFAM, citrate synthase, and PDK4) were evaluated, and methylation was found to be lower following acute exercise. Muscle-specific transcription factors remained unchanged [Physical activity (PA) is a modifiable lifestyle choice, with enormous health and fitness benefits. Regular PA is a known requirement to maintain a healthy cardiovascular system  and has nchanged .in vivo instead of in vitro stress models) by prescribed exhaustive exercise or by the use of acute models of physical inactivity may provide much more valuable and conclusive evidence in relation to the body's response to stress. Among the first to employ global DNA methylation pattern analysis was a study investigating the effects of a six-month exercise intervention on DNA methylation in adipose tissue of healthy men. Results demonstrated genome-wide DNA methylation changes, and 17,975 individual CpG sites exhibited altered levels of DNA methylation in response to exercise [Rather than taking baseline single point blood sample measurements, stressing the cardiovascular system  . This coBlood vessels are readily capable of remodelling/altering themselves in response to hemodynamic prompts associated with variations in blood flow. Significant variations of flow pattern occur, such as that produced in response to exercise or sedentary behaviour. In some cases, these may be pathological flow types that lead to aberrant remodelling and development of disease status. Calls were recently made for deeper investigation into possible correlation between sedentary behaviour and vascular function, particularly in children . A numbeThe complex composition of blood as a challenge to biomarker efficacy, discussed previously, was examined in a recent publication by Yuan et al. , where wThere can be no doubt that markers of endothelial dysfunction and epigenetic screening may be of prognostic value in detection of disease. In the immediate future, however, microvesicles (MVs) may represent a more feasible opportunity to translate potential prognostic biomarkers to clinical practice. These secreted vesicles permit reprogramming of recipient cells and are easily obtained in bodily fluids .Different types of MVs are released by mammalian cells into the extracellular space, including blood. Critically, they carry membrane and cytosolic components  and mainα stimulation. Neither hydrogen peroxide (H2O2) nor cigarette smoke extract  treatment of the same cell type yielded a similar result. Furthermore, the relationship between expression levels of particular antigens and levels detectable on MPs did not appear to directly correlate.Due to their origin from different cell types, their differential mechanism of biogenesis, and resultant variable composition, MV effector function is believed to be similarly varied. As such, it is believed that microvesicles represent detectable units involved in intercellular exchange. Indeed, particular biomarkers have been associated with particular types of vesicles; for example, the presence of CD80 and CD86 is indicative that exosomes found in plasma may be of antigen presenting cell (APC) origin . In termExaminations on circulating endothelial- and platelet-derived MPs have been conducted. Findings suggest that levels of circulating endothelial and platelet MPs correlate to the size of myocardium at risk in patients with ST-elevation myocardial infarction. This is indicative of the fact that they reflect the severity of endothelial injury and platelet activation during myocardial ischemia .As with cytokines, discussed previously, the sheer number of MVs present in vasculature from all cell types may present an overarching masking effect that renders individual effects difficult to perceive. Of interest is research carried out by Garnacho-Montero  on the r Caenorhabditis elegans [Perhaps one of the most important components of MPs is miRNA. These are short noncoding RNA molecules, approximately 22–25 nucleotides in length , the act elegans , the fie elegans . Target  in vitro and in vivo models has revealed mir-155 to be involved in a negative feedback loop. Impeding mir-155 resulted in downregulation of inflammatory cytokines, IL-6 and TNFα. Absence of these cytokines served to impede atherosclerotic development. Significantly, mir-155 is postulated to be responsible for posttranslational regulation of the MAPK pathway by targeting MAP3K 10 [Nevertheless, a number of potential circulatory miRNA markers have been determined. Of those investigated thus far, mir-155 has been revealed as an important herald of inflammatory response. Analysis of bothMAP3K 10 .μm sized anucleate fragments of precursor cells called megakaryocytes. Their production is arguably the most elegant and distinct developmental process in eukaryotes [Circulating blood platelets are 2–4 karyotes . An obvikaryotes .Typical biomarkers of platelet health include identification of primary platelet function changes in activation, aggregation, and secretion. A multitude of techniques, most notably flow cytometry , aggregometry, and Enzyme-Linked Immunosorbent Assay (ELISA) have previously been used to identify these changes. Lack of standardisation between labs and preactivation of platelets due to methodological pitfalls are major problems when analysing platelet-derived biomarkers, warranting a pressing requirement for reliable platelet biomarkers .It has been well documented that activated platelets also release the most abundant cell-derived MVs, accounting for 70–90% of circulating peripheral MVs. These MVs harbour an array of cell components including protein, DNA, and RNA and can act as a network of communication by selectively interacting with target cells. Platelets release increased levels of MVs in response to various stimuli and under a plethora of disease states including CVD, cancer, and neurodegenerative diseases.The modern “omics” revolution enables simultaneous quantification of hundreds of molecules from a single sample and may be more reflective of platelet function changes, providing novel biomarkers for a range of diseases . PlateleFurthermore, studies by Laffont et al.  and GidlPlatelets play a key role in CVD progression where the normal platelet response is modified by enhanced proaggregatory processes and decreased antiaggregatory mechanisms thereby generating a condition of platelet hyperaggregability on an acute and chronic level. Following atherosclerotic plaque rupture in severe CVD states, platelets adhere to the disrupted plaque and aggregate to form a prothrombotic surface, subsequently encouraging thrombosis and vascular occlusion. Platelet miRNAs have been differentially implicated in the presence and progression of a range of diseases, including but not limited to various CVDs such as Acute Coronary Syndrome (ACS), Atrial Fibrillation (AF), and coronary artery disease (CAD) .Platelet miRNA profiles are differentially expressed in ACS. In a study by Ward et al. , blood wCirculating levels of the platelet enriched miR-328 have also been linked to AF in 2445 offspring in the Framingham Heart Study . This miInvestigation of metabolic function in platelets for disease diagnosis and prognosis is a new area of translational research . Platelem) is reduced in a subtype of platelets known as Collagen and Thrombin Activated (COAT) platelets. Dual activation of platelets with collagen and thrombin results in striking alterations in function and structure. COAT platelets differ from typical “activated platelets” by exhibiting a myriad of features such as phosphatidylserine exposure due to cytoskeletal reorganisation, high microparticle release, and increased levels of fibrinogen on the platelet surface. In addition, mitochondria have been noted to be involved in ATP-controlled thrombotic signalling and platelet apoptosis [The traditional role of mitochondria in the platelet was considered to supply energy in the form of ATP for primary platelet functions: adhesion, activation, and aggregation. However, novel functions for mitochondria continue to emerge. Mitochondrial membrane potential (ΔΨpoptosis .MT-MD5), cytochrome c oxidase , tRNA leucine 1 (MT-TL1), and ATP synthase (MT-ATP6 and MT-ATP8) amongst CVD patients and healthy individuals by bisulfite PCR coupled with pyrosequencing. CVD patients had significantly higher mtDNA methylation than healthy individuals in MT-CO1, MT-C02, MT-CO3, and MT-TL1, genes involved in ATP synthesis.Platelets have a higher degree of ATP turnover than resting mammalian muscle which holds high levels of mitochondria, suggesting a critical role for mitochondria in platelet function and huge requirement for energy during platelet activation. As with nuclear DNA, mitochondrial DNA (mtDNA) has the potential to be methylated by factors such as disease, ageing, and environmental exposure, moderating control of mitochondrial gene expression. In tandem with platelet miRNA biomarkers, simultaneous understanding of epigenetic regulation of mitochondrial genes in platelets is proving crucial to understanding their implication in CVD development. Recently, Baccarelli and Byun  investigDNA methylation in platelet mitochondria could be a potential contributor to CVD development through regulation of platelet function. The role of mtDNA methylation in platelet function has yet to be fully determined, but this study is the first to portray platelet mtDNA methylation as being implicated in, and a possible biomarker of, CVD.Initiation of vascular inflammation is a key event in endothelial dysfunction and CVD as a whole. While a small group of markers  have been generally accepted by the scientific community, questions remain regarding the overall biological relevance, as both reporters and effectors of pathophysiological processes.Epigenetic biomarkers may yield far-reaching prognostic results but, as a technique, is in its infancy. Nevertheless, epigenetic alterations in vasculature have potential to provide novel therapeutic targets for vascular-associated chronic illnesses and initiate novel drug-development pipelines. in vivo is required. A great deal about MVs remains unknown; even their biogenesis is yet to be completely understood. A concerted effort must be made in the direction of MV profiling. Knowledge of miRNA and their associated pathways will prove necessary in order to understand effects on cells and, importantly, unlock their diagnostic and predictive potential. The observation of proinflammatory miRNA upregulation in stimulated MPs suggests that MV miRNA currently presents perhaps the most compelling opportunity for significant progress in the field of vascular research. Platelet miRNA signature may well quantitatively reflect platelet activation in vivo and therefore could have huge potential as biomarkers of cardiovascular risk. In addition, correlating platelet physiology and function with its miRNA content may provide new insights into the changes in age and lifestyle-associated epigenetic drift. To this end, it is essential to identify and subsequently interpret miRNA functionality and hence distinguish which of these are most relevant in a clinical setting.Yet, significant challenges remain. Deeper investigation into the physiological roles of MVs


Pseudomonas aeruginosa causes severe and persistent infections in immune compromised individuals and cystic fibrosis sufferers. The infection is hard to eradicate as P. aeruginosa has developed strong resistance to most conventional antibiotics. The problem is further compounded by the ability of the pathogen to form biofilm matrix, which provides bacterial cells a protected environment withstanding various stresses including antibiotics. Quorum sensing (QS), a cell density-based intercellular communication system, which plays a key role in regulation of the bacterial virulence and biofilm formation, could be a promising target for developing new strategies against P. aeruginosa infection. The QS network of P. aeruginosa is organized in a multi-layered hierarchy consisting of at least four interconnected signaling mechanisms. Evidence is accumulating that the QS regulatory network not only responds to bacterial population changes but also could react to environmental stress cues. This plasticity should be taken into consideration during exploration and development of anti-QS therapeutics. Pseudomonas aeruginosa is a ubiquitous, gram-negative bacterium that thrives in diverse habitats and environments. Usually a commensal on the host body, P. aeruginosa is capable of transforming into an opportunistic pathogen when there is a breach of host tissue barriers or a suppressed immune system  as therapeutics, since bacteria routinely encounters adverse environmental conditions when infecting host organisms. This review will provide an overview on the QS systems in P. aeruginosa, focusing on a recently discovered integrated quorum sensing system (IQS), and on the interactions between all the four QS systems and how environmental cues could affect the QS hierarchy.Given its importance as a human pathogen, P. aeruginosa was an extension of the studies based on the prototype luxI-luxR system in Vibrio fischeri, in which luxI encodes the biosynthesis of an acylhomoserine lactone (AHL) signal N-(3-oxohexanoyl)-L-homoserine lactone (OHHL), and luxR encodes an AHL-dependent transcription factor  was characterized as N-(3-oxododecanoyl)-homoserine lactone (OdDHL)  , some with BHL specificities (e.g. rhlAB), and some equally well to both signals   , highlighting the important roles of this new QS system in modulation of bacterial pathogenesis. Importantly, under phosphate depletion stress conditions, IQS was demonstrated to be able to partially take over the functions of the central las system , and other virulence genes that are part of its regulon  was also shown to participate in the inhibition of biofilm formation, c-di-GMP signal degradation and repression of the type III secretion systems . Remarkably, PhoBR is indispensable for coordinating the las-independent, phosphate-dependent IQS signalling activation, wherein the “IQS phenotype” would be abolished in a phoB mutant , exoenzymes that could cleave iron-bound host proteins  and other redox enzymes and toxins (exotoxin A)  in Castric, . The finCastric, . AdditiohcnABC cluster, exhibiting a synergistic effect brought upon by oxygen limitation stress. Further, the PRODORIC promoter analysis programme predicted the FNR/ANR binding consensus sequences in up to 25% of the predicted QS-controlled promoters, implying that ANR might be an important co-regulator of the QS-dependent virulence genes in anaerobic environments  family of transcriptional regulators and is the main transcriptional regulator that acts in parallel with the QS systems for the expression of hydrogen cyanide biosynthesis genes  are prematurely produced and PQS synthesis inhibited , an intracellular signal that enables the bacteria cell to self-perceive their inability in synthesis of proteins . This in turn activates the rhl QS system and substantially enhances the expression of lecA and production of its encoded virulence protein, galactophilic lectin. Pyocyanin, an additional QS-regulated virulence factor, was also found to be up-regulated in the presence of IFN-γ  that is produced by endothelial cells and used for maintaining body fluid homeostasis and blood pressure control, was demonstrated to have positive effects on P. aeruginosa QS and virulence profiles. When stimulated by exogenous LL-37 at physiological concentrations, P. aeruginosa exhibits heightened production of virulence factors pyocyanin, hydrogen cyanide, elastase and rhamnolipids. The PQS signal level is also elevated. LL-37 was also found to decrease the susceptibility of the bacteria to gentamicin and ciprofloxacin antibiotics. These phenotypes were suggested to be mediated by the quinolone response protein and virulence regulator PqsE (Strempel et al., Most recently, the human host defence peptide LL-37, the only cathelicidin class of cationic antimicrobial peptides synthesized by phagocytes, epithelial cells and keratinocytes, was revealed to exert a positive effect on Pseudomonas aeruginosa is one of the most notorious opportunistic human pathogens as it employs a variety of virulence factors and mechanisms during infection (Fig. P. aeruginosa. The findings from recent research progresses suggest that these effects could largely be through modulation of the bacterial QS network, which comprises at least four QS signaling mechanisms including las, iqs, pqs and rhl. In particular, the most recently identified IQS highlights how a bacterial QS system could integrate environmental cues with bacterial quorum information. These four systems interact closely with one another giving rise to an intricately linked intercellular communication network. Such a complicated and multi-component QS network may enable P. aeruginosa to accommodate various environmental cues and biostresses (Fig. ion Fig. . The typses Fig. .Figure 4las system (Borlee et al., las in conditions that closely mimics host infection (Lee et al., lasR typical of P. aeruginosa clinical isolates (Ciofu et al., las system is insufficient, and that the prevalence of IQS system in clinical isolates should be evaluated to ensure development of potent anti-QS therapeutics. Furthermore, we should also keep in mind that there are many unknowns that require further investigations for clear understanding of how the bacterial QS network could act on various environmental cues in regulation of bacterial virulence and biofilm formation. For example, it is not clear how IQS could regulate the downstream pqs and rhl signaling systems and what is the impact of iqs system on the virulence of clinical isolates. Similarly, much remains to be done in understanding whether and if yes, how environmental cues could modulate the las, pqs and rhl systems. Recognition of how the external stressors change the way the QS network is connected may generate tremendous impact on the perspective from which therapeutic interventions could be developed, especially those environmental cues almost always encountered by P. aeruginosa during infections of the host. For instance, successful establishment of an infection and colonization of the cystic fibrosis lung chambers would require P. aeruginosa strains to sense, withstand and respond to deprivation of iron, phosphate, and attacks by lung macrophage-derived factors (Campodonico et al., Previous efforts in the design of anti-QS therapeutics were focused primarily on inhibition of the


Premature infants (PI) are neonates born <37 weeks gestational age, with different PI subgroups being identified according to gestational age and birth weight.Prematurity is associated with increased morbidity and perinatal mortality, since yields increased risk for a number of pathological features and negative outcomes related mainly to the extent of all organs and functions' immaturity. Among these conditions, neonatal and post-natal infections play a major role. PI feature increased odds of infections and infections-related morbidity throughout their first months of life.Among all preventative, anti-infective strategies, active vaccination is a key point for PI.The most common vaccine-preventable diseases in PI are whooping cough, Haemophilus influenzae type-b (Hib) meningitis, invasive pneumococcal disease, rotavirus gastroenteritis, influenza.International guidelines recommend to deliver active immunization following the chronological timing , and not following the corrected gestational age (=counting the weeks of life starting from the expected moment of birth). However, vaccinations in PI are often performed later than recommended, or even less than expected, as shown by recent studies carried out in Italy assessing significantly decreased rates of active immunization in PI.This somewhat poor adherence to the international guidelines is related to concerns about weaker immune responses in PI, and possible adverse events.Nonetheless, several studies have shown that vaccines administered to PIs have excellent safety profiles, fully comparable to term infants.Transient, benign episodes of apnea, with or without associated bradycardia, have been occasionally described in PI occurring up to 48 hours post-immunization. Though no significant morbidity nor long-term sequelae has been associated with these events, it is advisable to monitor these episodes for 48 hours after the completion of vaccination.Vaccines are immunogenic, generally safe and well tolerated in all infants including PI. Noteworthy, only vaccines specifically authorized for use in premature infants should be used in PIs. The most immature neonates  should receive their first dose of vaccine during hospitalization, in order to allow such risky groups of infants achieving sufficiently protective immunization before their discharge. This strategy also allows for adequate monitoring of cardiorespiratory function, and ultimately improves adhesion to the vaccination programs.Strategies aiming at promoting education and awareness about vaccination practices and recommendations in PI should be reinforced. The ultimate aim is to increase delivery of effective protection against vaccine-preventable diseases to these vulnerable patients since their discharge from the NICU.All listed authors have no conflict of interest related to this article.


Salmonella enterica and Pseudomonas aeruginosa) and the influences of the phage on host gene expression. Understanding these interactions can offer insights into phage–host dynamics and suggest novel strategies for the design of bacterial pathogen biological controls.Complex interactions between bacteriophages and their bacterial hosts play significant roles in shaping the structure of environmental microbial communities, not only by genetic transduction but also by modification of bacterial gene expression patterns. Survival of phages solely depends on their ability to infect their bacterial hosts, most importantly during phage entry. Successful dynamic adaptation of bacteriophages when facing selective pressures, such as host adaptation and resistance, dictates their abundance and diversification. Co-evolution of the phage tail fibers and bacterial receptors determine bacterial host ranges, mechanisms of phage entry, and other infection parameters. This review summarizes the current knowledge about the physical interactions between tailed bacteriophages and bacterial pathogens (e.g.,  Bacteriophages (or phages) are the most abundant and most diversified microorganisms on Earth. Acting as obligate bacterial predators, phage can be found in all reservoirs populated by bacterial hosts, e.g., in soil , in aquaIn natural habitats, phages and bacteria are in a constant arms race that proceeds in continuous cycles of co-evolution . As bactEscherichia coli phages (or coliphages) and bacterial CRISPR/Cas immunity systems against phage genome replication. The focal points of this review, however, are physical interactions between tailed phages and other Gram-negative bacterial pathogens, specifically Salmonella enterica and Pseudomonas aeruginosa, and the influences of phage–host interactions on the gene expression of these clinically important bacteria and, more generally, on microbial diversity.Previous reviews of phage–bacteria interaction mainly focused on model organisms such as Caudovirales order . As the name implies, the adsorption machinery dedicated for specific host recognition is localized at the tail-end, varying from a simple tail tip to a complex base plate. Such mechanisms appear to be well-correlated with host adsorption strategies , including surface proteins, polysaccharides and lipopolysaccharides  of Salmonella phages are flagella proteins  creates high selective pressure, forcing diversification of the attachment-related structures and the emergence of various phage infection tactics  used for translocation of virulence factors into target cells: the same mechanism the phage uses to transfer its genome to the host (Pseudoalteromonas luteoviolacea can trigger metamorphosis of a marine tubeworm, providing novel insights into the intricate interaction between phage, bacterium and animal (Beyond superinfection exclusion, prophages can also enhance the fitness of their hosts in the environment. A recent study in rophages . In addirophages . These arophages . This phommunity . High seommunity . Anotherthe host . Moreoved animal .Taken together, it is clear that phages play key roles in shaping the evolution of bacteria and vice versa. Phages not only manipulate expression of bacterial genes but also provide competitiveness for their hosts to thrive in the ecosystem as that would mutually be beneficial for both parties. Substantial structural and mechanistic analogies among bacterial puncturing devices, pyocins and secretion systems indicate an intimate evolutionary relationship between phages and their hosts.in vitro characterization of host–phage relationships indicates short-term arms races are run between hosts and parasites (Extensive arasites ; howeverarasites . In a laarasites .S. enterica or lung infection by P. aeruginosa.In an attempt to better understand phage–bacteria population dynamics, extensive metagenomic studies have been performed to characterize microbial communities in various niches, including marine, soil, and animal hosts. Metagenomic studies of human feces have demonstrated high abundance and diversity of phages , 2013. BS. enterica, there have been reports of the use of phages as antimicrobial testing agents in areas ranging from food-producing animals to ready-to-eat foods (P. aeruginosa phages, many reports have suggested that phages can be used to treat P. aeruginosa infected wounds and to disrupt the bacterial biofilm (recently reviewed in As naturally designed predators to balance bacterial ecology, and at a time when multi-drug resistant bacteria are becoming widespread, phages have been identified as potential antimicrobial agents for biocontrol applications in various applications: medical therapy, agribusiness, and food safety. In the case of at foods . In the Figures 2A,B; Figure 2C; In terms of host–phage interaction, structural biology and microbiology are counterparts. Although several detailed structures of bacteriophages have been determined at near atomic resolution by cryoEM and image analysis (e.g., In addition, the cryoEM 3D reconstruction of phages and virus can be vastly improved by averaging taking advantage of the icosahedral symmetry of the virus . UnfortuMoreover, an electron-optical equivalent of “phase contrast” light microscopy called “phase plate” is being developed. Various prototypical versions of phase plate have been shown to improve contrast in EM significantly . GlaeserAdvances in metagenomics analyses and sequencing technologies have not only led to a much better appreciation of the abundance of uncultivable environmental microbial populations, but they have also enabled the monitoring of community dynamics . HoweverS. enterica and P. aeruginosa, which can be used as models, for other pathogens in the development of phage therapeutic strategy. The ongoing evolutionary battles between phages and bacteria continue to shape microbial communities. As the co-evolution continues, bacteria develop resistance and exploit phage machinery to fight other bacteria in order to increase environmental fitness while phages manipulate host behaviors by alteration of bacterial gene expression. Recent advances in metagenomic analysis open new doors to the vast resources of genetic diversity of viromes from various habitats, while cryoET gives much clearer pictures of how phages and hosts interact at the molecular level. Functional characterization of phages in the laboratory, combined with cryoET and subtomogram averaging, provide snapshots of different stages in phage infection. Furthermore, recent developments in direct-electron detection technology, phase plate and single particle EM provide routine protocols for determination of 3D structures of phages at near-atomic resolution. These could potentially revolutionize our understanding of the complex interplay between phages and hosts in their natural ecosystems, which is of fundamental importance in phage use in biocontrol and therapeutic strategies.In this review, we summarize the physical interactions between tailed phages and commonly found Gram-negative bacterial pathogens, specifically 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.


Oxytocin (Sintocynon) is considered an uncommon cause of severe allergic reactions during delivery. We have recently shown that allergic sensitization to latex might constitute an important predisposing risk factor for anaphylaxis after the first infusion of oxytocin during delivery.ex vivo models have shown that oxytocin, under pro-inflammatory cytokines stimulation, such as those occurring in asthma, may induce contraction of smooth muscle and airway narrowing.Some oxytocin cardiovascular activities such as lowering blood pressure, negative cardiac inotropy and cronotropy, parasympathetic neuromodulation, vasodilatation etc. can induce significant side effects mimicking cardiac anaphylaxis, and constitute an additional differential diagnostic problem in delivering women with suspected or real allergic background. Finally, some This background suggests that allergic sensitization to latex allergens constitutes a significant but underestimated risk factor for triggering severe systemic reactions after the infusion of oxytocin and, consequently, there is a need of particular attention in managing delivering women suffering from latex allergy and bronchial asthma. An accurate anamnestic, clinical and diagnostic evaluation, latex-free anesthesiological setting, use of oxytocin-alternative agents and, if necessary, a drug premedication are likely to reduce the risk of anaphylactic/broncho-obstructive reactions in these women. Oxytocin (Sintocynon) is usually considered an uncommon cause of severe allergic reactions during delivery. Very few documented reports on anaphylactic/anaphylactoid reactions as well as severe airway obstruction have been published-6. HowevOgata and Minami demonstrWe have recently described two life-threatening anaphylactic reactions with onset a few minutes after the infusion of oxytocin in two women sensitized to latex allergens. Both reDiagnostic procedures confirmed an IgE - mediated allergic response to both latex and oxytocin, no allergic response was found to other agents used before or during caesarian section such as local antiseptics, proton-pump inhibitors, antihypertensive drugs, low molecular weight heparin (enoxaparin sodium), human albumin.In view of the few data available in literature, we believe that this topic is underestimated because such adverse events might be easily attributed to latex allergy or to usual “side effects” or alternative/unknown causes. Moreover, this risk could be underestimated if we consider that female sex shows higher prevalence of both hypersensitivity reactions in the anesthesia setting and latex allergy than men.Among the possible “side effects”, it is important to outline the role of oxytocin on heart.It has been shown that both oxytocin and its receptors are found in the heart and large vessels, where aHowever, several other cardiovascular activities such as lowering blood pressure, negative cardiac inotropy and cronotropy, parasympathetic neuromodulation, vasodilatation etc. which may be beneficial in some clinical conditions, can induce significant side effects during delivery. These sOxytocin induces uterine contractions during delivery and milk ejection during lactation through activation of a specific G protein-coupled receptor. The expInterestingly, it has been shown that the expression of oxytocin receptors plays a role not only in uterine but also in other human tissues such as kidney, ovary, heart, vascular endothelium etc. Labour and inflammation increase the expression of oxytocin receptors in human amnion, inflammex vivo model, oxytocin, under pro-inflammatory cytokines stimulation, may induce contraction of smooth muscle and airway narrowing suggesting that oxytocin serves as a bronchoconstrictor[Recently oxytocin receptors have been found also in human airway smooth muscle. Moreovestrictor. As a costrictor.Taken together, these data suggest that inflammatory conditions of airways such as those found in asthmatic women might constitute an independent (from anaphylaxis) risk factor for airway obstruction after infusion of oxytocin during delivery. The role of oxytocin receptors could also explain the well known worsening of asthma control in about one-third of pregnant women suffering from asthma-28.Finally, Gonzalez-Perez et al. have shown that women suffering from severe asthma are at higher risk of anaphylaxis than men, as a coSince oxytocin causes the alveoli in the breasts to contract causing milk let-down as the milk ejection reflex, there is some controversy over whether or not a woman can be “allergic to breastfeeding”. In fact there are women who have allergy-like symptoms associated with the milk ejection reflex during breastfeeding. These symptoms can include itching, redness, rash or hives on the trunk, arms or legs, anaphylactic reactions as they have been also shown. It has in vivo tests with oxytocin have not been standardized, a diluted/undiluted oxytocin solution should be used by skin prick test/intradermal test. In our case report, both patients reacted after the use of skin prick test and, consequently, intradermal test was not necessary[in vivo (skin prick tests) and in vitro  tests.Although ecessary. Latex hIn conclusion, our findings suggest a particular attention in managing delivering women suffering from latex allergy and bronchial asthma. An accurate anamnestic, clinical and diagnostic evaluation, latex-free anesthesiological setting, use of oxytocin-alternative agents and, if suffering from asthma, a drug premedication are likein vitro studies are necessary to establish the occurrence of an immunological cross-reaction between latex and oxytocin as well as the role of oxytocin and its receptors in heart and airway. Finally, further clinical studies should be designed to a better understanding/management of respiratory and cardiac effects of oxytocin administration.Further Oxytocin may constitute a risk factor for anaphylaxis, bronchial asthma and cardiologic side effects in delivering women.All authors declare that they have no conflict of interest and that the study has been carried out without any financial support.


Falling is a significant health issue among elderly adults. Given the multifactorial nature of falls, effective balance and fall risk assessment must take into account factors from multiple sources. Here we investigate the relationship between fall risk and a diverse set of biochemical and biomechanical variables including: skeletal muscle-specific troponin T (sTnT), maximal strength measures derived from isometric grip and leg extension tasks, and postural sway captured from a force platform during a quiet stance task. These measures were performed in eight young and eleven elderly adults, along with estimates of fall risk derived from the Tinetti Balance Assessment. We observed age-related effects in all measurements, including a trend toward increased sTnT levels, increased postural sway, reduced upper and lower extremity strength, and reduced balance scores. We observed a negative correlation between balance scores and sTnT levels, suggesting its use as a biomarker for fall risk. We observed a significant positive correlation between balance scores and strength measures, adding support to the notion that muscle strength plays a significant role in postural control. We observed a significant negative correlation between balance scores and postural sway, suggesting that fall risk is associated with more loosely controlled center of mass regulation. Falling poses a significant health risk for the elderly. As the baby boom generation moves into the over age 65 demographic, the number of age-related falls will increase, highlighting the need for accurate and effective fall risk assessment tools. Unlike other age-related health problems that are often organ- or system-specific, fall risk is influenced by a complex range of factors involving neurological, cardiovascular, musculoskeletal, somatosensory, and vestibular components -4. Risk Quiet standing is a balance-related activity that involves contributions from both the musculoskeletal and proprioceptive systems, and represents an effective model in which to study their combined influence on balance regulation and fall risk. More specifically, we selected a quiet standing task for the present study because the postural sway outputs from the proprioceptive system are likely related to upper and lower extremity muscle strength and to serum biomarkers for muscle health, skeletal muscle-specific troponin T, sTnT.2+) and subsequent actin-myosin crossbridge formation. Age-related changes in connective tissue surrounding the muscle can impair muscle integrity, resulting in leaking of troponins into the bloodstream and reduced muscle contraction efficacy. Therefore, the presence of skeletal muscle Troponin T (sTnT) in the blood is a potentially valuable biomarker for muscle health and sarcopenia, which is an important risk factor for falls , mean medial-lateral COP displacement , and sway area  .Correlation analyses by age group revealed that sTnT is negatively correlated with TCOP, DML, DAP, and ACOP , gait (TGait), and total  scores from the Tinetti Balance Assessment.While the etiology of falls is multifactorial, research on risk tends to be reductionist. To our knowledge, to date, no others studies have simultaneously looked into several potential contributors or risk factors for falls ranging from strength to balance to a newly proposed serum biomarker. To address this gap, we conducted the simultaneous measurements of sTnT, left and right hand grip strength, leg extension strength, and postural sway to evaluate their relationship to fall risk as assessed using standing balance  that is well documented in the aging literature , 22, desStand, TGait, TTotal) were significantly lower among older adults in comparison to young. The older adults in our study had an average overall balance score  of 22.4 out of 28, which corresponds to the “Moderate” fall risk category. Young adults scored, on average, 27.9 out of 28, a nearly perfect score that corresponds to the “Low” fall risk category. While this clearly indicates an age-related increase in fall risk among our participants, care must be taken when interpreting this result: nearly all young adults received perfect scores, resulting in very small variation in comparison to older adults that may have contributed to a detection bias of the U tests and correlation analyses used.All balance scores (TCOP), average medial-lateral center of pressure displacement (DML), and sway area (ACOP). The findings for VCOP, DML, and ACOP collectively represent an age-related increase in center of pressure magnitude, which has been observed elsewhere [ML, which represents the difference between medial-lateral center of mass and center of pressure movements. In essence, older adults use a postural control scheme that is characterized by larger oscillations of the center of pressure around the center of mass , only the medial-lateral ones exhibited an age-related increase. This result suggests that, given the lack of age effects in the anterior-posterior directional COP parameters, the differences in sway velocity and area are attributed mainly to age-related changes in medial-lateral components of sway. This finding carries implications for lateral stability among older adults, particularly given the emphasis in the literature on laterally-directed stability and falls -30.Interestingly, we did not observe any significant age effects on sTnT, although there was a trend towards an age-related increase. One explanation for this observation relates to possible differences in activity levels between young and older adults. While we did not measure activity levels, anecdotal evidence from speaking with participants suggests that elderly adults in our study were active. If the young adults (who were college-aged) were relatively sedentary, this may have minimized the differences in sTnT between age groups. Future studies should include information about participants' activity levels in order to control for their potential confounding effect. Furthermore, while we apply a strict statistical comparison to detect age-related changes in sTnT, one might also extrapolate from the cardiac troponin literature that a 7% increase in serum levels would in fact be considered pathological. Given that we are defining and proposing a new biomarker, and until several studies are conducted and a more detailed behavior for sTnT is determined, the possibility remains that a 7% difference could be of high physiologic significance.Gait) and sTnT, suggesting that increased serum levels of troponin are associated with a decline in balance performance (at least on the gait portion of the TBA). Further analysis revealed that this effect was only present among older adults, as they were the only subjects who scored less than perfect on the gait portion of the TBA. This result has exciting implications for using sTnT as a biomarker for fall risk in older adults. However, care should be taken in interpreting this result: the variability in sTnT contributed to overlap between age groups, meaning there were some young adults with higher levels of sTnT who still received perfect scores on the gait portion of the TBA. This clearly suggests that age-related processes apart from sarcopenia play a role in older adults' increased fall risk. Although we did not measure these in the current study, age-related declines in proprioception and vision may have also contributed to older adults' lower gait scores. The variation in sTnT also demonstrates the need for future studies to have larger sample sizes.We observed significant correlations between balance scores and measures of strength, sway, and sTnT that largely support our second hypothesis. Although not reaching statistical significance, we found a negative correlation between gait balance score  and eight young adults, all female,  participated in the current study. Elderly adults were recruited from community sites at which the researchers had previously worked, and four had a previous history of fall. Young adults were recruited from the University of Missouri-Kansas City (UMKC) student body. All participants denied having any significant musculoskeletal, neurological, or cardiovascular conditions that would impair their ability to comply with study tasks, defined as the ability to stand or walk without assistance for 30 seconds at a time. UMKC's Institutional Review Board approved all study procedures.Each participant completed a single visit to UMKC's Human Motion Laboratory and, after signing informed consent, participated in the following procedures: (1) phlebotomy; (2) upper extremity grip strength test; (3) lower extremity knee strength test; (4) balance assessment.To determine serum levels of sTnT, a phlebotomist drew blood from participants into serum separator tubes. The blood was allowed to clot for two hours at room temperature and centrifuged at 1000 x g before serum samples were collected and stored at −80°C until sTnT levels were determined using a commercially available ELISA kit  following the manufacturer's instructions.Grip,R, FGrip,L) was assessed by asking each participant to perform three maximal grip contractions with their right and left hands, with three minutes of rest between contractions, using a grip strength dynamometer . For each hand, grip strength was determined as the highest force value of the three trials.To evaluate upper body strength, grip force (FLeg) was assessed by asking each participant to perform maximal isometric leg extensions with the dominant leg while positioned on a multi-mode dynamometer . Participants performed three contractions of three seconds duration each; the maximum measured torque among the contractions was recorded as TLeg.To measure lower extremity strength, leg extension torque (TStand), gait (TGait), and total  for each participant. Force plate data was captured at a sampling rate of 1000 Hz, and a separate center of pressure (COP) profile was recorded for each component of the standing balance portion of the TBA. While all components of the TBA were included in the aggregate fall risk score, only the eyes-open, quiet stance portion of the TBA was included in the quantitative sway analysis as stationary conditions are required to extract the biomechanical parameters of interest.Balance was assessed with the Tinetti Balance Assessment (TBA), a clinical tool used to measure balance impairment based on performance in several activities including sitting, standing, turning around, and gait . With thCOP), medial-lateral (DML) and anterior-posterior (DAP) average sway displacement, and COP area (ACOP). Additionally, we generated transverse plane center of mass (COM) time histories for each trial using a zero-point-to-zero-point integration algorithm [ML) and anterior-posterior (EAP) COM and COP time histories.All force platform data was processed with MATLAB V14 . We extracted several different measures of postural sway from COP time histories, including average sway velocity  with age group as the independent variable.To test the first hypothesis that age-related changes would be observed in sway, sTnT, strength, and TBA score, Mann-Whitney p < 0.05.To test the second hypothesis that a correlation existed between TBA score and measures of strength, sTnT, and sway, Spearman correlations were conducted to explore potential relationships among the variables. Spearman correlations were also conducted to examine this relationship by age group. All statistical analyses were conducted using the statistical software SAS version 9.3, with significance set at


Diabetologia, McAuley and colleagues (DOI: 10.1007/s00125-016-3981-9) report on the impact of a 50% reduction of basal insulin delivery before, during and after moderate-intensity aerobic exercise. Results from this study may contribute to a better understanding of the effects of basal insulin delivery manipulation and may aid in devising therapeutic approaches for glucose management during exercise.Exercise poses significant challenges to glucose management in type 1 diabetes. In spite of careful planning and manipulation of subcutaneous insulin administration, increased risk of hypoglycaemia and glycaemic variability during and after exercise may occur as a result of inherent delays in insulin action and impaired counter-regulatory hormone responses. Various strategies to mitigate this issue have been advocated in clinical practice, including ingestion of supplementary carbohydrate prior to exercise, reducing background and pre-meal insulin bolus and performing bouts of resistance/high intensity exercise before aerobic exercise. Insulin pump therapy, considered the most physiological form of insulin replacement for type 1 diabetes allows modulation of basal insulin delivery before, during and after exercise. However uncertainty remains regarding the optimal strategy to reduce basal insulin delivery and its efficacy. In this issue of  Insulin pump therapy is widely considered to be the most physiological form of insulin replacement for people with type 1 diabetes. It facilitates flexible modulation of basal insulin requirements and allows the user to temporarily change basal insulin delivery before and during periods of enhanced or reduced insulin sensitivity, such as in exercise or intercurrent illness. Despite perceived benefits, the frequency of insulin pump therapy varies widely in different European countries, from below 5% to up to about 25%  and Exercise poses several challenges to individuals with type 1 diabetes. Manipulating externally administered insulin, with inherent delays in absorption and action can be difficult and requires careful planning. Impaired counter-regulatory responses and inappropriately high insulin levels increase the risk of hypoglycaemia during or after exercise. Fear of hypoglycaemia may cause overcompensation by ingesting excessive amounts of carbohydrate or reducing insulin doses to a greater degree than necessary, leading to high glucose levels during or after exercise. However, a number of alternative strategies have been developed to limit the hypoglycaemia associated with exercise in type 1 diabetes (see text box) .StrategInsulin pump therapy allows the user to temporarily reduce or suspend insulin delivery, providing greater flexibility. This may be particularly useful for individuals undertaking unplanned physical activity and exercise, as well as those who carry out routine physical activity. As a result of delays in the absorption of subcutaneously delivered insulin, however, the effect of insulin delivery rate adjustment is not immediate and, therefore, adjustment prior to exercise related activity is usually recommended. However, clinical advice and guidelines on insulin pump management during exercise varies because of the low level of evidence available.In spite of current guidelines and diabetes technology such as insulin pump therapy and continuous glucose monitoring, avoiding exercise related dysglycaemia remains a challenge for many individuals with type 1 diabetes. Factors contributing to such complexity include inter- and intra-individual variability in insulin requirements and absorption, timing, exercise type , intensity and duration of exercise, amount of active insulin on board, antecedent hypoglycaemia, counter-regulatory hormone responses and the changes in insulin sensitivity induced by exercise itself. The effects of varying insulin pump adjustments in type 1 diabetes have been previously reported , but uncThe study by McAuley and colleagues investigated the impact of a 50% reduction of basal insulin delivery 1 h before, during and after moderate-intensity aerobic exercise (30-min of stationary cycling) in the fasting state, in a randomised crossover study design  Table 1 of the article [The second finding relates to the primary endpoint of the study, namely, circulating free insulin levels. These are increased, albeit transiently, during moderate-intensity exercise, in spite of halving basal insulin delivery 1 h pre-exercise. A plausible explanation is provided by the authors for this finding: increased cutaneous blood flow. Augmented absorption of subcutaneous human and analogue insulin during exercise and at varying ambient temperatures have been reported . Most st article ), with sHow does this study influence our understanding of exercise and glucose management in insulin pump users, and what are its implications for clinical practice? The rise in circulating free insulin observed in this study needs further evaluation to increase our understanding of the potential mechanisms associated with this outcome and to determine the clinical implications. It is also important to establish whether the same pharmacokinetic findings occur with different exercise modalities  and in various populations (e.g. different age groups and ranging participant characteristics) to determine the generalisability of these study outcomes. The authors hypothesised that, despite increased circulating free insulin levels, the modest rise in glucose with exercise may have been due to the effects of counter-regulatory hormones and that sustainment of glucose levels during recovery may have been due to the decline in insulin levels post-exercise. Combining the glucose level results with data on counter-regulatory hormone and glucose turnover (neither of which were assessed in this study) may help to elucidate the hormonal and metabolic interactions between the three and indicate clinical implications, especially in physically active insulin pump users with impaired counter-regulation. A recent study reported that during intermittent high-intensity exercise in type 1 diabetes without insulin delivery adaption, reduced exogenous glucose requirements may occur secondary to decreased glucose uptake and disposal, rather than increased hepatic glucose output or muscle glycogen utilisation . This poFuture exercise studies in type 1 diabetes following insulin delivery adjustments should include evaluation of blood glucose levels over longer periods post-exercise since delayed hypoglycaemia, particularly overnight, remains a clinical concern. Such studies should recruit participants with impaired hypoglycaemia awareness and antecedent hypoglycaemia events, who have an increased risk of hypoglycaemia. In addition to pre-exercise insulin delivery interventions, studies comparing or combining this strategy with novel hypoglycaemia avoidance strategies, such as threshold/predictive suspended insulin pump management , intermi


Maillard reaction produces flavour and aroma during cooking process; and it is used almost everywhere from the baking industry to our day to day life to make food tasty. It is often called nonenzymatic browning reaction since it takes place in the absence of enzyme. When foods are being processed or cooked at high temperature, chemical reaction between amino acids and reducing sugars leads to the formation of Maillard reaction products (MRPs). Depending on the way the food is being processed, both beneficial and toxic MRPs can be produced. Therefore, there is a need to understand the different types of MRPs and their positive or negative health effects. In this review we have summarized how food processing effects MRP formation in some of the very common foods. The Maillard reaction has been named after the French physicist and chemist Louis Camille Maillard (1878–1936) who initially described it. It is often defined as nonenzymatic browning reaction. While foods are processed or cooked at high temperature, a chemical reaction occurs between amino acids and reducing sugars which generate different flavours and brown colour . So it iBased on literature, Hodge 1953) first described the steps involved in Maillard reaction products (MRPs), also known as advanced glycation end-products (AGEs), formation. The whole process of MRPs formation can be divided into three major stages depending on colour formation. At the first stage, sugars and amino acid condense, and following condensation, Amadori rearrangement and 1-amino-1deoxy-2 ketose form. In the second stage, dehydration and fragmentation occur in the sugar molecules. Amino acids are also degraded in this stage. Hydroxymethylfurfural (HMF) fission products such as pyruvaldehyde and diacetyl are formed in this intermediate stage. This stage can be slight yellow or colourless. In the final stage, aldol condensation occurs and finally the heterocyclic nitrogenous compounds form, melanoidins, which is highly coloured 953 first–6.There is an ever-increasing preference for instant meal rather than traditional cooking, especially among the new generation of people. It has been reported that people consuming high amount of processed meat, pizza, or snacks develop insulin resistance and metabolic syndrome compared to people having high intake of vegetables and low processed food . MRPs thSoybean is widely used as flours, grits, flakes, isolates, concentrates, and textured soya proteins and also as cooking oil. Soybeans play important role in cardiovascular diseases, osteoporosis, and cancer . So procMilk is a beverage that is consumed throughout the world. At present, a large percentage of the milk consumed by people, especially in the western countries, is processed rather than raw milk. Ultra high temperature (UHT) treatment or conventional sterilization process is often used to process milk for improving quality and safety. Milk is rich in protein and sugar. So it is obvious that processing of milk at high temperature may lead to the formation of MRPs. Several methods have been approached to determine the extent of MRPs during milk processing. Both initial and advanced staged MRPs have been used as indicators of browning reaction which occurred in milk , 12. MRPIt has been reported that conventional sterilized bottle milk has different chemical composition compared to the UHT treated milk . HMF levPasta is considered one of the healthy foods suited to balanced diet; and its consumption is becoming increasingly high throughout the world. Pasta has different varieties and shapes. Till 2009, 310 different types of both dried and fresh pasta have been documented . AlthougMRPs like heterocyclic amine (HCA) level increases with elevated cooking temperature; and this phenomenon is more pronounced in meat than fish . Meat isIn processed food, more than 25 types of heterocyclic amines (HCA) have been identified . A studyf] quinoline was higher in well-cooked steak and hamburger patties. Like duck and chicken roast, roasting beef did not contain any of the 5 HCAs, but the gravy made from the drippings from well-done roasts had two types of HCAs [In another study, beef steak and hamburger patties were processed by pan-frying, oven-broiled, and grilled or barbecued to four levels of doneness  . Beef ro of HCAs . From thIn recent days, people consume more ready-to-eat food due to lack of time. Puangsombat et al. (2011) assessed HCAs level in some ready-to-eat products. They found that HCAs were higher in rotisserie chicken skin. In the other assessed food, HCA level was found in the order as follows: rotisserie chicken meat, deli meat products, and pepperoni . However Streptococcus mutans which is considered to be the major causative agent of dental caries in humans [As a beverage, coffee is an important item in the lives of billions of people and is also one of the most traded food products in the world. The brewed coffee has emerged as the second most consumed drink after water , 46. Then humans .Consumption of diets rich in fruits and vegetables renders many health benefits to us. However, processing method plays an important role in dictating the magnitude of the beneficial health effects obtained from fruits and vegetables. Depending on treatment temperature, furoylmethyl derivatives (FM) have been found in processed vegetables and fruits like orange juices  and procWhen vegetables are treated at low temperature, prooxidants are generated, whereas treating at high temperature decreases the prooxidants and increases antioxidant properties due to the production of MRPs . Such anMRPs can prevent the enzymatic browning reaction caused by polyphenol oxidase (PPO) . Plant d in vitro [Angiotensin-I converting enzyme (ACE) is the regulatory enzyme for upregulation of blood pressure. ACE inhibitory peptide lowers blood pressure by inhibiting ACE enzyme . Rufián-in vitro . Recentlin vitro . Panax species plant comprising ginsenoside Re or ginsenoside-derived saccharide treated with amino acid at temperatures between 100 and 130°C can either prevent, improve, or treat a renal disease [It has been claimed that administration of a Maillard browning reaction product obtained from an extract of disease . Pseudomonas aeruginosa isolates, one multidrug-resistant Pseudomonas aeruginosa (MDRP), one Escherichia coli, one methicillin-susceptible Staphylococcus aureus, and one methicillin-resistant Staphylococcus aureus (MRSA) compared to mikacin, ciprofloxacin, imipenem, and levofloxacin [Food derived versatile MRPs can act as bactericidal for a wide number of pathogens. For example, aminoreductone can act as a more effective bactericidal for fourfloxacin . MRPs hafloxacin .2 during extrusion process helps to reduce the level of acrylamide [Maillard reaction products have both positive and negative impacts on health. Diverse MRPs act as antioxidants, bactericidal, antiallergenic, antibrowning, prooxidants, and carcinogens. Most of these properties depend on processing of food. High temperature heating makes some food nutritious, whereas some of the foods lose their nutritional value. Many strategies are employed in the food industries to reduce the production of MRPs. For example, acrylamide has been classified as a probable carcinogen to humans by the International Agency for Research on Cancer . During rylamide .This review was aimed at summarizing our current knowledge regarding the changes in food mediated by Maillard reaction during the food processing steps. This may provide useful insights for those related to food processing facilities.


Some alternative control strategies of currently emerging plant diseases are based on the use of resistance inducers. This review highlights the recent advances made in the characterization of natural compounds that induce resistance by a priming mechanism. These include vitamins, chitosans, oligogalacturonides, volatile organic compounds, azelaic and pipecolic acid, among others. Overall, other than providing novel disease control strategies that meet environmental regulations, natural priming agents are valuable tools to help unravel the complex mechanisms underlying the induced resistance (IR) phenomenon. The data presented in this review reflect the novel contributions made from studying these natural plant inducers, with special emphasis placed on hexanoic acid (Hx), proposed herein as a model tool for this research field. Hx is a potent natural priming agent of proven efficiency in a wide range of host plants and pathogens. It can early activate broad-spectrum defenses by inducing callose deposition and the salicylic acid (SA) and jasmonic acid (JA) pathways. Later it can prime pathogen-specific responses according to the pathogen’s lifestyle. Interestingly, Hx primes redox-related genes to produce an anti-oxidant protective effect, which might be critical for limiting the infection of necrotrophs. Our Hx-IR findings also strongly suggest that it is an attractive tool for the molecular characterization of the plant alarmed state, with the added advantage of it being a natural compound. Plants are subjected to a variety of external factors that adversely affect their growth and development, and are often divided into biotic  and abiotic  stresses. Adaptation to these environmental stresses is essential for survival and propagation . Among tFigure 1). The “primed” state has been related to increased, more efficient activation of the defense response and enhanced resistance to challenging stress , which triggers SAR , and benzo thiadiazole-7-carbothioic acid gers SAR . A wide gers SAR . Non-progers SAR . This isgers SAR .Many natural compounds have been claimed to be plant growth promoters, plant activators or plant defense inducers, among other names. A large portion of them has already been related to priming, including oligosaccharides, glycosides, amides, vitamins, carboxylic acids, and aromatic compounds. In general, natural compounds tend to be better tolerated by plants than most of the synthetic compounds tested, but there is still concern about toxicity . Most miArabidopsis thaliana consists in redox active compounds. Among them, thiamine  gene and peroxidase (cprx1) genes implicated in the phenolic metabolism has also been observed in sugar beet (both) and rice (only PAL) after riboflavin application and challenging with Rhizoctonia solani. Phenolics play a role in cell wall fortification, and also show antimicrobial and antioxidant activity  is a cyclic amino acid belonging to the vitamin B group. Field experiments have proven that it is capable of enhancing resistance against Cucumber mosaic virus and nt yield . This coBrassica napus) from the fungus Leptosphaeria maculans by stimulating ROS production, but without inducing PR1. Other authors have shown that MSB has a systemic effect  is a vitamin K3 derivative known to be a growth regulator . Borges c effect  and reponduction . Borges nduction , 2004 alon day 3 . In theion day 3 . It is ron day 3 .Chitosan is a polymeric deacetylated derivative of chitin that is naturally present in some fungi cell walls, and has various deacetylation degrees and molecular weights. Although it performs several antimicrobial activities, its main contribution to reduce plant disease is to enhance plant defenses . ChitosaThere is evidence to support that the wound signal from the local attack site is transmitted to systemic undamaged regions, where priming or the direct induction of defense responses takes place. Signal transmission can occur either internally, probably through the phloem and xylem, or externally via VOCs . In the D-galacturonic acid with a degree of polymerization between 10 and 25, which can be methyl-esterified or acetylated depending on the source plant. They are considered endogenous elicitors, and the degree of methylation and acetylation has been found to affect the activation of defense responses -mediated signaling. A microarray analysis has shown that about 50% of the genes regulated by OGs display a similar change of expression during Botrytis cinerea infection  are plant cell wall pectin-derived oligosaccharides which consist in linear chains of α-(1-4)-linked esponses . OG treaesponses . Interesesponses . Exogenot manner . In Arabnfection .Azelaic acid (AA) has been suggested to be a phloem-mobile signal that primes SA-induced defenses . The AA Arabidopsis plants for more rapid SA biosynthesis, phytoalexin camalexin accumulation and defense gene expression  a Lys-derived non-protein amino acid has been recently implicated as pivotal regulator of SAR, and possibly as the long-distance phloem-mobile SAR signal compound . Amino apression . It alsopression .Another carboxylic acid with demonstrated inducer activity is hexanoic acid Hx; . We focuVolatile organic compounds play key roles in plant–plant communication as they act as airborne signals by enhancing disease resistance in the plant itself and in neighboring plants, and by also attracting parasitic or predatory insects, these being the enemies of attacking herbivores . This muCis-Jasmone (CJ) is a highly volatile compound product of further catabolism of JA, which is known to induce the release of defense VOCs that attract predatory/parasitic insects . A transBotrytis cinerea . However, the plants from Castlemart are protected by Hx in the absence of callose priming  was also impaired in Hx-IR in three different backgrounds , transcriptional factors, and signaling and hormone-related genes. Oxylipins-, ethylene-, and auxin-related genes were induced, which corroborate the involvement of these pathways in the early response to Botrytis. Remarkably a set of redox-related genes was also induced, which evidences the involvement of oxidative stress mechanisms in this plant–pathogen interaction.We recently analyzed the gene expression profile of analysis . The resBotrytis-induced genes  and Dox1 aplha-dioxygenase, were induced and primed by Hx  in responses to different abiotic and biotic stresses, including Botrytis cinerea infection. Among the Hx-induced WRKYs, there are orthologs of Arabidopsis WRKY18, WRKY33, WRKY40, WRKY53 and WRKY75, which form part of the Arabidopsis defense response, especially against Botrytis cinerea . Other genes, like GST and glutaredoxin, are induced only in treated plants and represent specific targets of the inducer treatment. The early boosting of detoxifying and redox-balance-related genes achieved by Hx supports the direct effect of this inducer on the control of these processes. The analysis of oxidative stress markers confirmed that Hx treatment protects plants by providing a less oxidized cellular environment after infections (2-) and peroxide hydrogen (H2O2) accumulation reduced and was more restricted around the infection site. The ascorbate and glutathione reduced/oxidized ratios rose in treated plants at 72 hpi, while the activities of glutathione reductase and catalase remained closer to those of healthy plants. No changes were detected in Hx-treated, but not-infected, plants. Therefore, Hx primes the transcription of the genes controlling the redox metabolism, which is fully activated and shown only after pathogen recognition . Thiamine can modulate the cellular redox status by activating the NADPH oxidase and promoting early ROS generation, which confers resistance against Sclerotinia sclerotiorum in Arabidopsis  protects plants from salinity and non-ionic osmotic stress by altering the redox machinery in a similar way than Hx . TreatmeIn conclusion, microarray data of Hx-treated plants have revealed the induction of many genes that help characterize the Hx priming effect, especially those related with defense, the signaling network and oxidative stress control, which are over-induced in Hx-treated plants upon fungal infection. The activation and priming of different defense genes responding to the SA and JA pathways match the broad-spectrum action of this natural inducer. This agrees with the present conception that the effectiveness of the plant response against biotrophic and necrotrophic pathogens is much more complex than the classical dichotomy between the SA and JA pathways antagonism. Finally, Hx priming of redox-related genes produces an anti-oxidant protective effect, which might be critical for limiting necrotroph infection. These findings back the importance of controlling oxidative stress to improve plant protection against different pathogens, and suggest that this natural inducer is an attractive tool to further study this topic.Botrytis cinerea growth at higher concentrations than those which allow the priming of plant defenses  on tomato plants prior to fungal inoculation reduces the diameter of necrosis by ∼60%. Application on previously infected plants further reduces necrotic expansion by around 30% .The study into natural plant inducers has helped unravel the complex mechanisms underlying the IR phenomenon . It can early activate broad-spectrum defenses by inducing callose deposition, in addition to the SA and JA pathways. Later it can prime pathogen-specific responses in each particular case according to the pathogen and its lifestyle. Interestingly, Hx primes redox-related genes and has an anti-oxidant protective effect, which might be critical for limiting the infection of necrotrophs.We propose Hx, a potent natural priming agent in a wide range of host plants and pathogens, as a model tool in this field of research (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.


Summary: As the volume of patient-specific genome sequences increases the focus of biomedical research is switching from the detection of disease-mutations to their interpretation. To this end a number of techniques have been developed that use mutation data collected within a population to predict whether individual genes are likely to be disease-causing or not. As both sequence data and associated analysis tools proliferate, it becomes increasingly difficult for the community to make sense of these data and their implications. Moreover, no single analysis tool is likely to capture all relevant genomic features that contribute to the gene’s pathogenicity. Here, we introduce Web-based Gene Pathogenicity Analysis (WGPA), a web-based tool to analyze genes impacted by mutations and rank them through the integration of existing prioritization tools, which assess different aspects of gene pathogenicity using population-level sequence data. Additionally, to explore the polygenic contribution of mutations to disease, WGPA implements gene set enrichment analysis to prioritize disease-causing genes and gene interaction networks, therefore providing a comprehensive annotation of personal genomes data in disease.Availability and implementation: wgpa.systems-genetics.netContact:enrico.petretto@duke-nus.edu.sgSupplementary information:Supplementary data are available at Bioinformatics online. In order to integrate these scores over sets of genes, we provide a gene set enrichment analysis (GSEA) implementation ; GRP, gene set (*.grp); GMX, gene matrix (*.gmx); GMT, gene matrix transposed (*.gmt); WGCNA, weighted gene co-expression network analysis output (*.wgcna); Variant-Level: manual data entry; list of protein substitutions (*.txt); list dbSNP identifiers (*.txt); Network-Level: manual data entry; list of gene identifiers for STRING (*.txt); list of gene pairs (*.txt)Analyses – RVIS, EvoTol (can be stratified by gene expression), GCS (user-selected); RVIS, EvoTol, GCS combined with variant-level consequence predictions (PolyPhen2 (Gene-Level inputs)Outputs – genes ranked by their genic intolerance or constraint scores ; GSEA results for gene sets ; gene pathogenicity annotation using both the predicted ‘functionally damaging’ mutations and genic intolerance (or constraint) scores  ; gene interaction network annotated according to RVIS, EvoTol or GCS allowing zooming out of a particular gene and visualizing its connections to other genes .In order to facilitate the annotation of personal genomes data with respect to disease pathogenesis, we have developed a unified web-based tool for pathogenicity analysis of individual genes, gene sets and gene interacting networks. To this aim, we developed an intuitive graphical user interface that will make the available prioritization methods  and integrated analysis tools  easy to access and use by the general scientific community. The type of input data, integrative analyses components and outputs are schematically summarized in de novo mutations from trio sequencing projects. For instance in the Epi4K project de novo mutations impacting 176 different genes. By cross matching the RVIS, GCS and EvoTol scores and focusing on the genes from the top 25 percentile, we identify a set of 17 genes of interest . Using WGPA it was also possible to perform a GSEA of each of the measures of intolerance using the Epi4K mutated genes as the gene set of interest, and show that in each case the Epi4K mutated gene set is significantly enriched for predicted pathogenic genes .An example of where WGPA will be useful is to prioritize the set of genes with The Duke-NUS Graduate Medical School Signatures Research Program (Program in Cardiovascular and Metabolic Disorders).Supported by Conflict of Interest: none declared.


Adult hippocampal neurogenesis is a process involving the continuous generation of newborn neurons in the hippocampus of adult animals. Mounting evidence has suggested that hippocampal neurogenesis contributes to some forms of hippocampus-dependent learning and memory; however, the detailed mechanism concerning how this small number of newborn neurons could affect learning and memory remains unclear. In this review, we discuss the relationship between adult-born neurons and learning and memory, with a highlight on recently discovered potential roles of neurogenesis in pattern separation and forgetting. The pioneering work demonstrating the continuous generation of adult-born neurons in the hippocampus by Altman in the 1960s  has led Increased adult neurogenesis in the dentate gyrus (DG) of the hippocampus has been shown to improve in memory acquisition, memory formation –14, and Many neurodegenerative and neurodevelopmental disorders involving cognitive impairments are possibly linked to hippocampal dysfunction, which is, at least, partly attributed to dysregulated adult neurogenesis . TherefoHere we discuss the relationship between adult neurogenesis and hippocampus-dependent learning and memory. We also discuss the recent progress showing how increased neurogenesis improves learning and memory through its essential role in forming pattern separation among similar events, as well as its newly identified role in forgetting past memories.The hippocampus, named for its structural resemblance to a seahorse, is a crucial component of the limbic system and is suggested to be indispensable for various functions, particularly memory acquisition and consolidation and spatial navigation , 29. TheThe hippocampus plays a crucial role in converting short-term memories to long-term memories , 33 by pAdult neurogenesis in adult mammals occurs in the subgranular zone (SGZ) of the DG , 43, 44.Using retroviral labeling in combination with rabies virus, Vivar et al. indicated that newborn neurons received synaptic input from mature granule neurons with primary innervation from the lateral entorhinal cortex which is responsible for the processing of cued and contextual information , 48. NewIn addition to synaptic inputs from the lateral and caudomedial parts of the EC, and the perirhinal cortex, newborn neurons also receive back-projections from CA3 pyramidal neurons . Such syAlthough the estimated daily quantity of integrated adult-born neurons is only ~9000 of the total granule cell population in young rats  and ~700As previously discussed, the hippocampus has been considered actively involved in memory formation  as well Levels of neurogenesis in the DG can be modulated by various factors, among which the most frequently employed strategies include environment enrichment  and phys in utero lipopolysaccharide induction, interruption of rearing, exposure to lead, or insufficiency of vitamin A have all been documented to reduce hippocampal neurogenesis. Such decreases in neurogenesis were coincident with deficits in novel objection recognition, reference memory performance, and formation of fear memories [In contrast to the positive regulators mentioned above, a variety of negative regulators have been found to impair learning and memory performances. As an example, surgical lesion of the cholinergic septohippocampal pathway reduced hippocampal neurogenesis and significantly compromised spatial learning in a reference memory paradigm . Similarmemories –87. Genememories , could smemories . Hyperacmemories , 90. It memories . Exposinmemories , all of memories . Moreovememories . Spatialmemories , 95, whimemories , 97.In order to tender more compelling evidence showing the important role of neurogenesis in learning and memory, three different strategies with the same aim of specifically eliminating neural progenitor cells have been employed, including application of antimitotic agents, X-ray irradiation, and genetic manipulations. Following the ablation of neurogenesis, behavioral assays such as the MWM, eight-arm Radial Maze (RAM), and Barnes Maze tasks, as well as the working memory test using the delayed matching to sample (DMS) or delayed nonmatching to sample (DNMS) protocol have been conducted to measure subsequent learning and memory performance. Given that the intervals required for a newborn neuron to fully integrate into existing neural circuits are distinct in mice , 59, 98 Shors et al. provided the first evidence showing that inhibiting neurogenesis by the antimitotic agent methylazoxymethanol (MAM) leads to the impairment of trace, but not delay eyeblink conditioning or contextual fear conditioning , 99. ThrMore recently, X-ray irradiation has been introduced  and has T2 transgenic mice, whose neuronal progenitors in the neurogenic regions are eliminated by diphtheria toxin after tamoxifen administration, showed deficits in reference memory retention in the Barnes Maze test and impaired contextual fear conditioning [In the past decade, genetic techniques have been adopted to generate transgenic mice with more restricted targeting to neural progenitor cells. Garcia and colleagues established an inducible glial fibrillary acidic protein- (GFAP-) thymidine kinase (TK) strain where administration of ganciclovir could eliminate neurogenesis . Experimitioning . Deng etitioning . This reitioning .Notably, conclusions obtained by different groups sometimes differ. For example, compromised contextual fear conditioning following suppression of adult hippocampal neurogenesis is reported by some , 115, buAs a gateway for information's entry to the hippocampus where memories are retained in associative networks , the DG With the aforementioned features of the DG, the pattern-separation function of adult-born hippocampal neurons has emerged as the neurobiological basis mediating the influence of adult hippocampal neurogenesis on learning and memory , 127. NaComputational models have proposed that pattern separation processing within the hippocampus involves the DG-CA3 circuit . ThroughA recent human study has reported that following six weeks of physical exercise training, individuals performed better in a hippocampus-dependent visual pattern-separation task, and a lower depression score was recorded compared with those who did not exercise . On the Mounting evidence over the past decade has shown that alterations in adult neurogenesis are a form of plasticity that improves hippocampus-dependent learning and memory formation. Of note, a recent study has unveiled a new role for adult neurogenesis in promoting forgetting of old memories . In memoFeng et al. 2001 have shown that environmental enrichment prior to learning increases hippocampal neurogenesis and improves performance in both contextual and cued fear-conditioning tests in forebrain-specific presenilin-1 knockout (PS1-KO) mice with impaired neurogenesis . However2+ channels and NMDA receptors, suggesting that excitation of the local neural network may regulate the neurogenic process [Evidence supporting both theories of neurogenesis contributing to the clearance of old memories or the formation of new memories has been reported. For example, in a previous study exploring how neurogenesis modulates hippocampal network activity to enable memory storage at different levels, Deisseroth and colleagues demonstrated an activity-sensing property of hippocampal neural progenitor cells via Cav1.2/1.3 (L-type) Ca process . In fact process . Their s process . Togethe process . Based o process . Future Akers and colleagues tested if the performance in memory retention could correlate the level of hippocampal neurogenesis , by compBased on the hypothesized role of adult neurogenesis in memory clearance, it is reasonable to speculate that suppression of this process, though impairing the formation of new memories, may facilitate the preservation or reconsolidation of previously formed memories. Kitamura and colleagues have demonstrated that inhibition of neurogenesis by X-ray irradiation prolonged the maintenance of LTP in DG, as well as preservation of old memories (up to 30 days after learning) in the contextual fear-conditioning task; this suggests that the hippocampus-dependent memory retention could be extended by inhibiting neurogenesis . Althoug vice versa [Computational models predict that encoding new information will not only remodel neural networks, but also weaken neural connections that have already been established for storing old memories . Increasce versa .Based on the evidence from both animal and human studies, investigations on adult neurogenesis have been answering the critical questions concerning how learning and memory are formed and regulated in adult mammalian brains. Pattern separation and forgetting induced by adult neurogenesis may be the way in which the brain normally learns and retrieves memory. A computational model used by Weisz and Argibay  demonstr


A fundamental initiative for evolutionary biologists is to understand the molecular basis underlying phenotypic diversity. A long-standing hypothesis states that species-specific traits may be explained by differences in gene regulation rather than differences at the protein level. Over the past few years, evolutionary studies have shifted from mere sequence comparisons to integrative analyses in which gene regulation is key to understanding species evolution. DNA methylation is an important epigenetic modification involved in the regulation of numerous biological processes. Nevertheless, the evolution of the human methylome and the processes driving such changes are poorly understood. Here, we review the close interplay between Cytosine-phosphate-Guanine (CpG) methylation and the underlying genome sequence, as well as its evolutionary impact. We also summarize the latest advances in the field, revisiting the main literature on human and nonhuman primates. We hope to encourage the scientific community to address the many challenges posed by the field of comparative epigenomics. Methylation of nucleotide bases is the only covalent modification of DNA commonly found across many different taxa. So far, three types have been described: N6-methyladenine (6mA), N4-methylcytosine (4mC), and 5-methylcytosine (5mC). While 6mA and 4mC are restricted to prokaryotes and certain eukaryotes –3, 5mC iThe distribution of CpG methylation is uneven across mammalian genomes. While the majority of CpGs ~60%–80%) are methylated , regions0%–80% arNon-CpG methylation, the methylation of cytosines followed by a base other than guanine, has also been reported. Indeed, non-CpG methylation is abundant in plants ,18 and rDespite being a key regulator of genomic function, the role of DNA methylation in species evolution is only beginning to be explored. More than four decades ago, it was first proposed that regulatory changes could lead to species-specific adaptations as well as phenotypic variability ; howeverThere is a close interplay between DNA methylation and the underlying nucleotide sequence. The mutagenic nature of 5mC is one of the primary players in this crosstalk, as methylcytosines are prone to spontaneous deamination to thymine, resulting in a CpG-to-Thymine-phosphate-Guanine (TpG) mutation rate that is ~10-fold higher than for other dinucleotides . This isIn contrast to the mutagenic effect of 5mC, many genomic regions with high densities of CpGs are constitutively unmethylated. Recent studies suggest that this CpG richness can be solely accounted for by low CpG deamination rates, rather than being a byproduct of purifying selection . About 8On the other hand, several lines of evidence indicate that the genome harbors the information necessary to establish local DNA methylation patterns. How this genomic information is encoded and read is still not fully understood, but recent work is beginning to shed light on the regulatory mechanisms. Fragment-insertion experiments have demonstrated that both a high Guanine-phosphate-Cytosine (GC) content and a high density of CpG dinucleotides are necessary and sufficient requirements to induce an unmethylated state ,32. FromHAR1A gene, which plays a crucial role in cortical development [Therefore, evolutionary changes in CpG dinucleotides and GC content could contribute to alter the methylation landscape of a species . With thelopment .Different mechanisms govern methylation patterning at CpG-poor regions. Some compelling studies have revealed that methylation patterns at such regions are likely driven by the binding of transcription factors (TFs), as the presence of specific motifs suffices to create unmethylated domains ,32,38,39Evolutionary studies in human populations have identified genetic variants that associate with methylation levels at nearby CpGs, termed methylation quantitative trait loci (mQTL) –45. ConsRB1 imprinted gene, which results from the differential methylation of a processed pseudogene [Finally, it has also been suggested that retrotransposition events coupled with gradual nucleotide changes could lead to the accumulation of novel CpG sites . These reudogene .The emerging picture is that the joint actions of all the above-discussed mechanisms throughout the evolutionary scale have modeled the architecture of the methylome .Comparative epigenomics, the interspecies comparison of DNA and histone modifications, is a promising research field that can enlighten our understanding of mammalian epigenome evolution . FurtherImportantly, co-localization of different epigenetic marks, including DNA methylation, several histone modifications, and H2A.Z, has been used to study the evolution of the epigenome, the transcriptome, and the genome in humans, mice, and pigs . This stThough methylomes are overall quite well-conserved between primate species ,57, seveIn sum, from primate epigenetic comparative studies, we have started to gain insight into the molecular mechanisms through which evolutionary novelties may have arisen in the human lineage. These studies suggest that epigenetic modifications in regulatory elements could have altered expression patterns of key genes, leading to evolutionary adaptations.HOXD cluster, a key regulator of limb development [HOXD9 promoter and HOXD10 gene body are hypomethylated in humans, both archaic species were hypermethylated at this locus, while the gene body of HOXD9 was also hypermethylated in the Denisovan genome. As a result, the authors postulated that this differential methylation in the HOXD cluster could account for some of the anatomical differences between archaic and present-day humans [Genome sequencing of extinct hominids has provided a better understanding of the population history and genome evolution of our species ,72. Howeelopment . Whereasy humans . HoweverFew studies have addressed DNA methylation patterns across human populations ,75. FrasThere is now accumulating evidence indicating the influence of different environmental factors on methylation patterns . AlthougDNA methylation patterns can be influenced by stochastic events or environmental factors creating a source of epigenetic variability ,78. WhetThe notion of transgenerational epigenetic inheritance is very appealing, particularly when suggested that it could respond to environmental challenges . In suchOur understanding of DNA methylation and evolution has substantially grown over the past years. Nonetheless, the field of evolutionary epigenomics is still in its infancy, and several steps are required to complete an accurate and detailed picture of the human epigenome.Despite remarkable achievements accomplished through the parallel survey of genetic, epigenetic, and transcriptional information ,94,95, aFuture studies also need to determine which genomic regions are more susceptible to experiencing epigenetic changes induced by environmental factors, since such regions could represent confounding factors when studying interspecies epigenetic differences. Studies of methylation patterns in populations with similar genetic backgrounds but different environments would be a good starting point. On the other hand, non-CpG methylation as well as 5mC derivatives have only been recently reported, and they have not been studied in an evolutionary context.Finally, the major challenge for the next years is to move beyond mere comparative descriptions and offer insight into phenotypic consequences. To that end, experimental assays are required. In this regard, humanized mice have proved useful resources ,98. Howe