Malassezia has received a fair amount of attention. Since the middle of the 19th century, researchers have linked these fungi with skin maladies such as dandruff and eczema Malassezia is the sole genus within the fungal order Malasseziales, contained within the proposed subphylum Malasseziomycetes . Although Malassezia is sister to the so-called “smut” plant pathogens, they are markedly divergent in ecological terms. A hallmark of Malassezia species is their incomplete fatty acids synthesis metabolic pathway, and reliance, instead, on a suite of extracellular lipases, phospholipases, and acid sphingomyelinases M. pachydermatis, is able to survive in axenic culture lacking lipid amendment As the dominant component of the mycobiota on human skin Malassezia evolved into a specialized and narrow niche associated with the skin of mammalian hosts. However, culture-independent studies of fungi from environmental samples show that Malassezia are exceedingly widespread and ecologically diverse Malassezia-like organisms, providing exciting opportunities to explore the ecology, evolution and diversity of this enigmatic group.Until recently, it was assumed that Malassezia are readily detected in environmental DNA samples using standard fungal “barcoding” approaches. Scanning GenBank and the scientific literature, therefore, is useful for approximating occurrence patterns. DNA sequences identical to M. globosa and M. restricta, which are both well characterized as human skin associates, appear to be cosmopolitan. M. restricta may be particularly widespread, and DNA sequences similar to these species have been detected in habitats as diverse as deep-sea sediments Anguilla japonica) gut and muscle tissue Malassezia in these studies are nearly identical to those of human associates, suggesting either a very recent divergence in habitat or else that these organisms are highly tolerant to some of the planet's most extreme environments. Unsurprisingly, Malassezia sequences are not uncommon in studies of human dwellings Despite being difficult to cultivate, putative Malassezia-like organisms are abundant on living marine hosts. Pollock and colleagues Malassezia dermatitis in captive pinnipeds. Two recent studies of marine biotrophic fungi show that Malassezia-like organisms can numerically dominate fungal communities on invertebrates. A cultivation-independent study of marine sponges from Hawaii Malassezia-like sequences, and indicated that a subset of these differed from the adjacent water column. The authors' analysis further suggested that some of these putative Malassezia taxa are host specific at the species level. In a study of the scleractinian coral Acropora hyacinthus, Amend and colleagues Malassezia-like DNA sequences comprised the majority of fungi on apparently healthy colonies. A single taxon, most closely resembling Malassezia globosa, was significantly more abundant amongst corals located in warmer water.Both putatively familiar and novel Malassezia and their relatedness to better- characterized terrestrial species is a matter of speculation. A phylogeny compiled from environmental samples and sequenced isolates  Malassezia that group with presumably free-living taxa in various marine and terrestrial habitats. The relatively long branch lengths separating some of these isolates from their sister taxa suggest either a particularly rapid diversification, or, alternatively, that intermediate taxa remain to be sampled and sequenced.The evolutionary origins of marine isolates  demonstrMalassezia shows that marine taxa are interdigitated amongst those from human hosts and other terrestrial substrates and do not form a single monophyletic clade. This topology suggests repeated transitions between marine and terrestrial habitats, a pattern not atypical of marine fungi within other Dikarya lineages, including yeasts A phylogeny of environmental sequences from putative Malassezia-like organisms are found suggests that marine species of this group may incorporate a spectrum of trophic strategies ranging from saprotrophy to biotrophy. Resident Malassezia-like organisms on seemingly healthy coral and sponge hosts may be commensals, latent pathogens awaiting host immunosuppression, or both, depending on host and environmental context. A study of crustose coralline algae around Palmyra Atoll found that a Malassezia phylotype was abundant in banding disease lesions 2 and temperature. Despite efforts, the authors were unable to cultivate the fungus, and it remains to be tested if Malassezia is the cause or merely a symptom of the banding disease. Nevertheless, the study presents the possibility that a putative Malassezia may act as a pathogen in nonmammal hosts under certain environmental contexts. The high incidence and virulence of the disease raises the possibility that when combined with environmental perturbations, marine Malassezia may even exert bottom-up control on reef community structure.The tremendous diversity of habitats in which Malassezia species on human skin Malassezia-like organisms as true marine residents. Edgcomb and colleagues Malassezia-like sequences in deep-sea sediments detected by sequencing environmental RNA. Because single stranded RNA degrades quickly in situ, its presence supports the notion of active growth as opposed to DNA “contamination” in this habitat. Furthermore, the RNA sequences were distinct from those of any organism known to associate with mammalian hosts, excluding the possibility of lab contamination. A follow-up study using even more stringent protocols and negative controls to exclude exogenous nucleic acids detected Malassezia-like sequences in samples located at depths of 1.6 and 45.1 meters below the sea floor Malassezia genomes Malassezia-like organisms are frequently found in remote marine locations far from humans  lifestyle evolved independently?The remarkable environmental plasticity of Malassezia has much to offer: three sequenced genomes, M. globosa, M. restricta, and M. sympodialis, contain fewer than 9 Mb and 5,000 genes Malassezia contains a rich and potentially novel suite of enzymes and metabolites As a model system, the genus Malassezia is in obtaining axenic cultures, and no marine isolates, to my knowledge, have been recovered to date. Even among species associated with the better-characterized terrestrial mammalian hosts, lab cultivation can be a hit-or-miss affair, involving media that requires a variety of specialized and exotic fatty acids Arguably, the greatest challenge to studying marine Malassezia-like organisms independent of culturing. Some of these questions can be addressed using microscopy in conjunction with labeling techniques such as fluorescence in situ hybridization (FISH). Even basic questions about where Malassezia-like organisms reside on corals remains to be answered. Are Malassezia-like organisms associated with coral mucus, for example, or are they more closely associated with the dinoflagellate symbionts? Does their exclusion affect host fitness? Is there evidence of host specificity or co-evolution?Nevertheless, there is much to learn about the basic biology and physiology of host-associated marine Malassezia lineage likely rank among the most widespread fungi on the planet. They are found in a startling diversity of habitats and locations, from polar regions to deep-sea vents. Malassezia-like species appear to dominate certain marine habitats, which should most certainly be the focus of future research into the diversity and distribution of this enigmatic group. Clearly, considering Malassezia a mere epidermis-commensal is a definition that is only skin deep.Analysis of environmental sequences demonstrates that putative members of the


In recent years, the treatment options for patients with severe cardiorespiratory failure have been extended by the implementation of mechanical circulatory support (MCS). Identification of patients that benefit most from this cost-intensive treatment modality is of central importance, but is also challenging. Previous studies unravelled certain patient characteristics that should be taken into account, such as age, weight, and underlying pathology, and also the delay until MCS implementation as well as tissue hypoxia as prognostic factors. Relevant comorbidities included neurologic, renal, and hepatic disorders. Of note, baseline liver function tests predicted outcome in patients on extracorporeal life support (ECLS), including short-term and long-term mortality. Most strikingly, increased levels of alkaline phosphatase and total bilirubin indicated unfavourable short-term and long-term survival even after adjustment for age, gender, left ventricular function, and relevant known comorbidities such as impaired renal function and diabetes. Therefore, the assessment of liver function tests may be regarded as another piece in the complex puzzle of our efforts perceiving the ideal ECLS candidate with positive long-term outcome. Over the last decade, the treatment of patients with severe cardiorespiratory failure has witnessed a dynamic evolution of new treatment options , 2. ThisThe central dilemma of the quick decision as to who should or should not receive ECLS therapy is the lack of data. No randomised trials exist, and thus available data are mainly derived from retrospective registries or anecdotal cases. Still, there is agreement that contraindications to ECLS present conditions associated with particularly poor outcome. These have been recently reviewed in this journal . Poor ECBesides some smaller registries indicating that the duration  and exteIn their article, Roth et al. confirm that, in patients undergoing veno-arterial ECLS therapy following cardiovascular surgery, liver function tests are strong predictors of short-term and long-term mortality. The majority of the patients suffered from circulatory failure or cardiogenic shock defined as hypotension despite optimal fluid and catecholamine therapy and signs of anaerobic metabolism. Although this cohort of postoperative patients differs from patients that receive ECLS support in more urgent situations, the study indicates the usefulness of evaluating liver function before ECLS initiation. Increased levels of alkaline phosphatase and total bilirubin indicated increased short-term and long-term mortality even after adjustment for age, gender, left ventricular function, and relevant known comorbidities such as impaired renal function and diabetes. The authors conclude that these values could possibly help guide therapy decisions.These data are in line with recent findings stressing the importance of liver function and dysfunction in cardiovascular patients. This applies for different patient cohorts. Murata et al. provided convincing evidence that the assessment of liver dysfunction using the model of end-stage liver disease (MELD) scores can be used for predicting postoperative morbidity and mortality in patients undergoing cardiac surgery . In paraAnother publication sheds additional light on the prognostic role of the liver in patients treated with ECLS. Mazzeffi et al. were able to show that 8 % of ECLS patients develop acute liver failure during ECLS, while patients with chronic liver disease or acute liver failure prior to ECLS initiation were excluded. In this study, the median ECLS duration for developing acute liver failure was 5 days and these patients has distinctly elevated mortality rates .These recent findings stress the clinical relevance of inter-organ crosstalk mainly either on the basis of haemodynamic interplay or due to humoral soluble factors. Furthermore, these results deem the treating physician to take hepato-cardiac comorbidities into account during risk stratification and therapy planning. The findings of Roth et al. may be regarded as another piece in the complex puzzle of our efforts perceiving the ideal ECLS candidate with favourable long-term outcome.


DNA vaccines have several attractive development potential, such as producing cross-protection towards different virus subtypes, enabling the possibility of mass manufacture in a relatively short time and a better safety profile. The biggest obstacle to DNA vaccines is low immunogenicity. One of the approaches to enhance the efficacy of DNA vaccine is to improve DNA delivery efficiency. This review provides insight on the development of intranasal DNA vaccine for respiratory infections, with special attention paid to the strategies to improve the delivery of DNA vaccines using non-viral delivery agents.Intranasal delivery of DNA vaccines has become a popular research area recently. It offers some distinguished advantages over parenteral and other routes of vaccine administration. Nasal mucosa as site of vaccine administration can stimulate respiratory mucosal immunity by interacting with the nasopharyngeal-associated lymphoid tissues (NALT). Different kinds of DNA vaccines are investigated to provide protection against respiratory infectious diseases including tuberculosis, coronavirus, influenza and respiratory syncytial virus (RSV)  Majority of the current licensed vaccines for the prevention of infectious diseases are live-attenuated vaccines, inactivated vaccines, or subunit vaccines. Each of them has its pros and cons. The live-attenuated vaccines can stimulate both cellular and humoral immune responses, and induce prolonged immunity that closely resembles natural infection. However, there are safety concerns associated with the use of the live attenuated virus or bacterial vaccines as they may revert to disease causing forms. It is also difficult to target multiple viral subtypes or pathogens using live-attenuated vaccines. Inactivated and subunit vaccines are safer options as they cannot replicate and do not cause disease. They confer protection mainly through humoral immune responses with little or no cellular immunity. The induced immunity lasts for a shorter period of time; therefore, supplemental doses are always required.in situ production of target antigens. Plasmid DNA encoding antigenic proteins is delivered to the appropriate tissues in the body, leading to the expression of the desired antigens, eliciting specific immunogenic responses and thereby inducing immune protection against the pathogens. Since the host cells are responsible for antigen production, the natural glycosylation and folding of the protein are warranted. Plasmid DNA encoding different bacterial and viral antigens have been tested for their immunogenicity and protective efficacy in vivo, confirming their clinical potential [In recent years, DNA vaccines have attracted considerable attention as an alternative vaccination method against infectious diseases, with the potential to provide broad immune responses similar to the live-attenuated vaccines without the risk associated with the replicating micro-organisms. DNA vaccine approach relies on the otential ,2,3,4. IThe field of DNA vaccination is developing rapidly. DNA vaccines are currently approved for veterinary use against various infectious diseases ,6,7,8. HSeveral strategies have been introduced to optimize DNA vaccines . One of + T-cells, the cytotoxic T lymphocytes, which are important in controlling infections [+ T-cells by DNA vaccines can occur in two major ways: (i) direct DNA transfection of the APCs such as dendritic cells (DCs); (ii) cross-presentation which occurs when somatic cells such as myocytes are transfected with DNA and the expressed antigens are taken up by the APCs, or when the transfected apoptotic cells are phagocytosed by the APCs. The mechanisms of DNA vaccines are illustrated in Typically, DNA vaccines are administered by intramuscular injection although other administration routes including intradermal, subcutaneous, oral and intranasal routes are also investigated. Upon administration, somatic cells  and professional antigen presenting cells (APCs) are transfected. As the antigens are expressed intracellularly, both humoral and cell-mediated immunity can be activated to offer broad immune protection. The host-synthesized antigens become the subject of immune surveillance in the context of both major histocompatibility complexes (MHC) class I and class II molecules of APCs. Antigen expressed APCs then travel to the draining lymph nodes where they present the antigenic peptide-MHC complexes and stimulate T cells. Alternatively, B cells are activated, initiating the antibody production cascades. The major advantage of DNA vaccines is their ability to activate CD8fections . This aci.e., the ear) was rapidly removed after immunization before antigens were expressed, indicating that mobile cells are important in elaborating immunity. In another similar study, surgical removal of the injected muscle within 10 min of DNA vaccines administration did not affect the magnitude or longevity of antibody responses to the encoded antigen [It is interesting to note that immunization may occur rapidly before a DNA vaccine expresses the antigens, and antigens expressed in somatic cells may not qualitatively be the major player in eliciting immune response. Comparing to the secondary role of somatic cells, bone marrow derived antigen presenting cell (APC) activation is an important indicator for successful induction of DNA vaccine immune response, as evidenced by the van Gogh mice experiment . In this antigen . Again, STING) [Early studies showed that DNA vaccines are poorly immunogenic with low levels of antigen expression. To improve the immunogenicity of DNA vaccines, CpG motifs are commonly employed in the construct of the plasmid backbone . BacteriSTING) , activatAlthough persistent antigen expression of DNA vaccine is usually expected to provide long-term immune protection against infectious diseases, the effect of sustained expression of antigen must be carefully examined and controlled. It has been reported that prolonged expression of antigen may lead to the switch of type-1 IFN from an antiviral cytokine to an immunosuppressive cytokine , or may in vitro serum antibodies and ELISpot assays are the direct detectable indicators of the clinical potential of a vaccine formulation. At later stage of development, morbidity and mortality  in animals upon target pathogen challenge is a more certain way to confirm the protective efficacy of vaccines [To evaluate the efficacy of DNA vaccine in humans, serum antibody titer or the enzyme-linked immunoSpot (ELISpot) assays are the commonly employed methods to measure the immunogenic response, although the induction of antigen-specific immune effectors by an immunization process does not imply that these antibodies or cytokines represent surrogates or correlates of vaccine efficacy. In early stage of vaccine development, vaccines ,29, as tvaccines ,31 or chvaccines . HoweverSafety is always a primary concern with any vaccine products. DNA vaccines are generally considered to be safer than conventional vaccine approaches as they lack the risk of reversion to a disease causing state or secondary infection. Similar to other gene therapy, the major safety issue related to DNA vaccines is the risk of integration of the plasmid DNA into the host genome, causing insertional mutagenesis, which may lead to the inactivation of tumour suppressor genes or activation of oncogenes, resulting in devastating adverse effects. According to preclinical and clinical studies, there is little evidence of genomic integration following DNA vaccines administration, and the risk of integration is found to be significantly lower than the spontaneous mutation rate ,34,35,36E. coli can effectively prevent pathogenic anti-DNA antibody production [Another safety issue of DNA vaccines is related to the development of anti-DNA immune responses. Animal studies showed that there is no increase in anti-nuclear or anti-DNA antibodies after DNA vaccination. In clinical trial studies, signs and symptoms of autoimmunity, and the markers of autoimmunity are sometimes monitored in the human subjects. There has been no evidence that autoimmunity is associated with DNA vaccines ,37,38,39oduction ,40. Antioduction . Anotheroduction ,43,44.+ or CD8+ lymphocyte counts occurred. Another early experiment conducted on pigs showed that electroporation of DNA vaccines was more efficient in enhancing immune response, but also stimulated inflammatory response and accompanying cellular infiltration, whereas the conventional intramuscular injection of DNA vaccines only showed low gene expression and low inflammatory cell infiltration [In recent years, there has been an increasing concern that vaccination in general may induce harmful systemic inflammation, which may lead to increase of cardiovascular risk ,47,48,49ltration . It was Overall, many recent preclinical studies and clinical trials have indicated that DNA vaccines are generally well tolerated with good safety profile, and no systemic inflammation was reported ,52,53,54Conventional vaccines are usually administered by parenteral injections which mainly target the systemic immune system, eliciting only weak mucosal immune response. When the vaccines are delivered directly to the mucosal site, mucosal immune response can be more efficiently potentiated. In particular, nasal mucosa has attracted considerable attention as the site of vaccination in recent years, including DNA vaccines, due to several distinct advantages. However, there are also some formidable barriers that need to be overcome to allow successful development of intranasal DNA vaccines.The intranasal route of drug administration has been frequently used to treat local conditions such as nasal congestion and allergy. Intranasal administration is characterized by easy administration, rapid onset of action and avoidance of first-pass metabolism. The needle-free administration route is non-invasive and can avoid the risk of spreading blood-borne infections, which is a particular problem in developing countries. These desirable features lead to the exploration of the systemic delivery of polar drugs or biomolecules including vaccines that are not feasible in other administration routes.®, a live-attenuated vaccine that targets influenza types A and B [®, a live-attenuated vaccine that targets H1N1 influenza virus [Intranasal vaccination has been investigated for over a decade. The majority of currently available vaccines are administered by intramuscular, subcutaneous or intradermal injection. Although these parenteral routes of administration are effective in inducing systemic immune responses, they are ineffective in inducing local immunity at mucosal sites. As many as 70% of pathogens infect human through the mucosal surfaces . Mucosal A and B ; and NASza virus . Apart fza virus , and a rza virus . With thIt is well established that mucosal vaccination can induce humoral and cell-mediated immune response systemically as well as at mucosal sites ,57. ImmuUpon B cells activation following nasal vaccination, the production of antigen-specific secretory immunoglobulin A (sIgA) is triggered. sIgA is a critical component in the mucosal immune system. It is protease resistant, and can effectively bind and neutralize pathogens and their toxic products on the nasal mucosa surface despite the protease rich environment, thereby preventing the pathogens from breaching the mucosal barrier . Local iOverall, cells of NALT are involved in the regulation of both humoral and cell-mediated immune responses locally and also systemically, offering a broad immune response. Since the nasal mucosa is an important portal of entry for respiratory pathogens, the nasal route has become attractive for the administration of vaccines by reinforcing the nasal mucosal immune response.The defense mechanism of the nasal cavity presents a significant barrier to the entry of pathogens and potentially harmful substances; however, it has also become an important barrier to intranasal DNA vaccines. The nasal mucosa, which constitutes the outmost layer of the nasal passage, consists mainly of ciliated columnar cells, non-ciliated columnar cells, goblet cells and basal cells. The proportions of these cells vary in different regions of the nasal cavity. Nasal mucus, which is produced by goblet cells and submucosal glands, provides a protective physical barrier to foreign materials. It is a highly viscous, gel-like heterogeneous mixture that contains glycoproteins, enzymes, immunoglobulins, salts, proteins and lipidic components . DNA vacTo overcome these barriers, a safe and efficient DNA delivery system must be developed. Ideally such delivery system should target the mucosal APCs for antigens processing that lead to specific B and T cell activation. The ultimate goals of DNA delivery systems are to facilitate the uptake of DNA to the target tissues and cells, protect DNA from enzymatic degradation, increase the residence time of the formulation in the nasal cavity, enhance the expression of the antigens and to increase the immune response without compromising safety. The different DNA vaccine delivery systems currently being investigated for intranasal administration are discussed in detail in E. coli as mucosal adjuvant. Since parenteral administration of inactivated influenza vaccine did not confer an increased risk of Bell’s palsy, nor the natural influenza virus infection, it was soon concluded that adjuvant LT from E. coli. was the culprit of these cases. For this reason, FluMist® and NASOVAC®—both are intranasal live-attenuated influenza vaccines without enterotoxin as adjuvants—do not appear to confer an increased risk for this condition.Despite the numerous merits of intranasal immunization, the potential hazard of nasal vaccines must not be overlooked. The concern of the safety of intranasal vaccination was raised after an intranasal inactivated influenza vaccine called NasalFlu, developed by Berna Biotech, was found to be associated with Bell’s palsy, a temporary neurological paralysis of one side of the face . NasalFlUnderstanding the pathogenesis of the Bell’s palsy that was connected to NasalFlu has become an important research focus for vaccine development. Following intranasal administration in mice, enterotoxins were found in the olfactory nerve and the olfactory bulb for an extended period. Since the olfactory epithelium is the only part of the central nervous system (CNS) that is exposed to the external environment, drugs and nanoparticles, including intranasal vaccines, may bypass the blood brain barrier and enter the CNS through olfactory transmission. There is a reason to concern the neurotoxic effects of vaccine containing enterotoxin adjuvant for intranasal administration. While the nasal delivery of neuronal-binding LT-derived adjuvants is inadvisable , other tMankind has been haunted by respiratory infectious diseases for aeons. They have created public health concerns since ancient times. With the emergence of new or drug-resistant strains, it is becoming a challenge to protect the public from infections using conventional vaccine methods. DNA vaccines have huge potential for the prevention of respiratory infections due to their ability to offer broad immunity, the relatively rapid process of designing new DNA vaccine construct and the possibility of large-scale production in a short period of time. In this section, four pathogens that cause severe diseases in the airways are highlighted, including tuberculosis, coronavirus, influenza and respiratory syncytial virus, with a brief discussion of current DNA vaccine development against these infections.Mycobacterium tuberculosis which is transmitted by respiratory aerosols. TB has become a major public health problem that threatens the progress made in TB care and control in the world. The only TB vaccine currently available is an attenuated strain of Mycobacterium bovis, Bacillus Calmette-Guérin (BCG), developed in the 1920s. However, its efficacy against adult pulmonary TB remains controversial [Mycobacterium tuberculosis requires cell-mediated immune responses, including both CD4+ and CD8+ T cells [Mycobacterium tuberculosis [Mycobacterium leprae (hsp65) [Tuberculosis (TB) is a bacterial infectious disease caused by oversial . With th T cells ,76,77. Srculosis  and the  (hsp65) . Other d (hsp65) ,83,84,85Coronaviruses (CoV) are potentially lethal pathogens, characterized by the presence of spike proteins on the viral surface. Two new strains, severe acute respiratory syndrome CoV (SARS-CoV) and Middle East respiratory syndrome CoV (MERS-CoV), have been identified. Both of them could cause acute respiratory distress syndrome (ARDS) and are associated with high mortality rates . CoV vacInfluenza is caused by orthomyxoviruses which are RNA viruses that affect mainly the upper respiratory tract. In recent years, zoonotic or variant influenza has become a serious threat to human health, including the avian influenza virus H5N1 and H9N2, and the swine influenza virus H1N1 and H3N2. Although these animal viruses are distinct from human influenza viruses and do not usually transmit between humans, they may still occasionally infect humans and cause severe pneumonia and even death. Furthermore, if such a virus acquired the capacity to spread easily among people, it could start an epidemic or even a pandemic ,93. To pDNA vaccines can potentially solve the mismatch problem by shortening the lag time , which iParamyxoviridae family. It accounts for one of the leading pathogeneses of lower respiratory tract infections and hospitalization in infants and young children [+ T cells responses have been shown to reduce disease severity [Respiratory syncytial virus (RSV) is a single stranded RNA pneumovirus which belongs to children , as wellchildren . Immunitseverity . Althougseverity ,110,111.Delivery is one of the major barriers to DNA vaccine. Administration of naked DNA is usually inefficient with only a small fraction of DNA being taken up by the cells and subsequently expressed . This isThe elicitation of immune responses of DNA vaccines mainly relies on professional APCs that present antigens to both B cells and T cells. To ensure good immune response, the DNA vaccine delivery systems should be able to target APCs. In addition, M cells in NALT, which is a major site of pathogen entry, are also a target of DNA vaccine.APCs are a heterogeneous group of immunocompetent cells that mediate immune response by processing and presenting antigens to the T cells. T cells recognize only antigenic peptide fragments on the surface of APCs through the T cell receptors. Helper T cells recognize antigen in association with class II MHC proteins, whereas cytotoxic T cells recognize antigen in association with class I MHC proteins. An additional co-stimulatory signal is then produced by the APCs, leading to the activation of T cells. Non-professional APCs lack the co-stimulatory signaling, and therefore do not simulate T cells sufficiently. There are three types of professional APCs, namely DCs, macrophages and B cells. Among them, DCs have the broadest range of antigen presentation and are considered as the most efficient cells for induction and regulation of immune responses. They play a central role in bridging the innate immune system with the adaptive immune system ,117,118.DCs express a large number of surface receptors such as C-type lectin receptors (CLRs) and TLRs, which are engaged in the recognition of pathogens. It has been reported that targeting antigens to receptors on DCs can significantly enhance immune responses . These r+ progenitors, including DC progenitors. In vivo treatment of Flt3 ligand is found to up-regulate the number of DCs, but not their activation [There are also several molecules being investigated to target APCs in general. One of the most widely studied molecules is the Flt3 ligand. Flt3 ligand is a growth factor that stimulates the proliferation of hematopoietic cells. It binds to the fms-like tyrosine kinase receptor Flt3. Flt3 expression is, in hematopoietic tissue, restricted to CD34tivation ,126. Furtivation . It has tivation . In additivation ,130.As discussed in Salmonella typhimurium [Mycobacterium tuberculosis [et al. reported an M cell targeting DNA vaccine delivery system consisting plasmid DNA and the recombinant protein σ1 as targeting ligand which was covalently attached to poly-l-lysine (PLL) for intranasal vaccination in mice. The results showed significant mucosal sIgA production as well as enhanced cell-mediated immunity [Apart from controlling particle size to achieve specific targeting passively, inclusion of targeting ligand could also increase uptake by M cells ,137. A nhimurium  and Mycorculosis  target Mrculosis . One exaimmunity . Other limmunity , Claudinimmunity ,143 and immunity  have beeimmunity . TherefoHigh versatility is one of the attractive features of polymer-based DNA delivery systems. Cationic polymers can form complexes (polyplexes) with nucleic acids through electrostatic interaction. Polymer synthesis is relatively cheap and is easy to scale-up. Particle size and surface properties of polymeric particles can be controlled by using different polymers and fabrication methods in order to optimize their cellular uptake and transfection efficiency. The polymeric particles can also be modified to include specific function groups or ligands to enhance immune responses.et al. described the use of a simple method to prepare PEI (25 kDa)–DNA complexes for vaccine delivery [et al. demonstrated the utilization of PEI (in vivo-jetPEI®) as DNA vaccine carrier for intranasal administration [Polyethylenimine (PEI)  is one odelivery . Plasmidstration . Plasmidet al. has developed a PEI derivative, deacylated PEI (dPEI), as DNA vaccine delivery agent [Although PEI appeared as a promising vector for gene delivery including DNA vaccines, the cationic PEI is highly charged and non-biodegradable, and it often encounters toxicity problems which cannot be ignored. In this regard, many groups are developing low toxic or biodegradable PEI derivatives for gene delivery application. Mann ry agent . dPEI isChitosan  is a natet al. reported the exploitation of chitosan nanoparticles to deliver DNA vaccine against acute respiratory syncytial virus (RSV) infection [A number of studies have described the use of chitosan nanoparticles to deliver DNA vaccine formulation for intranasal administration. Kumar nfection . A cocktnfection . It was et al. investigated a sophisticated DC targeted chitosan nanoparticle system for nasal DNA immunization against SARS-CoV [In a recent study, Raghwanshi SARS-CoV . The chiTo enhance the transfection efficiency of chitosan for intranasal administration, thiolated chitosan derivative has been introduced. Thiolated chitosan derivative has strong mucoadhesive properties due to the formation of disulphide bonds between thiol groups of the modified polymer and cysteine-rich subdomains of glycoproteins in the mucus layer, leading to an improvement in mucoadhesion of up to 140-fold when compared to unmodified chitosan . Improveet al. first employed the use of micro-particles consisting PLGA and PEI as DNA vaccine carrier for injection [et al. reported the use of chitosan coated PLGA nanoparticles to deliver plasmid DNA encoding FMDV (foot-and-mouth disease virus) capsid protein together with IL-6 as mucosal adjuvant for intranasal vaccination [Poly(lactic-co-glycolic acid) (PLGA)  is a synnjection . Later ocination . Chitosacination , followeLiposomes are vesicles comprised of phospholipid bilayers. They have been extensively investigated for delivering DNA into mammalian cells as well as vaccine adjuvants. The duo functions make them an excellent carrier system for DNA vaccine ,170,171.® V (influenza vaccine) and Epaxal® (hepatitis A vaccine). Both formulations employed the virosomes technology /cholesterol cationic liposome-DNA complexes (JVRS-100), in which an empty plasmid DNA vector was incorporated, were able to induce high levels of antibody and T cell immunity in mice and non-human primates [Currently, there are at least two approved liposomal vaccine formulations on the market for antigen delivery, including Inflexalchnology , in whicprimates . Since lprimates .et al. reported the use of a cationic and neutral co-lipid formulation, GAP-DLRIE:DOPE (aminopropyl-dimethyl-bis-dodecyloxy-propanaminium bromide-dioleoylphosphatidyl-ethanolmine) as a carrier of DNA vaccine against TB for intranasal administration [A number of studies described the use of liposomes as DNA vaccine carrier for intranasal administration. D’Souza stration . Plasmidet al. developed another liposome-based formulation of DNA vaccines against TB [sn-glycero-3-phosphoethanolamine/1,2-dioleoyl-3-trimethylammonium-propane) was used. Plasmid DNA encoding the 65 kDa mycobacterial hsp65 was either entrapped inside or complexed with the cationic liposomes, and the intramuscular and intranasal routes of administration were compared in mice. When administered intramuscularly, both liposomal formulations were ineffective in preventing tuberculosis infection in mice even after two doses. On the contrary, the complexed liposomal formulation of DNA vaccine was able to offer protection against infection even after a single dose, with a significant reduction of colony forming unit (CFU) in lungs after the immunized mice were challenged with Mycobacterium tuberculosis. However, four doses of intranasal administration of naked DNA vaccines failed to offer any protection. The authors reasoned that the intranasal vaccination enhance the immune response by stimulating the mucosal immunity, but the naked DNA failed to cross the mucosal barriers in the nasal cavity, demonstrating the importance with the use of delivery carrier for intranasal DNA vaccination.Rosada ainst TB . The nonApart from TB vaccine, there are studies that reported the use of liposome to deliver influenza DNA vaccine ,178. Catet al. modified the surface of liposomes by coating with glycol chitosan [To improve the DNA vaccine delivery efficiency of liposomes for intranasal administration, Khatri chitosan . The majchitosan . In the A number of studies have already demonstrated the potential of liposomal DNA vaccine system for intranasal administration, and some could offer considerable immune protection against respiratory infections in animals. However, the lipid composition of different liposomal systems varied greatly, and currently there is a lack of knowledge of how the composition may affect the immune response. To enable the utilization of liposomal DNA vaccine for clinical application and approval, a better understanding of how these factors govern the efficacy and immunity of the liposomal delivery system must first be sought.To further enhance the immune responses of the DNA vaccines, adjuvants are included in the formulation in many studies. Adjuvants are generally defined as agents that could enhance the immune response of the vaccinated subjects to an antigen. In DNA vaccine, since the delivery of DNA is a major hurdle, DNA carrier system using bacterial, viral or non-viral vectors, as well as the cell specific targeting ligands, which are discussed above, are also considered as DNA vaccine adjuvants. The summary of DNA vaccine adjuvants being investigated are shown in E. coli and cholera enterotoxin (CT) are very potent adjuvants but they are too toxic to be used in human. Therefore, detoxified mutants of enterotoxins have been produced by site-directed mutagenesis and they are extensively investigated as adjuvants for mucosal vaccine including intranasal vaccine. Intranasal antigen immunization with LT mutant adjuvants can provide effective protection against infectious diseases in animals [Enterotoxins are protein exotoxin released by pathogens that infect the gut. Enterotoxin-based mucosal adjuvants are the most potent and well-established strategy for the induction of both mucosal and systemic immunity to co-administered protein antigens . Heat-la animals ,184,185. animals . Mutants animals ,186,187, animals . It is e animals . The maj animals ,71,188. Salmonella minnesola. MPL is thought to interact with Toll-like receptor 4 (TLR-4) on APCs. It has been demonstrated that MPL could activate macrophages, increase their cytokine secretion and hence immune response [Lipopolysaccharides (LPS) are the major component of the outer membrane of Gram-negative bacteria, and could elicit strong immune response. However, they are also highly toxic. In order to make them safe and suitable as vaccine adjuvants, LPS derivatives are produced to reduce the endotoxic effect but to retain their immunostimulatory function. Monophosphoryl lipid A (MPL) is one example of LPS derivative that is investigated as vaccine adjuvant. MPL is prepared by removal of a phosphate and fatty acid group from the lipid A of response ,219,220.response . MPL hasresponse . MPL hasresponse ,223,224.et al. demonstrated that intranasal administration of pneumococcal polysaccharide conjugate vaccine in the presence of IL-12 was able to enhance systemic and mucosal immune responses to pneumococci in mice [in vivo and poor stability. They are also expensive and are associated with dose related toxicity. Therefore, these molecules may not be suitable for use as adjuvants in vaccines designed to protect against infectious diseases [in vivo and to reduce the cost of production [Cytokines are small proteins that are important in regulating immunological response by recruiting and stimulating T cells, or by directly acting on infected cells. They have potential to be natural adjuvants for DNA vaccines . Cytokin in mice . Howeverdiseases . Neverthdiseases . Alternaoduction ,198.With the emergence of new or antimicrobial-resistant bacteria and viruses, and the ease of transmission, especially the respiratory pathogens, respiratory infections are becoming serious threats to human health. Safe and effective vaccines are important to safeguard public health. Intranasal DNA vaccination appears to be a promising non-invasive approach to provide protection against various infectious diseases. Evidence shows that intranasal DNA vaccine could elicit strong and long-lasting humoral as well as cell-mediated immune responses in many animal models. DNA vaccines are already successfully used in veterinary products for protection against infections, but their immunogenicity needs to be further enhanced to make them suitable for human use. Improving DNA delivery and formulation is one of the several strategies to enhance the immune response. Various studies have demonstrated that significant improvement of immune response that could be achieved by the employment of DNA carrier system, or to target the DNA vaccines to APCs. DNA vaccines generally have good safety profile, but the potential toxicity associated with DNA delivery systems, especially when they are used at high concentration, must not be neglected. DNA vaccines may circumvent many problems associated with conventional vaccines such as high costs of protein vaccine purification and bacterial/viral inactivated or attenuated process, the incorrect folding of antigen and viral mutation risk, thereby offering a safer alternative to benefit humans. In addition, mass manufacture of DNA vaccine is easier and faster, and DNA product is usually highly stable. Once an effective intranasal DNA vaccine delivery system is identified and optimized, a delivery technology platform could be established to allow the development of DNA vaccine formulations for different infectious diseases in the future.


The neurovisceral integration model  posits t When the mind is strongly excited, we might expect that it would instantly affect in a direct manner the heart; and this is universally acknowledged and felt to be the case. Claude Bernard also repeatedly insists, and this deserves especial notice, that when the heart is affected it reacts on the brain; and the state of the brain again reacts through the pneumo-gastric nerve on the heart; so that under any excitement there will be much mutual action and reaction between these, the two most important organs of the body.emotion regulation  information  information , they primarily utilized LSF information. In contrast, when people were asked to discriminate whether the stimuli were expressive or not, they utilized HSF information. Thus, the utilization of HSF information facilitates the discrimination of expressiveness  or not  endogenous attentional orienting  are presented as cues in the task, people are faster to detect targets in valid trials  and slower to disengage attention away from cues in invalid trials , suggesting hyper-vigilant responses to threatening stimuli primilarly tapping into subcortical mechanisms. However, with longer SOAs, (960 ms), people with lower resting HRV showed significantly slower attentional disengagement from HSF fearful faces relative to people with higher resting HRV, suggesting the failure to inhibit attention from threatening stimuli primarily tapping into cortical mechanisms. These findings provide initial evidence that individual differences in resting HRV are associated with top-down and bottom-up emotional attention. People with higher resting HRV – associated with highly functional emotional and cognitive self-regulatory systems – show more adaptive top-down and bottom-up emotional attention, which may facilitate effective emotion regulation. In contrast, people with lower resting HRV – associated with poor emotional and cognitive self-regulatory systems – show maladaptive top-down and bottom-up emotional attention, which may be detrimental to emotion regulation.collicular  and (b) cortical mechanisms, including the temporo-parietal cortex and anterior cingulate cortex, which are typically associated with attentional control. We examined whether individual differences in HRV would predict the IOR to emotional and neutral face cues  and cortical mechanisms, respectively . In genesuppression or enhancement is associated with the exertion of self-regulatory effort may depend on the context in which phasic HRV changes occur (There is a growing body of evidence suggesting that changes in cardiac activity indexed by phasic HRV are linked with self-regulatory effort . Howeveres occur . Phasic es occur . For exaes occur , asked tes occur , or engaes occur , phasic es occur . For exaes occur . Additioes occur . Thus, ees occur .We recently examined the extent to which individual differences in self-regulatory capacity, indexed by resting HRV, were associated with the exertion of self-regulatory effort, indexed by phasic HRV, in response to emotional versus neutral distractors under different levels of cognitive load . ParticiResults of extensive research have indicated that low resting HRV is typically observed in people with various psychopathologies, including generalized anxiety disorder, panic disorder, depression, bipolar disorder, and schizophrenia . FurtherFurthermore, several studies provided evidence that resting HRV is associated with the ability to control autonomic responses . People The current review presented evidence of an underlying interaction between individual differences in cardiac vagal tone and top-down and bottom-up cognitive processing of emotional stimuli, which promotes further regulatory behaviors and autonomic flexibility. To further dissociate top-down and bottom-up mechanisms, we utilized stimuli with different spatial frequency ranges designed to tap into either top-down or bottom-up neural mechanisms of emotional processing. These studies provide evidence that higher resting HRV is associated with flexible and adaptive top-down and bottom-up cognitive processing, which facilitates effective emotion regulation. In contrast, lower resting HRV is associated with hyper-vigilant and maladaptive bottom-up and impaired top-down cognitive response to emotional stimuli, which is detrimental to emotion regulation. The results of these studies raise the possibility that maladaptive cognitive processing of emotional stimuli observed in people with lower HRV may be detrimental to emotional and physical health, which explains why people with a wide range of psychopathologies and health issues exhibit lower HRV.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.


Inflammation plays an important role in cardiac dysfunction under different situations. Acute systemic inflammation occurring in patients with severe burns, trauma, and inflammatory diseases causes cardiac dysfunction, which is one of the leading causes of mortality in these patients. Acute sepsis decreases cardiac contractility and impairs myocardial compliance. Chronic inflammation such as that occurring in Duchenne muscular dystropshy and myocarditis may cause adverse cardiac remodeling including myocyte hypertrophy and death, fibrosis, and altered myocyte function. However, the underlying cellular and molecular mechanisms for inflammatory cardiomyopathy are still controversial probably due to multiple factors involved. Potential mechanisms include the change in circulating blood volume; a direct inhibition of myocyte contractility by cytokines (tumor necrosis factor (TNF)-α, interleukin (IL)-1β); abnormal nitric oxide and reactive oxygen species (ROS) signaling; mitochondrial dysfunction; abnormal excitation-contraction coupling; and reduced calcium sensitivity at the myofibrillar level and blunted β-adrenergic signaling. This review will summarize recent advances in diagnostic technology, mechanisms, and potential therapeutic strategies for inflammation-induced cardiac dysfunction. Inflammation is a defending biological process to fence off injurious stimuli such as pathogens, damaged tissues, or irritants. Temporally it can occur acutely or chronically. In term of the scale of inflammation, it can occur locally confined within local tissues/organs or systemically (whole body). Acute and/or systemic inflammation often accompanies with a lot of diseases. Systemic inflammatory response always has noninfectious causes such as trauma and burns or infectious causes such as viral or bacterial infection. Though appropriate extent of inflammation is protective and beneficial to maintain homeostasis of the organism, excessive and chronic inflammation is detrimental in that it alters many vital organs. As we know, burn injury can lead to global immunological changes including suppressed immune function and increased susceptibility to infection. Sepsis aInflammation is an active biological process involving multiple components to clean pathogens , injured tissue/cells , or to respond to abnormal systemic function such as autoimmunity and lipoprotein deposition. It intenBoth acute and chronic inflammation has local or systemic response. In some severe infectious diseases, especially when the bacteria or pathogen were expanded in the vascular system, systemic inflammation response occurs. When activated immune cells, cytokines/chemokines, and tissue debris during acute inflammation are produced to such a scale that they are present in the circulation at high concentrations, acute inflammation becomes systemic and septic. Chronic inflammation happening in the vasculature, the fat tissue of obesity, and skeletal muscles of muscular dystrophy is also systemic. Local inflammation in vital organs such as the heart in ischemia/reperfusion injury impairs the function of the organ; systemic inflammation in sepsis may depress organ function remote from the inflammatory sites.2 from the tissues. Cardiac function is constantly regulated by the sympathetic and parasympathetic nervous systems (SNS and PSNS) to meet different needs of the body under different physiological and pathological conditions.2+ handli disease. also sti disease.In summary, the mechanisms for cardiac dysfunction induced by inflammation have not been fully understood and it seems that multiple underlying mechanisms for cardiac dysfunction for both acute and chronic inflammation are overlapping. There could be difference in the relative contribution to each inflammatory condition and to different stages of inflammation.et al.’s study, they found increased circulating levels of TPO after burn injury, as much as two-fold of that in the healthy. Additionally, they also found increased monocyte platelet aggregation in burn patients with sepsis, which can be a potential indicator for developing sepsis, microvascular dysfunction, and multiorgan failure.i that can only be reached during systole, prolonging the interaction of myosin and actin filaments through the inhibition of troponin I.[+ channels and mitochondrial ATP-sensitive potassium (mito.K(ATP)) channels in cardiac myocytes and vascular smooth muscle cells.[Levosimendan is a new pharmacological agent used in the management and treatment of acute and chronic heart failure. Unlike other inotropic agents, as a myofilament calcium sensitizer, it can improve cardiac performance without activating the SNS and changing oxygen consumption. Levosimendan binds to the N-terminal of troponin C with high affinity at high [Caoponin I. Levosimeoponin I. Levosimele cells. This inole cells. Moreoverle cells. Doorduinle cells. found lef. If is important for generating the pacemaking current, and responding to β-adrenergic stimulation of the SAN cells. The (If) blocker ivabradine reduces heart rate and improves systolic function without causing apparent direct hemodynamic effects. Contrary to β-blockers, ivabradine has no hypotensive character. When body suffers stress including sepsis, burn, and trauma; the heart rate is accelerated causing tachycardia.[Ivabradine is a selective inhibitor of the hyperpolarization activated cyclic-nucleotide-gated Ihycardia.,118 Lotshycardia. Reductiohycardia. In a dyshycardia.2+ and β-adrenergic system are underlying both the detrimental and salutary effects of inflammation. Anti-inflammatory clinical trials have not succeeded thus far. Current treatments for cardiac dysfunction after inflammation focus on cardiovascular support, limiting cardiac injury and adverse remodeling with inhibition of b-adrenergic and the reninangiotensin-aldosterone system (RAAS) systems, and reducing heart rate. A better understanding of the underlying mechanisms for both detrimental and protective effects of inflammation under different conditions in the heart is needed for future development of effective therapy to prevent cardiac injury caused by inflammation while at the same time to preserve necessary and protective aspects of inflammation.Both acute  and chronic inflammation  can cause cardiac dysfunction, which is an often cause of mortality and morbidity in medical conditions causing inflammation. However, inflammation may also exert protection on the heart. Many mechanisms including signaling pathways involving cytokines and chemokines, NO, and Ca


Bile acids were first proposed as carcinogens in 1939. Since then, accumulated evidence has linked exposure of cells of the gastrointestinal tract to repeated high physiologic levels of bile acids as an important risk factor for gastrointestinal cancers. High exposure to bile acids may occur in a number of settings, but most importantly, is prevalent among individuals who have a high dietary fat intake.A rapid effect on cells of high bile acid exposure is the generation of reactive oxygen species and reactive nitrogen species, disruption of the cell membrane and mitochondria, induction of DNA damage, mutation and apoptosis, and development of reduced apoptosis capability upon chronic exposure. Here, we review the substantial evidence of the mechanism of secondary bile acids and their role in colon cancer. Bile Acids (BA) are normal components of the lumenal contents of the gastrointestinal (GI) tract, where they enable absorption of lipids, cholesterol, and fat-soluble vitamins. In essence, they act as a physiologic detergent and regulator of intestinal epithelial homeostasis in the gastrointestinal tract.However, BAs, specifically lithocholic acid (LCA) - a secondary BA - also constitute a rare example of toxic endobiotics. In factAt high physiologic concentrations, BAs can cause oxidative/nitrosative stress, DNA damage, apoptosis, and mutation. FurtherAnd since BAs are normal components of the luminal contents of the GI tract, finding the exact mechanism of their carcinogenic effect has become intriguing. Several factors have been found to increase levels of BAs: most importantly, a high dietary fat intake.Our aim is to explain the correlation between the concentration of fecal secondary BAs -mainly deoxycholic acid (DOC) and LCA - and the colorectal cancer incidence that was highlighted by several epidemiological studies but whose molecular mechanism remain far from clear.Furthermore, BAs were also found to be etiologic agents of other GI tract cancers, namely that of the esophagus, stomachPrimary BAs (cholic acid and chenodeoxycholic acid) are derived from cholesterol by a sequence of enzymatic reactions occurring mainly in the liver. Synthesis of a full complement of BAs requires 17 individual enzymes and occurs in multiple intracellular compartments that include the cytosol, endoplasmic reticulum (ER), mitochondria, and peroxisomes. After sDOC is partly absorbed in the colon and enters the enterohepatic circulation, where it is conjugated in the liver and secreted in the bile. LCA is BAs are amphipathic and many of their properties are related to their amphipathic nature. Their aAlmost 40 years ago, Berg was the first to make the observation that CRC  risk was higher among descendants of individuals in low-risk populations after moving to developed countries and converting to a Western-type diet that is rich in red meat and saturated fatty acids. This waet al.[et al.[Prolonged, high consumption of red meat and saturated fatty acids were found to increase CRC risk. In their case control study, Bayerdorffer et al. confirme[et al. found th[et al.. Populat[et al.,13. Resu[et al.. Add to [et al..Conversely, diets rich in vegetables and fruits are linked to a decreased CRC incidence. Dietary fibers (from vegetables and fruits) can bind to LCA and aid in its excretion in stool; as suchLCA activates vitamin D receptor, which may activate a feed-forward catabolic pathway that leads to the detoxification of LCA. Whereas, high dietary calcium leads to the formation of insoluble calcium soaps, this in turn decreases the concentration of free BAs in the intestinal lumen that ultimately may protect against formation of colon cancer.An essential constituent of plasma membrane is cholesterol, which rigidifies the membrane and is an important structural component of membrane microdomains. Due to In the colonic epithelium, high secondary BA concentrations induce cell proliferation by activating epidermal growth factor receptors (EGFRs) and post-EGFR/ERK  signaling. In addiNuclear receptors (NRs) are transcription factors that act as sensors of dietary and endogenous molecules, translating nutritional and hormonal stimuli into transcriptional programs. RecentlVitamin D deficiency is a known major risk factor for colorectal cancer. When viInterestingly, the enteric NR transcriptome has been found to be downregulated in both mouse and human models of CRC when the tissues are progressing from normal intestinal epithelia to dysplastic lesions. This suin vivo status of HLA in response to LCA, however, is still lacking.Reduced or lost expression of HLA (human leukocyte antigen) antigens is seen in human colon carcinogenesis. LCA wasOne of the most important cytotoxic effects of BAs is the increased production of reactive oxygen/nitrogen species (ROS/RNS). ActivatThe balance between whether BAs induce proliferation or cell death in colonic epithelial cells is finely tuned by activation of NF-kB. The ability of BAs to induce activation of NF-kB largely determines intestinal epithelium cell fate. CloselyA report has highlighted the mechanism of DOC-induced NF-kB activation in HCT116 cells by showing the significant correlation that binds NAD (P) H oxidase, Na+/K + −ATPase, cytochrome P450 isoform 1B1, calcium levels and the terminal mitochondrial respiratory complex IV.BAs induce DNA damage in colon cells with oxidative DNA damage being a likely component. DefectiGenetic instabilities and chromosomal alterations are frequently found in CRC. BAs canet al. recently reported a biphasic effect of cytoprotection and induction of apoptosis by hydrophobic BAs in the HCT116 cell line[in vivo prevents tumor incidence by eliminating damaged cells[BAs have the potential to induce cell death both through nonspecific detergent effects and receptor-mediated interactions. Elevateell line, with inell line. In a phed cells. Severaled cells. In addied cells.Since the CRC incidence and mortality rates are expected to increase in the coming decades, in-depth understanding of the molecular and cellular mechanisms underlying CRC becomes crucial. Focusing on the role of secondary BAs in developing CRC would be of great significance since altering the risks that increase secondary BAs would alter the incidence of CRC. Not only that, but secondary BAs may become an effective screening marker. Secondary bile acids solve the puzzle of colorectal cancer because they sit at the crossroad of nutritional and hormonal signals modulating the tangled interactions between the environmental factors, such as diet, and the nuclear receptors such as VDR. The pivotal question that emerges from the current evidence is to specify what epigenetic abnormalities are required in the crypt stem cell to render them prone to secondary BA. The next challenge is to unravel how potential stem cell specific mutations render them more prone to environmental factors, which in turn, renders them more prone toward either normal or tumorigenic phenotypes.BA: bile acids; CRC: colorectal cancer; DOC: deoxycholic acid; EGFRs: epidermal growth factor receptors; ER: endoplasmic reticulum ERK, extracellular signal-regulated kinase; GI: gastrointestinal; LCA: lithocholic acid; NR: nuclear receptors; PKC: protein kinase C; PXR: pregnane X receptor; RNS: reactive nitrogen species; ROS: reactive oxygen species; SXR: steroid xenobiotic receptor; VDR: vitamin D receptor.The authors declare that they have no competing interests.HA drafted the manuscript, AS and DM participated in the design and coordination of the study. All authors read and approved the final manuscript.


EnSCs comprise of a population of epithelial stem cells, mesenchymal stem cells, and side population stem cells. When secreted in the menstrual blood, they are termed menstrual stem cells or endometrial regenerative cells. Mounting evidence suggests that EnSCs can be utilized in regenerative medicine. EnSCs can be used as immuno-modulatory agents to attenuate inflammation, are implicated in angiogenesis and vascularization during tissue regeneration, and can also be reprogrammed into induced pluripotent stem cells. Furthermore, EnSCs can be used in tissue engineering applications and there are several clinical trials currently in place to ascertain the therapeutic potential of EnSCs. This review highlights the progress made in EnSC research, describing their mesodermal, ectodermal, and endodermal potentials both  Cells in the earliest developmental stages in the embryo can generate embryonic and extra-embryonic tissues . The abiThe human endometrium is an extraordinary model of controlled tissue remodeling, unparalleled in other organs, which grows about 7 mm within one week in every menstrual cycle . At this11 cells from a single cell, with the ability to differentiate into large cytokeratin-expressing structures when cultured in Matrigel [in vivo and in vitro. These cells are not tumorigenic in nude mice and could be passaged approximately 40 times in vitro.The progenitor cells in the endometrium has a high proliferative potential that can generate 6 × 10Matrigel , comprisThe fact that about 600,000 hysterectomies are performed yearly in the United States creates a potential source of endometrial cells . StudiesEndometrial stem cells may be derived from fetal stem cells or bone marrow stem cells, including haemopoietic SCs, MSCs and endothelial progenitor cells -38. The Flow cytometric analysis of important endometrial stem cell markers are indicated in Figure Utilizing ES cells in regenerative therapy is associated with many hazards. Without adequate ES cells screening procedures, patients would face tissue rejection similar as rejection of organ transplants. A solution to this problem is to create patient specific stem cells. Unlike ES cells, adult stem cells are not rejected by the immune system . The humThe main features of mesenchymal stem cells are their ability for self-renewal, colony-forming ability, and differentiation into mesoderm-derived lineages including adipogenic, chondrogenic and osteogenic differentiation -65. EndoResearch pertaining to stem cell biology in the female reproductive tract is still in its infancy, and although surface markers of prospective isolation of human endometrial stromal colony-forming cells  have recently identified ,44, therThe final step for processing stem cells before transplantation would be the identification and isolation of the differentiated cells. Recognition and selection of desired cells can be done via reverse transcriptase polymerase chain reaction, flow cytometry or fluorescence-activated cell sorting (FACS). Unfortunately, these methods would destroy many of the differentiated cells, thereby rendering them unusable. Also, flow cytometry and FACS is dependent on the presence of specific markers on the cell surface, which are not always displayed on every type of differentiated cells . Thus, ain vivo. It was shown that EnSCs produced collagen type II and proteoglycans when cultured on nanofibrous polycaprolactone material, suggesting their cartilaginous tissue differentiation potential [-. Mature bone marrow MSCs are a heterogeneous population of cells that are able to support haematopoiesis, and capable of differentiating into different lineages such as neural cells [in vitro. It has been shown that bone marrow MSCs contain more mesenchymal progenitor cells when compared to umbilical cord blood stem cells. Indeed, the proportion of MSCs within umbilical cord blood is extremely low and difficult to detect [Genetic profiling of EnSCs and bone marrow MSCs revealed that they had similar  cytokine production, miRNAs, and gene expression . It was otential . It was otential . Umbilicotential . Consideal cells ,75, muscal cells -78, liveal cells ,80, amono detect . Neverthin situ differentiation might have been involved, in part due to the observation that the transplanted EnSCs homed to peri-muscle fibre regions and promoted angiogenesis. The concept of angiogenesis was also supported by another study, where EnSCs were improved critical limb ischaemia induced by femoral artery ligation [To date, EnSCs have been used in several pre-clinical and small animal studies. The regenerative potential of EnSCs was first seen in an experimental model of Duchenne muscular dystrophy in immunodeficient mdx mice, whereby EnSCs that were transplanted into atrophied skeletal muscle fibres contributed to muscle repair . Althougligation .+, troponin+ striated cardiac muscle cells [The use of EnSCs to treat myocardial infarction in a murine model was also seen. In this study, EGFP-labelled EnSCs were grafted into the infarct area of nude rat hearts, which subsequently differentiated into α-actininle cells . Furtherin vivo, on an animal skin wound repair model [Gargett et al., the first group that reported the existence of EnSCs in 2004 , are curir model ,85. ResuOur group has also advocated the tissue engineering approach to the application of EnSCs. To this end, we have fabricated nanofibrous silk-collagen fibres that were seeded with EnSCs for the reconstruction of urinary bladder wall for women . We had Intraperitoneal delivery of EnSCs were also studied in a murine model of encephalomyelitis . It was in vivo part of the study was done in a murine model of ischaemic stroke, whereby injection of EnSCs resulted in significantly lower histological and behavioural impairments. It was reported that EnSCs exerted a trophic effect, releasing factors that promoted survival of neural cells.EnSCs were also used to demonstrate neural regenerative capabilities, whereby reduced neuronal cell death was seen when oxygen-deprived primary neuronal cell cultures were exposed to EnSCs . It was The use of EnSCs to treat glioma was observed in a murine model. In this study, EnSCs were administered intravenously in a murine model of intracranial glioma. Results revealed a reduction of tumour size of almost 50%, possibly due to its anti-angiogenic effects .The applications of EnSCs have also been reported in several human studies. The first reported use of EnSCs was demonstrated by Zhong et al. . ClinicaAnother human study involved a patient suffering from Duchenne muscular dystrophy that was given intramuscular injections of EnSCs. Follow-up observations reported no adverse effects even after 3 years, and increased muscle strength and decreased respiratory infections was also reported .The third reported use of EnSCs in human was a patient with congestive heart failure, who was given intravenous administration of EnSCs. Results revealved that ejection fraction of the patient increased from 30% to 40%, decreased basic natriuretic peptide values (Pro-BNP), and decreased Minnesota Living with Heart Failure Questionnaire score at 1-year follow up .EnSCs are an attractive source of stem cells for regenerative therapeutics as they are easily obtainable and easily expandable in culture, as has been demonstrated to be safe for clinical use. Protocols and methods for extraction and isolation of EnSCs are well established, as purified EnSCs can be obtained using magnetic bead sorting using the W5C5/SUSD2 marker. In addition, clinical-grade good manufacturing practice (cGMP) are currently being developed for culture expansion of EnSCs, and have been tested in animals. However, there is a lack of published information on the exact cGMP protocols in place for the production of EnSCs. This is compounded by the fact that there is no general scientific consensus regarding specific MSC markers to detect EnSCs; rather, researchers rely on the ability of MSCs to adhere to plastic. Hence, the purity of EnSCs is not guaranteed as the cultures could potentially contain fibroblasts.EnSCs can be obtained from menstrual blood; hence no invasive procedures are needed to harvest these cells. A menstrual cup is used to collect menstrual blood over several hours on days 2 to 3 of the menstrual period. Although there is a potential risk of infection via vaginal contact, there have been no reports of any complications after antibiotic use.in vivo has been demonstrated, there is a theoretical risk that endometriosis could develop from using EnSCs. However, none of the animal model studies have reported this. Nevertheless, it is an aspect of EnSC application that warrants attention. Indeed, transdifferentiation (sometimes referred to as adult stem cell plasticity) is a controversial topic in the field of stem cells. It is highly probable that nuclear reprogramming and altered transcriptional activity of important developmental genes are in part responsible for transdifferentiation. Transdifferentiation can be seen as a form of metaplasia due to alterations in the extracellular environment, and often happens during tissue damage [+E+-Cadherin cells, CK-E-Cadherin- stromal cells, and an invasive CK+E-Cadherin-N-Cadherin+ epithelial cells (similar to carcinoma micrometastasis) are found in endometriotic lesions [Although the ability of EnSCs to re-integrate into tissue e damage . Hence, e damage . It was e damage . Changese damage . Corresp lesions . In lineIn terms of regenerative medicine applications, it is envisioned that bone marrow MSCs are more suited for osteogenic differentiation and therefore used for bone tissue engineering, while EnSCs are more suited for soft tissue engineering, such as bladder reconstruction. Due to the intense interest generated with using EnSCs for regenerative medicine, we believe that EnSCs were introduced too quickly into human tests without adequately assessing its impact on large animal models. Furthermore, the exact mechanism of how EnSCs exert is regenerative potential is not clearly understood. Hence more robust large animal studies are needed to fill this information gap that currently exists.Although EnSCs would form a non-invasive and steady supply of autologous stem cells for women, we must not forget that this means that 50% of the population  is excluded. Furthermore, non-invasive harvesting of EnSCs would not be possible from post-menopausal women . In addition, the issue of EnSCs storage could also develop into the same debates we are facing regarding the storage of cord blood; is there enough scientific evidence to justify storage of EnSCs for future use?clinicaltrials.gov database reveals that several clinical trials regarding EnSCs are underway, with applications ranging from treating critical limb ischaemia (NCT 01558908), liver cirrhosis (NCT01483248), type I diabetes (NCT01496339), and in vitro fertilization (NCT01649752). Hence, it can be envisioned that data from human patients would be available in due course to evaluate the clinical impact of EnSCs.Nevertheless, given the significant progress made in the application of EnSCs in regenerative medicine, it is envisioned that human clinical trials would be conducted in the near future. Indeed, the RECOVER-ERC trial was recently launched by Medistem Inc , which aims to evaluate the potency of EnSCs in 60 heart failure patients in a double-blind placebo controlled Phase II clinical trial . Furtherin situ role highlight these cells as being important for cell and tissue regeneration. This work highlights crucial features of an interesting population of mesenchymal progenitors isolated form endometrial tissue. Cells grown in vitro were characterized by a high clonogenic potential and a long-term survival. Advantages in comparison with MSC from other sources include a greater ease of supply and the protracted availability during a woman’s lifetime with the additional benefit of deriving stem cells from a waste tissue, thus avoiding critical ethical issues.In conclusion, the discovery of EnSCs represents a paradigm shift for the use of adult stem cells by offering an “off the shelf” therapeutic application regenerative medicine. The EnSCs niche and In the light of their capacity to differentiate into mesenchymal tissues and their propensity to undergo genetic manipulation, EnSCs hold considerable promise for novel therapeutic approaches for diseases in the wider field of regenerative medicine.The authors declare that no competing interests exist.JV, AT, AS, AMS conceived and planned the structure of the manuscript. JV, AT, AS, AMS collated and analyzed the data. JV, AT, AS, AMS wrote the manuscript. All authors read and approved the final manuscript.


This study quantified the nutritional and stress physiology of invasive silver carp across broad spatial and temporal scales to define how these fish interact with their environment. Results demonstrated that nutritional properties varied significantly across rivers and sampling time. Stress levels varied only across time periods. Hypophthalmichthys molitrix) inhabiting four large rivers throughout three distinct time periods in the Midwestern USA. Examining the basic biology and ecology of an invasive species is crucial to gain an understanding of the interaction between an organism and its environment. Analysis of the physiological condition of wild-caught silver carp across broad spatial and temporal scales is essential because stress and nutritional parameters can link individuals to their habitats and vary among populations across environments. During each time period, we collected blood samples from individual silver carp in the Illinois River and portions of the Mississippi, Ohio, and Wabash rivers in Illinois. We tested for relationships between silver carp nutrition and stress across rivers, reaches within rivers, and time periods. Principal component analyses separated physiological parameters into a stress component (cortisol and glucose) and two nutritional components representative of short-term feeding  and body energy reserves (cholesterol and protein). Akaike's information criterion suggested that time period had the greatest influence on stress. Stress levels were consistent in all four rivers, and declined across time periods. Akaike's information criterion also suggested that interactions of time period and river had the greatest influence on short-term feeding and body energy reserves. There was no specific pattern across time periods within each river, nor was there a pattern across rivers. Our results provide a better understanding of nutritional and stress conditions in invasive silver carp across a broad landscape and temporal scale, with implications for managing and predicting the spread of this species.We quantified nutritional and stress parameters  in invasive silver carp ( Riverine ecosystems are spatially and temporally dynamic, and this variability may influence habitat quality for fishes. For example, hydrological regimes, nutrient loading, seasonal variation, and human activity can interact, or act independently, to generate lotic habitat characteristics that vary across an extreme range of conditions . Warmer Access to habitat of varying quality may influence decisions by individual fish that relate to reproduction and/or movement, which may ultimately influence abundance, distribution, and mortality. Resource allocation is often shaped by habitat quality, with high-quality habitat providing sufficient food accessibility (maximizing fitness) and low-quality habitat potentially forcing an animal to disperse and find sufficient resources . For exaPhysiological tools are an integral way to quantify organism–environment interactions, particularly when considered over broad spatial scales , and canHypophthalmichthys molitrix) are relatively recent invaders to North America . We selected four reaches with established populations of silver carp in each of the Illinois, Mississippi, and Ohio rivers, and five reaches within the Wabash River for fish collection. These four rivers are all part of the Mississippi River Basin and have extensive agricultural land use, which has increased nutrient levels basin-wide  and the Long-Term Illinois, Mississippi, Ohio, and Wabash River Fish Population Monitoring Program  electrofishing according to methods outlined by −1. The cortisol EIA kit we used has been identified as accurate and precise when used for fishes , cholesterol (in milligrams per decilitre), alkaline phosphatase , and cortisol (in nanograms per millilitres) using the following commercially available kits: EnzyChrom Triglyceride Assay Kit (ETGA-200), EnzyChrom Cholesterol Assay Kit (ECCH-100), and QuantiChrom Alkaline Phosphatase Assay Kit (DALP-250), respectively , and Cortisol EIA Kit . Plasma glucose (in milligrams per decilitre) was determined enzymatically with a microplate spectrophotometer  following the procedure of a priori. Reaches were nested within rivers  to reduce the dimensionality of the data and to quantify relationships between nutritional and stress parameters of silver carp among rivers, among reaches within a river, and over time ; our sampost hoc analysis to aid with visualization of trends in the data  to quantify and rank the best approximating model . Based othe data . We perfP < 0.05, r2 = 0.90); therefore, only total length was included as a variable in analyses. Out of the 504 cortisol readings for silver carp, 297 fish had cortisol values below the detection limit of the kit  = 8.33, P < 0.0001]. Post hoc analysis showed that, in the late summer, silver carp PC1 scores peaked in the Ohio River, with negative short-term feeding scores in the other two time periods  = 7.40, P < 0.0001]. Post hoc analysis showed that silver carp sampled from the Mississippi River had PC2 scores that peaked in the early autumn relative to sampling periods earlier in the year  = 14.50, P < 0.0001]. Post hoc analysis determined that across all rivers, silver carp had the lowest stress scores during the early autumn time period relative to samples collected earlier in the year  during fasting, but recovered after feeding was resumed. Micropterus dolomieu) declined because of fasting during the parental care period of spawning. Mechanisms explaining our observed feeding variability across time periods may include changes in food availability, energetic constraints, or both  across time periods. Optimal metabolic conditions, food intake, growth, and nutrient use for fishes are affected by water temperature; fish generally consume more food during periods of elevated temperatures to balance metabolic costs and optimize growth . For silGadus morhua), while Similar to what was observed for indices of short-term feeding, time period considered independently for each river was the strongest predictor of variation in silver carp body energy reserves . Previous studies have shown that plasma protein concentrations respond to changes in body condition, with nutritionally deprived individuals showing reduced protein reserves relative to feeding individuals . CholestSeasonal differences in food availability and cyclical changes in energy allocation may explain variation in energy stores in silver carp. River temperature can fluctuate greatly, potentially influencing feeding conditions, food availability and quality, and subsequently fish body composition through the allocation of energy resources . Fishes Values of PC3 for silver carp (cortisol and glucose in plasma) were highly influenced by time period, independent of river or reach. Given that PC3 was composed exclusively of stress-related variables, our results indicate that stress levels change seasonally, and in synchrony, across broad spatial scales. Plasma corticosteroid (cortisol) is produced during the primary stress response to various physical and psychological stressors for vertebrate animals. Increased plasma glucose levels provide energy as part of the secondary stress response . SeveralMorone saxatilis), lipid reserves were greater in larger individuals, which was attributable to an increase in lipid mass throughout the growing season  than by size.Interestingly, variables previously shown to influence fish nutrition and stress were not significant drivers of variation for silver carp in our study. Principal components describing short-term feeding (PC1), body energy reserves (PC2), and stress (PC3), were not affected by total body length or by reaches within rivers. Previous studies have shown that total length can influence short-term feeding and body energy reserves in fish (g season . Direct Coregonus clupeaformis) sampled across several locations in Lake Michigan varied across years. Additionally, the rivers we sampled have been greatly altered by anthropogenic influences , potentially homogenizing the effects of reach-scale factors at the spatial scales examined.Likewise, our results also demonstrated that variation in PC scores did not change across reaches within the same river. Previous research has shown that species richness may vary from upstream to downstream locations in rivers, with species richness generally being greater downstream , and thiDespite the findings of our study, there are a number of avenues for future studies to explore related to this topic. For example, the influence of fine-scale habitat features, along with community structure and species diversity and richness, on the health and condition of fishes should be investigated. Examining specific, quantifiable habitat and community characteristics and relating them to nutritional and stress parameters may provide a better comprehension of the link between physiology and invasive species, community structure, and competition, and how interacting factors may influence movement. Other prospective research could address inter-annual variability by continuing similar work over several years for a more comprehensive understanding of silver carp nutritional and stress relationships at a broad spatial and temporal scale, similar to


Crop cultivation in controlled environment plant factories offers great potential to stabilize the yield and quality of agricultural products. However, many crops are currently unsuited to these environments, particularly closed cultivation systems, due to space limitations, low light intensity, high implementation costs, and high energy requirements. A major barrier to closed system cultivation is the high running cost, which necessitates the use of high-margin crops for economic viability. High-value crops include those with enhanced nutritional value or containing additional functional components for pharmaceutical production or with the aim of providing health benefits. In addition, it is important to develop cultivars equipped with growth parameters that are suitable for closed cultivation. Small plant size is of particular importance due to the limited cultivation space. Other advantageous traits are short production cycle, the ability to grow under low light, and high nutriculture availability. Cost-effectiveness is improved from the use of cultivars that are specifically optimized for closed system cultivation. This review describes the features of closed cultivation systems and the potential application of molecular breeding to create crops that are optimized for cost-effectiveness and productivity in closed cultivation systems. Plant factories are divided into the following three production types according to their light sources: (1) solar light, (2) combined solar and artificial light, and (3) perfection-artificial-light  can be achieved using high-pressure sodium lamps and metal halide lamps . If enerIn this review, molecular breeding of crops to enhance their suitability for cost-effectiveness in closed cultivation systems is discussed. Preferable crop traits for closed cultivation systems are also suggested in terms of the potential value.Figure 1). Profitability can be increased by producing crops with enhanced value.Two major considerations of closed cultivation systems are the initial costs and the running costs, with high energy costs of particular concern .Two of the main approaches to increasing the value of a food crop are (1) the accumulation of large quantities of a desirable intrinsic nutrient or reduction of undesirable compounds, and (2) accumulation of valuable compounds that are not normally produced in the crop . It has the unique ability to modify a sour taste into a sweet taste and has potential as a natural, safe, low-calorie alternative to artificial sweeteners for diabetics and people on restricted diets. However, despite its great potential, miracule fruit production is limited because it is a tropical plant that is difficult to cultivate . Moreover, closed cultivation systems that allow stable and uniform production of a crop are extremely suitable for the consistent production of pharmaceutical and industrial compounds using transgenic plants.Overall, molecular farming is an excellent system for utilizing crop characteristics to produce edible medicines with low purification costs . Therefore, many crops developed for cultivation in the field are unsuitable for closed cultivation system. If agricultural crops can be adapted for growth in closed cultivation systems, such as with lettuce, modification of growth patterns to allow high productivity will enhance cost-effectiveness. Furthermore, the specialized cultivars developed for use in closed cultivation systems can be used as host plants for genetic introduction of high-value traits.Features of closed cultivation systems include basal multistage-rack cultivation, soil-free cultivation, optional continuous photoperiod growth, limited light intensity, and low-stress conditions . Plants need to be relatively small to suit the closed system, although the importance of this factor depends on the crop. Field rice and wheat cultivars acquired semi-dwarf traits during the “Green Revolution” (Reduced height1 (Rht1), which is responsible for the semi-dwarf trait in wheat, encodes a negative regulator of the GA response (semi-dwarf1 (sd1), which is responsible for the semi-dwarf trait in rice, encodes the GA synthesis enzyme GA 20-oxidase (uzu is GA-independent. Semi-dwarf barley does not respond to brassinosteroids (BRs), a type of plant steroid hormone. The uzu phenotype is caused by a missense mutation in the BR receptor protein, barley (Hordeum vulgare) HvBRI, a homolog of Arabidopsis BR-insensitive 1 , which has the plant-specific transcriptional repression domain SRDX fused to the C-terminus of AtIBH1, induced a dwarf phenotype in Arabidopsis and tobacco plants, with reduced cell size  is a key gene that affects the developmental phases and architecture of Arabidopsis  gene, which is a homolog of TFL1, regulates switching from vegetative to reproductive phase  cultivar Micro-Tom, which was bred for home gardening purposes, has a miniature growth phenotype. Although Micro-Tom fruits have poor flavor, the cultivar exhibits small features (height approximately 10–20 cm) and can be easily transformed, and it is therefore widely used as a tomato research model (dwarf (d) in the gene encoding the BR biosynthetic enzyme, and miniature (mmt) in a GA signaling-related gene (SPgene. Micro-Tom has a short life cycle (70–90 days from seed germination to fruit maturation) compared with most cultivated tomato varieties (90–110 days) .The tomato (ch model . The dwated gene . Moreove10 days) , exhibit10 days) . Selecte1, Genes that Make Tomatoes2, Tomato Genetics Resource Center3).Luxuriant foliar development and fruit setting under weak light, as well as adaptability to nutriculture systems, contribute to balanced growth in closed environments; however, the factors that determine that these traits are not well-understood. Cultivars or mutants with desirable traits might be identified by screening genetic resources obtained from past breeding programs or from mutagenized populations developed using ethyl methane sulfonate (EMS) or gamma irradiation  analysis, genetic linkage maps, and transgenic techniques . Whole gFigure 1). For example, parthenocarpy, which is the production of fruit without fertilization, could reduce workloads and labor costs by reducing inputs needed for consistent pollination. Parthenocarpy is also valuable in the containment of transgenic plant materials. The jointless trait, where no abscission zones are formed on leaves, flowers, or fruit, would reduce product loss through dropping. The free-standing trait, which allows plants to grow without additional support structures, would be conducive to reducing time and material costs. The jointless and free-standing traits may be particularly useful for cultivation systems that employ moving shelves.In this review, breeding targets were discussed that are likely to be of greatest utility in optimizing plant growth in closed cultivation systems. Several additional traits could also be of use (Since the beginning of agriculture, numerous plants have been developed through artificial selection for field cultivation. Modern closed cultivation systems allow environmental conditions to be tightly controlled and ensure the production. The systems offer great potential for production of specialized crops which have additional value. However, crop breeding specifically for closed cultivation systems had been limited to date despite its potential availability. Novel cultivars are required to sympathize with the challenges of agricultural technology. Breeding an indoor suitable cultivar can be accomplished through complex molecular techniques, which property benefits will familiarize closed cultivation systems.All authors listed, have made substantial, direct and intellectual contribution to the work, and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.