Chronic diseases are the leading causes of morbidity and mortality inEurope, accounting for more than 2/3 of all death causes and 75 % of the healthcarecosts. Heart failure is one of the most prominent, prevalent and complex chronicconditions and is accompanied with multiple other chronic diseases. The currentapproach to care has important shortcomings with respect to diagnosis, treatment andcare processes. A critical aspect of this situation is that interaction betweenstakeholders is limited and chronic diseases are usually addressed inisolation.Health care in Western countries requires an innovative approach toaddress chronic diseases to provide sustainability of care and to limit theexcessive costs that may threaten the current systems. The increasing prevalence ofchronic diseases combined with their enormous economic impact and the increasingshortage of healthcare providers are among the most critical threats. Attempts tosolve these problems have failed, and future limitations in financial resources willresult in much lower quality of care. Thus, changing the approach to care forchronic diseases is of utmost social importance. Chronic diseases are by far the leading causes of mortality andmorbidity in Europe . TogetheA significant contribution to the burden is the co-concurrence ofmultiple chronic diseases (co-morbidities), which are increasingly frequent and are and represents a frequent co-morbidity in patients withother primary diseases. Heart failure care is often further complicated by generalfrailty and a need for highly organised and integrated care. Considering itscomplexity, heart failure care involving multiple specialists is an ideal,representative model to change the strategy of care for multiple chronicdiseases.Heart failure is a chronic, debilitating disease The incidence  of hearDespite improved treatment, heart failure is still associated withdebilitating symptoms, high hospitalisation rates and poor prognosis . This coinreality, this is not the case. Current practice hardly addresses anintegrated management of co-morbidities. Interaction and communication betweenpatients, relatives and caregivers as well as alignment between caregivers indifferent settings are often poor, which may be one of the most important reasonsfor insufficient care [Risk factors for heart failure are overlapping with other chronicdiseases, and therefore, patients with heart failure frequently have manyco-morbidities. More than 40 % of heart failure patients have five or moreco-morbidities , while almost none isfree of any co-morbid condition ,18. Theent care .Aspects such as diagnosis, treatment, follow-up and organisationall play important roles in the care process to optimise outcome with similardifficulties observed across different countries in Europe.Inaccurate or late diagnosis of heart failure, underlyingreason(s) and complications as well as co-morbidities result in poor quality oflife and outcome due to poor prediction of personalised needs, resulting ininadequate treatment and lack of preventing progression. Hence, there is a needfor guidance in determining which diagnostic tests should be performed and inadequately interpreting results.Symptoms of heart failure, like dyspnoea, are often non-specificand do not discriminate heart failure from other diseases. Especially in primarycare where patients present at early stages and symptoms are mild, heart failureis difficult to diagnose .ApproxiTreatment plans for heart failure patients are not adequatelypersonalised and do not consider co-morbidities. This may cause under- orovertreatment, resulting in side effects and poor outcome. There is an urgentneed for personalised treatment plans taking all patient’s characteristics intoaccount.Effective therapies for heart failure are often not utilised in asafe, timely, equitable, patient-centred and efficient manner . This isThe increasing complexity, assisted by developments in theinformation and communication technology (ICT) sector, resulted indecision-support systems for heart failure care. The main goal of such systemsis to improve the clinical decision making process by providing the necessaryknowledge to procure patients with optimal care. Algorithms generaterecommendations to support healthcare professionals. Still, their usage ishampered by different challenges for successful implementation. (1) Integrationinto clinical setting is difficult since care pathways and “ownership” arehardly defined; (2) comprehensibility in terms of ease of use and language isoften not given; (3) systems are mostly used as add-ons to existing care insteadof substitution of care; (4) semantic and technical interoperability of existingsystems as well as with electronic patient records is a major challenge that isnot yet resolved; (5) involvement of patients (or their family) regardingself-management is basically absent. Importantly, these systems are usuallylimited to one specific disease, failing to address co-morbidities andpersonalised needs of patients. Not surprisingly, there is no evidence yet thatsuch systems would improve outcome and there is currently no implementation ofdecision support for personalised treatment. Capabilities and effectiveness ofvarious remote monitoring systems have been reviewed recently, showing theselimitations .Lack of follow-up strategies and patient involvement results inpoor outcomes despite increasing costs. Hence, there is a need to develop andimplement personalised patient support strategies.Once a patient is diagnosed and treatment started, follow-upstrategies involving patients should be developed. Patients have importantresponsibilities including adherence to treatment, making significant lifestylechanges, monitoring themselves and reacting in case of problems. The importanceof this is often recognized, but seldom implemented , and intDifferent programmes have been set up to support patients such asstructured telephone support, self-management support programmes andtele-monitoring, but implementation is limited . These pLacking interactions between stakeholders leads to poorcomprehensibility of the care process. There is a need to facilitatecommunication including patients to personalise management.All stakeholders of an individual patient should collaborate andinteract appropriately. In the care for patients with chronic diseases, manystakeholders are involved Fig. .HoweverCommunication between patients and caregivers is often poor andnot oriented at the individual patient . PatientUnfortunately, interaction between different caregivers involvedin the care for different co-morbidities is often absent. Sometimes specialistsare unaware of the presence of co-morbidities and the co-medication a patient istaking . MoreoveTo improve these shortcomings, there has been a shift in heartfailure management towards a multidisciplinary approach in recent years. Thisshift has led to several randomised controlled trials of multidisciplinary,organised and/or managed care .Most ofAttempts to implement multidisciplinary heart failure care haveonly been partly successful in a few European countries. An important reason forthe poor implementation might be that most programmes are stand-alone, focusingon limited aspects of heart failure care. Lack of standardisation, limitedevidence of cost-effectiveness as well as lack of flexibility and acceptance byhealthcare providers are additional reasons for poor implementation.Clinical evidence retains patients with co-morbidities fromoptimal care. There is a need for evidence-based personalised care to addressco-morbidities of individual patients.To facilitate care for heart failure patients, the EuropeanSociety for Cardiology (ESC) and other authorities developed evidence-basedguidelines . These gMultiple treatment trials have been performed resulting inevidence-based clinical guidelines. These trials applied precisely definedinclusion and exclusion criteria usually not including patients withco-morbidities. Thus, the majority of patients are not properly represented inthe large treatment trials .Hence, Table From all these issues in diagnosis, treatment, chronic care,interactions between stakeholders and available evidence for heart failurepatients with co-morbidities, major underlying issues can be pointed out:firstly, there is lack of interaction, sufficient communication and knowledgeexchange between patients and their caregivers as well as among caregivers withdifferent expertise. Secondly, care is largely unidirectional from individualcaregivers to patients, with virtually absent personal responsibility by thepatients. Thirdly, the lack of clinical evidence for patients with multiplechronic diseases heavily hampers the care of these patients. Fourthly,ICT/tele-medicine plays so far an isolated, limited role and is used in additionto other care instead of as substitution of parts of care, and data exchange isnot integrated in care Fig. .UnfortuCollaboration, interaction and communication between allpersons involved in the care of chronic diseases needs to be substantiallyimproved. The patient must be central in his/her management system, asperson-centred care in heart failure seems to be promising . All invPatient involvement for the development of new ICT platformsis crucial to define the requirements in view of the fact that the typicalpatient with chronic diseases is elderly with difficulties to understand anduse standard ICT equipment. In order to support acceptance and lateradoption of the new system, the innovation process should include not onlythe perspective of the patient, but also the needs of the other stakeholdersin the care of chronic diseases. Constant feedback during the developmentprocess is important.Analysis and integration of multiple guidelines are crucial.Therefore, multidisciplinary committees should be formed to address thisissue. Moreover, multidisciplinary prospective cohorts with (long-term)follow-up are needed.Prediction of outcome should be improved by addressingmultiple chronic diseases and focusing on short-term changes that may beable to predict long-term outcome and respond to personalised treatment.This may allow steering the individualised care process more effectively andefficaciously.Prospective treatment trials addressing multiple chronicdiseases are required.Progress requires not only novel and innovative technologybut also an innovative and novel vision of care and health.Based on the considerations regarding current gaps in personalisedtreatment of patients with heart failure and multiple co-morbidities, severalaspects of health care should be differently addressed in the future . This maObviously, this list cannot be exhaustive in covering all futureneeds. Also, various initiatives and projects are required to address all theseissues as it is impossible to address them all together. Moreover, it may takeyears until significant progress is being made. However, very little attempts arebeing made so far in this regard and a paradigm shift is required in health care.Only then, the imminent problems in health care can be tackled to provide goodquality care at affordable costs for all.Health care in Western countries requires a new innovative approachto address chronic diseases such as heart failure to provide sustainability of careand to limit the excessive costs that may threaten the current system. Theincreasing prevalence of chronic diseases together with their enormous economicimpact and the increasing shortage of health care providers are amongst the mostcritical threats. Attempts to solve these problems have failed so far. Futurelimitations in financial resources will result in a significant reduction in thequality of care. Thus, changing the approach to care of chronic diseases is ofutmost social importance. This needs not only adoption and smarter use of moderntechnology, but also new a vision on both care and health.


Models outlining the neural organization of speech perception  has been associated with increased activation in auditory processing/speech production areas, including left Inferior Frontal Gyrus (IFG), left insula, bilateral ventral Premotor Cortex, right Pre- and Post-Central Gyrus, left anterior Superior Temporal Sulcus and Gyrus (STS/STG), left Planum Temporale (PT), left superior temporal parietal area (Stp), left Supramarginal Gyrus (SMG), and cerebellum bilaterally  is confounded with cognitive load. Note that such confounds also exist for other distortions of the speech signal, such as background noise. Listeners process speech in an unfamiliar accent slower and less efficiently  motor areas during perception of distorted speech signals is not related to actual articulatory processing, but reflects the recruitment of general cognitive processes, such as increased attention, or decision processes (Rodríguez-Fornells et al., Third, The results in Table The neural mechanisms responsible for processing accent variation in speech are not clearly outlined, but constitute a topic of active investigation in the field of speech perception. However, to progress our understanding in this area, future studies should meet several aims to overcome previous design limitations.First, experiments should be designed so that contributions from processing accented speech and effortful processing can be teased apart (Venezia et al., 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.


Many individuals enter the U.S. prison system with a history of high-risk drug and sexual practices, which are two primary contributors to the high rates of HIV and hepatitis cases among this population. As these individuals return to the community, they are likely to continue high-risk behaviors, making it critical that decisionmaking programs designed to reduce risky behaviors be instituted during incarceration close to their release. Prior research on Texas Christian University (TCU) Mapping-Enhanced Counseling  has identified its utility for improving decisionmaking, communication, problem exploration, and personal planning through the use of graphical representations designed to help clients “see” the links and relationships among thoughts, ideas, feelings, and behaviors. It serves as an important tool for helping move traditional, public health-focused educational efforts toward a more comprehensive approach that addresses the challenges of personal decisionmaking around sex and drug use behaviors.WaySafe, designed to increase positive decisionmaking skills among offenders for healthy living, including skills for reducing disease risk behaviors. WaySafe uses TCU Mapping-Enhanced Counseling to focus on the cognitive aspects of risky sexual and drug use behaviors during re-entry to improve problem recognition, commitment to change, and strategies for avoiding behavioral risks of infections. WaySafe includes six 1-hour, highly interactive sessions to increase motivation and planning. A total of 1398 incarcerated offenders from eight different correctional facilities in two states participated in the study during the last phase of their in-prison substance abuse treatment prior to transition back to the community.The TCU Disease Risk Reduction (DRR-1) project developed and tested an intervention, WaySafe intervention or treatment as usual (TAU). At the completion of the 6-week WaySafe curriculum, participating offenders completed follow-up surveys. Baseline and follow-up surveys included measures of knowledge, confidence, and motivation regarding general HIV information, risky sex and drug use, what to do if exposed, and general life skills. Results supported hypotheses that offenders in WaySafe improved significantly more than TAU clients on knowledge, attitudes, and behavioral intentions to avoid risks. Finding new methods to reinforce the lessons learned in the community is a critical next step, because these changes in attitudes and intentions will degrade over time with the stress and pressures of transitioning from prison to the community. Reinforcing lessons learned while offenders are currently facing risky situations may be helpful.All participating offenders completed baseline surveys and then were randomly assigned to the Clinicaltrials.gov NCT01900210


Dear Editor,Although it has been gradually recognized that a range of low-dose (or low-dose-rate) radiation effects on living cells are possible key factors in evaluating ‘low-dose radiation risk’, there remains little of the coherence required among robust data that can be used with confidence in risk assessments . It is tAt present, residents of Fukushima, especially those in Hamadōri, the eastern part of Fukushima, continue to be gripped by fear of contamination by radioactive substances; however, regardless of difficulties, medical personnel continue to cultivate close relationships with the patients. From previous statistical data, the long-term risk of cancer from exposure to radiation clearly increases at doses exceeding 100 mSv; therefore, following Japan's worst-ever accident, at the Fukushima Daiichi nuclear power plant (FNPP), radiation doses have been measured among Fukushima residents through the use of both personal dosimeters  and whole-body counters   ) . A numbeV gene]) . PatientV gene]) . As showV gene]) –14. AccoV gene]) , suggestIn conclusion, given the influence of precision medicine in this matter, we are concerned about the use of radiation exposure levels as the sole criterion in evaluating cancer risk among residents living near the FNPP accident site. The potential risk of radiation-induced cancer in Fukushima could be small at the population level due to the scarcity of patients and heterozygous carriers with radiosensitivity disorders; however, it should not be ignored . We hereFunding to pay the Open Access publication charges for this letter was provided by Hisanori Fukunaga.


Transradial access has now become a standard of care for percutaneous coronary angiography and interventions. We are currently dealing with a switch from transfemoral procedures towards transradial procedures because of several advantages related to the transradial catheterisation procedure, such as the lower number of access-related complications and early mobilisation. Recently, Valgimigli et al. suggested that radial access should become the default approach in patients at higher risk such as those with an acute coronary syndrome, with or without ST-segment elevation, undergoing invasive management. They observed that radial as compared with femoral access significantly reduces the rate of net adverse clinical events, defined as the composite of major adverse cardiovascular events or major bleeding. Moreover, the benefit of radial access is pronounced in highest-volume radial centres .Radial artery occlusion (RAO) is considered the most common complication of the radial access, affecting 1.5–33 % of patients early after the procedure . The truZwaan, Koopman and colleagues provide a systematic review and meta-analysis about the current knowledge on the incidences of access-site complications and upper extremity dysfunction following TR-PCP. From a selection of 176 eligible papers, the authors reported a mean incidence of known complications of up to 9.6 %, which mainly consists of radial occlusion, radial spasm, swelling and haematoma. The incidence of upper extremity dysfunction after TR-PCP is up to 0.8 % even though it is rarely or not sufficiently investigated. Therefore, this reported low incidence could be underestimated due to the under-reported data in the literature. Moreover, the incidence of complications is often published by centres with more experience in using the transradial approach, where there are usually less access-site complications .p = 0.014), probably related to major bleeding caused by the higher dosage of anticoagulants during PCI. Furthermore, the trend is not beneficial for early radial occlusion (4.9 %) and transradial coronary artery angiography (p = 0.060), as an effect of multiple catheter switches [Additionally, the reported meta-regression analyses showed a significant positive correlation between transradial PCI and compartment syndrome p = 0.01, probablThe authors emphasised that access-site complications might have a direct relationship with upper extremity dysfunction. Although few data have been reported about it, the authors have hypothesised that transitory ischaemia of the nerve due to radial spasm and irreversible nerve damage with a potentially debilitating outcome could occur. So, early detection and referral for any complications could be important to reduce their effect on upper limb function .Despite the above-mentioned accurate analysis, it is interesting to consider what Van Leeuwen and colleagues recently reported. They investigated the effect of the transradial procedure on upper limb function demonstrating that there was no significant difference between radial and femoral access with respect to upper limb function, cold intolerance, or other procedure-related extremity complaints. It is an important finding, especially for those patients for whom optimal upper extremity function is essential .. Moreover, RAO may reduce the opportunities to use transradial access in further procedures, losing the advantage of the radial artery as a superficial access site for cardiac catheterisation [Nevertheless, a limitation of the radial approach is still RAO, which may become symptomatic in some patientsrisation .The ability to recognise and treat complications of radial access is part of radial practice. It would be opportune that RAO is assessed at discharge and during follow-up, preferably by ultrasound. If unnoticed, complications could cause severe disability, with possible significant socioeconomic consequences.Imaging data detected by optical coherence tomography and intravascular ultrasound as well as histological and functional studies have demonstrated significant structural and functional changes to the radial artery post transradial catheterisation, resulting in significant endothelial cell dysfunction. Minimising radial injury and occlusion should be an important component of transradial procedures and consists of endothelial protection, minimisation of endothelial wall trauma and minimisation of thrombus formation. Therefore, radial injury may be minimised through the use of smaller diameter guide catheters or through the use of sheathless guide catheters, reduction of catheter exchanges and minimisation of radial spasm periprocedurally, either through the use of spasmolytic drugs or through the use of coated sheaths/guide catheters. Thrombus formation within radial artery post transradial procedures may be reduced by routine anticoagulation with heparin periprocedurally and with non-occlusive radial artery compression postprocedurally .The growing experience of transradial procedures and interventions is associated with a lower rate of access-site bleeding and vascular complications, improving clinical outcomes. Therefore the radial artery is an advantageous site of access and to use it in a safe way it is necessary to know its anatomy variability, the physiology and the eventual complications.None.None declared.


The correct functioning of the nervous system depends on the exquisitely fine control of neuronal excitability and synaptic plasticity, which relies on an intricate network of protein-protein interactions and signaling that shapes neuronal homeostasis during development and in adulthood. In this complex scenario, Kinase D interacting substrate of 220 kDa/ankyrin repeat-rich membrane spanning (Kidins220/ARMS) acts as a multi-functional scaffold protein with preferential expression in the nervous system. Engaged in a plethora of interactions with membrane receptors, cytosolic signaling components and cytoskeletal proteins, Kidins220/ARMS is implicated in numerous cellular functions including neuronal survival, neurite outgrowth and maturation and neuronal activity, often in the context of neurotrophin (NT) signaling pathways. Recent studies have highlighted a number of cell- and context-specific roles for this protein in the control of synaptic transmission and neuronal excitability, which are at present far from being completely understood. In addition, some evidence has began to emerge, linking alterations of Kidins220 expression to the onset of various neurodegenerative diseases and neuropsychiatric disorders. In this review, we present a concise summary of our fragmentary knowledge of Kidins220/ARMS biological functions, focusing on the mechanism(s) by which it controls various aspects of neuronal activity. We have tried, where possible, to discuss the available evidence in the wider context of NT-mediated regulation, and to outline emerging roles of Kidins220/ARMS in human pathologies. NTR [PSD-95/Disc large/Zonula occludens-1 (PDZ)-binding motif]  is a membrane protein preferentially expressed in the nervous system, where it modulates several crucial aspects of neuronal physiology such as cell survival, differentiation into axons and dendrites, and synaptic plasticity  elicited by acutely applied BDNF  synapses was increased in 3–6-month-old ARMS+/− mice  kinase pathway  receptors  was associated with GABAergic hippocampal neurons, but apparently absent in glutamatergic neurons. Inhibitory post-synaptic currents (IPSCs) of −/−Kidins220 neurons recovered considerably faster from synaptic depression than those recorded from wild-type neurons  target of proteolytic degradation mediated by Ca2+-dependent calpain proteases, the activation of which is triggered by neuronal activity. Calpains do not degrade their targets completely, unlike typical cellular proteases, but function to regulate their activity by partial cleavage, thereby contributing to synaptic plasticity and neurotoxicity  receptor (SNARE) proteins, glutamate receptor subunits, protein kinases, cytoskeletal and other scaffold proteins, just to name a few  channels 293 cells. Recordings of sodium currents mediated by heterologously expressed Nav1.2 further revealed dramatically slowed channel kinetics and shifted voltage-dependence in Kidins220-coexpressing cells, indicating that Kidins220 association can have unexpectedly strong effects on both Nav channel activation and fast inactivation processes pathologies. Although the literature concerning this aspect of Kidins220 function is still limited, several studies have started to investigate the genetic and molecular pathways linking Kidins220 to the onset of various diseases. For example, Kidins220 is overexpressed in human samples of melanoma  samples  patients revealed that Kidins220 levels were significantly decreased compared to controls  and as an immediate down-stream target of NT receptor kinases, has emerged as a novel player in the control of neuronal activity. However, our understanding of this role is currently far from being complete, and it is also unclear how it relates to the recent identification of Kidins220 in several neurological diseases. This summary may serve to focus on gaps in our knowledge on the subject and possibly to guide future investigations.de facto part of TrkB/BDNF signaling events and which are not. There is a multitude of mechanisms by which BDNF impacts on synaptic transmission and membrane excitability and which might also involve Kidins220. In fact, the list of ion channels targeted by the BDNF signaling pathway has grown considerably in recent years and now includes, apart from the above-mentioned NMDARs .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.


Inhibitory circuits play an important role in synaptic plasticity during development and adulthood. Changes in interneuronal activity induce structural and synaptic rearrangements of inhibitory interneurons, network oscillations, and homeostatic plasticity. In addition to epileptic seizures, deficits in the inhibitory system lead to aberrant information processing and cognitive impairment in various neurological disorders. Studies exploring the structural and functional plasticity of interneurons are essential, not only to understand the mechanisms underlying normal development and behavior but also, to identify the etiology of different psychiatric and neurological disorders to pursue new therapies. Here, it will be discussed the role of inhibitory circuit in the synaptic plasticity, and how cellular replacement strategies can remodel changes in circuit function and homeostasis in the context of brain repair.Most of the gamma-aminobutyric acid-containing (GABAergic) interneurons in the cerebral cortex and hippocampus originate from three progenitor regions in the embryonic subpallium: caudal ganglionic eminence (CGE), medial ganglionic eminence (MGE), and preoptic area (POA) –4. Each In the cortex and hippocampus, interneurons subtypes differ in their functional connectivity and generate differentially timed inhibition at distinct sites of postsynaptic cells –8. InterBesides the direct control of inhibition, synchrony and plasticity on principal cells, different interneuron subtypes exhibit a wide range of responses to different neuromodulatory inputs, leading to changes in net inhibition, synchronization, and synaptic plasticity . The innInterneuron subtypes differ in their functional connectivity to the principal cells in cortex and hippocampus , 8, provAs mentioned above, GABAergic interneurons targeting dendritic domains control the efficacy and plasticity of excitatory inputs onto principal neurons. This dendritic remodeling of inhibitory neurons, either in normal or pathological conditions, affects activity-dependent modulation of neuronal connectivity within local circuits . ReductiA receptor kinetics are likely to be needed to achieve precise timing gamma oscillations in cortex and hippocampus  (Ccnd2−/−)  has been performed in animal models. The studies had shown that MGE-derived cells are able to survive, differentiate in mature interneurons, migrate, and functionally integrate through the host brain parenchyma with low risk of promoting brain tumors. The integrated mature interneurons modify the neuronal network, rescuing the normal functional inhibition and the synaptic plasticity –44. SomeDlx1−/−)  and cyclcnd2−/−) . Dlx1−/−hibition , with coenotypes .A receptors containing the α1 subunit, abundant at synapses mediated by PV-expressing interneurons (The maturation of GABAergic interneurons, specifically PV- and SOM-expressing subtypes, has been strongly implicated in critical period of plasticity, such as the development of visual cortex , 46. It rneurons , 51.Changes in GABAergic function mediated by PV- and SOM-expressing interneurons impair synaptic plasticity and disrupt network organization and normal brain oscillations. However, by temporal and selective coordination of interneuronal activity, it is possible to modulate interneuronal network and increase GABA release on different subcellular domains of target cells to maintain or rescue normal network oscillations and synaptic plasticity. The investigation of PV- and SOM-expressing interneurons, using strategies that modify interneuronal network and regulate inhibitory input/output of such interneurons at different time points, as selective target them or using precursor cells grafts, are helping us to better understand the role of these specific interneuron subtypes in controlling and restoring synaptic plasticity and brain oscillations during development and adulthood.The author confirms being the sole contributor of this work and approved it for publication.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


MicroRNAs (miRNAs) are small non-coding RNA molecules that modulate expression of the majority of genes by inhibiting protein translation. Growing literature has identified functional roles for miRNAs across a broad range of biological processes. As such, miRNAs are recognized as potential disease biomarkers and novel targets for therapies. While several miRNA-targeted therapies are currently in clinical trials , no therapies have targeted miRNAs in respiratory diseases in the clinic. In this mini-review, we review the current knowledge on miRNA expression and function in respiratory diseases, intervention strategies to target miRNA function, and considerations specific to respiratory diseases. Altered miRNA expression profiles have been reported in a number of respiratory diseases, including asthma, chronic obstructive pulmonary disease, cystic fibrosis, and idiopathic pulmonary fibrosis. These include alterations in isolated lung tissue, as well as sputum, bronchoalveolar lavage fluids and peripheral blood or serum. The observed alterations in easily accessible body fluids  have been proposed as new biomarkers that may inform disease diagnosis and patient management. In a subset of studies, miRNA-targeted interventions also improved disease outcomes, indicating functional roles for altered miRNA expression in disease pathogenesis. In fact, direct administration of miRNA-targeting molecules to the lung has yielded promising results in a number of animal models. The ability to directly administer compounds to the lung holds considerable promise and may limit potential off-target effects and side effects caused by the systemic administration required to treat other diseases. More than 1000 unique miRNAs are present in the human genome  in length  of mRNA transcripts -induced asthma model  entered Phase II clinical trials in 2010 for the treatment of hepatitis C virus infection . These molecules are often comprised of locked nucleic acid backbones with specific modifications to increase stability in vitro and in preclinical animal models. However, this approach also requires delivery of a transgene-encoding vector to achieve stable inhibition . This approach provides the advantage of being able to tailor dosing and withdraw treatment if complications arise. In respiratory diseases, direct tissue administration by aerosol provides targeted delivery to lung tissue, with minimal dissemination into the circulation and systemic tissues. This is likely to limit side effects, compared to systemic delivery. However, this also means that these therapies will only be effective if altering miRNA function specifically within the lung is sufficient to dampen disease processes. Even with direct administration into the lung, efficient delivery to target cell populations remains difficult, as seen in siRNA-based therapies  add additional complications relating to the expression vector itself. Viral delivery platforms have been associated with direct toxicity, increased risks associated with genomic integration events and the inability to discontinue treatment if problems do arise (Ibrahim et al., Increasing numbers of publications have identified differences in miRNA expression and functional roles for miRNAs across a range of respiratory diseases. MiRNAs have been proposed as biomarkers of disease and potential novel therapeutic targets for treatment. In respiratory diseases, direct administration of miRNA-targeting drugs into the lungs may allow tissue-specific targeting, while limiting side effects resulting from systemic delivery. Continued mechanistic studies are required to optimize drug delivery systems and demonstrate acceptable efficacy and safety for translation into the clinic. While miRNA-targeting approaches hold promise, no studies have yet assessed impacts in human respiratory diseases.SM, MP, HT, AC, and PF wrote and edited the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


MicroRNAs comprise a large family of short, non-coding RNAs that are present in most eukaryotic organisms and are typically involved in downregulating the expression of protein-coding genes. The detailed mechanisms of miRNA functioning in animals and plants have been under investigation for more than decade. In mammalian cells, miRNA guides the effector complex miRISC to bind with partially complementary sequences, usually within the 3′UTR of mRNAs, and inhibit protein synthesis with or without transcript degradation. In addition to these main mechanisms, several other modes of miRNA-mediated gene expression regulation have been described, but their scale and importance remain a matter of debate. In this review, we briefly summarize the pathway of miRNA precursor processing during miRNA biogenesis and continue with the description of the miRISC assembly process. Then, we present the miRNA-mediated mechanisms of gene expression regulation in detail, and we gather information concerning the proteins involved in these processes. In addition, we briefly refer to the current applications of miRNA mechanisms in therapeutic strategies. Finally, we highlight some of the remaining controversies surrounding the regulation of mammalian gene expression by miRNAs. MicroRNAs (miRNAs) constitute a large family of short, non-coding RNAs (ncRNAs) (~22 nucleotides long) that are common in single-celled eukaryotes, plant and animal cells , and havMicroRNAs play a crucial role in the post-transcriptional regulation of gene expression, mostly involving gene silencing. These tiny RNAs are generated in cells through a biogenesis pathway that involves the two-step enzymatic processing of genome-encoded primary miRNA transcripts into short miRNA/miRNA* duplexes. During assembly of the miRNA-induced silencing complex (miRISC), the miRNA/miRNA* duplex is loaded into the Argonaute protein (Ago), and one of the strands  is released and degraded. Consequently, the bound miRNA strand (the “guide strand”) guides miRISC to interact with partially complementary sequences in target transcripts  and mainly triggers mRNA deadenylation and degradation or translation inhibition. However, several other mechanisms of gene expression regulation by miRNAs have also been described. The key attribute of miRNA is imperfect base pairing within miRNA/miRNA* duplexes as well as within duplexes formed by the miRNA and its target mRNA. It is estimated that most protein-coding genes are regulated by miRNAs , 6. ThusIn this review, after gathering the relevant information on miRNA biogenesis and miRISC assembly, we focus on the recent understanding of cellular mechanisms of gene expression regulation by miRNAs. This topic still arouses researchers’ curiosity because of the controversies around a detailed scheme of miRNA-mediated mechanisms.The canonical biogenesis pathway of mammalian miRNAs Fig. a is a twRNase III enzymes (Drosha and Dicer) cleave miRNA precursors imprecisely and generate heterogeneous products that often differ in length , 34–36. Apart from isomiRs, the pool of miRNAs may be varied by the competitive processes of post-transcriptional, non-templated oligo-adenylation , 40–42 oDrosophila melanogaster cells are sorted into different Ago paralogs ( a a188] anThe mechanism of deadenylation is being carefully studied, including the structural aspects of the protein-protein interactions, to create a detailed model of deadenylation-dependent repression and better understand this process . IDeadenylation may be continued by transcript degradation either via decapping by DCP1–DCP2 complex activity Fig. c and theD. melanogaster and human cells, which indicated that miRNA-mediated translational repression and degradation are mechanistically linked through the interactions of GW182 proteins with PABP and deadenylases [According to the previous suggestions , 233 andenylases . It coulFollowing the trend of epigenetic research, increasing attention has recently been paid to short non-coding RNAs  that may regulate transcription . ISurprisingly, a recently published work described a TNRC6A paralog that possesses both a nuclear localization signal (NLS) and nuclear export signal (NES) . These sTaken together, these results strongly suggest the importance of Ago protein localization and activity in the nucleus Fig. g. Thus fBased on the results described above and the widespread effect of chromatin remodeling on the regulation of gene expression, we anticipate that future research will reveal more examples of miRNAs participating in nuclear processes.The complexity of the abundant miRNA-mRNA interactions arises from the fact that one mRNA may harbor binding sites for numerous different miRNAs , 251; alThe precise modulation as well as reversal of such miRNA alterations is not only a challenge for fundamental research but also a promising strategy for miRNA-based therapy. Such therapy might be achieved by introducing miRNA blockers  or miRNA mimics  that may target dysregulated miRNA pathways. Recently, an efficient miRNA-based drug against hepatitis C  was tested in the second phase of clinical trials , 254.The knowledge of miRNA-mediated mechanisms is also very useful for designing improved RNAi-based therapeutic tools. A popular trend is the use of pri-/pre-miRNA-based shRNA expression cassettes for more efficient and longer lasting gene silencing . AAlthough important advances have been made in miRNA research over the past several years, we still remain unable to profoundly comprehend the miRNA-based cellular processes. Numerous detailed questions remain unanswered, and among them are the following: Which factor is most important in determining the effect of miRNA activity? Is it the position of miRNA/mRNA mismatches? Is it the localization and number of miRNA-binding sites? How miRNAs within miRISCs find their binding sites on the transcript sequence is also poorly understood. Furthermore, it is not clear whether we already know all of the critical factors involved in mature miRISC formation and miRNA functioning in the cell.As new mechanisms  are discovered, the question of whether they are widespread is of great importance. The results indicate that there is no single pathway of miRNA-mediated regulation common to all miRNAs in human cells. Moreover, the contradictory findings concerning the specific steps of translation that are inhibited via miRNAs remain very controversial. It seems that the variety of systems used to perform miRNA research might strongly contribute to the observed discrepancies.Nevertheless, in agreement with recent results, the prevailing opinion concerning miRNA-mediated mechanisms is that silencing is mainly caused by transcript decay, which follows mRNA deadenylation. Thus, much effort has been made to identify the proteins involved in this process as well as interactions that take place between these proteins. The relevant information is collected in Table More generally, we consider that multiple miRNA-mediated mechanisms may operate in cells depending on, i.e., the nature of the miRNA-mRNA interactions and the type of cell or its physiological state. In a single cell, there are abundant interactions among mRNAs, miRNAs and their variants as well as various protein factors competing for interactions with miRISC. Therefore, the notion that the specific miRNA/mRNA duplex may activate not one but two or more different silencing mechanisms concomitantly  cannot be ruled out.Over the last few years, a growing body of evidence has formed that supports the existence of non-canonical miRNA-binding sites, in a sense of: miRNA pairings with targets, localization within the CDS and interactions with noncoding RNAs. This notion needs to be implemented into next-generation MRE prediction algorithms and taken into account in analyses of relevant deep sequencing data. Also worth highlighting is the fact that miRNA binding to non-canonical targets can modulate the levels of miRNAs and their canonical functions. Together, it is also likely that additional new functions of miRNAs await to be disclosed.


Aggression is a core feature of borderline personality disorder (BPD). Well-replicated results from the general population indicate that men engage in aggression more frequently than women. This article addresses the question of whether gender also influences aggression in BPD, and whether the neurobiological mechanisms underlying aggressive behavior differ between male and female BPD patients. Data show that most self-reports, interviews and behavioral tasks investigating samples of BPD patients do not find enhanced aggressiveness in male patients, suggesting that BPD attenuates rather than aggravates gender differences usually present in the general population. Neurobiological studies comparing BPD patients with gender-matched healthy controls, however, reveal a number of interesting gender differences: On the one hand, there are well-replicated findings of reduced amygdala and hippocampal gray matter volumes in female BPD patients, while these findings are not shared by male patients with BPD. On the other hand, only male BPD patients exhibit reduced gray matter volume of the anterior cingulate cortex, increased gray matter volume of the putamen, reduced striatal activity during an aggression task, and a more pronounced deficit in central serotonergic responsivity. These neurobiological findings point to a particular importance of impulsivity for the aggression of male BPD patients. Limitations include the need to control for confounding influences of comorbidities, particularly as male BPD patients have been consistently found to show higher percentages of aggression-predisposing comorbid disorders, such as antisocial personality disorder, than female BPD patients. In the future, studies which include systematic comparisons between females and males are warranted in order to disentangle gender differences in aggression of BPD patients with the aim of establishing gender-sensitive treatments where needed.The online version of this article (doi:10.1186/s40479-015-0028-7) contains supplementary material, which is available to authorized users. Aggression may be defined as any behavior directed toward another individual with the intention to cause harm . Dysregud) depending on the method of data collection and the form of aggressive behavior: The effect size is stronger in real-world settings and varies as a function of the seriousness of the aggressive act, i.e., the more dangerous the behavior, the stronger the male preponderance . Similar to the female-only association between BPD traits and verbal aggression reported above, this finding marks the importance of considering the specific characteristics of the studied behavior when evaluating gender differences in aggression.Several studies using semi-structured interviews investigated whether male and female BPD patients differ in the prevalence of comorbid disorders that predispose them to aggression, such as ASPD. Male BPD patients were more frequently found to meet the diagnostic criteria of ASPD than female patients, irrespective of whether symptomatology was assessed dimensionally  or categStudies have repeatedly found more aggressive responses on the PSAP in patients with BPD compared to healthy individuals ,29,45. HTaken together, results from self-report and interview-based measurements and behavioral tasks in BPD patients show a different pattern than in the general population. In the general population, men have been found to engage in more aggression than women, whereas in BPD, most studies did not find such a gender difference in aggression. Findings reporting more verbal aggression in female subjects scoring high on BPD traits than their male counterparts emphasize the importance of differentiating between specific forms of aggression. Male BPD patients have been consistently found to show higher percentages of aggression-predisposing disorders such as ASPD. Research on intimate partner aggression has concentrated on male-to-female aggression and indicates a predisposing effect of BPD on intimate partner aggression, which might stem from different mate retention tactics. However, as the number of studies is limited, and as results are often confounded by comorbidity and grouping together of different forms of aggression, further research is urgently needed.Aggressive behavior is understood to result from biological as well as environmental vulnerabilities  than female patients . No diffInterestingly, in a previous analysis of the same data by Perez-Rodriguez and colleagues , no gendFunctional neuroimaging studies that analyzed only male BPD patients with comorbid ASPD found increased amygdala activity exclusively in response to high, but not to neutral and low emotionally salient stimuli . This isNeurochemically, the central serotonin system seems to differ substantially between male and female BPD patients. Using d-fenfluramine or m-Chlorophenylpiperazine to test the responsivity of the serotonin system, compared to gender-matched healthy volunteers, only male but not female BPD patients showed a diminished serotonergic response in most ,73,74 buInterestingly, a recent study found increased serotonin-2A receptor binding in female BPD patients compared to gender-matched controls and male BPD patients , which iSerotonergic x environment interactions may additionally be moderated by gender. For instance, the association between the short allele of the serotonin transporter gene and laboratory measured aggression was more prominent in men than in women . ContrarOther neurochemical systems might additionally play a role in the differential regulation of aggression between male and female BPD patients. In a sample of personality-disordered subjects, including BPD patients, a positive correlation was found between the concentration of vasopressin in the cerebrospinal fluid and aggression, which was stronger in male than in female subjects . Again, In the general population, a weak positive association between concentrations of testosterone in different body fluids  and antisocial, dominant, or competitive behaviors has been suggested . TestostIn sum, results from neurobiological studies suggest a different pattern of alterations between male and female BPD patients. Male but not female BPD patients when compared to gender-matched healthy volunteers exhibited reduced gray matter volume of the anterior cingulate cortex, increased gray matter volume of the putamen, reduced striatal activity during an aggression task, and a more pronounced deficit in central serotonergic responsivity. Specific alterations found in female BPD patients are rare. The few findings that exist raise the question of whether dysfunctions in the amygdala are more pronounced in female than in male BPD patients.This article addressed the question of gender differences in BPD patients’ aggressive behavior. In the general population, men show enhanced aggression compared with women. By contrast, most results from self-reports, interviews, and behavioral tasks do not indicate any gender differences in the patients’ aggressiveness. Taken together, these results suggest that BPD attenuates rather than aggravates the gender difference in aggression normally present in the general population.This article also reviewed gender differences in the neurobiological underpinnings of BPD patients’ aggression. Conclusions from these findings require a conceptual background: Aggression is a multi-faceted social behavior ,85, whicWithin this framework, the findings reviewed above may point to a particular importance of the biobehavioral dimension of impulsivity for male BPD patients’ aggressiveness. Most of the neurobiological alterations that have been reported in male but not in female BPD patients in comparison to gender-matched healthy volunteers, namely the volume loss of the anterior cingulate cortex ,87, the As mentioned above, specific alterations in female BPD patients are rare. Hypothesizing that dysfunctions of the amygdala might be more prominent in female than in male patients, one could speculate that affective dysregulation  may playThe following limitations should be noted. First, studies analyzing mixed-gender samples are scarce. To exclude a potential confounding influence of gender, the majority of the neurobiological mixed-gender studies reported here compared the patients with gender-matched healthy controls, e.g. male BPD patients with male controls. To our knowledge, only two neuroimaging studies ,63 have Comparing male directly with female BPD patients may also be of importance in terms of gender-specific treatments. For instance, psychopharmacological substances targeting the serotonin system may lead to different effects in male versus female BPD patients. However, the – at least to our knowledge – only study to date to have used the serotonin reuptake inhibitor fluoxetine to treat male and female patients with intermittent explosive disorder, and in part comorbid BPD, did not find a significant drug x gender interaction . PsychotUpcoming studies may also benefit from considering results of neurobiological gender differences found in precursor syndromes associated with aggression and BPD, such as conduct disorder and attention deficit hyperactivity disorder. Male patients with conduct disorder exhibited larger volumes of the anterior insula than their female counterparts . Men affSecond, results in the general population emphasize the importance of considering different contributions to gender differences depending on the form of aggression under focus ,15. UndeThird, the potential confounding effect of comorbidities needs to be stressed. As pointed out throughout the article, male BPD patients frequently suffer from higher prevalence rates of aggression-predisposing disorders such as ASPD. Future studies should control this influence, e.g. by analyzing matched samples. The confounding influence of comorbidities also implies the use of a dimensional rather than a categorical approach to BPD psychopathology, as has been proposed by the alternative DSM-5 model of BPD  and recewww.kfo256.de; Schmahl et al., 2014: He 2660/12-1; He 2660/7-2) as well as financial support by the German Research Foundation and Ruprecht-Karls-Universität Heidelberg within the funding program Open Access Publishing.This work was supported by two grants of the German Research Foundation to S. C. H. on borderline personality disorder (Clinical Research Foundation on Mechanisms of Disturbed Emotion Processing in Borderline Personality Disorder,


Despite advances in HIV commercial therapies, including fixed-dose combinations, unprecedented proportions of HIV patients remain without therapeutic options due to lack of access to treatment and epidemiologically significant proportions of multi-class drug resistance that are driving morbidity levels in both developed and resource-poor settings. The need to emphasize the development of therapeutic options with the potential to abrogate these correlating trends are receiving insufficient dedication of resources, as new clinical strategies do not address these enduring disparities. GenePro, a therapeutic vaccine candidate, has shown preclinical efficacy via publicly sponsored research that demonstrates the potential to address a current therapeutic issue.The GenePro therapeutic construct includes strong lentiviral promoter and 7 out of 9 HIV genes: Gag, pol, tat, rev, vpu, env and nef. In a preclinical study, kinetic analyses on splenocytes of BALB/c mice that were immunized by a single injection with a GenePro were performed. IFN-gamma-ELISPOT and multiparametric FACS analysis were conducted to characterize the induced CMI response. Animals immunized with a single high dose of GenePro were monitored longitudinally for vaccine-induced IR using multiparametric flow cytometry-based assays.Comprehensive analysis demonstrated the capacity of a single high dose of HIV DNA vaccine alone to induce long-lasting and polyfunctional T-cell responses in the nonhuman primate model. In preclinical studies, GenePro has been found to generate the HIV-specific memory cells found in elite controllers and reconstitution of the immune system. The TriGrid delivery system increases DNA delivery efficiency by up to 1,000 fold. This approach has less toxicity and no drug resistance and may allow for STIs from ART.Data from initial investigations of GenePro indicate improvements in immune response and generation of HIV-specific memory cells that induced immune responses by altered HIV DNA constructs, which synthetically mimic virologic responses of elite controllers. Advancement of GenePro into human studies will serve as a hallmark in continuing commercial efforts to discover a universal address for the global AIDS crisis.


PLOS Genetics, uses complementary in vivo and in vitro experiments to challenge this view and show that endocytosis of the Roundabout (Robo) receptor is a key component of receptor activation and precedes the recruitment of its effectors to the receptor cytoplasmic domain [Proper guidance and wiring of axons during development is essential for the normal function of the nervous system. Despite the identification, over the last 20 years or so, of many axon guidance cues and their cognate receptors, we still do not fully understand how their intracellular signal transduction is elicited. Current models often depict that ligand binding to its receptor at the surface of the growth cone leads to the activation of the receptor, recruitment, and/or activation of signaling molecules near the cytoplasmic tail of the receptor, resulting in local signaling emanating from the subcellular compartment where the receptor was located upon binding of its ligand . An artic domain .Drosophila embryonic ventral nerve cord, Slit is expressed by midline glia, which creates a repulsive zone for growth cones expressing Robo at their surface. In robo mutants, axons that normally do not cross the midline now ignore the presence of Slits and cross or re-cross the midline—as in a roundabout, hence giving the name to the Robo family of receptors.In both invertebrates and vertebrates, Robo receptors comprise a family of axon guidance receptors that mediate repulsion in response to their Slit ligands. In the Endocytosis of axon guidance receptors has previously been shown to modulate axon guidance receptor activity and signaling. For example, endocytic trafficking of Robo by Commissureless prevents its delivery to the growth cone surface. In addition, ligand-dependent receptor endocytosis can be an essential component of growth cone desensitization to guidance cues . This phDrosophila genetics to demonstrate an interaction between Slit-Robo signaling and endocytic pathway components. Specifically, they show that mutants for genes involved in Clathrin-dependent endocytosis , or for genes that regulate entry into the early (Rab5) or late (Rab7) endosome, enhance Slit-Robo signaling defects. These results are consistent with endocytosis contributing to Robo activation, as opposed to a change of Robo surface levels available to bind Slit. Interestingly, these endocytic pathway components do not appear to genetically interact with the Netrin receptor Frazzled/DCC, indicating specificity.In this study, Chance and Bashaw first use Drosophila cells. Upon Slit ligand stimulation, these cells rapidly undergo spreading of numerous processes in a Robo-dependent manner. In parallel to these morphological changes, Robo is rapidly internalized from the plasma membrane to Rab5-positive early endosomes. Preventing this Slit-dependent trafficking of Robo by either manipulating the endocytic trafficking machinery  or mutating conserved AP-2 Clathrin adapter binding sites in the Robo receptor cytoplasmic domain blocks Slit-dependent changes in cell morphology.To investigate the molecular mechanism underlying the endocytosis of Robo and, importantly, whether Robo is itself endocytosed in a Clathrin-dependent manner, they next turned to an in vitro assay using These results are consistent with a role for Clathrin-dependent endocytosis in Robo’s ability to elicit signaling, but what effector protein mediates this signaling? The Ras/Rho guanine-nucleotide exchange factor (GEF) Son of Sevenless (Sos) was previously shown to be recruited to Robo in response to Slit stimulation in vitro and to be required for Robo-mediated midline repulsion in vivo . Using trobo mutant embryos, they show that Robo mutants missing their AP-2 binding motifs are unable to rescue the midline crossing defects of robo mutant embryos.The next important step was to test whether Robo endocytosis is important for its activation in vivo. To assay this, the authors used two genetic assays. First, using a gain-of-function assay, they show that Robo mutants that lack AP-2 binding motifs are unable to induce ectopic repulsion from the midline. Second, testing the ability of Robo transgenes to rescue the loss of repulsion defects in Together, these data strongly support the model that Clathrin-dependent endocytosis of Robo in response to Slit is a critical step in transmitting Robo’s repulsive signal across the plasma membrane. It is remarkable that their data indicates that rather than acting to modulate responses by regulating surface levels, endocytosis of Robo is a key component of receptor activation and precedes the recruitment of Sos to the receptor cytoplasmic domain . TestingIn addition to Sos, Robo also has other downstream effectors, such as Enabled, Dock, and Pak. Future studies will be required to determine if these signaling molecules are also recruited to Robo in an endocytosis-dependent manner or not, and from which subcellular compartment they elicit their signaling activity.Drosophila also has two additional family members, Robo2 and Robo3. It will be interesting to determine whether Robo2 and Robo3 also require endocytosis for their signaling activity. The use of different intracellular and/or endocytic compartments for signaling might impart specificity to distinct family members and thus contribute to fine-tune the development of the nervous system.Drosophila Robo are partially conserved in human ROBO1, raising the possibility that endocytosis might also be an important step in vertebrate Robo signaling. Consistent with this idea, Clathrin-dependent endocytosis was found to be required for Slit-induced growth cone collapse of Xenopus retinal ganglion cells [Similarly, in vertebrates, four Robo receptors have been identified: Robo1, Robo2, Robo3/Rig-1, and Robo4/Magic Roundabout. The two endocytic motifs found in ROBO3 cause horizontal gaze palsy with progressive scoliosis (HGPPS) [ROBO receptors have been associated with dyslexia, autism, and schizophrenia [Mutations in human  (HGPPS) . Additioophrenia –8. It wi


A number of observations have led researchers to postulate that, despite being replication-defective, human endogenous retroviruses (HERVs) may have retained the potential to cause or contribute to disease. However, mechanisms of HERV pathogenicity might differ substantially from those of modern infectious retroviruses or of the infectious precursors of HERVs. Therefore, novel pathways of HERV involvement in disease pathogenesis should be investigated. Recent technological advances in sequencing and bioinformatics are making this task increasingly feasible. The accumulating knowledge of HERV biology may also facilitate the definition and general acceptance of criteria that establish HERV pathogenicity. Here, we explore possible mechanisms whereby HERVs may cause disease and examine the evidence that either has been or should be obtained in order to decisively address the pathogenic potential of HERVs. A surprising finding from the sequencing of the human and mouse genomes was the overall proportion, around 45%, of transposable elements Exogenous retroviruses in many host species and replication-competent ERVs in certain species exhibit well-established pathogenic potential, through either insertional mutagenesis or interference of their products with host physiological processes. Therefore, human endogenous retroviruses (HERVs) have attracted greater attention with respect to association with disease. However, the potential mechanisms of HERV pathogenicity might not be identical with those of exogenous viruses. For example, no HERV has been shown to date to produce infectious virus. Thus, insertional mutagenesis by HERVs is an unlikely contributor to host pathology. Nevertheless, our increasing understanding of HERV biology is uncovering novel interactions with the host that could lead to pathology. Here, we review the mechanisms by which HERVs might cause or contribute to disease, assess the current evidence linking HERVs to pathogenesis, and propose a set of criteria that should be considered in establishing HERV pathogenicity.gag), protease (pro), polymerase (pol), and envelope (env). The order of these is vital to the correct level of gene expression, subsequent cleavage and product formation, and is hence completely conserved amongst known retroviruses Retroviral particles contain dimers of the single-stranded positive-sense linear RNA genome, which carries the coding sequences: group-specific antigen , which can be attracted to the LTRs, may augment expression of proto-oncogenes.Several types of cancer exhibit global DNA hypomethylation, which is considered critical in the activation of proto-oncogenes. Studies of methylation status in human cancers supported a model of widespread loss of epigenetic silencing also of REs, and in particular of more recently acquired groups MER11CIn addition to influencing transcription of adjacent genes, HERV transcripts may also be part of the large and diverse group of regulatory RNAs. Several types of RNA can modulate gene expression through a variety of different mechanisms. These include microRNAs (miRNA) and small interfering RNAs (siRNA), both of which act through RNA interference, but can also affect target gene methylation. Long intergenic non-coding RNAs (lincRNAs) are non-protein coding transcripts thought to contribute to a complex regulatory network of gene expression. Accumulating evidence suggests that REs may be essential components of this system, providing an additional mechanism by which REs influence host gene transcription. A comprehensive study highlighted the intertwining nature of REs and human lincRNAs azoospermia factor a (AZFa) region of the Y chromosome, leading to male infertility eyes absent 1 (EYA1) gene in branchio-oto-renal syndrome and recurrent unbalanced translocations leading to intellectual disability have been shown to result from recombination between HERV-H copies HERVs may additionally impact genome regulation and stability simply by providing regions of homology for DNA recombination. Chromosomal rearrangements have established causative roles in a variety of human conditions, including cancer, and can be caused by many different mechanisms. Non-allelic homologous recombination is a frequent cause of deletions and duplications, causing genomic disorders Infections with exogenous pathogens, including retroviruses, elicit innate immune responses, which are initiated by recognition of pathogen-associated molecular patterns by specialized receptors 3′ repair exonuclease 1 (TREX1), a DNA exonuclease that cleaves ssDNA fragments, and subunits of the ribonuclease H2 (RNASEH2) trimer, an RNA nuclease that specifically degrades the RNA of RNA:DNA hybrids. They also include adenosine deaminase, RNA-specific (ADAR), which catalyzes the hydrolytic deamination of adenosine to inosine in double-stranded RNA (dsRNA), and SAM domain and HD domain 1 (SAMHD1), an enzyme that hydrolyzes intracellular deoxynucleoside triphosphates (dNTPs). Importantly, all these enzymes have been demonstrated to affect distinct steps in HIV-1 replication, and it is speculated that they might play similar roles against HERVs. Indeed, analysis of Trex1-deficient mice has revealed accumulation of cytosolic DNA fragments derived from reverse-transcription of endogenous REs Studies of human autoimmune or autoinflammatory conditions, including systemic lupus erythematosus (SLE) and the rare Aicardi-Goutières syndrome (AGS) have implicated a number of genes primarily involved in nucleic acid metabolism env genes with full coding capacity Despite the accumulation of replication-inactivating mutations, many HERVs still retain the potential to express retroviral proteins, which may affect cellular function and reactivity of both the innate and adaptive system. There is also evidence for a potential physiological role for some of these retroviral proteins, suggested by the retention in the human genome of more than a dozen Similarly to all other host proteins, expressed HERV proteins will inevitably provide antigenic epitopes for recognition by B or T cells. Mouse studies have revealed that ERV-encoded self-antigens can, in certain cases, mediate positive selection of developing thymocytes with specific T cell receptor (TCR) reactivity, and promote their peripheral response to cognate antigen It is also clear, however, that endogenous retroviral antigens do not cause complete immunological tolerance and both T and B cell responses are frequently detected in both mice and humans env sequences may also encode superantigens – proteins that stimulate T cells expressing specific TCR Vβ families. In particular, HERV-K18 env has been suggested to cause stimulation of Vβ7-expressing T cells in humans CD48 gene, encoding a lymphocyte-expressed member of the CD2 subfamily of immunoglobulin-like receptors, transcription of a copy of HERV-K18 can be induced by several stimuli that activate B cells, including EBV infection In addition to providing antigenic epitopes for lymphocyte recognition, certain HERV env of HERV-E syncytinsExogenous present-day retroviruses encode an envelope protein that has been demonstrated to exert potent immunosuppression env genes has been suggested for many neuroinflammatory diseases syncytin appears selectively upregulated in multiple sclerosis (MS) lesions syncytin, as well as of HERV-H and HERV-K env, is upregulated during monocyte activation and differentiation, suggesting that it might represent a consequence of increased immune activation in these diseases, rather than a cause of neuroinflammation. However, syncytin protein expression in astrocytes induces production of proinflammatory cytokines and an increase in the oxidation of cellular proteins syncytin.Evidence also exists to suggest that certain HERV envelope proteins can be immune-activating. Although elevated expression of several HERV Emerging data suggest that in addition to nucleic acid and envelope protein, the retroviral capsid can also trigger innate immune responses and production of IFNs in particular. Incoming retroviruses are targeted by TRIM5α, which leads to capsid disassembly rec gene, an alternative splicing product of the HERV-K(HML2) env gene  ene Fig. , when exThe immune system is able to recognize and respond to products of nearly all steps of the retroviral life cycle, although there are certain immune pathways that are triggered more efficiently, if not exclusively, by virions and associated structures. For example, TLR7-mediated recognition of retroviral genomic RNA might be more efficient when virions reach the endosome, where nucleic acid-recognizing TLRs reside Evidence for virus-like particle production has been obtained in samples from many different autoimmune, inflammatory, hematopoietic, or neoplastic diseases In addition to potentially triggering cascades of innate and adaptive immune responses, the most obvious pathogenic mechanism that can be envisaged for a HERV is the production of infectious virions. These could then cause de novo infection and associated pathology via mechanisms that are relatively well understood for exogenous human and murine retroviral infections. However, none of the HERVs that have been studied to date have demonstrated the potential to produce infectious virions. Furthermore, genome sequencing approaches that have revealed novel somatic long interspersed nuclear element (LINE-1) integrations, failed to find HERV integration that would be consistent with re-infection Replication-competent retroviruses could also arise from a series of recombination events between two or more replication-defective HERVs, which would restore their respective defects. Due to the random nature of recombination between co-packaged retroviral genomes and the degree of genetic defects acquired by HERVs, the frequency of such an event would be vanishingly low. However, recent studies in mice have demonstrated that this frequency is increased considerably in certain types of immunodeficiency HERV-Fc1 locus and MS Due to their considerable copy number in the genome, causality or even contribution may be more difficult to establish for many HERVs than for single etiologic agents. Fulfillment of Koch's postulates is a generally accepted criterion for establishing the causal relationship between an infectious microbe and a particular disease. However, two observations are not reconcilable in this frame of reference: firstly, most HERVs exist in the genome of all individuals; they are thus not uniquely present in disease. If such HERVs are considered infective agents, Koch's postulates are violated. Secondly, with the possible exception of an as-yet-undiscovered replication-competent member, the inability of all HERVs to replicate would also violate the second of Koch's postulates. HERVs might be viewed as genetic parasites, rather than transmissible infectious microbes, and, as such, investigation of their potential pathogenicity might require a modification of the original Koch's postulates. The molecular Koch's postulates are a set of criteria formulated by the microbiologist Falkow The first phase would involve detection of HERV expression in a manner consistent with disease involvement. This might involve an all-or-none effect (i.e. expression only in the disease group) or a quantitative effect (disease group expresses more than controls) though the degree of the difference might perhaps affect the enthusiasm for pursuing a viral etiology for the disease. Whether or not the provirus is expressed in normal subjects or tissues will have an important bearing on any subsequent models for disease involvement. Every effort should be made to detect expression levels of both RNA and protein.Proviruses encoding expressed viral genes can be identified by sequencing the expressed transcript, provided this is accomplished using a technique like single genome amplification The third phase of this analysis would comprise construction of a detailed, specific model for HERV involvement in disease. Such models are frequently missing in studies making the case for HERV involvement based simply on analogies with replicating exogenous retroviruses. One important property of such a model would be a clear statement of whether or not the proposed mechanism involved virus replication. Virus replication implies the generation of novel integration sites; any mechanism dependent on new proviral DNA must document such sites or risk the embarrassment of an eventual discovery of a non-human origin for such sequences Lastly, these models might be tested directly. It may be possible to set up animal models by expressing HERV sequences under the appropriate conditions, mimicking exposure from an endogenous element. Alternatively, one might consider attempts to modulate disease in humans by blocking viral protein function with antibody A pathogenic role for HERVs has long been hypothesized, but has remained difficult to prove conclusively. This difficulty may be partly due to the immense complexity of HERV biology, such as their repetitive nature and overabundance in the genome. However, difficulty may also stem from an expectation that HERV pathogenicity should involve mechanisms that are shared with fully-infectious exogenous retroviruses. Replication-competent HERVs may exist as very rare alleles or polymorphic insertions in the population, or emerge in pathological conditions, and evolving sequencing methodologies will certainly facilitate their discovery. However, HERVs can cause or contribute to disease via a variety of other mechanisms not involving infection that warrant further investigation. Study of the complex involvement of HERVs in regulating host gene expression will undoubtedly provide novel insights into their biology and potential pathogenicity. Particularly informative will be the dissection of the direct effects of HERVs on the transcription of adjacent genes, as well as the indirect effects on gene regulation through modulation of regulatory RNA networks. Current and future research is also expected to uncover more players in the interaction between HERVs and innate immunity. The replication-defective nature of HERVs may allow for the accumulation of replication intermediates that would not normally be found in abundance, and triggering of yet-undiscovered innate sensors. With increasing understanding of HERV gene structure and polymorphism, new genetic analysis of disease susceptibility can be designed taking into account the often multi-copy nature of HERVs. Also, the biological activity of proteins produced by HERVs, their interaction partners and pathways and potential effect on physiology and pathology is an area of considerable interest. Finally, inducibility of HERVs by external factors, such as microbial stimulation, also provides a potential mechanistic link between the environment, including commensals and pathogens, and disease development that would be important to explore.


The incidence of cutaneous melanoma has more than doubled over the last decades making it one of the fastest rising cancers worldwide. Improved awareness and early detection of malignant moles now permit earlier diagnosis aiming to decrease the likelihood of recurrence. However, it is difficult to identify those patients initially diagnosed with localized melanoma who subsequently develop metastatic disease. For this group, prognosis remains poor and clinical outcomes are variable and challenging to predict. Considerable efforts have focused on the search for novel prognostic tools, with numerous markers evaluated in the circulation and in tumor lesions. The most reliable predictors of patient outcome are the clinical and histological features of the primary tumor such as Breslow thickness, ulceration status, and mitotic rate. Elevated serum levels of the enzyme lactate dehydrogenase, likely to indicate active metastatic disease, are also routinely used to monitor patients. The emergence of novel immune and checkpoint antibody treatments for melanoma and increasing appreciation of key roles of the immune system in promoting or halting cancer progression have focused attention to immunological biomarkers. Validation of the most promising of these may have clinical applications in assisting prognosis, assessing endpoints in therapy, and monitoring responses during treatment. The incidence rates of melanoma are rising constantly, faster than for any other malignancy over the last two decades . CurrentCutaneous melanoma is classified into four types: (1) lentigomaligna melanomas with a papular or nodular structure; (2) superficial spreading  melanomas with large flat irregular pigmented lesions that grow laterally before invading the dermis; (3) nodular  melanomas with rapidly growing nodules that tend to ulcerate and bleed; (4) acral lentigo  melanomas mainly present at sites of friction, such as the sole, palm, or under nails.Similarly, staging of melanoma is determined according to defined criteria , until 2011 the only standard therapy, has a low objective response rate (<20%) in metastatic disease . NumerouV600 gene on the long arm of chromosome 7 is found in approximately 50% of melanomas , relapse-free survival (RFS) but only marginal improvements for OS , 28, 29.Ipilimumab is an IgG1 monoclonal antibody that recognizes the immune checkpoint cell surface molecule CTLA-4 on T cells. The antibody blocks the binding of the immunomodulatory molecule CTLA-4 to CD80/CD86 on the surface of antigen-presenting cells, preventing negative CTLA-4-mediated signals. This can result in universal activation of T cells, including some capable of specifically recognizing melanoma cells. Ipilimumab has shown efficacy in several clinical trials in untreated and treated metastatic patients in combination with a range of therapies such as peptide vaccines and as a mono-therapy , 31. SomBased on the promise demonstrated by anti-CTLA-4 treatment, antibodies against other immune regulatory proteins such as PD-1, PD-L1, CD137, OX40, and CD40 are being investigated , 33. AdmThe emergence of these new targeted and immune therapies, their success in only subsets of patient groups, as well as the need to assess patient benefit with combination treatments, all highlight the need to develop a range of tools, not only for prognosis but also to assist prediction and monitoring of treatment responses .The rapidly evolving clinical landscape of melanoma management and therapy mandates the search for new candidate biomarkers. Clinical trials traditionally rely on broad clinical groups and long-term objective outcomes, for example, assessment of OS in patients with previously treated stage IV melanoma. Patient stratification is partly served by co-diagnostic tools, such as in the case of the BRAF mutation test, which can help stratify patients who may receive pathway inhibitor drugs; however, these co-diagnostics do not always provide prognostic or predictive information. Biomarkers could revolutionize the process of drug development and those predictive of response could rationalize entry to trials for those patients most likely to benefit . New relDisease-relevant candidate markers for melanoma have traditionally been derived from dissecting melanoma disease pathways. A number of candidates have been studied in sera and tumor specimens Table  and someThe enzyme LDH is a clinical serological biomarker in melanoma. It remains to date the strongest prognostic indicator found to be elevated with tumor burden . LDH is Tyrosinase is part of the biosynthesis process of melanin and is constitutively expressed in melanocytes and melanoma cells. Tyrosinase mRNA levels are detectable in the blood of melanoma patients with advanced metastatic disease detected by nested RT-PCR , 43. IniGrowth factors in combination with interleukins are major regulators of inflammatory conditions in the tumor microenvironment. Vascular endothelial growth factor (VEGF) is known to support tumor-associated angiogenesis, and to contribute to inflammatory conditions, which promote immunosuppression and redirection of effective anti-tumoral immunity. Ugurel et al. reported that VEGF was an independent prognostic marker for overall and progression-free survival in their cohort (125 stage I–IV patients) . UnfortuOsteopontin, a secreted integrin-binding glycol-phosphoprotein, has been described to reduce apoptosis, enhance tumor growth, and be a major component for the recruitment of tumor-promoting stromal cells from the bone marrow , 77. In YKL-40 is a glycoprotein secreted by many cells including cancer cells and by immune cells such as macrophages and activated neutrophils –80. The The small protein melanoma-inhibitory activity (MIA) is secreted by melanoma cells and is involved in cell–cell contact by interacting with the extracellular matrix. It is also thought that MIA promotes tumor cell invasion and metastasis . EquallyThe family of S100 proteins has been of special interest as diagnostic markers in melanoma over the last decade, since the expression and secretion of these proteins is much higher in malignant compared to healthy tissues –87. S100Interleukin-8 (IL-8), also known as CXCL8, is a chemokine produced by malignant cells and associated with inflammatory responses; it can induce neutrophil chemotaxis and is also thought to be a potent promoter of angiogenesis. A recent study has shown that IL-8 correlated with tumor burden, stage, survival, and response to therapy such as BRAF inhibitors . These dTissue specific biomarkers are molecules demonstrated to be over-represented in cancer lesions, and may facilitate diagnosis, early detection, prognosis of disease progression, and patient stratification.Cyclooxygenases (COX1–3) are a group of proteins that are important modulators in the human body, affecting essential pathways such as the catabolic metabolism. COX1–3 also converts arachidonic acid into prostaglandin. Of this group, COX-2 can be induced in tumor cells , 93. BecThe Galectin-3 molecule is mainly secreted by inflammatory cells and is associated with both tumor progression and metastasis in melanoma . HoweverMatrix metalloproteinases play an important role in remodeling the tumor tissue microenvironment as they are responsible for proteolytically breaking components of the local tissue architecture, promoting tissue remodeling, and facilitating tumor cell migration , 96. ForCancer-associated cell adhesion proteins are proteins that adhere to different cells, show altered expression levels in cancer, and may participate in tumor cell migration, immune cell evasion, and angiogenesis . MoleculThe chondroitin sulfate proteoglycan 4 (CSPG4), also known as the high molecular weight melanoma-associated antigen (HMW-MAA) or the melanoma-associated chondroitin sulfate proteoglycan (MCSP) is a glycoprotein–proteoglycan complex expressed on the surface of melanoma, glioma, neuroblastoma, certain breast carcinomas, and acute leukemias . CSPG4 iCancer development and progression is associated with dysregulated molecular pathways in tumor cells, but it is now accepted that it may also be affected by cross-talk with the immune response. The latter could lead either to immune cell activation and destruction of cancer cells or to immune response redirection and suppression to support cancer cell survival and metastasis . Differe+FoxP3+CD25high) among TIL, or the presence of myeloid-derived suppressor cells (MDSC) may correlate with poorer survival in many cancers and with lower clinical response rates to anti-CTLA-4 antibodies  (immune score) may be more accurate prognostically than standard methods such as TNM stage , 111. Futibodies , 112–114tibodies . These f6 tumor cells, which translate to very low or undetectable serum levels of potential biomarkers. Unless actively secreted, molecules released from these few cells may not be easily detectable in the circulation, thus monitoring such markers could be challenging.Early disease stages are characterized by mostly minimal disease burden with no more than 1 × 10B cells can undergo class switching to produce affinity matured antibodies in response to tissue damage and to the presence of antigens, including cancer antigens, and this may happen at early stages of cancer development. An individual B cell can produce 5,000–20,000 antibodies/min, and B cells can go through mitosis every 3 days, which can maintain or further enhance antibody production –118. Theex vivo by recognizing tumor antigens on the surface of these cells (Until recently, the nature and functional significance of the humoral response in cancer were poorly understood. Moreover, whether and how the humoral response may be “immunoedited” by tumors has remained unknown. We have reported a tumor-reactive mature memory B cell compartment in the circulation of patients with melanoma. By isolating tumor-reactive antibody clones from patients’ mature B cells, we found that patient-derived antibodies can activate effector cells to destroy tumor cells se cells . These fHints that immune escape mechanisms may operate to suppress tumor-reactive humoral responses came from our findings that this tumor-reactive memory B cell compartment appears more prominent at earlier disease stages and is reduced with disease progression. It has also been demonstrated that both the broad and the tumor-reactive mature memory B cell compartments are reduced in patients with advanced disease .In melanoma, early studies indicated dysregulation and polarization of IgG antibody subclasses in the serum of patients with melanoma, but the biological relevance and functional implications of these findings were unclear . More reTumor antigen recognition may be another attribute of antibodies that could render them promising potential biomarkers. This may be facilitated by emerging screening platforms such as “immunosignaturing,” which is designed to identify reactive antibody signatures against panels of antigens, including those found in cancers. The “signatures” resulting from this process could allow for the identification of antibody reactivity patterns that may predict disease course. The potential for this approach has been demonstrated in the diagnosis of Alzheimer’s disease  and to dDifferent facets of antibodies, including tumor specificity and reactivity, as well as antibody class and subclass bias may therefore be examined in the context of cancer including; as measures of the potency of adaptive immunity in cancer, as readouts of tumor-induced immunoediting and as powerful therapeutic agents to activate patient immune cells against cancer . Host huBiomarkers that move beyond the current clinical pathological and radiological parameters, helping to identify those patients with early disease at high risk of relapse and guiding therapy choices for patients with metastatic disease, are still needed. A number of potential candidate biomarkers, including immunological markers warrant further evaluation in melanoma. Circulating or tumor-resident immune cells, including those associated with immunosuppressive forces in melanoma such as Treg MDSC, IgG4+ B cells, and also cytokines, chemokines, checkpoint molecules, and antibodies may point to yet unexplored biomarker signatures associated with particular clinical outcomes. Despite the considerable progress made in immune monitoring technologies, it has been challenging to draw accurate correlations between immunological parameters and clinical outcomes or patient responses to therapeutic agents. The reasons might include complex interactions between immune and tumor cells and the variable patient immune responses, making it difficult to account for all the interactions required for adequate prognostic readouts. Even when associations with melanoma are demonstrated, there is significant variability among patients, possibly reflecting the heterogeneity of individual tumors and of individual patient immune responses.Other important challenges for biomarker discovery and validation remain. As most biomarkers are normal cellular or immunological proteins, they can be detected in both healthy and disease states, reducing their specificity. Additionally, many studies report relatively small sizes of patient samples, and this may indicate a limitation potentially preventing biomarker selection and routine use. Future directions may include larger cohorts assessed in multi-center studies and also assessing combinations of multiple putative markers to identify specific prognostic footprints. Finally, complex statistical analyses and comprehensive algorithms will be needed in order to integrate these multiple lower specificity biomarkers.In conclusion, the potential of biomarkers to contribute toward better clinical monitoring and management will most likely be aided through further refinement of emerging design, statistical, bioinformatics, and analytical methods , as wellThe 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.


Shigella flexneri and the autophagy machinery were first discovered in 2005. Since then, work has shown that multiple autophagy pathways are triggered by S. flexneri, and autophagic responses can have different roles during Shigella infection. Here, we review the interactions between S. flexneri and the autophagy machinery, highlighting that studies using Shigella can reveal the breadth of autophagic responses available to the host.Autophagy, an intracellular degradation process, is increasingly recognized as having important roles in host defense. Interactions between  In this review, we discuss canonical and non-canonical autophagy pathways, their interplay with invasive bacteria, and highlight what has been discovered from their interactions with Shigella flexneri.The process of autophagy degrades intracellular material via fusion with lysosomes, and is evolutionarily conserved  proteins to an isolation membrane called a phagophore  is the best-studied example of a non-canonical autophagy pathway , autophagy receptors such as p62 (sequestosome 1) and NDP52  act as cytosolic sensors, bind ubiquitinated substrates and LC3 family proteins to selectively target bacteria to degradation by canonical autophagy , a needle-like apparatus used to inject bacterial effector proteins into the host cell and enable an intracellular lifestyle  and N-WASP via IcsA , which act synergistically to promote bacterial escape from LC3-positive vacuoles.LAP can also occur during s Figure , where Ls Figure . HoweverShigella. These mechanisms can be viewed as a crucial aspect of host defense to control bacteria at the onset of bacterial interactions with the host cell.Together, different sensing mechanisms including NOD1/2 and LAP trigger autophagic recognition of invading S. flexneri can exit the phagocytic vacuole and gain access to the cytosol. In epithelial cells, rupture of the phagocytic vacuole by Shigella can initiate autophagy by inducing nutrient starvation . From this, it has been proposed that Shigella-induced amino acid starvation can be sensed by the host cell to initiate an immune response.Shortly after invasion of host cells, Shigella ruptures its phagocytic vacuole in epithelial cells, membrane remnants can be ubiquitinated  recognized by galectins, a family of beta-galactoside-binding proteins Figure . StudiesS. flexneri can subvert the host actin cytoskeleton by expressing the outer membrane autotransporter protein IcsA to recruit N-WASP and the actin related protein 2/3 (ARP2/3) complex, inducing actin-based motility to spread from cell-to-cell  can increase Shigella replication and decrease zebrafish survival. Overall, these data indicate that host survival depends on the appropriate autophagic response to control Shigella infection in vivo (Mostowy et al., Shigella (Lassen et al., Septins, GTP-binding proteins essential for cell division (Saarikangas and Barral, Shigella can help to investigate the breadth of autophagy responses available to the host.Originally discovered as a bulk degradation process important for cellular homeostasis (Yang and Klionsky, S. flexneri infection model can be used to study key issues in cell-autonomous immunity and host cell biology, including the ability of host cells to sense and defend against intracellular bacteria. The host cell has evolved multiple sensing mechanisms that together enable a multi-tiered defense network to counteract Shigella invasion (Table Shigella has co-evolved a variety of protection mechanisms, and in some cases can interfere with host cell biology to circumvent autophagic processes (Table Shigella infection model, many outstanding issues remain. For example, what is the role of the host cytoskeleton in autophagy and its ability to restrict or promote bacterial replication? Actin, microtubules, intermediate filaments, and septins are four main cytoskeletal components of mammalian cells, yet their precise roles in autophagy are not understood (Mostowy, Shigella will require in depth experimentation to be performed in vivo. Though mammalian models remain poorly suited to image the cell biology of Shigella infection in vivo, the natural translucency of zebrafish larvae enables non-invasive in vivo imaging at high resolution throughout the organism (Kanther and Rawls, in vivo role of Shigella interactions with the autophagy machinery can be examined at the subcellular, cellular, and whole-animal level (Mostowy et al., The on Table . Howeveres Table . DespiteMostowy, . What isMostowy, . FinallyS. flexneri and the autophagy machinery in vitro using tissue culture cells and in vivo using animal models can help to better understand fundamental mechanisms of host defense. They could additionally suggest the development of novel therapies for infectious diseases.Taken together, outcomes generated from studying the interplay between 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.