To address major societal challenges and enhance cooperation in research across Europe, the European Commission has initiated and facilitated ‘joint programming’. Joint programming is a process by which Member States engage in defining, developing and implementing a common strategic research agenda, based on a shared vision of how to address major societal challenges that no Member State is capable of resolving independently. Setting up a Joint Programming Initiative (JPI) should also contribute to avoiding unnecessary overlap and repetition of research, and enable and enhance the development and use of standardised research methods, procedures and data management. The Determinants of Diet and Physical Activity (DEDIPAC) Knowledge Hub (KH) is the first act of the European JPI ‘A Healthy Diet for a Healthy Life’. The objective of DEDIPAC is to contribute to improving understanding of the determinants of dietary, physical activity and sedentary behaviours. DEDIPAC KH is a multi-disciplinary consortium of 46 consortia and organisations supported by joint programming grants from 12 countries across Europe. The work is divided into three thematic areas: (I) assessment and harmonisation of methods for future research, surveillance and monitoring, and for evaluation of interventions and policies; (II) determinants of dietary, physical activity and sedentary behaviours across the life course and in vulnerable groups; and (III) evaluation and benchmarking of public health and policy interventions aimed at improving dietary, physical activity and sedentary behaviours. In the first three years, DEDIPAC KH will organise, develop, share and harmonise expertise, methods, measures, data and other infrastructure. This should further European research and improve the broad multi-disciplinary approach needed to study the interactions between multilevel determinants in influencing dietary, physical activity and sedentary behaviours. Insights will be translated into more effective interventions and policies for the promotion of healthier behaviours and more effective monitoring and evaluation of the impacts of such interventions.The online version of this article (doi:10.1186/s12966-014-0143-7) contains supplementary material, which is available to authorized users. Health is a key driver of Europe’s growth and prosperity and European governments are struggling with the growing social and economic consequences of an alarming increase in lifestyle related diseases, including obesity, cardiovascular disease, diabetes and cancer . A healtTo address major societal challenges and enhance cooperation in research across Europe, the European Commission has initiated and facilitated ‘joint programming’. Joint programming is a process by which Member States engage in defining, developing and implementing a common strategic research agenda, based on a shared vision of how to address societal challenges that no Member State is capable of resolving independently.Such joint programming across European countries can help to pool resources to enable more comprehensive and larger-scale research with more variation in exposures and outcomes, avoid unnecessary overlap and repetition and enable and enhance the development and use of standardised research methods, procedures, tools, infrastructure and data management necessary for high-quality research. It should thus contribute to improving and aligning the research infrastructure, including human capital, so that this can be used more effectively and efficiently. In addition, joint programming not only involves aligning or defining joint research agendas, but also combining and/or pooling research funding from the participating countries.Within the EU Committee for Scientific and Technical Research, the High-Level Group on Joint Programming has identified and substantiated various themes for Joint Programming Initiatives (JPIs), one of which is ‘A Healthy Diet for a Healthy Life (HDHL)’. The JPI HDHL was then adopted by twelve Member States that set up a management and development structure and produced a strategic research agenda . In thisThe first act of this JPI is to realise joint programming, collaboration and harmonisation to further research on the determinants of diet and physical activity. Therefore the Determinants of Diet and Physical Activity (DEDIPAC) Knowledge Hub (KH) was started to establish a European transdisciplinary research network programme on determinants of diet and physical activity and their relationship to best-practice implementation strategies for long-term behaviour changes.In this paper, we share this new European initiative in the field of behavioural nutrition and physical activity by describing the focus, organisation, aims, management and dissemination of the DEDIPAC KH and discussing the expected pros and cons and the challenges of European Joint Programming in this field.The overarching goal for DEDIPAC is to understand and be able to enact the most effective ways of improving public health through interventions targeting motivation, ability and opportunity to adopt and maintain healthy dietary, physical activity and sedentary behaviours . In ordeEnabling a better standardised and more continuous pan-European ‘needs analysis’ , i.e. to monitor dietary, physical activity and sedentary behaviours and changes in these behaviours across the life course and within populations to identify targets and target populations for (policy) interventions;Exploring the main correlates and determinants of these behaviours in and across populations to help to tailor policies and interventions to target these determinants;Learning from successes and failures of previous and on-going interventions and policies in order to improve evaluation and increase effectiveness of future interventions and policies and to identify and benchmark best practices across Europe and compare these internationally.The DEDIPAC KH aims to contribute to building the research network and infrastructure and perform preparatory work to be able to better face this challenge. More specifically, the DEDIPAC KH aims to contribute to:TA 1: Assessment and harmonisation of methods for future research, surveillance and monitoring and evaluation of interventions and policies;TA 2: Determinants of dietary, physical activity and sedentary behaviours across the life course and in vulnerable groups;TA 3: Evaluation and benchmarking of public health interventions and policies aimed at improving dietary, physical activity and sedentary behaviours across the life course.The DEDIPAC KH comprises three Thematic Areas (TAs) that will be strongly interlinked and will work in parallel to cover the logical path from assessment and surveillance to public health interventions and policies via the exploration and modelling of determinants of dietary, physical activity and sedentary behaviours. The three TAs are characterised as follows:www.dedipac.eu). The three TAs and their backgrounds are described in more detail below.Each of the TAs consist of different Work Packages (WPs) that constitute specific tasks that will be conducted by collaborating experts from different research groups within the DEDIPAC KH. Each of the TAs will deliver harmonised or aligned infrastructures, methodologies and/or evidence that will be compiled in a ‘toolbox’ available to scientists, health promotion professionals and policy makers. This toolbox will be made accessible via the DEDIPAC website . The . The 7])Physical activity and sedentary behaviours have traditionally been assessed by self-reporting in large-scale cohort studies and surveillance systems. However, objective assessment methods, such as accelerometry, heart rate monitoring and combined accelerometry and heart rate monitoring have emerged as potential methods to assess physical activity and sedentary behaviours in large scale studies as well ,10. WhilSystematic literature reviews will help to identify relevant reliability and validation studies on assessment of dietary, physical activity and sedentary behaviours and their determinants. First, reference instruments will be identified  gold standards) for dietary behaviour, physical activity and sedentary behaviours. Then the performance of existing proxy measurements will be assessed in terms of validity and reliability and both consistency and sources of heterogeneity of the assessment methods will also be identified. Prototyping and piloting innovative assessment methods will be initiated by mapping databases of on-going research. In addition, existing pan-European datasets on reference methods  will be exploited to identify the main contributors to relevant variation  in dietary, physical activity and sedentary behaviours. This will result in the description of the European diversity and at the same time stimulate and elicit potential innovative short-cut measurement methods to be explored in a feasibility pilot study. The range of relevant dietary, physical activity and sedentary behaviours and their determinants is very broad and priorities must be set. To efficiently select the most relevant contents, we will make an inventory of topics covered in earlier research and prioritise specific areas for further research in the DEDIPAC consortium. Prioritisation of topics will assist in maintaining focus and relevance in terms of public health challenges in specific stages of the life course. For example, but without anticipating any decisions, diet related topics could include sugar-sweetened beverage or fruit and vegetable intake in schoolchildren, or vitamin D and cobalamin supplementation in community-dwelling and institutionalised older people. Similar considerations will shape the selection of topics for physical activity and sedentary behaviour.Dietary, physical activity and sedentary behaviours are determined by interactions between biological, psychological, sociological, economic, ecological, socio-economic and cultural factors, which are at least partly modifiable -13. AlthSeveral frameworks and models describe the theoretical relationship between the different levels and categories of determinants of behavioural nutrition and physical activity. For example, Glass and McAtee , have meInitially, we will explore, apply and enrich this framework for the three separate behaviours or behavioural categories and will progressively try to integrate insights across the three behaviours to generate knowledge on the interplay between common determinants of these behaviours. Cross-country comparisons and exploration of these multilevel determinants in pooled (secondary) data analyses will contribute to gaining further insight in this regard. This focus on combining and integrating behavioural nutrition and physical activity research closely matches with the focus of the International Society of Behavioural Nutrition and Physical Activity.With the Glass & McAtee model and ANGELO framework as points of departure, review and integrate the current state-of-the art determinant frameworks (uni- or multi-disciplinary).Proof-of-concept explorative secondary data analysis using state-of-the-art statistical methods (conducted within European transdisciplinary research networks) and explorative case studies of social inequalities and ethnic minorities.This TA will develop a European multi-disciplinary network and framework for determinant research in Europe by means of a two-stage process conducted in each of the work packages, each focusing on one of the behaviours, i.e. diet, physical activity and sedentary behaviour:A mix of established methods will be used, including systematic literature reviews and scoping reviews, a multi-disciplinary Delphi consensual process and expert workshops , sport and physical activity sciences, , psychology , biology, geography, public health,  economy, general/clinical medicine, informatics, engineering for mobile devices, statistics, medical biometry, sociology, policy, natural sciences, biomedicine, metabolic physiology, genomics, human ecology, environmental science, mathematics, Agro-Food Marketing etc.However, this multidisciplinary approach will generate a major challenge in achieving harmonisation and matching across data sources as well as data sharing across research groups. Novel ways to perform pooled analysis of individual-level data without actually sharing the data will be explored. For instance, EU BioSHaRE established a password-protected web portal that facilitated remote and federated analyses of the harmonised datasets conducted at each participating institution using DataSHIELD software . AnotherAs stated in the JPI-HDHL strategic research agenda, DEDIPAC’s ultimate goal is to contribute to improving public health through interventions, actions and policies targeting dietary and physical activity behaviours . TA 3 wiMultilevel or multi-component interventions are defined as theory-based interventions that use knowledge of the behavioural determinants at different levels  to improve dietary, physical activity and sedentary behaviours in individuals. We will focus in particular on multi-component interventions that take a social-ecological approach because literature reviews suggest these multi-component interventions are potentially the most effective [ffective -21. ThesSeveral frameworks for evaluating both multi-component interventions and public policies aimed at enhancing physical activity  or diet TA 3 will utilise the DEDIPAC- toolbox developed in TA 1 and the individual, socio-cultural- and environmental-level determinants of dietary, physical activity and sedentary behaviours described by TA 2 to develop an evaluation framework and open-access database in which policy and multi-component interventions will be described. This will include their evaluation in terms of reach, effectiveness, adoption, implementation and maintenance. This pan-European framework will then be pilot tested in ongoing interventions or policies in a number of DEDIPAC Member States and will be adapted accordingly. Eventually, the framework will outline evidence- and practice-based policy and multi-component intervention options.In order to fulfil the aforementioned aims, the draft evaluation framework will first be pilot-tested by 19 partners across nine countries using ‘natural experiments’ or policies that are currently being implemented or have recently been implemented. An expert consensus meeting with a variety of stakeholders will be organised to agree upon a final framework. Getting their input is an essential step in shaping the end-product. A final symposium will also be organised and held to inform and consult stakeholders –with separate and joint sessions for scientists, policy makers, health promotion professionals- about the results, conclusions, recommendations and continuation of the DEDIPAC Knowledge Hub.A consortium of 46 consortia, research institutes and groups from 12 EU Member States has been established , scientific publications and non-scientific publications in English and other languages, consortium meetings, as well as in a series of workshops specifically aimed at early-career researchers. The workshops will address the conduct of systematic literature reviews, secondary data analysis and/or advanced statistical methods, for instance.The progress and deliverables will be communicated and disseminated via the DEDIPAC website , covering three different lifestyle behaviours. DEDIPAC will make use of, but will also go beyond what has been done is separate EU funded projects, aiming to work towards agreed upon measurement standards for cross European research.A major strength of the DEDIPAC KH is its ability to facilitate the simultaneous study of dietary, physical activity and sedentary behaviours, their determinants and the policies and multi-component interventions that target these determinants and behaviours. Although these behaviours comprise sets of different specific behaviours influenced by a range of personal, socio-cultural and environmental determinants, they form the two sides of the energy-balance equation and thus need to be targeted simultaneously or at least in relation to each other in order to promote synergy and avoid negative compensation. The DEDIPAC KH will help to identify common ground between dietary, physical activity and sedentary behaviours with regard to assessment, surveillance, determinants and intervention strategies. Ultimately, this will facilitate better synchronisation in diet and physical activity research, policy and practice.As a potential downside, it should be acknowledged that working in multi-disciplinary teams from a multitude of research institutes across 12 countries , with 8 different main languages, with financial support from as well as reporting to 12 different funding agencies, is already a major organisational endeavour, to say the least. In the context of a complex organizational structure such as this, it is even more challenging to reach consensus about harmonisation of measures, benchmarking of policies, data sharing and other big issues. Nevertheless, the DEDIPAC consortium was able to write, submit and receive approval for the DEDIPAC proposal from the 12 countries, all in a matter of about four months’ time. Furthermore, all WPs have started and are working according to schedule in the first several months after the official start of DEDIPAC, i.e. 1 December 2013.The current DEDIPAC consortium has good coverage of the different relevant disciplines. Because DEDIPAC originates in the JPI HDHL, experts in human nutrition are very well represented. The focus of DEDIPAC was extended to also include physical activity and sedentary behaviour at an early stage and these areas of expertise are therefore also very much involved. However, DEDIPAC’s focus on multilevel determinants towards a more systems-level approach to behavioural determinants and including policy evaluation and benchmarking, requires involvement of a range of social and behavioural expertise that needs further strengthening within the consortium. From the very start and made explicit in the approved proposal, DEDIPAC will be open to other groups and will, wherever necessary, actively try to include new groups with expertise to further enrich the KH. This first DEDIPAC act will help to expand on existing collaborations and assist in building new ones while identifying expertise gaps within the KH. This may help the KH to stay relevant in the longer term. Another way to improve the long-term sustainability of the DEDIPAC approach and community are the dissemination and capacity building activities. These activities aim to expand the DEDIPAC community and to train early-career scientist in this multi-disciplinary approach.The DEDIPAC KH uses joint research programming and funding across different countries in Europe to work towards better research harmonisation and collaboration in the field of behavioural nutrition and physical activity research. As a first joint action in this joint programming initiative, DEDIPAC has already made an excellent start in setting up a complex, cross-country organisational structure.


Survival depends on an organism’s ability to sense nutrient status and accordingly regulate intake and energy expenditure behaviors. Uncoupling of energy sensing and behavior, however, underlies energy balance disorders such as anorexia or obesity. The hypothalamus regulates energy balance, and in particular the lateral hypothalamic area (LHA) is poised to coordinate peripheral cues of energy status and behaviors that impact weight, such as drinking, locomotor behavior, arousal/sleep and autonomic output. There are several populations of LHA neurons that are defined by their neuropeptide content and contribute to energy balance. LHA neurons that express the neuropeptides melanin-concentrating hormone (MCH) or orexins/hypocretins (OX) are best characterized and these neurons play important roles in regulating ingestion, arousal, locomotor behavior and autonomic function via distinct neuronal circuits. Recently, another population of LHA neurons containing the neuropeptide Neurotensin (Nts) has been implicated in coordinating anorectic stimuli and behavior to regulate hydration and energy balance. Understanding the specific roles of MCH, OX and Nts neurons in harmonizing energy sensing and behavior thus has the potential to inform pharmacological strategies to modify behaviors and treat energy balance disorders. Food and water are essential for survival, and organisms have developed physiological systems to ensure that the body maintains sufficient stores of these resources  in controlling energy balance for three reasons: (1) The LHA modifies intake of natural and pharmacologic rewards and physical activity, and such function via the LHA is required for survival, (2) The LHA receives circulating energy balance cues and projects to brain regions that regulate motivated behaviors, (3) Distinct neuronal populations within the LHA are “tuned” to specific energy cues (such as ghrelin or leptin) and induce cue-appropriate behavioral responses. Thus, understanding the precise neurochemistry, connectivity and function of the LHA neuronal subpopulations will suggest mechanisms by which to suppress or enhance feeding, drinking and energy expenditure as required to restore energy balance. Modifying action via the LHA therefore has potential to improve a spectrum of health problems.ability to move, eat and drink, but lost all motivation to do so: as a result they all died of self-inflicted starvation and dehydration  drive that can, in time, be sufficient to mediate survival  to permit their survival  fibers as well as axons of passage within the medial forebrain bundle (mfb), each of which terminate in brain centers associated with reward and motivation  or periventricular hypothalamic regions  suggests that these regions differ in neurochemistry and/or their anatomical engagement of brain systems that regulate behavior. Hypothalamic nuclei such as the VMH, arcuate nucleus (ARC), dorsomedial hypothalamus (DMH) and paraventricular hypothalamic nucleus (PVN) are compact, cell-dense and have well-defined projection targets throughout the brain. The LHA, by contrast, encompasses a large swath of tissue over the entire rostral-caudal extent of the hypothalamus. The sheer expanse of the LHA, coupled with the fact that it lacks obvious cellular architecture, complicated anatomical and functional studies. The LHA lies lateral to the DMH and PVN, and the area just above and surrounding the fornix is referred to as the perifornical area of the LHA. Pioneering work by the Swanson group utilized these anatomical landmarks in combination with neuronal tract tracing methods to determine the precise connectivity of LHA subregions with the rest of the brain , the orexins/hypocretins (OX) or neurotensin (Nts). Intriguingly, these neuronal populations are molecularly and spatially distinct, suggesting that each population may also differ in connectivity and functional output  in regulation of feeding, drinking and energy expenditure. The amount of energy expended (and thus weight), is influenced by the amount of physical activity, sleeping/arousal and thermogenic outflow, so each of these facets of energy expenditure will be discussed.Melanin concentrating hormone (MCH) is a 19 amino acid cyclic neuropeptide that was first documented in the pituitary of teleost fish, enabling them to change skin color and blend into their environment , which can be coupled to either Gpreferentially stimulate intake of palatable food or sucrose, suggesting that MCH regulates general consumption behavior, but not necessarily hedonic aspects of feeding , compared to normal controls  social interactions, and this has inspired interest in MCHR-1 blockade for treating anxiety disorders  sleep, while MCHR-1 antagonists reduce it  and to promote locomotor activities to obtain the food. OX neurons are activated in anticipation of feeding and couple arousal and feeding behaviors -expressing synaptic inputs, and in lean mice these CB1 terminals are primarily glutamatergic (excitatory). In the obese state the ratio of CB1-expressing inputs is predominantly GABAergic (inhibitory), but leptin treatment restores excitatory CB1-terminal bias similar to that of lean mice , which coordinates locomotor response during stressful experiences to facilitate survival, such as the ability to escape predators  the tuberonmamillary nucleus (TMN) and via regulation of LHA MCH neurons. The LC receives dense innervation from OX neurons, and OX increases activation of LC neurons, arousal and locomotor behaviors  and arousal , as well as to sympathetic response centers that are necessary to support such behaviors , midbrain  limbic system , forebrain , thalamus, and within the hypothalamus, particularly the preoptic area, PVN and the LHA  and NtsR-2,  permitted the facile identification of Nts neurons throughout the brain, including a large population of Nts neurons within the LHA  suppresses locomotor activity (Ervin et al., LHA Nts neurons additionally project within the LHA and modulate OX neurons (Leinninger et al., To date there are no direct reports concerning the role of LHA Nts neurons in arousal. Burdakov described a large population of non-MCH, non-OX GABAergic neurons in the LHA that are spontaneously active during waking and sleeping periods, and it is possible that these are Nts neurons (Karnani et al., Central Nts increases analgesia (Clineschmidt et al., The LHA has now long-been regarded as an essential brain region for promoting feeding, drinking and energy expenditure behaviors that inherently modify energy balance and weight. The LHA, however, appears to coordinate a diverse array of peripheral cues that range in nature from hormones, nutrient levels, osmolality, light and sound as well as neurotransmitter inputs from other brain centers. From an anatomical perspective, the large LHA contains multiple populations of neurons that project to virtually every region of the brain. Thus, the LHA seemingly manages the daunting task of receiving and interpreting cues from the body, and then matching appropriate behaviors and physiologic regulation to each cue via a broad network of neuronal regulation. The discovery of neuropeptide-specific populations within the LHA has catapulted the field’s understanding of how the LHA can manage such a formidable task: via specialized, cue-sensitive neurons that mediate actions via distinct neuronal projections. While the characterizations of MCH, OX and Nts neurons have begun to illuminate specialized functions of each population, these studies are only the tip of the iceberg, so to speak. As we drill down to understanding the precise molecular signature of OX, MCH and Nts neurons we are likely to identify even further specialized subpopulations of these neurons. It is tempting to speculate that, perhaps, specified subpopulations mediate the response to particular physiologic cues and match behavior to restore energy balance. Such parceling would enable targeting of therapies to either amplify or inhibit feeding, drinking, physical activity, antinociception and arousal, and accordingly ameliorate a number of health conditions. In reality, it is unlikely that the neuronal signatures will be easily identifiable or so conveniently specialized, as neuronal systems tend to overlap in function and projections. Indeed, the fact that MCH, OX and Nts neurons are neuropeptide-distinct but project to and regulate similar neuronal targets and sites within the brains speaks to this fact. Even so, MCH, OX and Nts neurons do contribute differential physiologic regulation. MCH neurons largely regulate physical activity and energy expenditure but do not contribute as abundantly to regulation of arousal. OX neurons are the predominant LHA neuronal mediators of arousal, and must integrally regulate feeding, activity and sympathetic outflow to support alertness. Inherently, both MCH and OX neurons may increase feeding to support energy-consuming physical activity and alertness. In contrast, the few studies to date suggest that LHA Nts neurons are required to restrain feeding and promote physical activity. Thus overall, there are likely to be distinct physiologic contributions of these neuropeptide-specific populations that permit the LHA’s impressive regulation of motivation, energy balance and survival. Just as the discovery of neuronal heterogeneity in the LHA prompted the idea of functional divergence, utilization of reagents to selectively identify and modulate MCH, OX and Nts neurons will reveal their selective circuitry and functional contributions to energy balance.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.


Streptococcus gallolyticus. Three-dimensional transesophageal echocardiography was performed and revealed multiple mitral valve vegetations, with leaflet perforation and important mitral regurgitation, as well as large aortic vegetation, with cusp perforation and severe regurgitation. Additionally, a small vegetation was observed on the tricuspid valve, which presented moderate regurgitation. Three-dimensional transesophageal echocardiography provides appropriate visualization of complications resulting from infectious endocarditis.A 42 year-old woman was referred to our hospital with a history of fever and poor general status for the last 30 days. She presented tachycardia and a systolic apical murmur. Laboratory tests revealed leukocytosis of 13,100/mL, hemoglobin of 8.4g/dL and positive systemic lupus erythematosus antibodies ; blood culture was positive for  The natural history of the disease varies and is characterized by periods of remission and acute disease. Women, especially those between 20 and 30 years of age, are more affected than men.Systemic lupus erythematous (SLE) is an inflammatory autoimmune disease with multiple organ involvement.The heart is frequently affected, specially the valves. Valvar lesions vary from mild thickening to large verrucouse lesions (Libman-Sacks endocarditis). The mitral valve is more frequently involved. Transesophageal ultrasound is a sensitive tool for diagnosis of valve impairment and can be used to determine the extension of complications and progression of valvar diseaseStreptococcus gallolyticus is a Gram-positive coccus and is part of the Streptococcus bovis (Streptococcus) and Streptococcus infantarius group. It is an important cause of infectious bacteremia and endocarditis in adults. Infections by this agent were associated with colon tumors and gastrointestinal tract lesions. However, the association between Streptococcus gallolyticus endocarditis and lupus causing valve leaflet perforation has not been described yet.. So far, three-dimensional echocardiography has not been extensively used for diagnostic investigation and identification of complications associated with infectious endocarditis. In this case report three-dimensional transesophageal echocardiography was used in a SLE patient presenting with infectious endocarditis.The three-dimensional echocardiography allows for multiplanar spatial observation of the heart and better understanding of the anatomy and geometry of cardiac structures. The use of three-dimensional echocardiography is recommended in the following situations: (1) to measure the volume of the left ventricle; (2) to determine the ejection fraction of the left ventricle; (3) to evaluate mitral valve anatomy; (4) to determine the degree of mitral valve stenosis; (5) to guide percutaneous procedures in the cath labStreptococcus gallolyticus. Transthoracic echocardiogram revealed an image of a large aortic vegetation with mild regurgitation, and images of multiple vegetations on the mitral valve with severe regurgitation. Abdominal CT revealed multiple kidney, spleen and liver infarctions. Colonoscopy showed sigmoid diverticulum and two hyperplastic polyps. Histology of the colonoscopy findings suggested no evidence of colon neoplasia. Three days later, the bidimensional and three-dimensional transesophageal echocardiograms revealed multiple vegetations on the mitral valve, perforation of the cusp and important reflux and also an imaging of a great vegetation with valvular perforation. In addition, the exam also revealed a small vegetation on the tricuspid valve and severe regurgitation . Blood cultures tested positive for gitation , 2, 3, 4Streptococcus gallolyticus from the blood was unexpected, because this agent causes important bacteremia and infectious endocarditis in elderly and adult men,6. The presence of Streptococcus gallolyticus is frequently associated with lesions of the gastrointestinal tract, which were also present in this case. The fast destruction of the valves (perforation of two leaflets) is not common in patients with Streptococcus gallolitycus infection and is associated with increased morbidity and mortality .Systemic lupus erythematous (SLE) is a pleomorphic condition that may pose a clinical challenge. In the case of this patient, the first manifestation of the disease was an infectious endocarditis, with multiple cardiac valve involvement, which is a very rare presentation. In addition, the isolation of .Despite advances in medical and surgical treatments, infectious endocarditis still leads to high morbidity and mortality rates. Early identification of patients with large vegetations, as in the case described, is important to define the best therapeutic strategy, because, in these cases, surgical excision of the valve and valve replacement are followed by low perioperative mortality ratesIn patients with endocarditis, the transesophageal echocardiogram is more suited for identification and determination of the degree of valve involvement than the transaortic echocardiogram. More recently, the three-dimensional echocardiogram added relevant pieces of information that contribute to determining the exact mechanism causing valve dysfunction. This is due to its higher spatial resolution and multi-angle visualization of the structures, which in the case described allowed for the exact determination of the site of leaflet perforation.


Individuals addicted to drugs of abuse such as alcohol, nicotine, cocaine, and heroin are a significant burden on healthcare systems all over the world. The positive reinforcing (rewarding) effects of the above mentioned drugs play a major role in the initiation and maintenance of the drug-taking habit. Thus, understanding the neurochemical mechanisms underlying the reinforcing effects of drugs of abuse is critical to reducing the burden of drug addiction in society. Over the last two decades, there has been an increasing focus on the role of the excitatory neurotransmitter glutamate in drug addiction. In this review, pharmacological and genetic evidence supporting the role of glutamate in mediating the rewarding effects of the above described drugs of abuse will be discussed. Further, the review will discuss the role of glutamate transmission in two complex heterogeneous brain regions, namely the nucleus accumbens (NAcc) and the ventral tegmental area (VTA), which mediate the rewarding effects of drugs of abuse. In addition, several medications approved by the Food and Drug Administration that act by blocking glutamate transmission will be discussed in the context of drug reward. Finally, this review will discuss future studies needed to address currently unanswered gaps in knowledge, which will further elucidate the role of glutamate in the rewarding effects of drugs of abuse. Rewards increase motivation to perform or repeat tasks and can be broadly classified as natural and drug rewards Schultz, . NaturalDrug rewards can be broadly classified into licit  and illicit  substances. These drugs can also be classified based on their effects in humans as stimulants (cocaine and nicotine) and depressants . Irrespective of the type of drug, the rewarding effects associated with drugs of abuse play a role in the initiation and maintenance of the drug taking habit Wise, . TherefoOver the last three decades, the role of the excitatory neurotransmitter glutamate has been extensively studied in several aspects of drug addiction, including drug reward. Interestingly, some recent studies have demonstrated that glutamate may be involved in mediating natural reward as well , and intracranial self-stimulation (ICSS). Drug self-administration is the most robust and reliable model to measure the rewarding effects of drugs of abuse  depending on their signal transduction pathways, sequence homology, and pharmacological selectivity  and vesicular glutamate transporters (VGLUTs). The EAATs are located on glutamate terminals and presynaptic glial cells and play an important role in glutamate homeostasis O'Shea, . Till daDrugs of abuse alter glutamate transmission via different mechanisms. The primary site of action of cocaine is the dopamine uptake transporter  and Group III (mGlu7 and mGlu8) mGlu receptors decreases glutamate transmission. In accordance, administration of the mGlu2/3 agonist LY379268 decreased self-administration of cocaine  and receptors for other neurotransmitters such as serotonin, dopamine, and GABA  in brain regions associated with reward . Amongst these regions, the NAcc is a major terminal region of dopaminergic neurons originating in the VTA. Systemic administration of cocaine, nicotine, alcohol, and heroin increase dopamine levels in the NAcc  in NAcc glutamate levels after heroin administration , location (synaptic vs. extrasynaptic), temporal resolution (transient vs. sustained), precise glutamatergic inputs activated] that can influence changes in NAcc glutamate levels in response to drug and/or drug-associated cues.Blockade of glutamate neurotransmission in the NAcc had a differential effect on rewarding effects of drugs of abuse  mixed with GABA and cholinergic interneurons. The medium spiny GABAergic neurons project to several brain regions, including the ventral pallidum and VTA, that are responsible for behavioral activity required to obtain rewards  of cocaine and nicotine self-administration (Peoples et al., Genetic manipulation of glutamate transmission has further strengthened our understanding of the role of both ionotropic and mGlu receptors in drug reward. For example, selective knockout of NMDA receptors located on VTA dopaminergic neurons in mice attenuated the acquisition of nicotine-induced CPP (Wang et al., Mice lacking mGlu2 receptors demonstrated increased alcohol consumption, thus supporting an important role for mGlu2 receptors in alcohol reward (Zhou et al., In summary, the rewarding effects of drugs of abuse play a crucial role in continued drug use and development of drug addiction. Over the years, there has been considerable progress in understanding the role of the excitatory neurotransmitter glutamate in drug reward. Drugs of abuse discussed in this review increase glutamatergic transmission in the VTA and facilitate the firing of mesocorticolimbic dopaminergic neurons. Significantly, blockade of glutamate transmission via ionotropic and mGlu receptors attenuates the rewarding effects of drugs of abuse. Further, blocking glutamate transmission in brain regions associated with reward, such as the NAcc and VTA, likewise attenuates drug reward. Finally, repeated exposure to drugs of abuse induces plasticity in several brain regions including the NAcc and VTA that leads to development of drug addiction. Taken together, these findings make glutamate transmission an alluring target for developing medications to treat drug addiction.The ubiquitous distribution of glutamate makes targeting glutamate transmission to decrease the reinforcing effects of drug rewards very challenging. Further, it must be emphasized here that glutamate transmission is involved in many other physiological functions such as learning, memory, regulation of normal behavior and reinforcing effects of natural rewards. Hence, there is a need to develop medications that selectively attenuate the reinforcing effects of drugs of abuse without affecting other physiological functions. Nevertheless, as described in this review, the FDA has approved several medications that attenuate glutamate transmission, which suggest that glutamate transmission remains a viable target for medication development. In fact, drugs targeting the mGlu receptors are in various stages of clinical development for several CNS disorders. In conclusion, while much has been understood about the role of glutamate in drug reward, more work needs to be done to fully exploit the therapeutic potential of glutamate in drug reward and addiction.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.


The Deepwater Horizon blowout, which occurred on April 20, 2010, resulted in an unprecedented oil spill. Despite a complex effort to cap the well, oil and gas spewed from the site until July 15, 2010. Although a large proportion of the hydrocarbons was depleted via natural processes and human intervention, a substantial portion of the oil remained unaccounted for and impacted multiple ecosystems throughout the Gulf of Mexico. The depth, duration and magnitude of this spill were unique, raising many questions and concerns regarding the fate of the hydrocarbons released. One major question was whether or not microbial communities would be capable of metabolizing the hydrocarbons, and if so, by what mechanisms and to what extent? In this review, we summarize the microbial response to the oil spill as described by studies performed during the past four years, providing an overview of the different responses associated with the water column, surface waters, deep-sea sediments, and coastal sands/sediments. Collectively, these studies provide evidence that the microbial response to the Deepwater Horizon oil spill was rapid and robust, displaying common attenuation mechanisms optimized for low molecular weight aliphatic and aromatic hydrocarbons. In contrast, the lack of evidence for the attenuation of more recalcitrant hydrocarbon components suggests that future work should focus on both the environmental impact and metabolic fate of recalcitrant compounds, such as oxygenated oil components. The Macondo 252 well, located ~45 miles off the Louisiana coast in the Gulf of Mexico, suffered a catastrophic blowout on April 20, 2010, which resulted in the tragic death of 11 workers. After continuing to burn for 2 days, the mobile offshore drilling unit, the Deepwater Horizon (DWH), sank on April 22, 2010. Over the next 3 months, an estimated 4.1–4.4 million barrels of crude oil  necessitates different microorganisms with specific biochemical mechanisms directed at the metabolism of the various classes of hydrocarbon compounds   can metabolize hydrocarbons, including the more recalcitrant aromatic compounds. Hydrocarbon activation is achieved through a variety of mechanisms, including hydrocarbon addition to fumarate, intra-aerobic hydroxylation, oxygen-independent hydroxylation, and carboxylation Figure .The ability of microbial communities to metabolize hydrocarbons is not surprising given the ubiquitous distribution of hydrocarbons in the environment resulting from both natural and anthropogenic inputs. The National Research Council  reportedsuccession of dominant bacteria present at any given location was reflective of the corresponding availability of hydrocarbon compounds , but also contained 16% aromatic hydrocarbons and 10% polar compounds .The Gulf of Mexico harbors abundant and diverse microbial communities, including aerobic and anaerobic hydrocarbon degraders, associated with diverse physical niches . Studies following the DWH oil spill established that these communities underwent shifts in composition that reflected the availability of specific hydrocarbon compounds  showed an overwhelming dominance of a novel, uncultured gammaproteobacterium in the order Oceanospirillales , leading to speculation that Colwellia played a crucial role in the in situ degradation of ethane and propane during this time period , and to a much lesser degree (<1 to 5–10%) Oceanospirillales  had all but disappeared and were replaced by a more diverse community structure. The post-capping plume community included previously undetected methylotrophic bacteria , in addition to Flavobacteria and Rhodobacterales  increased  that contained high concentrations of Although much of the oil and gas from both natural seeps and anthropogenic spills can persist in the water column or rise to the water surface as discussed above, it has also been shown that substantial amounts of water-soluble and/or particle-associated hydrocarbons also settle in deep-sea sediments  in the sediments located near the well (within 3 km) compared to a distant (128 km) control sample  were detected in deep-sea sediments near the wellhead compared to distant cores (~50 μg/kg), confirming a greater exposure of the resident microflora to aromatic hydrocarbons near the DWH well OSAT-I, . MetagenColwellia sp., in the sediments that demonstrated high levels of hydrocarbon contamination  revealed an increased abundance in gammaproteobacteria, particularly an uncultured gammaproteobacterium and a Methylococcus, Methylobacter, Actinobacteria, Firmicutes, and Chlorofexi  and BTEX/C3-benzenes (an order of magnitude lower levels) in all of the mousses  and droplets that settled in coastal sediments and sands , and to a lesser extent alphaproteobacteria (Rhodobacteraceae) groups, as the driving force behind the increased abundance , were able to proliferate at the expense of the large influx of labile hydrocarbons.Even a cursory look at the available evidence reveals that not only were the indigenous microflora present and capable of responding to the influx of the hydrocarbons released during the DWH incident, but they also responded quickly and efficiently. This is evident from the integration of isotopic labeling studies and genomic, transcriptomic, and metabolite profiling information Table  that werin situ temperatures, speculated to restrict natural hydrocarbon biodegradation processes, proved not to be a major factor limiting the fate of the oil  and nahA  genes (as well as many other genes) within the plume (Lu et al., n-alkane and naphthalene substrates, respectively. However, it should be noted that the naphthalene-1,2-dioxygenase is one of the most diverse enzymes known, and it can catalyze the transformation of up to 76 different substrates via dioxygenation, monooxygenation, desaturation, O- and N-dealkylation, or sulfoxidation (Resnick et al., bbs genes that encode proteins involved in beta-oxidation of benzylsuccinate. The latter is formed during anaerobic toluene biodegradation and used as a signature metabolite (Elshahed et al., One of the more unique features of the DWH incident was the formation of a hydrocarbon and dispersant plume within the water column. The succession of microorganisms in this amorphous habitat has been documented (Kostka et al., n-alkanes as well as low molecular weight aromatic hydrocarbons (Aeppli et al., Despite the complex microbial community succession patterns and recovery associated with the DWH incident, the metabolic patterns associated with this oil spill were not drastically different from other oil spills. Characteristic changes occurred to the spilt oil, including the loss of a diverse range of Despite the importance of microbial hydrocarbon degradation, it represents only a fraction of the complex oil weathering process Fingas, . A recenIn a world heavily dependent upon the use of fossil energy Odell, , 2013, iDuring the peer-review of this article, two additional reviews were published (Joye et al., The authors declare that the research was conducted without any commercial or financial relationships that could be construed as a real or perceived conflict of interest.


The role of alterations of mitochondrial DNA (mtDNA) in the development of human pathologies is not understood well. Most of mitochondrial mutations are characterized by the phenomenon of heteroplasmy which is defined as the presence of a mixture of more than one type of an organellar genome within a cell or tissue. The level of heteroplasmy varies in wide range, and the expression of disease is dependent on the percent of alleles bearing mutations, thus allowing consumption that an upper threshold level may exist beyond which the mitochondrial function collapses. Recent findings have demonstrated that some mtDNA heteroplasmic mutations are associated with widely spread chronic diseases, including atherosclerosis and cancer. Actually, each etiological mtDNA mutation has its own heteroplasmy threshold that needs to be measured. Therefore, quantitative evaluation of a mutant allele of mitochondrial genome is an obvious methodological challenge, since it may be a keystone for diagnostics of individual genetic predisposition to the disease. This review provides a comprehensive comparison of methods applicable to the measurement of heteroplasmy level of mitochondrial mutations associated with the development of pathology, in particular, in atherosclerosis and its clinical manifestations. Analysis of genetic predisposition to the development of chronic diseases in humans may be a significant and innovative addition to the existing approaches to evaluation of predisposition to different pathologies, including atherosclerotic disease and cancer. Mutations of mitochondrial genome represent the novel type of genetic markers to predict the development of disease. Advisability of studies on pathogenic role of such mutations is derived from molecular mechanisms, by which they can cause defects in the protein structure of some respiratory chain enzymes and in transfer RNA (tRNA), synthesized directly in the mitochondria. This, in turn, leads to reduction in concentrations of these enzymes and tRNA or their complete dysfunction in the mitochondria. As a result, oxidative stress occurs in the cells, which is likely to contribute to the emergence and development of pathology. Typically, mutations in mitochondrial genes coding tRNA and respiratory chain proteins lead to the disruption of protein synthesis in mitochondria, which decrease the efficiency of oxidative phosphorylation and ATP production.In contrast to the nuclear genome, mitochondrial DNA (mtDNA) is inherited strictly through the maternal line. The systems of mtDNA reparation act less efficiently than those of nuclear DNA . This reTo detect and quantify mtDNA heteroplasmy levels many methodologies may be employed, including Sanger DNA sequencing , high peIn general, there are two major pathologies, which provide the “lion's share” of morbidity and mortality in modern society and represent the key challenge to healthcare systems; these are atherosclerosis and oncopathology. The mitochondrial mutations may play mechanistic role in the development of both pathologies, and nowadays there is a growing body of evidence in support of a nonredundant role of mitochondrial factors in the pathogenesis of these diseases.During the process of aging, mtDNA increasingly accumulates somatic mutations through multiplication of damaged DNA molecules because of limited effectiveness of the reparative mechanisms. In experiments in mitochondrial DNA polymerase-deficient mice (mtDNA mutator mice), it was demonstrated that the premature aging is the result of the increase of mtDNA mutations, which primarily alter genes encoding respiratory chain proteins, thereby reducing the functions of mitochondrial complexes I, III, and IV . Thus, tNowadays, there is a limited number of studies on evaluation of mtDNA mutations in atherosclerosis. Since several early studies have demonstrated the monoclonal type of increase of vascular smooth muscle cells number in atherosclerotic lesions –21, a poIn two studies, del4977mtDNA mutation (deletion of ~5 KB mtDNA region containing five tRNA genes and seven genes of respiratory chain enzymes) was analyzed, but no significant differences in the heteroplasmy levels were found between control subjects and patients with coronary atherosclerosis , 23. On A mitochondrial disorder can result from the substitution, deletion, and duplication of mtDNA bases and depletion of mtDNA copies. Most of the data come from the studies on point substitute mutations. As an example, heteroplasmy level of mutation A3243G in the mitochondrial tRNA-Leu 1 gene (UUA/G) in diabetic patients with atherosclerosis was 4-fold higher compared to those age-matched nondiabetic subjects [Mutation T16189C located in the hypervariable D-loop region of mtDNA has attracted attention because of its probable association with a variety of multifactorial diseases. In the study, which included 482 patients with coronary heart disease (CHD), 505 patients with type 2 diabetes, and 1,481 healthy individuals, it was shown that the prevalence of T16189C mutation was significantly higher in patients with coronary artery disease and with diabetes .Recent studies have included a comprehensive analysis of mtDNA mutations in tissue of the atherosclerotic vascular wall. In two studies, 10 of 40 analyzed mutations differed in heteroplasmy level between atherosclerotic and unaffected autopsy samples of human aortic intima , 26. MutIn the cross-sectional clinical study the association between the level of heteroplasmy for the mutation C3256T in human white blood cells and the extent of carotid atherosclerosis, as well as the presence of coronary heart disease (CHD), was investigated . High-reAn association of mitochondrial genetic variation with the severity of carotid atherosclerosis, as assessed by carotid intima-media thickness and the presence of coronary heart disease, was analyzed . SignifiTaken together, these findings indicate that mutations of mitochondrial genome play a substantial role in the development of atherosclerosis. However, mechanistic explanations of this role are not available yet. As an example, C3256T mutation is located in coding sequence of the MT-TL1 gene (codon recognizing UUR) which encodes tRNA leucine and is expressed at the cellular level as a reduced amount of cellular organelles and impaired protein synthesis –32. MutaThere is also the growing evidence of the role of mitochondrial mutations in oncopathology. As an example, highly significant associations were found between mtDNA mutations and the risk of stomach cancer, colon carcinoma, lung cancer, and breast cancer –36.The above considerations raise an important methodological problem, since quantitative evaluation of a mutant allele of mitochondrial genome may be a keystone for diagnostics of individual genetic predisposition to the disease, including atherosclerosis and oncopathology. Routine methods of direct sequencing, like Sanger sequencing, are generally inappropriate due to erroneous results in the case of polynucleotide sequences, very often observed in mtDNA, although such method has been used earlier for the measurement of heteroplasmy level . As a ru(1) enzymatic approaches:restriction fragment length polymorphism (RFLP);amplified fragment length polymorphism (AFLP);cleavase fragment length polymorphism (CFLP);enzymatic mutation detection using the bacteriophage resolvase (EMD);analysis based on the ligase reaction ;invasive oligonucleotide cleavage (Invader);random amplification of polymorphic DNA ;PCR with direct termination (DT-PCR);allele-specific PCR (AS-PCR);(2) chemical methods:(i) chemical cleavage of heteroduplexes;(ii) chemical ligation;(3) methods based on the different electrophoretic mobility of DNA polymorphisms:(i) analysis of the conformation of single-stranded fragments (SSCP);(ii) heteroduplex analysis;(iii) DNA sequencing;(4) detection of the solid phase:(i) hybridization to oligonucleotide arrays;(ii) fiberoptic DNA hybridization analysis;(iii) elongation of immobilized primers (minisequencing);(iv) pyrosequencing;(5) chromatographic methods:(i) denaturing HPLC;(6) physical methods:(i) mass spectrometry;(ii) resonance fluorescence quenching using fluorescence resonance energy transfer (FRET).At present, all existing approaches to the detection of mtDNA allelic variants can be divided into several groups:Subdivision of these methods into above groups is rather nominal, primarily because in most cases a combination of different methods is used .The choice of the optimal approach depends on the specific objective of the study. Historically, widely applied methods for the detection of DNA polymorphisms employed restriction enzymes and were limited to detection of changes in the restriction sites. Later, other enzymatic approaches have been developed. Although they are currently implemented mainly in a high-performance design, a good portion of data indicates the possibility of their expansion and automation. The advantages of these methods are the relatively low cost of the analysis, which is independent of the number of samples, and relative easiness of implementation. However, such methods are time-consuming; that should be regarded as obvious flaw.Each of the above-mentioned strategies has its own advantages and disadvantages. For example, HRM, HPLC, and endonuclease method may be excellent for qualitative detection of heteroplasmy itself, but they have insufficient resolution for the quantitative measurement of mtDNA heteroplasmy level. Pyrosequencing and next generation sequencing provide high accuracy for the quantitative measurements of heteroplasmy levels but are extremely expensive, thus limiting the possibility of routine analysis of a large number of samples.Currently, the most versatile methodology for the quantitative measurement of mtDNA heteroplasmy level is qPCR employing fluorescence detection. To detect the presence of specific nucleotide sequences using fluorescence detection in real time, there are several techniques, the most known of which are the ones with intercalating dye SYBR Green I and methods with specific degradable oligonucleotide probes (TaqMan) and hairpin structures (Scorpions) .Summarizing the advantages and disadvantages of this approach, it should be noted that the use of DNA probes in some specific form is the most preferable to increase the specificity of analysis of mtDNA heteroplasmy. However, the disadvantages of the above described approach relate to a high cost of probes, and accordingly, the primers and amplification process are expensive as well. At the same time, the use of intercalating agents is very simple and cheap. The need for selection of specific primers and probes seems to be a limitation factor, but now the number of commercially available primers steadily increases, and their efficiency has already been validated. Thus, the use of intercalating agents becomes highly attractive.low specificity and frequent false-positive results for the intercalating dye SYBR Green I, the inconvenience of job associated with the need to carry out the test in two separate tubes;low efficiency of reaction and, consequently, low sensitivity of hairpin structures method (Scorpions).However, the use of intercalating dye SYBR Green I and hairpin structures (Scorpions) has certain limitations that prevent the wide use for quantitative measurement of mtDNA heteroplasmy, such asA widely recognized method of mutations detection is the use of the energy transfer through the fluorescent resonance phenomenon (FRET), which is used, for example, in analysis systems based on TaqMan-type and similar probes . TaqMan Considering the presence of two alleles  within the mitochondrial genome in the same sample in an unknown proportion, the most appropriate approach for the development of RT-PCR method of mtDNA heteroplasmy quantitative measurement is the use of TaqMan probes. As for SYBR Green method, it can be used for the cases when the multiplex measurement of heteroplasmy level of some mutations would be impossible.At present, it is clear that mitochondrial factor may play a pivotal role in susceptibility, induction, and progression of chronic diseases, including atherosclerosis and oncopathology. However, the impact of certain mtDNA mutations to the development of pathology is widely unknown. More studies focused on the assessment of new heteroplasmic mtDNA mutations affecting mitochondrial function in relation to atherosclerosis and oncopathology are definitely required. The deep knowledge about the pathophysiological relevance of each disease-associated mtDNA mutations is of great importance.Among the various modern methods for quantitative assessment of mtDNA heteroplasmy level, the qPCR method based on TaqMan technologies seems to be the most promising for the purposes of clinical diagnostics. The simplicity of sequences design, low price, wide availability of equipment, and the simplicity of the analysis itself can make it available for clinical practice. In parallel, an informative pyrosequencing method may be further considered as the “gold standard” for the measurements of mtDNA heteroplasmy for gaining reference values.


The amygdala is a structure of the temporal lobe thought to be involved in assigning emotional significance to environmental information and triggering adapted physiological, behavioral and affective responses. A large body of literature in animals and human implicates the amygdala in fear. Pain having a strong affective and emotional dimension, the amygdala, especially its central nucleus (CeA), has also emerged in the last twenty years as key element of the pain matrix. The CeA receives multiple nociceptive information from the brainstem, as well as highly processed polymodal information from the thalamus and the cerebral cortex. It also possesses the connections that allow influencing most of the descending pain control systems as well as higher centers involved in emotional, affective and cognitive functions. Preclinical studies indicate that the integration of nociceptive inputs in the CeA only marginally contributes to sensory-discriminative components of pain, but rather contributes to associated behavior and affective responses. The CeA doesn’t have a major influence on responses to acute nociception in basal condition, but it induces hypoalgesia during aversive situation, such as stress or fear. On the contrary, during persistent pain states , a long-lasting functional plasticity of CeA activity contributes to an enhancement of the pain experience, including hyperalgesia, aversive behavioral reactions and affective anxiety-like states. Many cerebral structures, constituting the pain matrix, participate in nociception and in the expression and modulation of pain. They transmit and decode nociceptive information, generate, amplify or reduce the pain sensation, allow expression of defensive or distress behavior, and influence the affect. Among these brain structures, the amygdaloid complex, or amygdala, appears as a key component of the pain matrix . Indeed,Pain includes a strong emotional component making it aversive and a reciprocal relation exists between pain and affective states. Stress can either inhibit or amplify pain; and anxiety as well as depressive states are often associated with more intense pain sensation, particularly when pain becomes chronic. In such chronic pain state, the amygdala could contribute to the hyperalgesia as well as to the anxio-depressive consequences of pain.A few publications reviewed the role of the amygdala in persistent pain , 6. The The amygdala is a nuclear complex with almond shape, located in the temporal lobe of mammals and identified by Burdach at the start of 19th century. It is an heterogeneous structure, grouping around a dozen of nuclei depending on the considered nomenclature, these nuclei being classically clustered in 4 groups: superficial, basolateral, central and medial , main component of the central group of amygdala nuclei, is considered as the output nuclei of the amygdaloid complex. It integrates information treated by the corticobasal group and it influences effector centers. The CeA is also reciprocally connected to more rostral forebrain structures, such as the lateral part of the bed nucleus of the stria terminalis (BSTL) and the dorsal part of the substantia innominata. This group of interconnected structures presents strong morphofunctional homogeneity and is designed as the central extended amygdala , 13. In The CeA has three subdivisions: capsular (CeLC), central (CeL) and medial (CeM), that are defined based on cytoarchitecture, neurochemistry and connectivity , 11. TheAmong the various CeA afferents, two main pathways are preferentially conveying nociceptive information Figure . A firstA second pathway conveys to the CeA more direct and raw nociceptive information. While spinal cord only sparsely sends direct projection to the amygdala, spinal nociceptive information is largely transmitted to the CeA via the parabrachial nucleus (PB). This pontine integrative center is a major target for superficial layers of the spinal cord, but also for deeper layers, for the trigeminal complex and for the nucleus of the solitary tract , 20. TheIn vivo electrophysiological studies allowed characterizing the response of CeA neurons to nociceptive stimulations [ulations , 22, 23.ulations  even thoulations , 22, 23.ulations , 19, 20.cfos  the type of pain ; (2) the measured parameters ; and (3) the emotional state of the subjects .Nociceptive tests, relying on the latency of appearance of avoidance behavior or on the stimulus threshold, are used to determine the analgesia or hyperalgesia . These tThese data therefore show that even if the CeA is not directly involved in the modulation of basal nociceptive thresholds in normal situations, it strongly influences analgesia processes Table .Stressful situations (restraint or noise) and fear, especially the experimental model of fear conditioning, cause analgesia as measured by the tail flick or the hot-plate tests. Several studies demonstrated that bilateral lesion or inactivation of the CeA diminishes or abolishes this analgesic effect , 80, 92 The amygdala thus can participate in adaptive processes leading to alleviation of pain sensation but its role is not limited to antinociception. While CeA lesion can block the antinociception induced in the tail-flick test by electrical shock pre-exposure, the affective hyperalgesia, measured by the latency to vocalize after the shock is also reduced by the same CeA lesion . InteresPersistent pain, such as inflammatory or neuropathic pain, has a different profile than acute pain. While animal studies often don’t address all the symptoms described in humans, the models that are used can display spontaneous nociceptive behaviors, hyperalgesia and/or allodynia, aversion as well as the emotional consequences of pain such as anxiety and depression .The role of the CeA in sustained pain has been examined after intraplantar injection of formalin (somatic inflammatory pain), intraperitoneal injection of acetic acid , intraarticular injection of kaolin and carrageenan (somatic inflammatory pain), and ligation/compression of the sciatic nerve (neuropathic pain) Table . GeneralThe affective and emotional dimensions of pain can also be under the influence of the amygdala. The effects of pharmacological manipulations of the CeA on vocalizations reported above (see also Table Finally, the CeA may be a part of the mechanism linking sustained pain to anxiety and depression-like states. It has been shown that the models of monoarthritic pain or of sciatic nerve ligation induce anxiety that is correlated with the activation of the CRF system in the CeA. The expression of this neuropeptide, which has a well established role in anxiety, is increased in the CeA of neuropathic animals , 40, whiIn humans, brain imaging studies  demonstrIn summary, the amygdala, particularly the CeA, may have a mainly antinociceptive influence in acute/phasic pain conditions associated with situations of stress or fear, and a mainly pronociceptive influence in persistent/tonic pain conditions which concerns the sensory as well as the emotional and affective dimensions.Some information is available on the cellular and molecular mechanisms by which the amygdala can exert its bidirectional effects on pain parameters. Projections from the PB to the CeA give direct information about the perceived nociceptive stimulation through glutamatergic and peptidergic synapses –102, whiIt is now clearly established that persistent pain causes long-lasting changes in the activity of the CeA that could account for its pronociceptive influence on sensory and affective components of pain , 6. In aAn intriguing observation is that the right CeA appears largely more implicated in persistent pain than the left CeA. Indeed, the activation of ERK after intraplantar formalin is restricted to the right CeA, and the blockade of ERK activation in the right CeA decreased mechanical hyperalgesia at both hindpaws, irrespective to the side of formalin injection , 31. In 2 adrenoceptors located on PB-CeA pre-synaptic elements which strongly regulate glutamate release [A number of neurochemical and molecular mechanisms involved in the sensitization of the CeA have been elucidated. They especially implicate the modification of pre-and post-synaptic glutamatergic transmission, with an impact on GABAergic transmission, along with neuropeptidergic modulation , 6 Figu. Followi release .Figure 3While we focused this review on the CeA, it is necessary to remind the contribution of BSTL and BLA to pain process.2-adrenoceptor agonist [The CeA belongs to the central extended amygdala; it is largely interconnected with the BSTL and shares similar afferents, including polymodal and nociceptive inputs from the BLA and the PB , 21. Whi agonist  or CRF1  agonist  decrease agonist –122 and  agonist . These d agonist .c-fos mRNA [The BLA, beyond its role as a mere input provider to the CeA and BSTL, also appears as an important actor in pain processes. Some observations suggest that the BLA and the CeA can have a parallel role in nocifensive responses. Stress- and fear-induced analgesia are reduced by intra-BLA infusion of cannabinoid CB1 receptors antagonists, of muscimol or of diazepam –127; mu fos mRNA , arthritfos mRNA  and neurfos mRNA . A functfos mRNA .The affective component of pain can also be modulated by the BLA. The formalin-induced conditioned place aversion is reduced by lesion of the BLA , as it iThese influences of BLA on sensory and affective/emotional aspects of pain can, at least in part, involve its outputs to the CeA and/or BSTL. However, the BLA also appears to be implicated in pain-related cognitive impairment. Indeed, in arthritic rats, blockade of pain-induced plasticity in the BLA with CRF1 antagonism reversed the deficit in decision making evaluated in a gambling task. This deficit has been shown to be dependent upon a deactivation of the medial prefrontal cortex driven by the hyperactivity in the BLA. Importantly, this mechanism appears independent of the CeA . Thus, tThe data from the literature show that the amygdala, and in particular the CeA, contributes to pain processes. Its anatomical and functional relations with pain ascending systems, with pain facilitating or inhibitory descending systems, and with affective and cognitive centers are placing the CeA in a critical position within the pain matrix. Acute pain reactions only require CeA if they occur in specific emotional or adaptive contexts. On the other side, persistent and chronic pains alter CeA activity, which influences the pain experience and the related emotional, affective and motivational states. Reciprocally, these states can modify the amygdala processing of information and its impact on pain.The amygdala has been mostly considered for its roles in emotion, especially in fear , 4. Whil


Genetics play a pivotal role in cancer. This is best exemplified in sporadic intestinal cancer development, which usually starts with mutations in APC then in Ras, p53 and TGFβ (Sears and Garrett, Pseudomonas aeruginosa and intestinal disease in patients with cancer (Markou and Apidianakis, Enterobacteriaceae are suspected contributors to intestinal inflammation and cancer, P. aeruginosa exemplifies the opportunistic nature of many bacterial species toward colonization and disease. The suggested guidelines therefore provide a simple framework within which clinical associations, experimental data, and improved outcomes upon treatment against suspected bacteria need to be taken into account in order to prove causation.The review and opinion articles accompanying this editorial describe key aspects of modeling the hologenome with an emphasis on intestinal infection, inflammation and cancer. One major issue discussed is the adaptation of Koch's postulates in order to assess causation between the human opportunistic pathogen Enterobacteriaceae and Bacteroides species contribute to disease progression in various mouse models. In addition, intestinal dysbiosis is influenced by diet, antibiotics, and an immune genetic background conducive to exacerbated adaptive and diminished innate immune response. The authors highlight the potential of targeting the dysbiosis-inflammation-tumorigenesis axis for the development of novel therapeutic strategies for IBD and colorectal cancer.Experimental data can be obtained with the various mouse models of intestinal inflammation and cancer described comprehensively by Gkouskou et al. . This reWhereas studies on bacteria dominate the literature on the role of dysbiosis in inflammation and cancer, viruses were historically the first microbes to be linked to cancer. A modern approach to this issue is described by Iacovides and colleagues who suggest that the interplay between cancer and cell stemness can be influenced by oncogenic viruses (Iacovides et al., Caenorhabditis elegans has provided valuable insights into the tight regulation of apoptosis during development, infection, and DNA damage (Arvanitis et al., Drosophila models of infection and cancer, as reviewed by Bangi (Drosophila hemocytes, describing the multifaceted roles of these innate immunity cells in development, immunosurveillance, and tumorigenesis (Wang et al., Drosophila Duox, an NADPH oxidase, the homologs of which mediate bacterial killing via oxygen radicals in mammalian mucosae and phagocytes (Kim and Lee, Duox in gut immunity, homeostasis of the intestinal epithelium, and stem cell proliferation. Complementarily, Ayyaz and Jasper put in perspective aging and three responses of Drosophila to intestinal microbes, namely, Duox, the Immune deficiency NF-κ B pathway, and the renewal of intestinal enterocytes (Ayyaz and Jasper, Drosophila as a model for the study of intestinal immunity, inflammation, and disease.Innate immune and stress responses lie at the intersection of apoptosis and cell proliferation during inflammation and cancer. In this regard the simple model organism by Bangi . This reby Bangi . LigoxygDrosophila and mammals may not necessarily emanate from tissue infiltrating blood cells (Panayidou and Apidianakis, Drosophila intestinal epithelium (Kux and Pitsouli, Drosophila model is that biochemical studies can be combined to in vivo genome-wide organ-specific genetic screens to identify ICN components.Regenerative and tumor-promoting cytokines in The authors declare that the study was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The respiratory disease field is changing because of recent advances in our understanding of the airway microbiome. Central to this is dysbiosis, an imbalance of microbial communities that can lead to and flag inflammation in the airways. The increasing momentum of research in this area holds promise for novel treatment strategies. In the past decade, a multiplicity of evidence has emerged in support of the central role of the human microbiome in the etio-pathogenesis of a wide variety of diseases. Work in this area has been aided by the increasing application of culture-independent microbial detection tools, such as high-throughput sequencing and metagenomics. Owing to the relative accessibility of the upper airway, previous attention has focused on sampling microorganisms from this region as representative of the airway microbiota. Recently, however, characterization of the lower-airway microbiota within the context of respiratory health and disease has prompted a shift away from the age-long credo of sterility of the lower airways, towards an overall consideration of the vital role of host-microbiota interactions in respiratory health.Concurrently, interest on the role of non-bacterial members of the microbiota — viruses and fungi — is beginning to emerge. Notably, a complex mixture of bacterial and fungal communities has been identified in the airways of patients with cystic fibrosis. In addition, human tissue surfaces, including the lungs, harbor diverse viral populations . NumerouThe human respiratory tract is not homogenous but is comprised of microhabitats that are distinguished by different constitutive physico-chemical properties, such as pH, temperature, oxygen concentration, mucus production and nutrient availability, all of which could potentially shape the composition of the airway microbiota . The preHaemophilus, Streptococcus, Corynebacterium, Staphylococcus, Moraxella, and Alloiococcus [Pseudomonas, Streptococcus, Prevotella, Fusobacterium, Haemophilus, Veillonella, and Porphyromonas [Streptococcus pneumoniae, which has been shown to predate invasive pneumococcal disease in children, is characterized by a dysbiotic precursory outgrowth of S. pneumoniae within the nasopharyngeal microbiota [The currently proposed concept of a ‘healthy’ airway microbiota in humans, consisting of several putative airway pathogens, contradicts established models of infection in sterile tissues. For instance, the six dominant genera in the upper airway microbiota in children have been shown to be iococcus , membersyromonas . In addicrobiota .Staphylococcus and Corynebacterium, replaced at one year by Alloicoccus and Moraxella. Importantly, these authors found significant associations between nasopharyngeal colonization by Haemophilus, Streptococcus and Moraxella and increased risks of virus-associated acute respiratory illnesses [Highlighting the role of the microbiota in shaping airway immunity, Teo and colleagues  recentlyllnesses . The subllnesses . There aPrevious studies have suggested a link between microbial dysbiosis, defined as a perturbation in the structure of complex microbial communities , and theLow diversity of the gut microbiota in early infancy has been shown to be an important risk factor for the development of allergy. Furthermore, epidemiological evidence suggests an association between growing up in a farm environment, with early life exposure to barn-derived microbes and unpasteurized milk consumption, and decreased risk of developing allergy later in life . EvidencA substantial body of evidence now supports the concept that dysregulation of host–microbiota crosstalk at body surfaces may underlie chronic inflammatory disorders. From a clinical perspective, there is growing interest in the prospects of translating the results of airway microbiome analyses into practical patient-management tools for lung diseases; but in order to accelerate translational prospects of airway microbiome research, several challenges will need to be addressed. First, a harmonization of methodologies for airway sample collection, processing and analysis is required to provide comparable datasets. In addition, more work is needed to elucidate the broader interactions between bacterial, viral and fungal components of the microbiota and how they impact airway disease pathogenesis. Finally, a functional characterization of the airway microbiota using animal models and proteomic, transcriptomic and metabolomic approaches should provide crucial insights into how differential characteristics of the airway microbiota impact respiratory health. Specifically, these tools may provide insights into the predominant transcriptional and metabolic pathways, small molecules and metabolites within the microbiota that mediate host–microbiota and microbe–microbe interactions. Microbiota-derived metabolic pathways and metabolites could serve as future therapeutic targets in airway diseases.A microbiota profile may serve as a future diagnostic or prognostic marker in airway inflammation. Similarly, changes in the microbiota could become indicators for monitoring airway disease progression and could guide therapeutic interventions. Approaches towards deliberate manipulation of the airway microbiota opens exciting possibilities for non-antibiotic-based treatment modalities in upper and lower respiratory tract infections. In addition, engineering the microbiota, by use of metabolites or probiotics during critical developmental windows in early life, may serve as an important prophylactic intervention to prevent chronic airway inflammation.


Nearly 40,000 women will die due to breast cancer annually (In 2015, the American Cancer Society estimates that 234,000 new cases of breast cancer will be diagnosed along with an additional 60,000 cases of carcinoma Frontiers in Oncology Research Topic is focused on providing information to practitioners in this quickly growing discipline. In this research topic, Mookadam et al. eloquently frames the current practice in cardio-oncology and provides future strategy and direction for this novice field.Cardio-oncology is a relatively new field which offers an interdisciplinary and integrative management approach to cancer patients with cardiovascular risks specifically designed to mitigate risks of oncologic therapies and provide early detection and treatment of those at greatest risk of cardio-toxicity. Given the growing clinical relevance of cardio-oncology, this Effects on the heart are a potentially significant and serious clinical problem in the treatment of breast cancer with RT. Over the course of the past 50 years, there have been great advances in the delivery of RT due to the development of new techniques, beam energy, improvement in imaging modalities, and development of image registration strategies. In this research topic, Yue et al. and Chen et al. present novel techniques to track cardiac motion during radiation treatments using fluoroscopy and cardiac MRI, respectively , 5. In aIt is hypothesized that cardiac damage from RT is correlated to the dose absorbed by the heart and differs between left- and right-breast radiotherapy. The damage to cardiac micro- and macro-vasculature is the pathophysiological cause of RT-related heart disease. Taunk and colleagues summarize the literature regarding the underlying pathologic abnormalities and mechanisms of RT-related heart disease .Other manuscripts in this issue reflect the eclectic nature of the field of cardio-oncology. Sharp and George review the potential benefit of stem cell therapy against cardio-toxicities from breast cancer treatments, while Tian et al. review the literature of serum biomarkers of cardiac toxicity after breast cancer treatments , 11. FinIt is our hope that through this research topic, we may continue the dialog of reducing collateral damage to the cardiovascular system by breast cancer therapies, both local and systemic. Guidelines regarding the prevention and treatment of cardiac toxicities are scant and collaborative efforts are needed to facilitate their development.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The prevalence of diabetes is rising globally and, as a result, its associated complications are also rising. Painful diabetic neuropathy (PDN) is a well-known complication of diabetes and the most common cause of all neuropathic pain. About one-third of all diabetes patients suffer from PDN. It has a huge effect on a person's daily life, both physically and mentally. Despite huge advances in diabetes and neurology, the exact mechanism of pain causation in PDN is still not clear. The origin of pain could be in the peripheral nerves of the central nervous system. In this review, we discuss various possible mechanisms of the pathogenesis of pain in PDN. We discuss the role of hyperglycaemia in altering the physiology of peripheral nerves. We also describe central mechanisms of pain. Diabetes affects 382 million people wordlwide and its prevalence is expected to increase to 592 million by the year 2035 . DiabetiPainful diabetic neuropathy (PDN) symptoms exhibit a symmetrical “stocking and gloves” distribution and are often associated with nocturnal exacerbation. It can be presented from a mild pins and needle sensation to stabbing, burning, unremitting, or even unpleasant electric shock sensation. There can be allodynia in the form of cutaneous hypersensitivity leading to acute distress on contact with an external stimulus, such as clothing . The paiPain is the body's perception of actual or potential damage to the nerve or tissue by noxious stimuli. The sensory afferent nerves carry sensations from the skin, joints, and viscera via large and small fibres. Large fibres, such as A-alpha, are responsible for limb proprioception and A-beta fibres carry sensations of limb proprioception, pressure, and vibration. Large A-delta myelinated fibres and small C unmyelinated fibres are mainly responsible for carrying nociceptive sensations. Superficial pain is often a sharp or pricking sensation and is transmitted by A-delta fibres. A deep-seated, burning, itching, aching type of pain is often accompanied with hyperalgesia and allodynia and is transmitted via slow, unmyelinated C fibres. Tissue damage results in the release of inflammatory chemicals, such as prostaglandins, bradykinins, and histamines, at the site of inflammation, which triggers the depolarization of nociceptors, thereby generating an action potential. The action potential transmits the nociceptive sensation, via the dorsal root ganglion (DRG), to the dorsal horn of the spinal cord. The release of glutamate and substance P results in the relay of nociceptive sensations to the spinothalamic tract, thalamus, and, subsequently, the cortex, where pain is interpreted and perceived .Nociceptive pain is the normal response to noxious insult or injury of tissues such as skin, muscles, visceral organs, and joints. Nociceptive pain usually subsides upon the healing of the tissue injury. On the other hand, neuropathic pain arises as a direct consequence of a lesion or disease affecting the somatosensory system without any noxious stimuli. Neuropathic pain is caused by damage or pathological change and is characterised by the activation of abnormal pathways of pain at the peripheral nerves and posterior roots  or spinal cord and brain  . NeuropaThe origin of pain in PDN is not fully understood. The abnormalities in the peripheral or central nervous system could be related to hyperglycaemia, as this is the key metabolic abnormality of diabetes. There are many other conditions that produce pain similar to that of PDN and they may also aid our understanding of the pathophysiology of PDN.Chronic hyperglycemic damage to the nerves can cause regeneration of nerve sprouts, called neuromas, at the stump. The sprouting of the new nerves in all directions causes collateral damage of otherwise undamaged nerves and expands the sensitized area . HyperexTreatment of induced acute neuropathy due to rapid glycemic control in the first month of the initiation of insulin or oral hypoglycemic agents has been reported in the literature as “insulin neuritis.” In 1992, Boulton  first deIn 1996, Tesfaye et al. , 19 Fig. SymptomThe expression of voltage-gated sodium and calcium channels and voltage-independent potassium channels in the DRG has a significant role in the generation of nociceptive sensation and peripheral sensitization that leads to central sensitization. Hyperexcited ectopic impulses are generated by the expression of various voltage-gated sodium channels, such as Nav1.3, Nav1.7, and Nav1.8 . The volMethylglyoxal (MG) is a reactive intracellular by-product of several metabolic pathways. However, the most important source of MG is glycolysis and hyperglycaemia . Studiesα-adrenoreceptor agonists found that it did not activate sympathetic neurons at nociceptor fibres under normal conditions [Nociceptive A-delta and C fibres are normally not directly connected to sympathetic nervous system. Several experiments using nditions , 34. It Neuropathy causes hypersensitivity in nerves as a result of an abnormal epinephrine-mediated transmission from one axon to another. This unusual connection is called ephaptic transmission or cross-talk . It was α-adrenergic antagonists with intravenous phentolamine [Several studies proved this hypothesis and showed dramatic improvement in pain relief after sympathetic blockage , sympathtolamine .In 1965, Melzak and Wall  describeCentral sensitization was first described by Woolf in 1983. Nonnoxious stimuli transmitted from the periphery with A-beta fibres (touch) were perceived as painful by patients with allodynia . A-deltaImpulses from the brainstem nuclei descend to the spinal cord and influence the transmission of pain signals at the dorsal horn. The periaqueductal grey matter (PAG), locus coeruleus, the nucleus raphe magnus, and several bulbar nuclei of reticular formation give rise to descending modulatory pathways. These pathways dampen or enhance the pain signal. The projections from the nucleus raphe magnus to the spinal cord are the major source of serotonin in the spinal cord. Exogenous opioids imitate the endogenous opioids and induce analgesia by acting upon the PAG, reticular formation, and the spinal dorsal horn . The antThe nociceptive hyperexcited impulse generated within primary afferent nerves is modulated and amplified not only at the DRG-spinal cord level but also at the thalamic ventral posterolateral (VPL) level, before being relayed to the cerebral cortex. This was experimentally proved in streptozosin rat model with PDN. The experiment demonstrated hyperexcitability in thalamic VPL neurons, with increased responses to phasic brush, press, and pinch stimuli applied to peripheral receptive fields. VPL neurons from diabetic rats also displayed enhanced spontaneous activity, independent of ascending afferent impulses, and enlarged receptive fields . SelvaraNeuroplasticity or plasticity of the brain is the term used to describe the adaptive change in structure, chemical balance, and function of the brain in response to changes within the body or in the external environment. In response to chronic neuropathic pain, neuroplasticity is associated with somatosensory cortex remodelling, reorganization, and hyperexcitability in the absence of external stimuli. A study of patients with chronic neuropathic and nonneuropathic pain using functional and anatomical magnetic resonance imaging found cortical reorganization and changes in somatosensory cortex activity only in the neuropathic pain group . ProvokeIn summary, the exact mechanism of pain in PDN is far from being clear. The source of pain could be anywhere in the pathway from the damaged nerves to the somatosensory cortex or it could be due to a combination of pathologies. PDN is a distressing condition and, as a result, adversely affects a patient's quality of life, both physically and mentally. Despite significant advances in therapeutics, the treatment of chronic symptoms of pain in PDN is still suboptimal and challenging for clinicians , 60. Thi