Major surgery and critical illnesses such as sepsis and trauma all disturb normal physiological fluid handling. Intravenous fluid therapy for resuscitation and fluid maintenance is a central part of medical care during these conditions, yet the evidence base supporting practice in this area lacks answers to a number of important questions. Recent research developments include a refinement of our knowledge of the endothelial barrier structure and function and a focus on the potential harm that may be associated with intravenous fluid therapy. Here, we briefly describe the contemporary view of fluid physiology and how this may be disrupted by pathological processes. The important themes in critical illness fluid research are discussed, with a particular focus on two emerging ideas: firstly, that individualising fluid treatment to the patient, their underlying disease state and the phase of that illness may be key to improving clinical outcomes using fluid interventions and, secondly, that fluids should be considered to be drugs, with specific indications and contraindications, dose ranges and potential toxicities. Disturbances of body fluid homeostasis are common in major surgery and a range of critical illnesses including sepsis and trauma. Administering appropriate intravenous fluid is a core part of medical care during these episodes. Despite years of research, there is still widespread debate about the best dosing strategy for these fluids and the optimum fluid composition for a given clinical situation. These are still important research questions, as there are clear signs from the current literature that fluid administration strategies have the power to affect clinical outcomes in a variety of areas.Our study of fluid therapy is being refined by recent advances in basic physiology research. In particular, our knowledge of the structure of the vascular endothelium and the way in which fluid is handled at a capillary level has recently been redeveloped. There has also been progress in pharmacokinetic modelling of the movement of exogenous fluids between the body's fluid compartments. At the other end of the research spectrum, clinical fluid trials are becoming ever larger in an effort to address the limitations of the preceding literature, which is based predominantly on relatively small, single-centre studies.Some key themes have emerged from this body of research. Firstly, it is increasingly clear that clinical context is key to the success or failure of a given fluid strategy. A fluid strategy which appears to reduce morbidity in one clinical population (e.g. patients undergoing major surgery) may actually be harmful to another (patients with established sepsis). Fluid therapies are being increasingly tailored to very tightly defined clinical subgroups and indeed to individual patients and the stage of their clinical course. A further theme is the emergence of toxicities related to fluid composition which have only become apparent through large clinical trials. Fluids are therefore increasingly being seen as drugs, with specific indications, contraindications and dose ranges .In this review, we will outline the contemporary model of normal physiological fluid handling. The effects of a range of extreme physiological situations will be described, including acute haemorrhage and systemic inflammation. We will then review the latest literature in relation to clinical fluid strategies in a number of clinical settings Table Water makes up approximately 60% of total body weight in the average adult. Due to the low water content of adipose tissue, this proportion varies widely with obesity (as low as 45%), age  and sex . Body water occupies functional and anatomical compartments. Fluid within cells—the intracellular compartment—is separated from the extracellular compartment by the cell membrane. This is impermeable to large hydrophilic molecules and charged particles, although these may cross it by specific transport mechanisms.A proportion of extracellular water is contained within the bone and dense connective tissue, although due to slow kinetics this is viewed as non-functional. The extracellular compartment is further divided into interstitial fluid between cells and in lymphatics, intravascular fluid within blood vessels, and transcellular fluid including gastrointestinal secretions, joint fluid, cerebrospinal fluid and pleural, peritoneal and pericardial fluid.The vascular endothelium is the key barrier between the intravascular and interstitial spaces. It is particularly relevant clinically as it can be damaged during pathophysiological states such as inflammation, allowing potentially harmful fluid accumulation in the interstitial space. The vascular endothelium in a typical capillary is composed of a single layer of endothelial cells with intercellular clefts closed by tight junctions. The cells sit on a continuous basement membrane. On the vascular aspect of the endothelial cells lies a continuous layer of glycosaminoglycan chains, membrane-bound proteoglycans and glycoproteins. These form the endothelial glycocalyx layer (EGL), which is approximately 1 μm thick.Water and electrolytes can pass freely across the glycocalyx and then beyond the endothelial cells via the intercellular clefts. Proteins and other large molecules however are unable to pass through an intact glycocalyx and are transported in relatively small quantities across the endothelial cells by active processes. The EGL is therefore still part of the intravascular space but contains up to 700–1,000 ml of almost protein-free fluid with the same electrolyte composition as plasma. This creates a large protein concentration gradient—and oncotic pressure gradient—between the plasma and the space immediately below the EGL. This opposes the hydrostatic pressure gradient at the arteriolar end of capillaries, reducing the volume of water and electrolytes filtered out of the capillary. Net filtration across a capillary is expressed by the modified Starling equation , gastrointestinal tract  or kidneys  without adequate replacement.Acute blood loss causes a sudden reduction in intravascular volume, cardiac filling and cardiac output. The transient reduction in arterial pressure is rapidly sensed by the high-pressure baroreceptors in the aortic arch and carotid sinus. The resultant sympathetic activation results in vasoconstriction, increased cardiac inotropy and increased heart rate. A range of other neurohumoral mechanisms are activated, including the renin-angiotensin-aldosterone axis and antidiuretic hormone release. These further contribute to vasoconstriction, reduction in glomerular filtration rate and renal retention of salt and water to compensate for the acute volume loss.These early mechanisms may initially result in apparently normal haemodynamic observations. However, these responses divert blood away from ‘non-essential’ organs  and towards essential organs . Healthy volunteer experiments have demonstrated signs of critical organ hypoperfusion, measured by a reduction in gut pH, despite normal-range blood pressure and heart rate after up to 30% blood volume loss ,5.Acutely reduced capillary pressure, seen in blood loss or due to widespread vasodilatation due to sepsis or anaesthesia, alters fluid movement at the capillary level also. In the initial phase, there may be an ‘autotransfusion’ of interstitial fluid into the intravascular space, although this is limited to approximately 500 ml . OngoingFluid homeostasis becomes even more disturbed if the endothelium becomes damaged and loses its barrier function. A range of insults has been associated with degradation of the glycocalyx, including inflammatory mediators  or even The clinical management of intravenous fluid administration therefore has the potential to influence organ function, morbidity and mortality in pathophysiological settings. If fluid replacement is inadequate during critical illness, inadequate organ blood flow and therefore inadequate delivery of oxygen and energy substrates may occur. This results in inefficient anaerobic cellular metabolism and in severe cases cell death and organ failure. Conversely, excessive salt and water administration, particularly in the setting of endothelial dysfunction, may worsen organ oxygenation and function through oedema formation.Again, the capillary handling of administered intravenous fluids is critical and has recently been re-examined. Free water contained in hypotonic solutions distributes evenly throughout all fluid compartments, leaving only a small proportion within the intravascular space. Traditionally, isotonic electrolyte solutions were thought to distribute evenly through the extracellular  space only, leaving approximately 20%–25% of their volume in the intravascular space. Colloids, due to their superior oncotic pressure, were thought to initially remain almost completely within the intravascular space and were seen as the best fluid for acute plasma volume expansion.It is now proposed that the ‘context’ of capillary pressure is important . At low For both fluid types, in the setting of normal or supra-normal capillary pressure, net filtration will occur, with loss of fluid from the intravascular space and the potential for oedema formation. Although in health the maintained oncotic pressure associated with colloids helps to limit this filtration, this effect is reduced in the presence of endothelial dysfunction. Here, there is the potential for large colloid molecules to pass freely into the interstitial space. However, as more crystalloid volume is needed to achieve similar resuscitation endpoints to colloids, patients resuscitated with crystalloid do ultimately gain a more positive fluid balance  Table 1.As a core part of medical treatment during acute illness and trauma, it is not surprising that a number of fluid administration strategies have been trialled in various clinical settings over the years. This evidence base has shown that fluid strategies have the potential to influence morbidity and possibly mortality outcomes after critical illness and major surgery. However, clear-cut answers which can be confidently translated to widespread clinical practice are rare. A summary of some of the critical illness fluid research areas is contained in Table There are a number of challenges which have hampered evidence-based practice in this area. Firstly, fluid therapy is a prime example of what the Medical Research Council terms a ‘complex intervention’ . This coOne example of this complexity leading to ongoing uncertainty is in fluid strategies around the time of major surgery. An approach to haemodynamic therapy, based on targeting supra-normal global oxygen delivery—monitored using the pulmonary artery catheter—was initially investigated in the 1970s ,40. DespEven at a basic physiological, pathological and pharmacological level, there is considerable complexity which may be important in fluid therapy. The mechanisms of action of fluid strategies at an organ and cellular level have been relatively under-explored in relevant human clinical settings. This gap in the knowledge base may help to explain the variable success of many fluid clinical trials. The ultimate goals of fluid therapy are to provide enough circulating volume and blood flow to organs, so that the quantities of oxygen and nutrients delivered are sufficient to meet their metabolic requirements, while avoiding iatrogenic harm through excess dosing of water, electrolytes or exogenous compounds. Yet this apparent simplicity belies the biological complexity of achieving these goals in diverse disease settings. The clinical benefit or harm brought by administering fluids may result from fluid composition, dose, physiological target, timing during the illness, or complex interactions between these factors, the patient's pre-morbid phenotype and the acute but ever-changing pathophysiological processes.Developing the evidence base behind critical illness fluid practices further therefore needs a combination of basic science, translational trials and clinical trials. Controlled trials with large sample sizes are necessary to definitively detect differences in outcome which may be small but important when applied to the huge number of patients undergoing major surgery or critical illness. But achieving this size often requires pragmatic trial design and heterogeneous groups  which potentially obscure the nuances of which individual patients might get benefit or harm and when. More mechanistic studies are needed to explore what aspect(s) of the intervention strategy brought the benefit or harm and to address apparently basic but as-yet unanswered questions, such as how the body handles the currently available range of exogenously administered anions.It is implausible that one fluid administration strategy will ever be beneficial to all patients in all clinical settings at all time points in their critical illness or physiological insult. There is an increasing realisation that trial findings in one group cannot be readily extrapolated to other groups. The theme of ‘individualisation’—tailoring fluid interventions to each patient and each setting—is gaining momentum and is seen in the following areas.Goal-directed haemodynamic therapy in patients undergoing major surgery is a longstanding example of adapting fluid dosing to a patient's baseline physiology. This is also true of contemporary algorithms aimed at optimising cardiac stroke volume. These involve a physiological ‘test’ in the form of a fluid bolus with ongoing monitoring of stroke volume. If the patient is fluid replete—or cardiac performance is beyond the optimum point of the Starling curve—there will be no further increase in stroke volume and fluid bolus administration can stop. This is in contrast to other dosing strategies based on patient weight or physiological signs which are poor at predicting a response to fluid.There is growing evidence that the timing of plasma volume expansion is important in its potential success. This may be seen even within the relatively short time frame of a surgical operation. One study showed that despite only modest differences in the overall volume of fluid given to patients undergoing surgery, the use of cardiac output monitoring to guide fluid input led to fluid being given earlier in the operation, which led to sustained increases in cardiac output not seen in the control group . This waThe differing physiological requirements from fluid therapy during the developing stages of critical illness and circulatory shock have been recognised in a new proposed treatment approach :• Salvage phase—the immediate phase where fluid resuscitation is used as a lifesaving measure, guided by immediately available information such as clinical assessment, blood pressure and heart rate• Optimisation phase—guided by more in-depth monitoring, and aiming to optimise global oxygen delivery, with resultant improvements in arterial lactate and mixed venous blood desaturation• Stabilisation phase—a more cautious approach to fluid administration, aiming to minimise iatrogenic harm caused by fluid overload and institute more general organ support where required• De-escalation phase—appropriate during the start of resolution of critical illness, aiming to reduce vasoactive treatments and achieve a negative fluid balanceA number of clinical syndromes  appear to have common features which may be addressed by similar fluid approaches. Yet the underlying disease process seems to be critical to the benefit or harm brought by these fluid strategies. In mixed critical care populations, including severe sepsis, there is growing evidence that starch-based colloids are associated with an increased requirement for renal replacement therapy ,18. This−1), which may have contributed to intracranial hypertension [Another example of this is the Saline vs. Albumin Fluid Evaluation (SAFE) study, a large multicentre trial comparing albumin with isotonic saline for fluid resuscitation in a mixed critical illness population . Overallrtension . Conversrtension . No diffLastly, the most basic example of matching fluid input to the pathophysiological situation has been summarised in the UK national guidance on intravenous fluid use . Here, c−1 but avoiding starches)• Resuscitation • Routine maintenance (25–30 ml.kg• Replacement due to ongoing losses and redistribution (fluid composition tailored to match the estimated water and electrolyte loss)A further layer of complexity in fluid treatments may come from the health-care setting in which they are used. Fascinating insights have come from the FEAST study, which took the principle of fluid bolus therapy early in the presentation of patients with sepsis but applied it in the setting of children presenting to hospital in Africa . In contAbnormal fluid handling is a central part of many critical illnesses, and exogenous fluid administration has become a core part of medical care in these settings. Despite apparently rational physiological goals, the reality of successful fluid intervention is extremely complex at a basic biological, clinical and health-care system level. As a result, many of the key research questions in this field have not yet been definitively answered despite years of research. Large trials and meta-analyses have raised the possibility of fluid-related toxicities long after these fluids have been brought to market. Intravenous fluids must be re-framed as drugs, with indications, contraindications, side effects and dose ranges. Lastly, the emergence of the theme of individualisation—to timing, patient, disease state and health-care setting—is an important step forward, but a combination of robust basic science, translational trials and large clinical trials will be required to progress this area further.Jv: net transcapillary flow; Pi: interstitial hydrostatic pressure; Pv: vascular lumen hydrostatic pressure; S: subglycocalyx layer.EG: endothelial glycocalyx; EGL: endothelial glycocalyx layer; ESL: endothelial surface layer; IIs: subglycocalyx oncotic pressure; IIv: intravascular oncotic pressure; MRE declares that he has no competing interests. MGM has received honoraria for speaking, or consultation and/or travel expenses from Baxter, B. Braun, Covidien, Fresenius Kabi, Hospira and LiDCO. He is the National Clinical Advisor for the Department of Health Enhanced Recovery Partnership, is a Smiths Medical Professor of Anaesthesia and Critical Care at UCL, has consulted AQIX , is the Director of Medical Defence Technologies LLC (‘Gastrostim’ patented) and is a co-inventor of ‘QUENCH’ (pump) IP being exploited by UCL Business. Professor Mythen's institution has also received charitable donations and grants from Smiths Medical Endowment and Deltex Medical. Professor Mythen is also a co-author of the GIFTASUP guidelines on peri-operative fluid management, the Chair of the National Institute of Academic Anaesthesia, a Board member of the Faculty of Intensive Care Medicine, a Council member of the Royal College of Anaesthetists of Great Britain and Ireland, the Editor in Chief of Peri-operative Medicine, on the Editorial Board of the BJA and Critical Care, a member of the Improving Surgical Outcomes Group, a member of the NICE IV Fluids Guideline Development Group (now an expert advisor) and the Co-Director of Xtreme Everest.MRE and MGM both conceived and planned this review. MRE drafted the manuscript. MGM helped to draft the manuscript and revised it critically. Both authors read and approved the final manuscript.


Mitophagy is an important mechanism in mitochondrial quality control through autophagic clearance of damaged mitochondria and has been considered to assume protective roles against some diseases, especially neurodegeneration, including Parkinson's disease (Haelterman et al., MitoTracker has emerged as a useful tool in evaluating mitophagy. Mitochondrial potential-independent MitoTracker is able to stain live mitochondria and enables demonstration of colocalization of mitochondria and autophagosome or lysosome with the aid of corresponding marker (Klionsky et al., Despite the promising role of MitoTracker, we would like to highlight potential limitations in its interpretation as some factors may significantly affect determination of mitochondrial mass by MitoTracker staining. First, MitoTracker Deep Red is actually a mitochondrial potential-dependent dye though this is usually not highlighted in the manufacturer's manual. It has been used as an index for mitochondrial potential (Lugli et al., To further elucidate if mitochondrion-bound MitoTracker dyes could retain in mitochondria regardless of mitochondrial potential, experiments were also carried out with MitoTracker loaded prior to drug treatments. We found that CCCP treatment reduced fluorescence intensity in both MitoTracker Deep Red and MitoTracker Green Figures , rather Changes in the fluorescence intensity of MitoTracker dyes can be caused by mitochondrial protein degradation mediated by mitochondria-derived vesicles (McLelland et al., In essence, our data clearly show that currently used mitochondrial dyes, including MitoTracker Deep Red or MitoTracker Green, may be affected by mitochondrial potential or ROS. Taking these limitations into account, we suggest that quantitative assessment of mitophagy using MitoTracker should be always interpreted along with other mitophagy assays. Meanwhile, we await future development of a more specific marker for precise measurements of mitophagy.BX and ET conceived and designed the experiments. BX, XD, and WZ performed the experiments. BX and ET analyzed the data and wrote the paper.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.


A large gap remains between the amount of knowledge in scientific literature and the fraction that gets curated into standardized databases, despite many curation initiatives. Yet the availability of comprehensive knowledge in databases is crucial for exploiting existing background knowledge, both for designing follow-up experiments and for interpreting new experimental data. Structured resources also underpin the computational integration and modeling of regulatory pathways, which further aids our understanding of regulatory dynamics. We argue how cooperation between the scientific community and professional curators can increase the capacity of capturing precise knowledge from literature. We demonstrate this with a project in which we mobilize biological domain experts who curate large amounts of DNA binding transcription factors, and show that they, although new to the field of curation, can make valuable contributions by harvesting reported knowledge from scientific papers. Such community curation can enhance the scientific epistemic process.http://www.tfcheckpoint.orgDatabase URL:  We show this by example, with our own efforts in curating the scientific literature for knowledge about DNA binding transcription factors (DbTFs) in the model species human, mouse and rat. DbTFs guide the RNA polymerase II transcription machinery to specific gene regulatory elements, and play a crucial role in the targeted unlocking of information in the genome. According to bioinformatics analyses, the genome-scale repertoire of DbTFs in humans may comprise around 1700–1800 proteins . We founAlthough many ongoing professional curation projects exist , it is bThis paper reports on an initiative of a group of domain-expert scientists teaming up with professional curators, to exhaustively curate experimental evidence about DbTFs from human, mouse and rat. This effort generates enhanced resources that will provide unique, computationally accessible data about mammalian transcription factors for the research community and will thereby boost genome-wide understanding of gene regulation. This result demonstrates that community curation can make a difference.sequence-specific DNA binding RNA polymerase II transcription factor activity (GO:0000981), or terms that are even more specific. The IntAct database in addition supports recording of the target genes experimentally shown to be regulated by a particular DbTF  and is enabled by the web app SciCura . Its curation results are exported to both GOC’s GO database (http://www.geneontology.org) and EMBL-EBI’s IntAct database (http://www.ebi.ac.uk/intact/).To take on the challenge of curating the remaining literature and archiving this knowledge into databases, a group of domain experts at the Norwegian University of Science and Technology (NTNU) teamed up with professional curators at the Gene Ontology Consortium (GOC) and at the European Bioinformatics Institute (EMBL-EBI). Our initiative builds upon NTNU’s earlier work with TFcheckpoint (Together we work towards three aims: (i) Protocol: we developed a detailed protocol for identifying, characterizing and qualifying knowledge about the DNA-binding and RNAPII regulatory functions of DbTFs in the scientific literature and made the protocol publicly available to serve as curation guidelines ; (ii) SuOur current aim is to complete the curation task for the remaining ∼500 human, mouse and rat DbTFs  for whicOur joint work essentially mobilizes ‘dormant’ knowledge. It gives the scientific community much needed access  to high all available functional annotations, regardless of the level of experimental support. Particular subsets of DbTFs or DbTF:target gene interactions can then be chosen, depending on how stringent the supporting evidence must be for a particular use case. For example, regulatory network building would often take into account only DbTF:target gene interactions meeting the highest confidence criteria, whereas the integration of genome-scale data sets for high-throughput hypothesis assessment may consider interactions supported by any confidence level. In this context, a central challenge is to provide full transparency of the suggested scoring criteria, and also to provide access to the detailed underlying evidence in a way that enables users to implement their own scoring or selection criteria. For example: IntAct records the specific version of an EMSA experiment used for establishing interaction of DbTFs and target DNA sequences. This allows a user to select only annotations based on high-confidence EMSAs that use purified DbTF protein, and to dismiss EMSA experiments performed with nuclear extracts, as the latter leave open the possibility that proteins other than the putative DbTF mediated DNA-binding.The added value of specifying the experimental context in which functional evidence was obtained should not be underestimated. Such experimental details can be specified with PSI-MI terms, for instance for DNA binding. This enables confidence scoring in a manner analogous to the MIscore protocol established for protein–protein interactions , availabOur work also leads us to contribute to the PSI-MI vocabulary. While we curate, we encounter opportunities to refine and extend the PSI-MI vocabulary with terms that allow for a more differentiated annotation of experimental evidence, and for documenting causal, transcription regulatory relations between DbTFs and their target genes. Work like this will further increase the power of scoring opportunities and the rich semantic depth of structured knowledge.Much work remains in harvesting valuable information and enabling knowledge from the scientific literature, not only about DbTFs but also protein-binding transcription regulators, chromatin modulators, etc., or indeed proteins and other biological components in any other biological domain. The experts most qualified for this task are out there, in the scientific community. All of us can significantly complement professional programs; participate in the development of curation protocols, ontologies, and annotation databases; and allow colleagues to benefit from cooperative efforts like the one described here.We encourage funding agencies to acknowledge our shared responsibility for taking care of knowledge generated in costly research activities, as current practices may lead to waste: discovered knowledge, or sometimes re-discovered knowledge which is not made commonly available in a format easily enabling computational retrieval. We hope that the scientific community as a whole can identify incentives and place increasing emphasis on various important curation endeavors. This includes continuing to support the professional curation programs that guarantee the necessary foundation for data governance, maintenance of standards, databases, access through web-interfaces and automated data exchange technologies. Only then can valuable results of public financing persist, become broadly available in formats practical for consumption, and increase the general efficacy of research projects. In addition, we call for the scientific community to explore new approaches for ‘curation at the source’. Perhaps efforts are needed for lowering thresholds to curation and for persuading or rewarding  the origWe are confident that the product of curation, including efforts described here, will serve as a reference for both small-scale assembly of regulatory pathways, and genome-scale analyses of gene regulatory networks [such as ENCODE for genome-scale DbTF function evidence ]. Our cuOur story provides evidence that joint action can make a difference. We learned that a community of volunteering domain experts can team up with professional curators and together develop specific and effective curation protocols. A community can more readily identify gaps and hurdles in ontologies that are needed to capture essential experimental context and biological relationships. A community can together make a significant impact on the information available from annotation databases. The impact of such actions grows larger as more of our responsible colleagues step up and mobilize their peers to take similar action. We welcome colleagues to get into contact, as together we can share and evolve the procedures and tools to get additional efforts accomplished.ST conceived the idea, participated in DbTF annotations, their quality check and manuscript writing. SV designed and developed a web app SciCura that enables community curation of DbTFs at NTNU and participated in manuscript writing. KC maintained the TFcheckpoint database and helped to coordinate the curation process. KRC, JAB participated in mouse DbTF annotations, quality check and manuscript writing. RPH participated in human DbTF annotations, quality check, submission to GO database and manuscript writing. SO, HH participated in DbTF - target gene annotation, submission to the IntAct database and manuscript writing. LT participated in DbTF annotation and their quality check. MK, AL supervised idea conception, participated in DbTF annotation, their quality check and manuscript writing. All authors read and approved the final manuscript.


By creating this web application, researchers can immediately obtain and survey numerous amounts of expression energy data from the ABA, which they can then use to supplement their work or perform meta-analysis. In the future, we hope to enable ABADV across multiple data resources.The Allen Brain Atlas-Driven Visualizations (ABADV) is a publicly accessible web-based tool created to retrieve and visualize expression energy data from the Allen Brain Atlas (ABA) across multiple genes and brain structures. Though the ABA offers their own search engine and software for researchers to view their growing collection of online public data sets, including extensive gene expression and neuroanatomical data from human and mouse brain, many of their tools limit the amount of genes and brain structures researchers can view at once. To complement their work, ABADV generates multiple pie charts, bar charts and heat maps of expression energy values for any given set of genes and brain structures. Such a suite of free and easy-to-understand visualizations allows for easy comparison of gene expression across multiple brain areas. In addition, each visualization links back to the ABA so researchers may view a summary of the experimental detail. ABADV is currently supported on modern web browsers and is compatible with expression energy data from the Allen Mouse Brain Atlas  Gene expression is the process by which a gene, a segment of DNA, is turned into a protein or RNA structure. When studying gene expression, researchers analyze changes in the expression of a particular gene or set of genes by quantifying the amount of its gene-specific transcript. The ability to measure and localize these gene-specific transcripts in the nervous system enables researchers to investigate a broad range of brain phenomena. Researchers use gene expression data in various ways, such as looking at profiles or patterns of expression across several genes, cross-species comparisons, searching for biomarkers, validating different data modalities, correlating gene expression to neuroanatomy, and other large-scale data analysis  images by applying Gene Ontology categories with the genomic set of mouse neural ISH images available in the ABA. Another group also systematically explored high resolution ISH images contained in the ABA by using a data mining tool they developed called Hippo-ATESC , which helped detect neuropil-encoded genes in the hippocampus that are known for their involvement in synaptic structure and plasticity , a growing collection of online public data sets that integrates extensive gene expression and neuroanatomical data complete with a suite of search and viewing tools , we created a web-based, free access and open source application called the Allen Brain Atlas-Driven Visualizations (ABADV). ABADV generates simple-to-analyze visualizations of numerous mouse gene expression data across brain structures using Data-Driven Documents (D3), a Javascript library that uses data to drive the creation and control of visualizations in web browsers   and expression intensity (sum of expression pixel intensity divided by sum of expression pixels). From these statistics, the ABA calculated the expression energy, defined as the sum of expression pixel intensity divided by the sum of all pixels in division. Expression energy is then computed for each brain structure delineated in the Allen Reference Atlas by unionizing grid voxels with the same 3-D structural label. Aligning the expression energy back to the coordinate space of the Allen Reference Atlas allows for cross-comparison of gene expression data between brain structures.The data retrieved by ABADV from the ABA API represents quantified gene expression energy values per 200 μm voxel of the mouse brain. To obtain these values, the Allen Institute developed an informatics data processing pipeline on top of their high-throughput RNA ISH data generation to enable the navigation, analysis and visualization of their massive dataset , the data must be further parsed and formatted so that it is compatible with the user's visualization choices.If multiple pie charts were selected, ABADV will use the appropriate D3 features to generate pie charts, one pie chart for each gene of interest with a pie slice within a chart representing a brain structure of interest Figure . MultiplIf grouped bar charts were selected, ABADV will appropriately create various bar charts of expression energy Figure . GroupedLastly, if heatmap was selected, ABADV will substitute each individual gene expression value into a 2D data matrix of colors Figure . HeatmapFor all three visualizations, ABADV can link back to the ABA experiment page by clicking on any pie chart, bar chart, or cell in heatmap so users may view a summary of its experimental detail, which includes an interactive 3-D representation of gene expression, a histogram of expression energy across major brain structures, probe and gene metadata, and an interactive image viewer that displays ISH images from the experiment. In addition, different color schemes are provided for pie and bar charts. Users can either select a color scheme based on the RGB values assigned to each brain structure according to the anatomic ontology derived from the Allen Reference Atlas Dong, , or a dehttp://www.socsci.uci.edu/~jkrichma/ABADV/Using the ABA API with D3, we designed ABADV to aid users in visualizing expression energy data across many genes and brain structures. This section presents an example of the type of output ABADV is capable of generating in comparison to the Brain Explorer, an application developed by the Allen Institute for visualizing expression data. These examples were performed using the ABADV web application available at: Drd1a, Drd2, Drd3, Drd4, Drd5), serotonin receptors , and various brain structures involved in reward: prelimbic area (PL), infralimbic area (ILA), hypothalamus (HY), hippocampal formation (HPF), striatum-like amygdalar nuclei (sAMY), nucleus accumbens (ACB), ventral tegmental area (VTA), and dorsal raphe nucleus (DR). Together, they form a significant part of the reward circuit, a network responsible for processing various aspects of positive emotional stimulus  is due to sequence-specific binding to mRNA and not with other targets within the cell. The ability to retrieve experimental details directly from the ABA allowed us to unveil this information about our results that would otherwise cause confusion in data interpretation.3 resolution. With Brain Explorer, users have the ability to display ISH expression data from multiple genes superimposed on each other in 3D, as well as fully interact with the Allen Reference Atlas. To obtain gene expression data for display in Brain Explorer, users can either perform a search on their website at http://mouse.brain-map.org/, select gene(s) of interest, then click on the “View in 3D” link in the search results list or search within Brain Explorer itself.ABADV complements the Brain Explorer, which is provided by the Allen Institute. Brain Explorer is a free-to-download desktop application for viewing brain anatomy and gene expression data in 3D . We first searched for our genes of interest through their website. After downloading each gene into Brain Explorer, we navigated through their structural ontology panel within the application, turning on all 3D polygonal brain structures associated with the reward circuit and turning off all other brain structures. Figure While the results from Brain Explorer are identical to ABADV because the data is the same, there are complementary differences. As with our dopamine receptor gene query using ABADV Figures , 5, the Superimposing multiple genes using Brain Explorer made it difficult to identify which dopamine or serotonin SectionDataSetId were expressing the most. In order to know which SectionDataSetId expressed the most in Figures Another key difference between Brain Explorer and ABADV is the type of expression data presented. In Brain Explorer, clicking on colored spheres reveals the expression density and intensity of that voxel, whereas ABADV displays expression energy across the entire brain structure. While the Brain Explorer offers detailed expression data at the voxel level, the ABADV allows quantitative analysis of expression energy across the entire brain structure.Aside from these key advantages of ABADV, Brain Explorer does offer features not available in ABADV. In particular, Brain Explorer enables 3D representation of gene expression with the original full resolution 2D tissue sections is a rich feature that may make data exploration convenient, easily mapping the localization of expression. By selecting a colored voxel like those in Figures http://www.socsci.uci.edu/~jkrichma/ABADV/, has been developed to extend the ABA by providing users with a quick and intuitive way to survey large amounts of expression energy data across multiple brain regions of interest. ABADV combines the ABA API with D3 to obtain and visualize expression energy data from various genes and brain structures using pie charts, bar charts, and heatmaps to display these quantified measurements. Our demonstration of querying the ABA for available dopamine and serotonin receptor genes showed that ABADV could help identify prevalent genes and brain structures that may otherwise be obscured in other types of visualizations, as we showed in our Brain Explorer example where their application superimposes data on top of one another. Using ABADV revealed heavily expressed brain regions such as the nucleus accumbens and dorsal raphe across dopamine and serotonin receptor genes, respectively, (Figures ABADV, which is available for use at:  Figures –7. This http://github.com/UCI-CARL/ABADV. Thus, ABADV is not limited to these summary-level visualization options we have provided. Presently, ABADV does not offer the same richness the ABA provides nor can it presently correspond with other resources available through the ABA. In the future, ABADV will be extended to include other neuroinformatic resource, such as the Allen Human Brain Atlas and the Allen Developing Mouse Brain Atlas, and also resources outside of the ABA such as Gene Expression Omnibus (Edgar et al., Future work may involve incorporating other forms of visualizations and different data sets onto ABADV. ABADV was designed in such a way that someone with some web technology experience could modify the existing code to extend its capabilities. The source code can be obtained at: 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.


Different approaches have been described to make myogenic progenitors either by gene overexpression or by directed differentiation through culture conditions, and several myopathies have already been modeled using iPSC-MP. However, even though results in animal models have shown improvement from previous work with isolated adult myoblasts, major challenges regarding host response have to be addressed and clinically relevant transplantation protocols are lacking. Despite these challenges we are closer than we think to bringing iPSC-MP towards clinical use for treating human muscle disease and sporting injuries.The use of adult myogenic stem cells as a cell therapy for skeletal muscle regeneration has been attempted for decades, with only moderate success. Myogenic progenitors (MP) made from induced pluripotent stem cells (iPSCs) are promising candidates for stem cell therapy to regenerate skeletal muscle since they allow allogenic transplantation, can be produced in large quantities, and, as compared to adult myoblasts, present more embryonic-like features and more proliferative capacity  A resident population of adult myogenic stem cells called “satellite cells” is the main player in the regeneration process. These cells reside in a quiescent state, located between the basal membrane and the plasmalemma of each myofiber. Upon signaling from the damaged myofibers, satellite cells become activated, undergo an asymmetric division to self-renew, and produce activated myoblasts that are able to proliferate, migrate to the site of injury, and fuse with the existing myofibers or to form new myotubes [in vivo they contribute to a much smaller extent than satellite cells [Skeletal muscle is a dynamic organ in which an efficient regeneration process ensures repair after damage. The process of muscle regeneration creates new myofibers after necrosis resulting from injury or a degenerative process. The myonuclei of multinucleated myofibers are post mitotic, arrested in the Gmyotubes . Besideste cells . Repeated cycles of myofiber necrosis and regeneration in muscle dystrophies (MD), such as Duchenne muscular dystrophy (DMD) and some limb girdle dystrophies, result in exhaustion of satellite cell regenerative capacity in humans . Similarin vitro expansion because myoblasts are limited to a few passages due to senescence and the decreased self-renewal capacity of the cells due to the expansion process [Transplantation of donor myoblast or satellite cells isolated from healthy individuals has been tried extensively in the past with somewhat positive but insufficient results and scarce references to functional improvement . In 1995 process . Therefo process , pericyt process , and mes process . FurtherClinical trials using myogenic cell therapy to treat muscular dystrophies started in the 1990s, showed some engraftment of the donor cells but no clear signals of disease recovery or symptom alleviation see . However, extensive preclinical and clinical work over the past few decades has helped to identify some relevant issues to address in order to improve cell therapy in muscular dystrophies. The main limitations of this therapy are transplanted cell engraftment and contribution to host myofibers, which seems to be highly dependent on survival—immunosuppression is thus required but other factors might be contributing as well—and migration out of the site of injection. The transplantation regime can also affect engraftment success . Taking all this into account, the ideal donor cell for skeletal muscle regeneration should be easily accessible and able to expand extensively without losing myogenic and engraftment capacity, have a great survival and fusion rate with host myofibers (high myogenic capacity), and be highly motile to spread within the muscle. Moreover, it should contribute to the satellite cell compartment, enabling indefinite muscle regenerative capacity. Finally, the ideal myogenic donor cell should have low immunogenicity, and be able to be delivered systemically, since intramuscular injection does not seem a feasible approach given the large volume of muscle tissue to be treated.However, extensive preclinical and clinical work over the past few decades has helped to identify some relevant issues to address in order to improve cell therapy in muscular dystrophies. The main limitations of this therapy are transplanted cell engraftment and contribution to host myofibers, which seems to be highly dependent on survival—immunosuppression is thus required but other factors might be contributing as well—and migration out of the site of injection. The transplantation regime can also affect engraftment success . Taking all this into account, the ideal donor cell for skeletal muscle regeneration should be easily accessible and able to expand extensively without losing myogenic and engraftment capacity, have a great survival and fusion rate with host myofibers (high myogenic capacity), and be highly motile to spread within the muscle. Moreover, it should contribute to the satellite cell compartment, enabling indefinite muscle regenerative capacity. Finally, the ideal myogenic donor cell should have low immunogenicity, and be able to be delivered systemically, since intramuscular injection does not seem a feasible approach given the large volume of muscle tissue to be treated.in vitro and can differentiate into myoblast-like cells, they represent an attractive source of myogenic donors for muscle regeneration. Induced PSC-MP also represents a highly valuable tool for in vitro drug testing and disease modeling for muscular genetic conditions that were so far limited because of the difficulties of obtaining large quantities of tissue.Embryonic stem cells (ESC) are pluripotent stem cells derived from the inner cell mass of a blastocyst that are able to self-renew and to be differentiated in all tissues in the body. Induced PSCs share most of the features of ESCs but are derived from adult somatic cells, e.g., dermal fibroblasts, by the transient expression of a defined set of reprogramming factors . The facin vivo but these protocols were lengthy and inefficient [et al. described how overexpression of MyoD1—a transcription factor that appears after Pax3 and Pax7 in muscle development and in activated satellite cells—alone could not induce myogenic commitment directly on hESCs, but concomitant overexpression of the chromatin remodeling complex component BAF60C overcame the mesodermal transition limitation [et al. describe hESC-derived myogenic progenitors by inducible lentiviral overexpression of MyoD1 directly on hESC cells, without a previous EB formation [Initially, human ESCs (hESCs) proved to be difficult to differentiate into myogenic progenitors, probably due to the fact that paraxial mesoderm and subsequently the myogenic program are not well recapitulated during embryoid body (EB)—three-dimensional aggregates of pluripotent stem cells—formation . The firfficient . It has fficient . Higher fficient . Albini mitation . In oppoormation .+ population from EB derived-paraxial mesoderm [+—satellite cell-like—population from differentiating mouse ESC-derived EB cultured in high serum [Other more efficient and genetic modification-free protocols have been described to obtain myogenic progenitors from hESCs, such as isolation of the PDGFRαmesoderm  or isolagh serum . et al. produced satellite cell-like progenitors by overexpression of Pax7—a transcription factor required for somite myogenesis in the embryo and a marker for satellite cells in the adult—in EB from mice (miPSCs) and humans (hiPSCs) [in vitro and much better engraftment as compared to myoblasts. Lentiviral inducible overexpression of Pax3 in iPSCs from dystrophin-lacking mice, which were gene corrected with a truncated version of dystrophin (μ-utrophin), produced in a similar fashion myogenic progenitors that engrafted, differentiated, and repopulated the satellite cell compartment and exhibited neuromuscular synapses [in vitro and survive in vivo, careful dosage and timing are necessary when using this transcription factor. Since the appearance of iPSCs, extensive work has been done to obtain myogenic progenitors with a vision to their clinical application and disease modeling . The fir(hiPSCs) ,39. The synapses . Goudenesynapses . Also, usynapses . A limitThough gene overexpression approaches are fast, efficient, and appropriate to generate myogenic precursors for disease modeling, the risk of undesired genetic recombination or reactivation makes them unsuitable for a future application in the clinic for regenerative cell therapy. Different ways to obtain transplantable myogenic progenitors that do not involve any genetic modification and are still efficient and fast have recently been described. Recently, several reports describe other protocols without gene overexpression that include high concentrations of bFGF and EGF on free floating spheres  and, fasin vitro-synthesized mRNA to overexpress the required transcription factors for myogenic conversion. It was recently shown as a proof of principle that transfection of MyoD1 mRNA in hiPSCs produced myogenic cells with the ability to fully differentiate [in vitro.Another way of avoiding introducing exogenous DNA is the transfection of rentiate  in vitroOther cells with myogenic potential that are not myoblasts have been derived from iPSCs: the group of Tedesco has developed mesangioblast (pericyte progenitors)-like cells that have been tested in animal models .in vitro, as the produced cells recapitulate the expression of markers observed in vivo. They are able to fuse to produce premature myofibers in the animal in vitro and in most cases they have been tested in animal models for engrafting and fusion with host fiber. Several reports describe the establishment of myogenic cell lines produced from iPSCs from patients with different types of muscular dystrophy. Human iPSC-MPs have been established using MyoD1 overexpression by a PiggyBac vector on hiPSCs: Miyoshi Myopathy, a distal myopathy caused by mutations in DYSFERLIN, patients’ fibroblasts [The different approaches published so far to make myogenic progenitors from hiPSCs are good models of myogenesis roblasts , and carroblasts . Duchennroblasts  and by iroblasts . The groroblasts . The creMyogenic progenitors made from iPSCs seem to be a promising candidate for stem cell therapy to regenerate skeletal muscle since they can be produced in large quantities and present more embryonic-like features, so are probably more motile and proliferative compared to adult myoblasts. However, even though results in animal models show an improvement from previous work with isolated myoblasts, in terms of fiber contribution and functional recovery ,41, a clin vivo engraftment and fusion with host cells, but greater extent is needed for a clinically relevant cell therapy protocol. Limited migration from the injection site, in part due to high mortality, but also to intrinsic capacity, is another major limitation that iPSC-derived cells must overcome to outperform myoblast therapy. Some authors describe iPSC-MP as resembling embryonic more than adult myoblasts [One of the major caveats of myoblast therapy was the massive death after transplantation. The inflammatory and immunological response to allogenic transplants probably played a role in the survival of the cells and also engraftment, migration, and differentiation . Howeveryoblasts . The useyoblasts . Also, byoblasts  and can Another major limitation to regeneration is dense fibrotic tissue. TGF-β1 induces collagen I deposition from myogenic cells with subsequent fibrotic tissue formation. Fibrosis limits myoblast engraftment as well as motility and this prevents axons from arriving to myofibers. Unfortunately, there are no drugs on the market that can overcome fibrosis in MD patients. However, there is a report that bone marrow-derived stromal cell transplantation in the muscle of an ischemia model reduced fibrosis due to paracrine effects . This inin vivo by using biomaterials  will conserve the engrafted cells’ homeostasis and allow asymmetric division and myogenic commitment [in vitro. To complete the niche, extracellular matrix components and signaling molecules to stimulate proliferation, migration, and angiogenesis should be included. Muscle flaps made with decellularized devices from large mammals and synthetic scaffolds complemented with an in vitro-produced extracellular matrix from cell cultures derived from the host provide suitable tools for translation to the clinic [in vivo, some issues have already been addressed, such as restoration of the muscular-tendon junction or vascularization, while others like reinnervation still need further work [Tissue engineering can also be of great help for the survival of transplanted myogenic progenitors in the hostile environment of a damaged tissue. Creating a three-dimensional niche for the transplanted myogenic progenitors that resembles satellite cells’ natural niche mmitment . The cele clinic . From thher work .When addressing genetic origin myopathies, the transplanted cells should contain the correct version of the gene. This can be achieved in two ways: by genetic correction of patient-derived cells or by allogenic transplantation of immunocompatible donor cells. One of the major features of iPSCs is the possibility of generating patient-derived tissues with minor invasion. Several groups have performed gene correction on patient iPSCs. iPSC-derived mesangioblasts, from a Limb-Girdle MD patient, in which the wild-type alpha-sarcoglycan gene had been restored by lentiviral delivery, engrafted, and fused with host fibers when transplanted in nude mice . Lamin AAnother approach is to transplant cells created from a healthy donor that are matched for the main antigens in the host immunological rejection, the HLA antigens. An HLA-typed bank of iPSCs could be created to provide a source of compatible donor cells for the individual patients. A relatively small number of donors can provide an acceptable match to a high percentage of the population . This apIt is necessary to take into account that in the case of genetic diseases that lack the native protein, its expression from the grafted tissue will most likely induce a considerable immune response that needs to be carefully addressed.Moreover, the desirable myogenic progenitor should be able to cross the blood barrier to allow for systemic delivery. Treatment of local damage could be done by local intramuscular injections or bio-engineered grafts, but for a cell therapy for MD, SMA, and ALS, in which all muscles in the body are affected, a systemic delivery is necessary. Very few reports show successful engraftment after intra-arterial delivery ,39,46. Tet al. described how, from several clones tested for in vivo engraftment and fiber contribution, those that performed better were the ones with a normal karyotype [For all the reported work in humans and animals models using muscle stem cells, neither adverse side effect has been described, nor colonization in other organs when systemically delivered . Also, faryotype . In thisaryotype  and cellaryotype  have beearyotype .Whatever the method of choice is for generating the myogenic progenitors, a clinical grade protocol must be designed for the cells to be used in patients. The generation process should not include any viral vector or exogenous DNA, should be free of animal products, and should use as far as possible defined media to increase reproducibility and comply with good manufacturing procedures. Such a protocol has not yet been described for either iPSC generation or the derivation of MP.in vitro, and (iii) better performance when tested in mouse models in vivo—possibly because of more embryonic-like features. In recent years, several protocols of derivation of myogenic progenitors from iPSCs have been described reaching very satisfactory efficiency in a short time. The use of transcription factors  overexpression or GSK3β inhibitors has contributed greatly in this direction. However, a clinical grade protocol still needs to be described, including the definition of safety and genetic stability requirements for clinical applications. Also, isolation of the MP presenting the most promising features for successful regeneration in vivo could improve the performance of the cell therapy, such as selecting cells that are more migratory and proliferative or with the possibility of systemic delivery. Other limitations relating to the host—for example, the inflammatory and immune response and the appearance of fibrotic tissue—present a major hurdle to a cell therapy approach. More research with selective inhibitors or modulators of these processes is needed, and the use of bioengineering to create a 3D protective niche for the transplanted cells would contribute to the long-term success of a muscle stem cell therapy strategy. The use of hiPSCs as a source of myogenic progenitors for cell therapy for the treatment of muscle degenerative diseases overcomes several of the limitations encountered in adult myoblast therapy: (i) easy non-invasive source of donor cells; (ii) unlimited proliferative capacity


Neonicotinoid insecticides are widely used for control of insect pests around the world and are especially pervasive in agricultural pest management. There is a growing body of evidence indicating that the broad-scale and prophylactic uses of neonicotinoids pose serious risks of harm to beneficial organisms and their ecological function. This provides the impetus for exploring alternatives to neonicotinoid insecticides for controlling insect pests. We draw from examples of alternative pest control options in Italian maize production and Canadian forestry to illustrate the principles of applying alternatives to neonicotinoids under an integrated pest management (IPM) strategy. An IPM approach considers all relevant and available information to make informed management decisions, providing pest control options based on actual need. We explore the benefits and challenges of several options for management of three insect pests in maize crops and an invasive insect pest in forests, including diversifying crop rotations, altering the timing of planting, tillage and irrigation, using less sensitive crops in infested areas, applying biological control agents, and turning to alternative reduced risk insecticides. Continued research into alternatives is warranted, but equally pressing is the need for information transfer and training for farmers and pest managers and the need for policies and regulations to encourage the adoption of IPM strategies and their alternative pest control options. Before taking any decision on pest control, harmful organisms must be monitored by adequate methods and tools, where available; tools should include observations in the field as well as scientifically sound warning, forecasting, and early diagnosis systems;Treatments may then be carried out only where and when the assessment has found that levels are above predetermined economic thresholds for crop protection;If economic thresholds are exceeded, agronomic solutions, mainly rotation, should be considered to avoid damage to maize crops including the interference of newly established pest populations with tillage timing and other modifications, choice and modification of sowing dates, and alterations of rotation sequences;If economic thresholds are exceeded and no agronomic solutions are available, biological control or physical treatment or any other non-chemical pest control method should be considered as a replacement for chemical treatment;If economic thresholds are exceeded and no agronomic solutions, biological control or physical treatments or any other non-chemical pest control methods are available, chemical treatments should be selected among those that pose the lowest risk to environment and human health, and they should be used in a way that minimizes the risk of pest resistance by limiting their use over space and time.Systemic neonicotinoid insecticides are used to protect a wide variety of crops. Based on their efficacy to control many insect pests and their systemic activity, they are used extensively in agriculture so that by 2008, neonicotinoids accounted for one quarter of the global insecticide market  treatment of maize crops, in which it was shown that there was a link between neonicotinoids and negative effects on honeybees  (http://www.venetoagricoltura.org/subindex.php?IDSX=120). This advisory bulletin is based on a low-cost area-wide pest and disease monitoring system that establishes when and where pest populations pose an economic risk to arable land. Where the risk actually occurs, it advises how the field evaluation should be carried out. Area-wide monitoring is low-cost since it is based on: (a) pheromone traps, which are user-friendly and inexpensive; (b) pest population models using meteorological information . In the last few years, research has provided useful information about the biological significance of pheromone trap catches and has demonstrated their range of attraction  which are then interfaced with agronomic and geographic variables, leading to improved analysis of risk and optimization of monitoring costs , as well as other soil characteristics are the main factors that influence larval population densities Furlan . GeneralData from studies in Italy indicate that the most important factor in influencing wireworm population levels is crop rotation , may also reduce wireworm populations Furlan , 2004. Ts Furlan , 2004. Ag Furlan . Therefog Furlan ..A range of other potential options are available for fields infested with damaging wireworm populations when planting the sensitive crop in non-infested fields has been ruled out Furlan . The mecIn fields where wireworm populations exceed economic thresholds and the agronomic and biological alternatives are not feasible, alternative insecticides to neonicotinoids, such as pyrethroids and phosphorganics, are available .Western corn rootworm (WCR) damage to maize in Europe is only a risk where continuous maize cropping is adopted, especially when cropping is prolonged for several years . These models have shown that 100 % rotation of maize is not necessary to keep regional WCR populations beneath economic thresholds, as, e.g. the interruption of continuous maize cropping after 3 years reduces the need for rotation to manage successfully WCR to below 60 % of the maize fields  does not appear to be viable for practical application at the moment  would also reduce WCR adult numbers leading to non-economic population levels in the following year. However, foliage treatments should be used with caution and only when other options under an IPM approach have not been successful or are not feasible because wide scale use of insecticides can lead to (i) resistance as already demonstrated in WCR larvae  against WCR beetles may sometimes (i) protect maize silks from beetle chewing if applied before flowering, but this is needed only with very high WCR populations  that should not be the case when IPM strategies are implemented; and (ii) actually reduce WCR population levels and the subsequent oviposition by females. The use of a development model  An IPM approach to managing BCW is based on a combination of large-scale pheromone trap monitoring to detect population levels, the analysis of southerly winds that may carry flying moths, and a development model  that ensures a guaranteed farm income in all cases.Agrilus planipennis (Coleoptera: Buprestidae), is a wood-boring exotic invasive insect pest that is increasingly threatening the health and survival of ash (Fraxinus spp.) trees in large regions of eastern North America  cutting and removing living ash trees in advance of the infestation, (ii) girdling living ash trees on the leading edge of an infestation, and (iii) the application of an effective systemic insecticide  has been successful in establishing a measureable population and has the potential for beginning to control emerald ash borer infestations .Three species of hymenopterous parasitoids (parasitic wasps) were found to parasitize emerald ash borer larvae or eggs in China, and these are being reared in the USA as potential biological control agents Lyons . The empPhasgonophora sulcata, Braconidae: Atanycolus hicoriae) have shown relatively high rates of parasitism on emerald ash borer and hold some promise as a native biological control agent  for farmers, in which the required insurance premium is usually lower than insecticide costs (Furlan et al. We recognize that the adoption of alternatives to neonicotinoids and moving agricultural practices to an IPM approach is particularly challenging where large-scale, cost-effective agricultural operations are on the landscape. Over the past two decades, the trend toward large, commercial agricultural operations has focused on scale economies and efficiencies (Morrison Paul et al. While some of the options for alternative pest control that we illustrate in these case studies have been successfully demonstrated and field-tested, others are under ongoing development. Continued research into alternatives is warranted, but equally pressing is the need for transfer and training of IPM technologies for farmers and other practitioners by public agencies and the need for policies and regulations to encourage the adoption of IPM strategies and their alternative pest control options.


People with an intellectual disability experience both acute and chronic pain with at least the same frequency as the general population. However, considerably less is known about the pain perception of people with Down syndrome. In this review paper, we evaluated the available clinical and experimental evidence. Some experimental studies of acute pain have indicated that pain threshold was higher than normal but only when using a reaction time method to measure pain sensitivity. However, when reaction time is not part of the calculation of the pain threshold, pain sensitivity in people with Down syndrome is in fact lower than normal (more sensitive to pain). Clinical studies of chronic pain have shown that people with an intellectual disability experience chronic pain and within that population, people with Down syndrome also experience chronic pain, but the precise prevalence of chronic pain in Down syndrome has yet to be established. Taken together, the literature suggests that people with Down syndrome experience pain, both acute and chronic, with at least the same frequency as the rest of the population. Furthermore, the evidence suggests that although acute pain expression appears to be delayed, once pain is registered, there appears to be a magnified pain response. We conclude by proposing an agenda for future research in this area. Until recently, the commonly held view was that individuals with intellectual disability have decreased sensitivity to pain , acquired cardiac disease (16%), chronic pulmonary changes (30%), osteoarthritic degeneration of the spine (32%), osteoporosis with resultant fractures of the long bones (55%) or vertebral bodies (30%), untreated atlanto-occipital instability (8%), eye problems (36%), celiac disease (11%), eczema (23%) , published bibliographies of related topics, and references provided by colleagues. We limited our review to publications in the years 1960–2014. For this brief review, we have not attempted to conduct an exhaustive systematic review, but instead have selected literature germane to focus of our paper. Wherever possible in this paper, we will focus on the evidence regarding Down syndrome specifically, although in many studies the population described is a more general intellectual disability group of which some have Down syndrome. We will make the distinction wherever the literature allows us to.In order to determine whether pain sensitivity is altered in people with Down syndrome, it is necessary to have methods by which pain sensitivity can be measured accurately. Sensitivity to pain is measured experimentally by introducing subjects to stimuli of ascending or descending order until the boundary of pain is reached, a procedure termed “method of limits”. Another psychophysical method is the procedure of repeatedly introducing stimuli of various intensities and determining the threshold within the range of the stimulus that can almost never be detected, and that which is almost always detected as painful (“method of constant stimuli”). In both methods, pain threshold is defined as the smallest amount of stimulus energy (or intensity) necessary to evoke pain  capacity to report pain quickly; (b) possible altered somatosensory processes in Down syndrome. Some studies suggest that median nerve conduction velocities,  but not scalp-evoked potential latencies, were slower and their amplitude lower among children and adults with Down syndrome when compared with controls. This suggests impaired peripheral somatosensory function in Down syndrome Kakigi, .In contrast, normal peripheral conduction velocity but prolonged latencies of somatosensory evoked potentials were recorded among infants or young adults with Down syndrome  and there may be facial indicators or physiological indicators (such as heart rate or respiration rate).Although frequently used in studies of general intellectual disability  studies reveal distinctive alterations in brain anatomy among individuals with Down syndrome. For example, there is evidence of smaller overall brain volume, disproportionately smaller cerebellar volume and relatively larger subcortical gray matter volume in people with Down syndrome compared to controls  and lower connectivity in dorsal executive networks . For example, a case report study of a person with a mild intellectual disability who had chronic pain indicated that psychological treatments may be of benefit  designed to be used by caregivers to assist people with intellectual disability in developing pain self-management strategies  also indicate delayed response to noxious stimuli compared to control mice  was abnormal  as well as structures involved in pain processing . Still, some inconsistency exists in pain threshold measurements that may reflect interruption in peripheral conduction and central processing of sensory signals, especially if pain threshold is measured with methods that include reaction time. While such alterations have been reported for innocuous stimuli, studies are needed to prove that such alterations indeed occur in nociceptive pathways. Due to the possibility of delayed reaction time, measuring pain threshold with methods that bypass this limitation, i.e., reaction-time free methods, is preferable. However, pain threshold measurement is suitable only for individuals with mild and perhaps moderate cognitive impairment. Thus, the use of indirect indices of pain is necessary. Additional studies are needed in order to explore which indices best reflect pain in Down syndrome.While individuals with Down syndrome are at increased risk to experience pain due to congenital and acquired abnormalities and environmental risk factors , they typically have difficulty in expressing their pain and their caregivers face great challenges in identifying and quantifying pain. Thus, from a clinical point of view, it is imperative to investigate pain processing and pain expression of individuals with Down syndrome in both the experimental and clinical setting. Until optimal tools are available for this purpose, caregivers should take into consideration unexpected and sometimes seemingly ambiguous responses to painful incidents. We have previously advocated the use of more than one source of information to identify pain, in order to increase the reliability of the information obtained  or derivation of pain , gastrointestinal, dental etc.).The perception of pain associated with self-injury in low functioning persons with Down syndrome.Evaluation of how the presence of dementia affects the manifestation of pain in people with Down syndrome.A greater emphasis on evaluating pain management interventions, including self-management .In the experimental domain:Further evaluation of observer-based and self-report pain assessment tools using calibrated noxious stimuli of varying intensities.Measuring physiological and electrophysiological reactions to experimental pain that may potentially replace self report, including but not restricted to heart-rate variability and electromyography (EMG) and electroencephalogram (EEG).Measuring conduction velocity and reaction time to noxious stimuli.Studying pain perception using event related potentials (ERP) and functional magnetic resonance imaging (fMRI) that enable the association between noxious stimulation and activation in specific brain regions of interest.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.


Dopamine neurotransmission has long been identified as a key component of prefrontal cortical function and flexible cognition; however, flexible behavior does not depend solely on dopamine. Together, multiple systems allow fast and flexible behavior in the face of a constantly changing environment. This Research Topic illuminates work by researchers who are leading efforts to understand how different neural systems yield flexible behavior.Yen et al.).The psychostimulant amphetamine is, paradoxically, a useful treatment for Attention Deficit Hyperactivity Disorder (ADHD) symptoms. Here, Yen and colleagues demonstrate in a mouse model of ADHD that the calming effect of amphetamine is due to decreased GSK3B, initiated through NMDA receptor signaling (Mizuki and Fujiwara).Stress is known to be under the partial regulation of oxytocin, and oxytocin levels in adults may be influenced by severity of childhood maltreatment. The inconsistent quality of results on this topic are tackled by Mizuki and Fujiwara, who were able to demonstrate a correlation of severity of childhood maltreatment with diminishing oxytocin levels in humans . This information is critical in developing individualized treatment plans for treating maladaptive behaviors.A key challenge in combatting drug addiction is the incentive motivational property of a stimulus. This is known to develop through Pavlovian Instrumental Transfer (PIT). Garofalo and di Pellegrino were able to show, for the first time in humans, that Sign-Tracker responses were biased by reward-paired cues while Goal-Trackers were not . These data suggest the usefulness in physical exercise to promote improved flexible cognition in adolescents.Physical exercise is known to promote better cognition, but the mechanism remains unknown. In their article Berse and colleagues identified a particular single nucleotide polymorphism in the Dopamine Transporter (DAT1/SLCA6A3) gene which contributed to improved cognition under physical exercise conditions in adolescent humans . These results fail to replicate an earlier and smaller experiment, and the authors propose that the difficulty in diagnosing ADHD may lie in a number of false-negative studies.Altered neurochemistry in the anterior cingulate cortex (ACC) is thought to be a root cause of the behavioral symptoms of ADHD. However, when Endres and colleagues conducted a large single voxel proton scale magnetic resonance spectroscopy study, they identified no neurometabolic differences in ACC or cerebellum between control and experimental groups (Bissonette and Roesch). Importantly, these populations of neurons also modified their activity based on feedback, significantly increasing activity for rewarded outcomes and significantly decreasing activity for non-rewarded choices. These results demonstrate how the mPFC signals the need for behavior to become flexible.Flexible behavior is known to depend on the medial prefrontal cortex (mPFC). Bissonette and Roesch demonstrate how neural correlates of rules and conflict are encoded and signaled by mPFC neurons as rats modify their behavior in the face of changing contingencies (Raver and Lin). This review serves focuses attention on how the BF salience system is a critical part of signaling attention towards relevant stimuli, promoting flexible behavior.Raver and Lin present a very cogent review of the role that the basal forebrain plays in signaling arousal, attention and decision-making. They discuss relevant literature focusing on how a population of BF non-cholinergic neurons encode motivational salience through ensemble burst firing . These findings are important for interpreting data that manipulates the mPFC and uses reward-based learning paradigms.Most animal research uses reward-based learning paradigms, requiring animals to consume their reward. In this research topic, Parent and colleagues demonstrate how cholinergic and ghrelinergic signaling—both working through KCNQ channels in the mPFC—are critical for regulating the duration of licking. These data suggest that natural variation among animals in their tendency towards either sign or goal-tracking may be correlated with drug addiction vulnerability.Sign-tracking in animals is associated with increased sensitivity to food cues. Nasser and colleagues demonstrate that sign-tracking, but not goal-tracking, rats are more sensitive to food associated cues and that the degree of sign-tracking correlated with a failure to suppress responses during devaluation . These results suggest a role for LC not during initial learning, but during situations which require flexible shifts in behavior.The locus coeruleus (LC), which provides noradrenergic projects to the cortex, is important for attention. Janitzky and colleagues demonstrate that optogenetically suppressing LC activity during a set shifting task impaired performance on reversal and extra-dimensional set-shifting but not on learning of intra-dimensional shifts or compound discriminations . This information is necessary to guide future research on how LHb activity may be important for flexible behavior.The lateral habenula (LHb) acts as a gatekeeper for information about negative outcomes being transmitted to the midbrain dopamine system. Baker and colleagues provide a thorough review of the LHb literature and present data to support a hypothesis whereby LHb neural activity is important for the maintenance of goal-directed actions to new response types during changing contingencies . These data provide further support that frontal cortical neural activity works best with a basal level of dopamine.Particular behavioral responses need to be organized over time, allowing appropriate actions to occur at the right time. Parker, Ruggiero and Narayanan demonstrate that blockade of D1-type dopamine receptors in the medial frontal cortex leads to altered interval timing performance by impacting ramping patterns and differentially field potential activity without impacting baseline firing rates of neurons (Hernandez and Cheer). This review summarizes the literature around how different aspects of the endocannabinoid system may modulate dopamine release.Many neural systems interact while animals engage in complex behaviors and make decisions. Hernandez and Cheer provide a thorough review of the literature surrounding the endocannabinoid system and how endocannabinoid and dopamine systems may interact while animals make decisions .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.


Clinicians often modify evidence-based psychotherapies (EBPs) when delivering them in routine care settings. There has been little study of factors associated with or implications of modifications to EBP protocols. This paper differentiates between fidelity-consistent and fidelity-inconsistent modifications and it examines the potential influence of two clinician characteristics, training outcomes, and attitudes toward EBPs on fidelity-consistent and fidelity-inconsistent modifications of cognitive behavioral therapy in a sample of clinicians who had been trained to deliver these treatments for children or adults.Survey and coded interview data collected 2 years after completion of training programs in cognitive behavioral therapy were used to examine associations between successful or unsuccessful completion of training, clinician attitudes, and modifications. Modifications endorsed by clinicians were categorized as fidelity-consistent or fidelity-inconsistent and entered as outcomes into separate regression models, with training success and attitudes entered as independent variables.Successful completion of a training program was associated with subsequent fidelity-inconsistent modifications but not fidelity-consistent modifications. Therapists who reported greater openness to using EBPs prior to training reported more fidelity-consistent modifications at follow-up, and those who reported greater willingness to adopt EBPs if they found them appealing were more likely to make fidelity-inconsistent modifications.Implications of these findings for training, implementation, EBP sustainment, and future studies are discussed. Research on contextual and protocol-related factors that may impact decisions to modify EBPs will be an important future direction of study to complement to this research. The past decade has seen an increase in efforts to implement evidence-based psychotherapies (EBPs) and psychosocial interventions in public sector mental health settings . ConsideDifferent modifications can occur in routine care settings, and distinguishing between types of modifications may be useful both conceptually and for the purpose of understanding whether different kinds of modifications have implications for clinical or implementation outcomes. As Fig. In conceptualizing modifications to evidence-based mental health treatments, it is important to consider the relationship between modification and treatment fidelity. Fidelity has been described as comprising adherence to the intervention components, competence with which the intervention is delivered, and differentiation from other treatments , 15. ImpAdding new, intervention-consistent elements that were not originally included in the intervention Removing intervention components Integrating components of other established interventions Tailoring aspects of the intervention  for an individual patient or population Loosening the structured aspects of the intervention Changing the length of the intervention without adding or removing aspects of the intervention Temporary drift or deviation from the core elements of the intervention as originally designed Some of these modifications can be considered inherently fidelity-consistent or fidelity-inconsistent based on several considerations, including implementation frameworks that differentiate between core and peripheral elements of interventions ) Although this study has a number of strengths, namely being the first prospective study to focus on individual-level factors associated with subsequent modifications to EBPs, there are limitations. First, we relied on interviews to obtain information about the types of modifications that clinicians make in usual care, introducing the potential for recall bias or failure to recognize specific modifications in their own practice. It is likely that modification is better captured when observed in audiotaped sessions by expert raters and in conjunction with fidelity ratings . It willThere is little doubt that clinicians modify EBPs in routine care settings. However, there are potential implications of modifying EBPs, and these are the subject of considerable debate in the literature , 35. PosUntil more is known about the ways in which modifications impact clinical outcomes in different contexts, developers of complex psychosocial interventions, implementation facilitators, and training consultants can either allow modifications to occur naturally and hope for good outcomes or attempt to facilitate an effective process of adaptation. The development and testing of highly flexible protocols that include guidance on selection of appropriate modules or interventions is a promising direction toward this goal . The res


Over the past decade, new diagnoses of HIV have increased eightfold among men who have sex with men (MSM) of other or of mixed ethnicity in the UK. Yet there is little intervention research on HIV among black and minority ethnic (BME) MSM. This article aimed to identify effective HIV and sexual health prevention strategies for BME MSM.We searched three databases PubMed, Scopus and PsychInfo using a combination of search terms: MSM or men who have sex with men and women (MSMW); Black and Minority Ethnic; HIV or sexual health; and evaluation, intervention, program* or implementation. We identified a total of 19 studies to include in the review including those which used randomised control, pre/post-test and cross-sectional design; in addition, we included intervention development studies.A total of 12 studies reported statistically significant results in at least one of the behavioural outcomes assessed; one study reported significant increases in HIV knowledge and changes in safer sex practices. In 10 studies, reductions were reported in unprotected anal intercourse (UAI), number of sexual partners, or in both of these measures. Six out of the 13 studies reported reductions in UAI; while seven reported reductions in number of sexual partners. Seven were intervention development studies.Research into the mechanisms and underpinnings of future sexual health interventions is urgently needed in order to reduce HIV and other sexually transmitted infection (STI) among UK BME MSM. The design of interventions should be informed by the members of these groups for whom they are targeted to ensure the cultural and linguistic sensitivity of the tools and approaches generated. Men who have sex with men (MSM) continue to be the group most affected by HIV infection . EstimatPublic Health England collect annual surveillance relating to diagnosis and routes of transmission; these figures indicate that men of white ethnicity comprise 84 %  of cases of newly diagnosed MSM with HIV in E&W with a route of exposure through sex with men ,–45,inters health  sought tack MSMW . Findingack MSMW . HealthMack MSMW . HOLA enmoderate to high efficacy of behavioural change interventions in African-American, Latino and Asian and Pacific Islander (API) men exclusively in the North American context. Six studies showed reductions in condomless anal intercourse, while seven studies showed a decline in the number of sexual partners: in the absence of other prevention methods, these behaviours place men at increased risk of HIV acquisition.The overall results of this review indicate A number of the studies reported extensive preparatory work to ensure the relevance of interventions, including the development of culturally sensitive approaches and materials. Many studies adopted a holistic approach exploring the context for men’s sexual behaviours within the realities of their everyday lives, which is also highlighted in psychotherapeutic approaches to HIV prevention . MoreoveMore specifically, these psychosocial factors may reduce the individual’s engagement with ability to negotiate safer sex practices, such as negotiating condom use, discussing HIV with partners, or adhering to pre-exposure prophylaxis (PrEP). Previous research has indicated that behavioural change interventions alone will not lead to reductions in HIV or sexually transmitted infections among BME MSM because recent increases in diagnoses do not seem to be attributable to an increased prevalence of at risk behaviours in comparison to other MSM . Rather,A number of factors contributed to the effectiveness of the interventions in our review in reducing HIV risk behaviours. Notably, studies were underpinned by theoretical frameworks and risk behaviours were addressed in the wider context of men’s lives. Thus, it appears that those interventions that integrate community-based knowledge in broader theoretical frameworks regarding risk-taking behaviour are more likely to be effective. Interventions formed an integrated programme working alongside community based organisations, some were concurrently conducted in multiple cities, studies were conducted over a 3 – 6 month timescale following the intervention, retention rates were high and studies used a range of incentives to minimise attrition. This suggests that future interventions can be conducted and evaluated over longer time periods during which actual behaviour change is most likely to be observable.A number of pilot studies and intervention development research projects were also recognised. There is a promising trend towards designing behavioural and psychosocial change approaches through a complex, iterative and multiple staged process involving individuals who are members of the communities for which the interventions are designed. This can help facilitate a culturally and linguistically appropriate perspective that takes into consideration the factors that are likely to promote positive engagement with the intervention. This cultural perspective is then integrated into theoretical models of behaviour change. Some of the IDS were focused on raising health awareness, providing HIV/STIs prevention or offering sexual health training through electronic means of communication. These IDS were pilot interventions planned and disseminated through contemporary methods of communication . This is consistent with the prominence of internet, mobile and application communication within young MSM communities with regard to gay in-group socialising, and sexual partner seeking . FinallyWhile this systematic review identifies a number of important factors that underpin effective sexual health interventions, there are some limitations. Specifically, the review focussed on papers published in peer reviewed journals and excluded grey literature. Although the grey literature may not be methodologically robust, it might indicate specific insights which may lead to new research directions. This focus may have biased the results of this review towards studies with significant findings and may have excluded studies with null findings. Moreover, the existing evidence base suffers from some methodological limitations, such as the existence of conflicting and limited evidence which can preclude the formation of robust interventions. For instance, there are inconsistent findings about the link between PrEP use and condom use. While PrEP is protective against HIV, its use without condoms could place MSM at risk of other STIs. The heterogeneity of populations included in studies may also be problematic given the distinct cultural and linguistic needs of particular BME MSM communities. Many of the interventions include only limited follow-up periods which can make it difficult to assess the robustness and duration of the intended behavioural changes. The changing nature of HIV risk-related behaviours and the diverse prevention methods employed by MSM, such as serosorting, strategic positioning and biomedical prevention methods, are not currently reflected and represented in the current interventions and as yet, evaluation of these strategies are poorly represented in the extant literature. Other limitations include the heterogeneity of intervention content and of the outcome measures. Despite these limitations, the review makes an important contribution to developing the knowledge base to inform future behavioural and psychological interventions for BME MSM. The review provides strong evidence that effective interventions for this previously overlooked group of men are underpinned by relevant theoretical frameworks, cultural sensitivity and the involvement of potential users in both the design and delivery of health education and behavioural change approaches. The limitations acknowledged here should be taken into consideration in the development of future interventions.Despite the relevance of HIV and sexual health risk prevention for BME MSM, this review has not produced any intervention studies conducted within a UK context. Research into the mechanisms and underpinnings of future sexual health interventions is urgently needed in order to reduce HIV and other STI infection among UK BME MSM, who remain a high risk group . There hART, anti-retroviral therapies; BME, black and minority ethnic; CPI, condom protected intercourse; IDS, intervention development studies; MSM, men who have sex with men; MSMW, men who have sex with men and women; PEP, Post-Exposure Prophylaxis (PEP); PICO, participants, interventions, comparators and outcomes; PREP, pre-exposure prophylaxis; RCT, randomised control trial; STI, sexually transmitted infection; UAI, unprotected anal intercourse; UAVI, unprotected anal and vaginal intercourse