Since the discovery of dendritic cells (DCs) by Ralph Steinman and Zanvil Cohn 40 years ago , the rolV600 mutant melanoma patients, and by Palombo et al. (We begin this research topic with an Opinion article by Rolf Zinkernagel  and a reo et al. , who desA recurring theme in many of the articles presented herein is that tumor immune evasion arises not only from a simple failure of the tumor microenvironment to support DC maturation but also from an active recruitment and exploitation by tumors of immature, tolerogenic DC that suppress adaptive responses. Seliger and Massa  review mex vivo-generated DC or in vivo-targeted DC. In addition to highlighting the need to better understand tumor microenvironmental factors that should be targeted to improve the efficacy of DC-associated immune stimulation in cancer patients, the authors identify a variety of other factors that must further be studied to optimize these therapies, including mechanisms of antigen delivery to endogenous DC; the frequency, route, and site of DC vaccination; methods of DC activation both ex vivo and in vivo; and the particular DC subsets that should be employed or targeted during immunization. These and other factors are considered more specifically in the context of DC-based therapies in acute and chronic myeloid leukemia patients by Schürch et al. (V600 mutation. Finally, Toubai et al. (Because of the potential of DC to serve as both targets of and delivery agents for tumor immunotherapies, significant efforts have been focused on how best to utilize these cells in the treatment of cancer. Gallois and Bhardwaj  review mh et al. , in ovarh et al. , and in h et al. . Additioh et al.  offer ini et al.  review tWe conclude this collection of articles with a review by Chmielewski et al. , who desAs the intricacies of DC biology and the influence of tumors on DC phenotype and function continue to be uncovered, additional insights into the role of these cells in the induction, regulation, and maintenance of anti-tumor immune responses will continue to shed light on mechanisms of tumor immune escape and inform the design of novel therapies to enhance anti-tumor immunity. It is our hope that the advances highlighted herein and the questions raised for future consideration will generate additional discussion, drive experimental inquiry, and bring into focus the significance of tumor/DC interactions and their impact on overall anti-tumor immunity. Such emphasis is sure to bring rapid advancements in this field and will ultimately lead to the development of more effective cancer immunotherapies and improved clinical outcome in cancer patients in the future.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Many studies have associated environmental exposure to chemicals with neurological impairments (NIs) including neuropathies, cognitive, motor and sensory impairments; neurodevelopmental disorders (NDDs) including autism and attention deficit hyperactivity disorder (ADHD); neurodegenerative diseases (NDGs) including Alzheimer′s disease, Parkinson's disease and amyotrophic lateral sclerosis (ALS). The environmental chemicals shown to induce all these diseases include persistent organic pollutants (POPs), the plastic exudates bisphenol A and phthalates, low molecular weight hydrocarbons (LMWHCs) and polynuclear aromatic hydrocarbons (PAHs). It is reported here that though these chemicals differ widely in their chemical properties, reactivities and known points of attack in humans, a common link does exist between them. All are lipophilic species found in serum and they promote the sequential absorption of otherwise non-absorbed toxic hydrophilic species causing these diseases. From 1990 to 2010, mental and behavioral disorders increased by more than 37%, Parkinson's disease increased by 75%, Alzheimer's disease doubled, autism increased by 30% and attention deficit hyperactivity disorder (ADHD) increased by 16% , which once absorbed can remain in the body's adipose tissue for up to 30 years or more and can be transferred to serum . As used here, total lipophilic load refers to the total concentration of all exogenous lipophilic chemicals found in serum, without specification of individual chemical species.As discussed below, exposures to POPs, plastic exudates, PAHs and LMWHCs have been found to be associated with neurological disorders. The POPs include polychlorinated biphenyls (PCBs), organochlorine pesticides (OCs), dioxins and furans and polybrominated diphenyl ethers (PBDEs). Plastic exudates include BPA and phthalates. LMWHCs include benzene, toluene, ethyl benzene, xylenes, C3–C8 aliphatics, gasoline, chlorinated methanes and ethanes and chlorinated ethylenes.et al.,et al., Escudar-Gilabert et al.,et al.,et al.,et al.,et al.,POPs, PAHs, LMWHCs, BPA and phthalates all have been shown to penetrate the blood-brain barrier (BBB) , as well as peripheral nervous system maladies (neuropathies)  – polychlorinated bipehenyls (PCBs) ;  ; ALS , phthalates, BPA and hydrocarbons. These chemicals come from a variety of chemical classes that include chlorinated and brominated hydrocarbons, esters, ethers, polynuclear aromatic hydrocarbons, mononuclear aromatic hydrocarbons and straight chain aliphatic hydrocarbons. These chemicals differ widely in chemical properties, reactivity and rates of metabolism and elimination from the body.et al.,et al.,et al.,et al.,et al.,et al.,et al.,The lipophiles associated with induction of neurological disorders can be long-lived POPs, which once absorbed can remain in the body's adipose tissue for up to 30 years or more and can be transferred to serum . Until now, however, no single mechanism that accounts for the induction of a broad spectrum of neurological diseases has been proposed. The association with the onset of widely differing NIs, NDDs and NDGs with exposures to POPs, BPA, phthalates, PAHs and LMWHCs, chemicals which differ widely from each other, yet all of them are able to penetrate the blood-brain barrier, strongly suggests a lipophile-dependent mechanism for the induction of CVDs. The concept of lipophilic chemicals serving to assist the penetration of hydrophilic therapeutic drugs through the blood-brain barrier is well established  from which they can pass to the blood and be transported around the body , absorbed even at toxic concentrations, are more rapidly metabolized/eliminated. Nevertheless, they persist in body serum for days to weeks , coronary heart disease , endocrine disruption  and lipophilic pharmaceuticals, examples of which are statins, taken regularly (Culver et al.,et al.,People are routinely exposed to many other lipophilic chemicals that are retained in body serum. These include mycotoxins produced by mold and found in wet environments and in comtaminated food (Brasel et al.,et al.,The prevalence of neurological diseases discussed here as well as the other diseases cited have all increased dramatically in the past half century. For example, the prevalence of the bipolar disorder and of schizophrenia has increased in the range of 40% from 1990 to 2010 (Murray et al.,et al.,et al.,As previously discussed, PAHs emanating from the combustion of fossil fuels and tobacco are considered to induce many environmental diseases. Several of the studies cited have made the association with inhalation of fine particles rather than with the PAHs (Costello As seen in et al.,et al.,et al.,et al.,et al.,It is to be noted that although the literature relating neurological disease to other exogenous lipophilic chemicals is scanty, mycotoxin exposures have been associated with neurological disease (Pestka The prevalence of neurological diseases, including NIs, NDDs and NDGDs is increasing rapidly throughout the world. The evidence presented here strongly suggests that this increase is due in large part to increased exposure to exogenous lipophilic chemicals which, though varying widely in structure, toxicology, chemical reactivity and retention time in the body, render the body susceptible to attack via subsequent exposure to low levels of hydrophilic toxins that would otherwise not be absorbed. The lipophilic chemicals can be POPs that are metabolized and eliminated slowly, or BPA, phthalates, PAHs, LMWHCs and other lipophilic species that are eliminated from the body more rapidly, but are constantly replenished in the body from polluted air and water and contaminated food. The accumulation of lipophilic chemicals in the body proceeds until a critical lipophilic load level is reached, at which point the body is vulnerable to attack by low levels of toxic hydrophilic chemicals that would otherwise not be toxic. Sequential absorption of lipophiles followed by hydrophiles provides a unified explanation of how low levels of rather different environmental pollutants are responsible for the alarming increase of neurological diseases.


Neurodegenerative disorders such as Parkinson's diseases (PD) and Huntington's diseases (HD), neuronal injuries following trauma and neuronal cell death following strokes are major debilitating affections that are often accompanied by motor and cognitive dysfunctions with limited treatment options. Cell transplantation therapies have been considered for the last three decades as a serious avenue to explore with the ultimate aim to replace lost neurons with “new ones” initially originating from fetal neuroblasts and recently deriving from various sources of stem cells.One of the many factors affecting the success of cell transplantation therapies is host immune response to the graft. This came as an evidence when attempts of cell therapy was undertaken with the use of human fetal neuroblasts or porcine fetal neuronal tissue for a time envisaged as a potential useful cell source for xenotransplantation in the human brain. However, and as Bonnamain et al.  describePauly et al.  focus onGiven that the use of fetal tissue as a cell source for neural transplantation is not without causing logistical and ethical problems, a significant research activity has been oriented toward finding alternative sources of neural cells. Among these, pluripotent stem cells seem to be an obvious choice as these cells are theoretically able to differentiate into any cell type of any organ. As Benchoua and Onteniente  put it iThe paper of Denham et al.  reviews in vitro. They detail the use of a combined photolithographic technique to create a topography of micropatterned substrates composed of extracellular matrix providing the cells with appropriate cues for their differentiation. Although these cellular models are far from mimicking the characteristics of neuronal cells in vivo, they can set the basis for new avenues to explore the differentiation of stem cells in vitro after adjustment of the proper microenvironment in order to obtain the requested specific neuronal subtype.García-Parra et al.  propose in vivo area, de Chevigny et al. (In the y et al.  present Following transplantation, axons derived from transplanted neurons need to find their way and innervate target areas. Our own findings showed that embryonic mesencephalic dopamine neurons transplanted in the substantia nigra in an animal model of PD are able to extend axons toward the striatum (Gaillard et al., The review by Saha et al.  is focusAlthough cell transplantation in the damaged brain is not likely to be routinely performed in the near future, the different paths that are evoked in this series of reviews should yield safer, more effective and physiologically relevant transplantation procedures.


Autism spectrum disorders (ASDs) are a group of developmental psychiatric disorders characterized by impaired social interaction and communication, and by restricted, repetitive and stereotyped behaviors and interests , has been found to be associated with autism, supporting genetic defects in cholesterol metabolism can cause autism   which encode key enzymes  in the cholesterol metabolic pathway and cholesterol concentrations are altered in the brain in a development dependent manner (Buchovecky et al., RTT is caused largely by mutations in the X-linked methyl CpG-binding protein 2 gene (Disturbances in cholesterol homeostasis in ASDs raise the question of how changes in cholesterol metabolism might be involved in the development of autism. Cholesterol is not uniformly distributed in biological membranes, but concentrated in lipid rafts, together with other lipids like sphingolipids (Pfrieger, Lipid rafts are specialized membrane structures that form an organized portion of the membrane with concentrated signaling molecules and link to the cytoskeleton (Simons and Toomre, Mecp2 mutant mice has identified that a nonsense mutation in SQLE, which is an obligate rate-limiting enzyme in cholesterol biosynthesis, diminished some of RTT related phenotypes (Buchovecky et al., Mecp2 mutant mice (Buchovecky et al., Interestingly, genetic or pharmacological manipulation of cholesterol metabolism has been found to alleviate ASD associated animal behaviors. In the aforementioned study, a mutagenesis screen in Noteworthily, the effect of statin treatment has also been observed in animal models of other ASDs, including fragile X syndrome and neurofibromatosis type 1 (Li et al., It is important to note, cholesterol may regulate protein function not only through altering lipid rafts but also by direct interactions with the proteins. However, it is very challenging to discriminate between these possibilities for any specific type of protein because it is unreasonable to test the effect of cholesterol on the function of a membrane protein in an environment without lipid rafts (Levitan et al., Despite the controversies in lipid raft research, it is now clear that lipid rafts which represent dynamic structural components of cellular membranes do play roles in neuronal signaling and that dysregulation of lipid rafts can lead to diseases (Allen et al.,


Interprofessional collaboration (IPC) is an integral part of the practice of Medicine and Family Medicine. The World Health Organization (WHO) defines IPC as occurring when “multiple health workers from different professional backgrounds work together with patients, families, carers and communities to deliver the highest quality of care” WHO, . To provThe delivery of responsive, effective, and high-quality patient care is indeed a complex activity. It demands health care professionals collaborate in an effective manner (Reeves et al., Competence, or the knowledge and skills base underlying a particular profession, can also be considered a profession's “cognitive map.” Good teamwork relies on a joint understanding of one's own as well as other team members' cognitive map (Drinka and Clark, Defining one's own professional competencies is therefore an important first step in effective IPC. One approach to gathering the necessary information to determine what competencies are required for a particular profession is to ask those already active and skilled in the profession itself. This approach was used by the CFPC in the development of the competency–based objectives in family medicine using practicing family physicians as their source of information (Allen et al., Another approach includes consultation with others. Realizing the importance of patients, physicians, and other health care professionals in the delivery of health care, research in the 1990's by the Ontario provincial government in Canada sought both public and other health care professionals input in an attempt to define the expectations of physicians performance. The results showed significant difference of opinion between physicians and other health care professionals on items referring to distribution of responsibility, control, and authority of physicians within the health care system at that time (Neufeld et al., Pharmacists represent the one health care profession all family physicians are most likely to collaborate with in providing optimal patient care. A recent UK study on the collaborative relationship between community pharmacy and general medicine, highlighted the dynamic nature of the relationship and the key components of collaboration. These included the importance of trust, communication, professional respect, and “knowing each other” when referring to roles, abilities and responsibilities (Bradley et al., Our research team is exploring pharmacist's expectations of the competent family physician using an adaptation of the CFPC's original competency questionnaire distributed to practicing family physicians (Allen et al., Continued research in this area is essential to deepen our understanding of the essential components of the interprofessional relationship. Understanding and clarifying role expectations is central to this and can provide valuable information when designing and reviewing professional competency frameworks and IPE curricula. As more research becomes available, the effectiveness of these programs on outcomes such as IPC in clinical practice and patient care should be reviewed.


The Hippo signaling pathway comprises a series of cytoplasmic tumor suppressor proteins including Merlin and the Lats1/2 and MST1/2 kinases, and is thought to play a critical role in determining the sizes of organs and tissues. The Hippo pathway is regulated upstream by extracellular mechanosensory signaling arising from cell shape and polarity, as well as by a variety of extracellular signaling molecules. When active, the pathway maintains the transcriptional activators Yes-associated protein (YAP) and TAZ in phosphorylated forms in the cytoplasm, preventing cell proliferation. When the Hippo pathway is inactivated, YAP and TAZ are translocated to the nucleus and induce the expression of a variety of proteins concerned with entry into the cell division cycle, such as cyclin D1 and Fox M1, as well as the inhibition of apoptosis. The failure of the Hippo pathway has been implicated in the development of many different types of cancer but there is limited information available as to its involvement in melanoma. We hypothesize here firstly that the Hippo pathway is involved in maintaining density of cutaneous melanocytes on the basement membrane at the junction of the epidermis and the dermis, and secondly, that its function is disturbed in melanoma. We have analyzed a series of 23 low passage human melanoma lines as well as cultured normal melanoma, and find that melanocytes, as well as all melanoma cell lines examined express TAZ. Melanocytes and most melanoma lines also express YAP. E-cadherin, an upstream regulator of the Hippo pathway, and Axl, a receptor tyrosine kinase regulated by the Hippo pathway, are expressed in melanocytes and in several melanoma cell lines. These observations, together with published evidence for the presence of Merlin, Lats1/2, and MST1/2 in melanocytes and melanoma cells, support the hypothesis that the Hippo pathway is an important component of melanocyte and melanoma behavior. Drosophila that together were found to control organ size. These genes specify a series of kinases and adaptor proteins including Hippo (Hpo), Warts (Wts), and Salvador (Sav), loss of function of which results in flies with enlarged, folded eyes and excess head cuticle, a “hippopotamus-like” phenotype , the homolog of Yorkie in Studies of the Hippo pathway suggest it has a three-dimensional “sense” that is communicated across the whole organ and controls both cell proliferation and apoptosis. A possible example of such organ size control is provided by liver; it has been known for many years that surgical reduction of liver volume leads to extensive cell division and regeneration until the liver approaches its original size. The Hippo pathway has been hypothesized to have a role in this process  constitutes one of the main causes of cell injury and death  (Tschop et al., To collect evidence for the involvement of the Hippo pathway in melanoma, a series of melanoma lines developed in this laboratory (Marshall et al., CTGF (Braig et al., GLI2 (Alexaki et al., It is known that from animal models of melanoma that proliferation and invasiveness are promoted by YAP-TEAD (Lamar et al., RASSF1 is frequently inactivated by promoter hypermethylation in a variety of human tumors including melanoma (Spugnardi et al., GRIN2 gene, which codes for a subunit of the glutamate ionotropic receptor, is mutated in approximately 25% of melanomas (Wei et al., NF2 gene, which encodes Merlin (Figure FAT4 gene is recurrently mutated in several types of human cancer including melanoma (Katoh, Our previous studies have suggested that two changes may be important to distinguish melanoma cell lines from cultured melanocytes: the partial loss of serum dependence of some intracellular signaling pathways (Kim et al., n Figure  have beea Katoh, .One of the fascinating features of the Hippo pathway is that it is able to integrate several different types of signaling, including those induced by cellular shape and adhesion changes, stress responses, and fluctuating concentrations of extracellular signaling molecules. It mediates cellular decisions on the control of proliferation, motility and cell death, and existing evidence indicates a complex and possibly redundant series of intracellular pathways (Halder et al., The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Since the first seminal reports of young infants' abilities to use both acoustic (e.g., Mandel et al., The goal of the current Research Topic is to provide a cutting-edge snapshot of this active research field integrating original research papers using both behavioral and neurophysiological techniques with review articles providing ideas for general frameworks capturing those findings.Three reviews and one methods article offer global and thought-provoking views on basic principles and computational mechanisms that are operative in early language learning. Seidl and Cristia  provide Two of the original research papers applied neurophysiological methods. Minagawa-Kawai et al.  report aAnother set of research articles focuses on the contribution of prosodic information to the perception of sentential structure. Wellmann et al.  evaluateThe largest group of papers deals with specific questions related to speech segmentation. Bosch et al.  investigFinally, our Research Topic contains one study which analyzes infant speech production during the second year of life. Yamashita et al.  study EnAs a compendium of current research efforts in the field of early language learning mechanisms we are confident that this Research Topic offers novel and stimulating ideas for those who are new to the field and would like to get a timely overview as well as for experts who are interested in current developments.


Qiliqiangxin (QL), a traditional Chinese medicine, was approved by China Food and Drug Administration for the treatment of heart failure in 2004 (Li et al., Recent progress in studying QL on improving exercise tolerance and patients' quality of life highlighted that QL may be through a variety of biological mechanisms to exert its beneficial effects. One possible mechanism may be the regulation of the balance between pro-inflammatory and anti-inflammatory cytokines in cardiomyocytes (Xiao et al., Traditionally, the adult mammalian heart is regarded as a post-mitotic organ without any regenerative capacity (Rosenzweig, In conclusion, we think QL may mimic a phenotype of physiological hypertrophy and induce heart regeneration, which is beneficial for heart failure.


A funding organization was incorrectly omitted from the Funding Statement. The Funding Statement should read: "This study was financially supported by Projects SAF2009-08570 and SAF2012-35474 from the Ministerio de Economía y Competitividad of Spain and Fondo Europeo de Desarrollo Regional (FEDER). EGS has a fellowship from the Gobierno de La Rioja of Spain, and CL has a fellowship from the Ministerio de Economía y Competitividad of Spain. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."


Dear Editor,et al.[Nocardia brasiliensisbrazilienses / Nocardia asteroides.[We read with interest the rare case of disseminated orbital actinomycetoma reported by Shanbagh et al. The authteroides. We wouldet al., which described a 26-year-old male who was found to have multiple discharging sinuses on the scalp, with orbital and intracranial invasion.[The authors’ claim to be the first in reporting orbital invasion of actinomycetoma is misplaced. A quick literature search revealed a report by Patil invasion.2et al. have described the characteristic moth-eaten appearance of the frontal bone along with orbital and intracranial invasion, in the case reported by them.[Figure 3 in the article is wrongly labelled as computed tomography (CT) scan of orbit while it shows pictures of magnetic resonance imaging (MRI). The authors have described the osteolytic changes seen in longstanding undiagnosed cases of actinomycosis. However, by them.et al.[et al. described a case of orbital actinomycosis in a patient with a history of carcinomas of the breast and kidney.[et al. described a case of orbital actinomycosis with a small ill-defined lesion in the posterolateral orbit.[It may be of interest to note that orbital actinomycosis has been also reported by Pagliani and Sullivan et al.4 Pagliand kidney. A CT scad kidney. Sullivan


To the Editor: Floret et al. argue convincingly that natural disasters, including severe floods and windstorms, tend not to result in epidemics of infectious disease  and 2002–2004 (≈870 cases). Sporadic cases of shigellosis were not seasonal.Two hurricanes resulted in heavy rainfall in Baltimore during the study period .


We approach this challenge through a network-centric perspective, as we believe that brain function is fundamentally determined by patterns of connectivity between neurons, and the resulting dynamics. This is in contrast to traditional computational neuroscience techniques that focus on models of individual neurons and compartments. Progress, in consequence, is essential on (at least) three major fronts: measurement front spans the range from multi-electrode recordings to whole-brain measurements using imaging. Several basic scientific questions arise: What do we need to measure in brain networks? Are there theoretical constraints that would dictate this? How do we design our experiments to generate the most meaningful data? How do we record from awake/behaving animals, or even from multiple animals interacting socially?The analysis front consists of creating network models from the measurements. Some promising techniques explore the estimation of networks using causality. However, several open questions remain: How do we define the fundamental units within the network? Are these units fixed or do they evolve dynamically? How do we infer connectivity between network elements? How do we identify functional clustering, based on the individual neuronal features? How do we quantify and interpret the activity of multiple neurons via multi-unit recordings, especially when there is no stimulus-response paradigm?The modeling front can proceed in several directions. From the extracted network we can identify topological regularities, such as motifs and cycles. An interesting research direction is to analyze the relationship between the structure of the network, as represented by its motifs, and its function. A growing body of work is examining the relationship between network structure and phenomena such as stability and synchrony. For instance, neurons in the hippocampus could be modeled as a network wherein hubs consisting of hub neurons promote synchrony, while cycles in this network may cause instability. The theme of synchrony as an important network phenomenon emerges in several articles in this research topic  subjects and normals have been reported, among other psychiatric and neurodegenerative conditions. Cortical network properties ultimately determine how different network oscillation states are established and maintained and defining these principles could explain why there is impaired synchronization between different brain areas in schizophrenics and Parkinson's patients. Overall, network-based measures capture better the dynamics of brain processes, and provide features with greater discriminative power than point-based measures.The articles in this research topic cover these different aspects of cortical networks. To guide the reader, we provide below a brief summary of each article, and relate it to the overall theme of the research topic.Rothganger et al.  present Ratnadurai-Giridharan et al.  develop Bhattacharya et al.  present Dey et al.  use RestAlonso et al.  test a sAlmeida-Filho et al.  study muin vitro. Their simulations support the proposed underlying biological mechanisms: the increase of neuronal response latency to ongoing stimulations and temporal or spatial summation required to generate evoked spikes.Vardi et al.  propose Tibau et al.  monitoreRiera et al.  describeFor the past several decades, theoreticians and experimentalists alike have focused on neuronal synchrony and on the important roles that it might play in brain function, from “The Binding Problem” in perception Gray,  to ConscCanavier et al.  address Konstantoudaki et al.  explore Nie et al.  make useCavallari et al.  investigThivierge et al.  investigChary and Kaplan  investigLatorre et al.  implemenKaplan and Lansner  address Eguchi et al.  use a deThe 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.


Early human development from late gestation to the neonatal period is a critical time in the individual's life span. Preterm birth or medical issues affecting the brain function during late gestation or the first months of life may lead to an avalanche of neurodevelopmental problems, including cognitive deficits and motor disability, with lifelong consequences for the individuals, their families, the health care system and the society  is a well-established diagnostic tool for fetal surveillance. Two original research papers describe the findings of studies performed with fMCG to estimate the development of the autonomic nervous system. It is known that, around 32 weeks of gestation, four fetal behavioral states (quiet and active sleep and awakeness) can be identified by combining fetal heart rate variability (fHRV) and fetal movements. Brändle et al.  confirmer et al.  proved tin utero.To understand the normal functional development of the fetal brain, periods of developmental vulnerability should be identified to assess the function of the fetal nervous system and open a window on novel prenatal diagnostics and prognostics. MRI has become increasingly feasible and clinically important in fetal brain studies due to its higher tissue resolution and better visualization of normal and pathological development of macroscopic anatomy or white matter microstructure and connections. Jakab et al.  employedDespite the advancements in monitoring techniques and perinatal care, clinicians still have limited means to predict neurodevelopmental outcomes and plan early intervention. In their opinion article, Giampietri et al.  offer a The development of functional connectivity patterns from infancy to childhood were studied also by Berchicci et al. , who invFinally, electroencephalography (EEG) is the most commonly used technique to non-invasively assess neonatal brain activity, but the main challenge in interpreting EEG signals is the quantitative characterization of the spontaneous “background activity” in sick neonates. Matic et al.  applied 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.


In this opinion article we examine the relationship between penetrometer resistance and soil depth in the field. Assuming that root growth is inhibited at penetrometer resistances > 2.5 MPa, we conclude that in most circumstances the increases in penetrometer resistance with depth are sufficiently great to confine most deep roots to elongating in existing structural pores. We suggest that deep rooting is more likely related to the interaction between root architecture and soil structure than it is to the ability of a root to deform strong soil. Although the ability of roots to deform strong soil is an important trait, we propose it is more closely related to root exploration of surface layers than deep rooting. However, roots which extend into burrows followed them to their end. Ehlers et al. . The pores were created by burying a two‐dimensional geotextile at 20 cm, with the different pore‐density treatments. Although no genotypic differences were found, this approach would seem to provide a method to assess genotypes. Either McKenzie et al. (et al. (et al.,In an interesting field study, McKenzie et al.  comparede et al.  were unl (et al.  that roo et al., will becet al. (et al. (The basis for the location of soil pores by roots seems to be a relatively unexplored area and given the increases in soil strength with depth Fig.  it wouldet al.  found th (et al.  also repet al.If we accept the thesis that deep root penetration is facilitated by exploiting existing pore networks, then the question arises of whether an ability to penetrate a hard layer is useful. Actually, we maintain that it is useful. Roots which deform soil are likely to have better root–soil contact and improved ability to extract water and nutrients from the soil in the shallower layers. At depth, roots in pores are less well connected hydraulically to soil, although White & Kirkegaard  show thaWhile the tendency for deeper roots to be found in pores is well reported (e.g. Lynch & Wojciechowski,


Analogical reasoning is a landmark of human cognition. Based on the realization that the elements of two situations are organized by similar systems of relations, analogical inferences allow the transfer of knowledge structures from a better-known situation  to a target situation that is relatively less understood .Experimental research has demonstrated that the retrieval of base analogs from long term memory in response to the proceesing of a target analog is infrequent in the lack of semantic similarities between both situations  the referendum on the independence of Quebec, Blanchette and Dunbar  obtainedh et al.  providedh et al.  failed th et al.  or sellih et al. , both exh et al.  expert mh et al.  novice pAnother explanation for the frequent use of distant analogies in naturalistic studies might arise from comparing the psychological constraints of naturalistic analogy generation against those of classical experimental studies. The standard experimental procedure comprises an encoding phase, during which participants learn the base analogs, and a transfer phase, where experimenters present participants with either a semantically close or a semanically distant target situation, and assess whether its processing elicits the spontaneous retrieval of the base analog. Based on this procedure, differences accross conditions were typically taken to demonstrate the centrality of surface similarities during analogical transfer. In contrast with this highly controlled environment—in which transfer can only originate in the retrieval of the critical base analog from memory—in naturalistic settings participants are free to generate analogies by means of retrieving their own base analogs (Blanchette and Dunbar, The fact that the core components of our retrieval mechanisms are invariably set to favor semantically close base analogs (Gentner et al., The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Beta-band oscillations in basal ganglia-cortical loops have been shown to be linked to motor function in both healthy and pathological states Brown, . As wellThe Journal of Neuroscience, Bartolo et al. (In a recent study published in o et al.  examinedThey found that LFPs in the beta band exhibit interval tuning and showed a preference for intervals with duration around 800 ms, similar to the preferred duration observed in medial premotor cortex neurons (Merchant et al., The present study and previous work from this group (Merchant et al., The present study only addresses internal timing in regular sequences and it is not yet known how temporal jitter would affect the putaminal beta-band activity. In irregular sequences, beta-band modulation may remain the same or change as a function of the amount of jitter. As natural sounds are rarely presented in a temporally regular context, future work needs to address how the brain perceives time in irregular sequences. Related to this question, Fujioka et al.  used magTemporal processing in irregular sequences has been shown to be preferentially mediated by a network based in the cerebellum whilst timing in regular sequences is carried out by a striato-frontal network (Teki et al., Bartolo et al.  only briIn summary, this stimulating work on the role of putamen during internally driven timing behavior significantly adds to previous work by Merchant and colleagues based on recordings from medial premotor cortex (Merchant et al., The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Production of biogas from agricultural biomass or organic wastes is an important source of renewable energy. Although thousands of biogas plants (BGPs) are operating in Germany, there is still a significant potential to improve yields, e.g. from fibrous substrates. In addition, process stability should be optimized. Besides evaluating technical measures, improving our understanding of microbial communities involved into the biogas process is considered as key issue to achieve both goals. Microscopic and genetic approaches to analyse community composition provide valuable experimental data, but fail to detect presence of enzymes and overall metabolic activity of microbial communities. Therefore, metaproteomics can significantly contribute to elucidate critical steps in the conversion of biomass to methane as it delivers combined functional and phylogenetic data. Although metaproteomics analyses are challenged by sample impurities, sample complexity and redundant protein identification, and are still limited by the availability of genome sequences, recent studies have shown promising results. In the following, the workflow and potential pitfalls for metaproteomics of samples from full-scale BGP are discussed. In addition, the value of metaproteomics to contribute to the further advancement of microbial ecology is evaluated. Finally, synergistic effects expected when metaproteomics is combined with advanced imaging techniques, metagenomics, metatranscriptomics and metabolomics are addressed. Over the past 10 years, conversion of biomass to methane in biogas plants (BGPs) has become a reliable source of renewable energy. In 2013, about 7500 BGPs produced 3.5% of the annual electricity demand in Germany  experimental design, (ii) sampling, (iii) protein purification, (iv) protein separation, (v) liquid chromatography (LC)-tandem mass spectrometry MS/MS), (vi) bioinformatics and (vii) examining the protein identification Figure .et al., et al., et al., et al., Reproducible scientific studies require carefully planned and documented experiments. In order to correlate metaproteome data from different BGPs with process data, at least a minimum set of meta-information has to be provided, comprising BGP design, process temperature, pH value, nitrogen content, inocula, gas composition and volume as well as feed composition. In addition, technical and biological replicates are required. However, for most industrial scale BGPs, no true replicates exist, because each BGP is individual concerning its construction and operational parameters. An acceptable workaround would be to sample at least two independent technical replicates at close time points. Otherwise, investigations using lab-scale equipment are required to complement studies. Here, critical process conditions can be applied without risking the crash of a full-scale fermenter. Depending on the scientific question, simplifying the complex microbial community by feeding a synthetic medium  sampling before feeding and at same time of the day, (ii) mixing the BGP before sampling and (iii) discarding sufficient material before sampling in order to flush the sample port.et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., Sample preparation includes cell lysis, protein extraction, protein quantification and separation. Main problems during cell lysis and protein extraction are the high amount of sample impurities and the different levels of microbial community organization, such as scattered microorganisms, biofilms on the substrates or granules  evolved in the proteomic community as a standard (Elias et al., The problem of metaproteomics in contrast to other proteomics with pure or defined mixed cultures is that the taxonomic composition of complex microbial communities is not known and the database cannot be reduced to keep the number of false positive hits low. In this context, the idea of Jagtap and colleagues  to repeaet al., et al., et al., de novo sequencing of peptides using acquired spectra (Frank and Pevzner, et al., de novo results requires manual inspection. Therefore, a more straightforward strategy is sequencing the metagenome of the analysed sample.In the next step, protein identification is achieved based on identified peptides (Bradshaw et al., et al., et al., et al., et al., et al., After successful protein identification, the importance of a single identification can be improved by acquiring meta-information concerning taxonomy and function from repositories, e.g. UniProt (Consortium, et al., et al., et al., et al., et al., For a better survey, taxonomic composition can be visualized in a Krona plot (e.g. Krona plot for a mesophilic/thermophilic BGP: Figure et al., et al., et al., et al., et al., Shifting to protein functions, overview plots, such as a Voronoi Treemap (Bernhardt et al., Tools for the characterization of microbial communities (Table Coprothermobacter and Methanothermobacter in a thermophilic reactor treating thermally pretreated sludge (Gagliano et al., et al., et al., et al., In microbial ecology, a wide mixture of different methods are commonly applied to investigate a specific problem. Hence, knowledge of how to combine these methods is important. First of all, a standardization of sample preparation is essential. With regard to the use of multi-omic approaches, the sample preparation workflow should comprise most analytes namely DNA, RNA, proteins and metabolites in an adequate manner (Roume es Table , metaproet al., et al., et al., et al., et al., et al., et al., So far, only a few metaproteome studies of BGPs were carried out Table  (Abram eet al., et al., et al., et al., et al., While in early metaproteome studies, only a few proteins were detected (Abram Actinobacter, Bacteriodia, Bacilli, Chlostridiales, Thermotogae and different Proteobacter groups. Archaeal proteins were dominated by the orders Methanobacteria and Methanomicrobia. A comparison of phylogenetic profiles derived from metaproteomics and molecular biological studies revealed significant differences in the relative abundance of methanogens [about 20–30% in metaproteome data compared with the 4% derived by metagenomics (Hanreich et al., et al., Gelria by metaproteomics, although it was highly abundant in the pyrosequencing data. In this case, the bias might have been introduced by the lack of protein entries for Gelria in the UniProt database.The majority of the identified bacterial proteins belonged to the orders et al., et al., et al., Besides proteins from Archaea and Bacteria, several proteins from plants and animals are frequently identified in samples from BGPs. They are originated from plant feedstock or manure and represent the incomplete usage of substrate. In addition, a few proteins were identified to belong to Fungi (Kohrs Caldicellulosiruptor and that celluloses were degraded by the cellulosome of Clostridium thermocellum. Surprisingly, Lü and colleagues (Clostridium proteolyticus, indicating its function as predator or scavenger of dead biomass. The observed proteolytic activity nicely confirmed a study (Binner et al., As already discussed, the main advantage of metaproteomics is the functional characterization of microbial communities together with the phylogenetic assignment. Lü and colleagues  showed, lleagues  also obset al., et al., et al., et al., et al., For the subsequent fermentation step, mainly proteins from sugar uptake, glycolysis and to some extent from the pentose phosphate pathway were identified. This is in accordance with carbohydrates as the major substrate of biogas production. Whether the Entner–Doudoroff pathway is of relevance for fermentation (Abram de novo synthesis of methionine, lysine and threonine surpassing their low proportion in maize protein (Ridley et al., Amino acids derived from feedstock proteins are fermentable substrates and precursors for synthesis of microbial biomass. Based on metaproteome data, Heyer and colleagues  reportedIn contrast to the previous steps of the biogas process, the investigation of acetogenesis is even more challenging. In fact, Lü and colleagues  were ablet al., et al., et al., et al., et al., et al., et al., et al., et al., Proteins involved in methanogenesis are highly abundant in metaproteomes. Nearly all enzymes of the hydrogenotrophic and the acetoclastic pathway were identified. Under psychrophilic and mesophilic conditions, the acetoclastic pathway is favoured (Abram et al., et al., Although MS-based metaproteomes of different samples show some similarities due to high abundance of methanogenic enzymes and also the presence of similar dominating phylogenetic groups, each BGP seems to have its own protein signature. Surprisingly, separation of proteins by SDS-PAGE is sufficient to produce individual protein patterns (Heyer In depth, analysis of microbial communities in BGPs is required to use their full potential for biogas production. The comparison of methods for the characterization of microbial communities and recent results regarding the functional and taxonomic composition of these microbial communities obtained by various research groups showed that metaproteomics is developing into a powerful tool for the exploration of the biogas process. Besides identifying major pathways of biomass degradation, it links single metabolic pathways with microbial taxa. Each BGP shows its own, time stable protein pattern. Strong alterations in this pattern can be linked with process disturbances, and some enzymes were identified as potential biomarkers for process monitoring and fault detection.Metaproteome analysis of BGPs is still hampered, however, by sample impurities, sample complexity, redundancy of protein identifications and a lack of genome sequences required for protein identifications. Nevertheless, the presented workflow overcomes at least parts of these problems. In the future key issues to be addressed include comprehensive sample preparation, a suitable protein separation, grouping of redundant proteins and the incorporation of meta-information from online repositories. In particular, more efficient protein extraction, improved MS and new algorithms for the verification of protein identification are urgently required to further improve this workflow and to exploit the full potential of metaproteomics. In addition, it has to be taken into account that metaproteomics is no stand-alone approach. For comprehensive analysis of microbial communities, metaproteomics should be applied in concert with microscopy, cytometry, metagenomics, metatranscriptomics and metabolomics.


Arabidopsis root development is coordinated via a localized auxin concentration maximum in the root tip, requiring the regulated expression of specific genes. However, little is known about how hormone and gene expression patterning is generated.• Patterning in • Using a variety of experimental data, we develop a spatiotemporal hormonal crosstalk model that describes the integrated action of auxin, ethylene and cytokinin signalling, the POLARIS protein, and the functions of PIN and AUX1 auxin transporters. We also conduct novel experiments to confirm our modelling predictions.aux1 mutant, the amounts of cytokinin, ethylene and PIN protein, and PIN protein patterning in wild-type and mutant roots. Modelling analysis further reveals how PIN protein patterning is related to the POLARIS protein through ethylene signalling. Modelling prediction of the patterning of POLARIS expression is confirmed experimentally.• The model reproduces auxin patterning and trends in wild-type and mutants; reveals that coordinated PIN and AUX1 activities are required to generate correct auxin patterning; correctly predicts shoot to root auxin flux, auxin patterning in the • Our combined modelling and experimental analysis reveals that a hormonal crosstalk network regulates the emergence of patterns and levels of hormones and gene expression in wild-type and mutants. Arabidopsis root development and response to varying environmental conditions involves a complex network of overlapping interactions between plant signalling hormones and gene expression known as ‘hormonal crosstalk’. Hormone concentrations in the cells are a function of multiple factors such as hormone biosynthesis, long- and short-range transport, rate of influx and efflux by carrier proteins, and hormone activation, inactivation and degradation  peptide function. The PLS gene of Arabidopsis transcribes a short mRNA encoding a 36-amino-acid peptide that is required for correct root growth and vascular development  Heynh have been described previously , 1% sucrose, and 2.5% Phytagel (Sigma)) at 22 ± 2°C as previously described  ecotypes and the −1 propidium iodide as previously described . Statistics were carried out using Excel (Microsoft). Results were visualized as average intensities with error bars representing SD of the mean.Confocal images (for GFP imaging) were taken with a Leica SP5 microscope  after counterstaining tissues with 10 mg ml−5 and 10−10, respectively. Much smaller convergence tolerances for both iterations are also tested, and the numerical results show that further reduction of convergence tolerances for both iterations does not improve the accuracy of numerical simulations.The set of partial differential equations, which describes spatiotemporal dynamics of hormonal crosstalk in the root Figs, is solvhttp://imagej.nih.gov/ij). The output of ImageJ is the intensity of each pixel in an experimental image. The relative intensity over the whole image shows the relative hormone response or protein concentration patterning for any measured component. The detailed method for using ImageJ to analyse experimental images is described in Supporting Information In this work, experimental images were analysed using ImageJ  follow experimental trends , the modelled cytokinin concentration in the vascular and pericycle cells is higher than that in the epidermal cells, a result similar to experimental observations. Nevertheless, the trend of cytokinin patterning along the longitudinal root axis still differs between the experimental images and modelling results . These rArabidopsis root is similar to the experimentally determined auxin IAA2::GUS response patterning, with an auxin maximum established at or close to the QC , showing that modelling results are robust to variations in parameter values. We then used the model to study the level and patterning of hormones and gene expression in pls mutant and pls etr1 double mutant show that auxin flux from shoot to root in the pls mutant is significantly lower than that for wild-type. This effect reduces the total amount of auxin in the root tip and reduces auxin responses in the pls root. The auxin flux into the root, and the root auxin content for the pls etr1 double mutant also recover approximately to the value of the wild-type . In the pls mutant, the suppression of X is relaxed owing to the loss of PLS function. This enhances the rate of PIN1 biosynthesis at the region near the tip and therefore PIN1 patterning shows an increasing concentration trend from the proximal to the distal region. In addition, in the pls mutant, the auxin concentration decreases  is similar to the corresponding experimental trend. However, a noticeable difference for the pls etr1 double mutant can be identified. This indicates the limitation of our model for analysing this double mutant.Therefore, the overall effects of auxin, ethylene and cytokinin result in opposite trends in PIN1 patterning in wild-type and Patterning of PIN2 protein was also analysed . ModelliPLS gene of Arabidopsis, which transcribes a short mRNA encoding a 36-amino-acid peptide  and PIN7 (which is positively regulated by cytokinin) are localized at similar positions  enzymes. While ARR5::GUS expression, as a proxy for cytokinin distribution; Werner et al., et al., et al., Our modelling results for cytokinin concentration patterning  are quanet al., ARR5::GUS images (Werner et al., ARR5::GUS images measure the activation of the ARR5 promoter by cytokinin, therefore indicating the activity of cytokinin signalling rather than cytokinin concentration. Bishopp et al. (ARR5 expression, is regulated by auxin in the xylem axis. This could indicate that ARR5::GUS images represent the effects of both cytokinin and AHP6 concentration, and therefore might not solely reflect cytokinin concentration. Third, it has been demonstrated that cytokinin is transported from the shoot to the root in the phloem (Bishopp et al., et al., This discrepancy leads to the following possibilities. First, the experimental data (Nordstrom p et al.  have diset al., et al., et al. (et al., et al., et al., et al. (Based on experimental results (Nordstrom , et al.  have sho, et al. .et al., et al., In root development, the complexity of hormonal signalling includes many aspects. This work has shown that the spatiotemporal dynamics of hormonal crosstalk, which integrates hormonal crosstalk at a cellular level with root structure, is able to explain two important aspects – the steady-state level and the patterning of hormones/hormone responses and gene expression. Recent studies have shown that growth and hormonal patterning can affect each other (De Rybel et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., All recent modelling and experimental work (Chickarmane


Bacillus subtilis the process involves the differentiation of cell fate within an isogenic population and the production of communal goods that form the biofilm matrix. Here we review recent progress in understanding the regulatory pathways that control biofilm formation and highlight developments in understanding the composition, function and structure of the biofilm matrix.Biofilm formation is a social behaviour that generates favourable conditions for sustained survival in the natural environment. For the Gram-positive bacterium  In the biofilm individual cells are held together by a self-produced extracellular polymeric matrix commonly comprised of polysaccharides, proteins and DNA  and the tapA-sipW-tasA (hereafter tapA) operon , it facilitates continued transcription of slrR. This means that cells accumulate high levels of SlrR in a self-reinforcing negative feedback loop . To date, only a subset of proteins encoded by the eps operon has been studied in any detail. EpsA and EpsB act as a tyrosine kinase modulator and tyrosine kinase, respectively, and both are required for biofilm formation . The prevalence of each sugar was largely dependent on the integrity of the eps operon  that prevented expression of this enzyme (Leiman et al., d-amino acid inhibition of biofilm formation was not observed (Leiman et al., et al., et al., B. subtilis completely lacks the biosynthetic pathway required for norspermidine synthesis and, consistent with this, detection of norspermidine within biofilm samples was not possible despite utilization of two separate detection methods (Hobley et al., et al., et al., B. subtilis remains a much-debated topic within the field. Further investigation will be required to determine whether the reduction in biofilm biomass observed in late-stage biofilms is the result of an organized disassembly process or simply the result of the onset of sporulation by the majority of the population after exhaustion of the nutrient supply.The second method of biofilm dispersal would involve degradation or disruption of the macromolecules in the extracellular environment. Consistent with this mechanism of dispersal, production of extracellular proteases has been correlated with late stages of biofilm formation (Marlow et al., The environmental signals and regulatory pathways that control entry into biofilm formation have been well studied and it is known that they largely converge to control the production of the biofilm matrix components (Vlamakis


Dictyostelium in response to nutritional starvation and stress. Here, the trypanosome signalling pathway is discussed in the context of these conserved pathways and the possible contributions of opposing ‘slender retainer’ and ‘stumpy inducer’ arms described. As evolutionarily highly divergent eukaryotes, the organisation and conservation of this developmental pathway can provide insight into the developmental cycle of other protozoan parasites, as well as the adaptive and programmed developmental responses of all eukaryotic cells.African trypanosomes, parasites that cause human sleeping sickness, undergo a density-dependent differentiation in the bloodstream of their mammalian hosts. This process is driven by a released parasite-derived factor that causes parasites to accumulate in G1 and become quiescent. This is accompanied by morphological transformation to ‘stumpy’ forms that are adapted to survival and further development when taken up in the blood meal of tsetse flies, the vector for trypanosomiasis. Although the soluble signal driving differentiation to stumpy forms is unidentified, a recent genome-wide RNAi screen identified many of the intracellular signalling and effector molecules required for the response to this signal. These resemble components of nutritional starvation and quiescence pathways in other eukaryotes, suggesting that parasite development shares similarities with the adaptive quiescence of organisms such as yeasts and  The ability of cells to sense their surroundings and respond to changes in the environment is fundamental to their survival. Monitoring cell density and its regulation is especially beneficial, ensuring the availability of nutrients and space and preventing the accumulation of toxic metabolic waste. This capacity of cells to sense their critical density, referred to as quorum sensing (QS), requires signal molecule/s that serve as reporters of the population density and signal relay components that transmit this information within the cell, causing it to generate a response. The response to increased cell numbers is often a switch to quiescence , a response that promotes survival of the cells under conditions of dwindling resources.African trypanosomes, the causative agents of sleeping sickness in humans and ‘nagana’ in livestock, are protozoan parasites that alternate their lives between a mammalian host and an insect vector, the tsetse fly. In the mammalian bloodstream, the parasites are ‘pleomorphic’ (i.e. of different morphologies) and undergo a developmental transition from rapidly proliferating long ‘slender’ forms to non-dividing short ‘stumpy’ forms, upon attaining a critical cell density Fig. . The slein vitro is linked to their cell density, and this is irrespective of their initial seeding density  before differentiation and proliferation arrest in G1/G0 as stumpy forms can occur  as auto inducers of QS signalling cascades. The AHLs are freely diffusible molecules that can bind to and activate a transcriptional activator, which in turn induces expression of target genes. Another class of molecules that mediate similar effects is the 4-quinolones. On the other hand, Gram-positive bacteria use small post-translationally processed peptides as signal molecules. These peptides have been shown to interact with a histidine kinase two-component signal transduction system  secrete a soluble, low molecular weight, heat stable factor/s termed stumpy induction factor (SIF). SIF is of parasite origin, and its accumulation in the culture medium/blood stream, with increasing parasite density, is proposed to act as the trigger for the slender to stumpy differentiation. This factor is believed to act on the same population that produces it, such that it induces an autocrine response. Monomorphs, in contrast, seem to be ‘signal-blind’ to the factor they produce, thus providing them with a growth advantage et al., et al., et al., et al., Lack of information on the chemical identity of SIF has made it difficult to understand the production of the signal, its turnover and the molecular cascade involved in relaying this signal. Early studies on slender forms noted a two- to threefold increase in cyclic AMP (cAMP) levels at peak parasitaemia followed by a decline in its levels during the transition to stumpy forms  or threonine (Thr) residues, making Ser/Thr protein phosphatases (PPs) key players in the eukaryotic cell. The Ser/Thr protein phosphatases of type 1 (PP1) are the most abundant and among the best characterised protein phosphatase family. The PP1 catalytic subunit can interact with a large number of regulatory proteins, resulting in a conformational change of the phosphatase and a specific dephosphorylation of the target. Protein tyrosine phosphatases (PTPs), which also include dual-specificity phosphatases, dephosphorylate proteins at the tyrosine residues and play a major role in signal transduction. The ability of phosphatases to interact with multiple substrates explains their involvement in a wide range of cellular activities although there is limited information on their role in trypanosome biology. Nonetheless, a tyrosine phosphatase in et al., et al., 1998; 2002et al., et al., T. brucei. For example, one could speculate that PP1 acts upstream of AMPK in the SIF pathway such that AMPK depletion by RNAi might prevent the cell-cycle arrest generated upon PP1 overexpression, although PP1 might also have other targets in the pathway. Dual specificity phosphatases (DsPhos) in yeast have also been shown to regulate cell growth via regulation of the protein kinase A (PKA) cascade  plays a vital role in glucose signalling by interacting with a large number of regulatory proteins (e.g. Reg1). Glc7 is then capable of inactivating 5'AMP-activated protein kinase (AMPK) (Snf1 in yeast), which in turn transmits the glucose starvation signal  being identified in the RNAi screen for stumpy inducers. The RNAi-mediated ablation of PKA-R (found in the flagellar matrix) in monomorphs inhibited the motility of the parasites  has a potentially comparable function in cell cycle inhibition, as its depletion via RNAi results in prolonged cell proliferation and maintenance of slender forms  act as major regulators of cellular energy homeostasis in eukaryotic cells. The AMPK cascade is regulated by the AMP:ATP ratio and hence acts as an energy sensor. A decline in ATP levels, along with a concomitant rise in AMP, activates this kinase, which then phosphorylates a range of downstream targets to bring about changes that reduce catabolic processes within the cell. AMPKs are thus capable of responding to a reduction in ATP levels (that may be brought about by nutrient depletion) by pushing the cell towards a state of reduced activity, in order to maintain the AMP:ATP ratio. Interestingly, AMPK is also known to be an inhibitor of the mTOR pathway  cascade in yeast is involved in stress responses as well as cell cycle regulation. This cascade, which involves a module of three protein kinases  that sequentially activate one another by phosphorylation, form a crucial part of intracellular signalling pathways in eukaryotic cells –related kinases or NEKs, a class of serine/threonine kinases, were originally identified in a genetic screen for cell division cycle mutants in et al., et al., In summary, both protein phosphatases and protein kinases have been identified as regulators of the transition between slender and stumpy forms, though the targets upon which they operate are unknown. Nonetheless, phosphorylation is an expected contributor to the regulation of the signalling molecules themselves and also the effector molecules for differentiation upon which they may act, such as RNA binding proteins. Phosphoproteomic datasets analysing cultured bloodstream and procyclic forms   drive stumpy formation suggests they are fundamental components of a conserved cell communication or cell quiescence pathway found in other kinetoplastids. One observation from Table  and HYP2et al., et al., et al., et al., et al.  in pleomorphs inhibited growth and also increased the rate of slender to stumpy transition et al., Combined, one can speculate that trypanosomes have a dual mechanism for controlling differentiation, comprising of a slender retention (SR) as well a SI arm Fig. . A set oet al., et al., et al., et al., et al., A less investigated but crucial aspect of the differentiation process is the timing of the onset of the commitment to differentiation from the long, slender to the short, stumpy form. Although asymmetric division was suggested to be the means by which trypanosomes underwent commitment and morphological transformation  of stumpy-like forms in vitro has led to the discovery of various components of the SIF pathway (Mony et al., in silico information, the existing literature as well as experimental data from other eukaryotes (Fig. et al., Plasmodium (Ikadai et al., T. cruzi and Leishmania spp. The evolutionarily ancient position of trypanosomes in the eukaryotic lineage also has the potential to inform understanding of the molecular basis of cellular quiescence in other cells including those of mammals, whose breakdown is a key contributor to the development of many cancers.With the development of tools such as the genome-wide RNAi (Alsford tes Fig. , validat


Imagine yourself walking down the street. The act of initiating the first step triggers an increased firing of some medium spiny neurons (MSNs) in the striatum. These MSNs are part of the direct pathway in the basal ganglia complex (for a review on the basal ganglia, see Parent and Hazrati, Nature, Cui et al. (in-vivo photometry to observe what is active in the striatum when mice are performing an operant task. The changes in fluorescence intensity of GCaMP3, a calcium indicator, correlated to the MSN activity. In particular, Cui et al. (In a recent publication in i et al.  describei et al.  observedThe Journal of Neuroscience. In their experiment, Freeze et al. (For every action, there is an equal and opposite reaction. The opposite reaction to the Cui et al.  paper ise et al.  investigNevertheless, it is critical that we should approach the conclusions of the Freeze et al.  paper wiIn light of the results in Cui et al. , it is pLet us return to that walk down the street. When we make those first steps during action initiation, both dMSNs and iMSNs are concurrently active to help funnel the coarse cortical afferents through the basal ganglia. This type of pruning allows the dMSNs to help activate the right sets of muscles to move forward while permitting the iMSNs to help inhibit other muscles that can disrupt our balance or the forward motion. Ultimately, it is a coordinated effort between the two pathways that gets us to the final destination.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.


In its recent edition the Archives of Toxicology have published a transcriptome-based classifier that identifies developmental toxicants acting as histone deacetylase inhibitors. The authors, Eugen Rempel and colleagues from Dortmund Technical University, demonstrate that based on only eight genes the gene expression pattern of developmental toxicants acting as histone deacetylase inhibitors can be identified. They used a test system that recapitulates generation of neuroectoderm from pluripotent stem cells (Balmer et al., 2014; Leist ein vitro overlapped with concentrations known to cause teratogenic effects in humans in vivo. A next challenge in test system development is to develop expression signatures which indicate that specific toxic mechanisms are active. In this respect the present work of Rempel et al., using a set of structurally not related compounds acting by a similar mechanism, represents an important proof of concept (Rempel et al., 20156]; Grinbal., 2011; Liu et al., 2011; Xi et aal., 2011; Oesch eal., 2011; Stapletal., 2011; Ejaz an


The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The following information is missing from the Funding section: This study was supported by an unrestricted grant from UCB Pharma (The authors have also provided an updated Competing Interests statement: This study was supported by an unrestricted grant from UCB Pharma. João Eurico Cabral da Fonseca received this grant as the head of Rheumatology Research Unit at Instituto de Medicina Molecular, Faculdade de Medicina da Universidade de Lisboa. This does not alter our adherence to PLOS ONE policies on sharing data and materials.


Possibilities in auxiliary technique combinations with small- and wide-angle X ray scattering are described, as well as more complicated sample environments used in X-ray and neutron scattering. via either simultaneous or independent measurements, has advanced understanding of the driving forces that lead to the structures and morphologies of materials, which in turn give rise to their properties. The simultaneous measurement of different data regimes and types, using either X-rays or neutrons, and the desire to control parameters that initiate and control structural changes have led to greater demands on sample environments. Examples of developments in technique combinations and sample environment design are discussed, together with a brief speculation about promising future developments.Small- and wide-angle X-ray scattering  are standard tools in materials research. The simultaneous measurement of SAXS and WAXS data in time-resolved studies has gained popularity due to the complementary information obtained. Furthermore, the combination of these data with non X-ray based techniques,  The earliest SAXS beamline was constructed in Hamburg, in order to elucidate the mechanism by which muscles contract (Rosenbaum q range  would be projected onto a limited number of detector pixels.A conventional SAXS/WAXS beamline has two detectors placed several metres apart to collect SAXS and WAXS data simultaneously. With the lower divergence of third-generation synchrotron X-ray sources, the compromise between under- and over-focusing of the beam between the two detectors is reduced to an acceptable level. While high collimation allows the collection of both SAXS and WAXS data on a single detector, this arrangement will be detrimental to the low-angle information content, where a very large et al., 2004et al., 2006et al., 2013et al., 1995et al., 2012For some time, the photon flux delivered by most beamlines exceeded the capabilities of the detectors available. However, with the introduction of efficient high count-rate semiconductor-based photon-counting detectors (Schmitt It is not practicable to provide here a comprehensive overview of the developments in online time-resolved techniques used in combination with SAXS/WAXS (‘hyphenated’ techniques) and the accompanying more complicated sample environments. Instead, we try to give a flavour of some of the developments, with an emphasis on materials science experiments. Where possible, hints as to the timeline of how certain areas have developed in the last 20 years are given.2.et al., 2007et al., 2009et al., 2011et al., 2012et al., 2010et al., 2013The combination of techniques is not always beneficial. If the timescale over which samples are modified is such that it is also feasible to perform independent experiments without introducing a notable error margin between the experiments, then one is better off carrying out optimized independent experiments. In combinations, often at least one of the experiments yields suboptimal, but still useable, data. The only advantage of combining techniques is the synergy that results when data sets are obtained simultaneously from a single sample with, for example, a well characterized thermal history or other provenance. The longer limit we define, somewhat arbitrarily, as being 1–2 min per time frame. The limit on the fast side is in general no longer set by the available X-ray flux or the detectors. Where the fastest of the older generation of CCD area detectors could collect data at rates of 15 Hz with a limited dynamic range  and interception by the X-ray beam can then be varied to provide time resolution. In this case the problem of radiation damage is overcome as well.Very fast reactions can be studied using microfocus techniques, where a miniature extruder, et al., 1998et al., 2000et al., 1998et al., 1998et al., 2003When combining techniques one has to be aware of the fact that there might be differences in sensitivity for certain aspects of the experiments. For example, in an extended debate on polymer crystallization that raged roughly between 1998 and 2005, a proposal was made that the distinction between a nucleation and growth crystallization model, and a spinodal-like phase-separation model that preceded crystallization, could be made by the sequential occurrence of scattering and diffraction intensity in the SAXS and WAXS regimes , if we assume that no heat exchange will take place between the irradiated area and its surroundings. This does not suggest that any sample will heat up appreciably, since the adiabatic approximation is somewhat rigorous. However, if the thermal sensor of the temperature controller is some distance away from the position where the beam hits the sample, it is not advisable to over-interpret a combination of online DSC and X-ray scattering, since a thermal gradient will almost certainly exist and indeed has been shown experimentally  , where q is as defined in §1et al., 2005When hierarchical length scales are involved, the simplest experimental option is to combine SAXS and WAXS in the conventional transmission mode. Realistically, on most beamlines this can render data over a continuous scattering vector range 3 × 10550-b-PFS50, in decane solution can be mentioned  image of a sample which is to be studied by SAXS is invaluable, since it narrows down the nearly limitless possibilities for data interpretation which are possible when relying only on first principles and SAXS data Fig. 3. Anotheret al., 1995stratum corneum, the top protective layer of human skin  tail organizations. Direct correlations between these parameters and the improper skin barrier function suffered by eczema patients were established in this way.The combination of SAXS with Fourier-transform IR spectroscopy (FT–IR) has been shown to be useful in determining the extent to which chemical reactions have progressed and what this means for structure formation  which was uniaxially oriented  in a quasi-simultaneous mode  muscles  and a mid-block of poly(methacrylic) acid, which exhibits a coil–globule transition as a function of pH, has been set up as a free-running motor system by applying a periodically changing pH. The idea was also to show that the cycling in length of the block copolymer reaction could be repeated The above two examples highlight some of the most challenging experiments known to the authors. However, it is clear that not every experiment requires a very complicated setup. To determine the phase diagram of a block copolymer or liposome solution, a simple heating stage and a night of beamtime might be sufficient. What it is important to realise is that such complicated experiments are feasible, and it only requires the will to do them and the perspiration of beamline users and staff to make them a reality.7.in extremis, is required. Most synchrotron laboratories have several beamlines and organization units dealing with extreme conditions. The parameters that are most often classified as ‘extreme’ in this context are temperature and pressure. The application of the most extreme conditions is not employed very often in SAXS. This is most likely due to the fact that moderate temperatures are already sufficient to modify the long-range structures that can be studied with SAXS, and extremes of temperature and pressure have a more profound effect on the shorter atomic or molecular length scales.Sometimes the variation of a single parameter, but then et al., 2008et al., 1996et al., 2008et al., 1999et al., 2010P-jump type experiments. The cells share common features, including custom PTFE (polytetrafluoroethylene) spring seals for work above 0.4 GPa. Recent improvements to cell design have included a dedicated sample-loading port which allows the windows, often diamond, to be left in place during experiments, thus alleviating the problems of background subtraction which were evident in previous designs. Scientifically, the cells have been used to study the distinction between pressure and thermal or chemical denaturation in biological proteins  of 3 × 10−4 Å−1  has been introduced, which transfers spin polarization from electrons to nuclei, thereby aligning the nuclear spins to the same extent that the electron spins are aligned (Abragam, 19618.3.et al., 2010et al., 2006et al., 2007et al., 2010et al., 2010et al., 2011Polymerization solutions are models of living systems, in that the species and content of the solutes change as time proceeds. Anionic copolymerization of styrene and isoprene monomers in a dilute solution, with deuterated benzene as the solvent, has been studied by means of combined time-resolved SANS, size-exclusion chromatography, NMR and UV–Vis spectroscopy (Zhao 9.et al., 2012Crystallization studies have been an important driving force for technique developments in time-resolved SAXS/WAXS. The technology for constructing sample environments to make such experiments possible, even at microsecond timescales, has been developed. The difference in sensitivity between SAXS and wide-angle diffraction/scattering events with respect to their ability to shed light on the earliest stages of crystallization, mentioned earlier, has evoked two different responses. The first was to develop very high count-rate single photon-counting WAXS detectors that were at least an order of magnitude more sensitive and efficient than the photon-counting detectors used for SAXS experiments. However, this is an approach that will only bring improvements until one reaches the intrinsic limits, dictated by physics, where differences in technique sensitivity become manifest. Beyond that point one has to revert to different approaches. One of these methods is the use of pair distribution function analysis (Tyrsted ad hoc and quite often open to debate. Attempts to utilize similar methods to those used for solution scattering for particle growth have been carried out, but the methods remain cumbersome (Shaw et al., 2002In data analysis great strides can still be made. Protein solution scientists have been well served by data packages that allow them to squeeze the last piece of information out of their low-information-content data sets consisting of the scattering curves of monodisperse proteins. However, when dealing with time-resolved data from polydisperse particles and high matrix backgrounds, most analysis methods are still et al., 2011et al., 2014When dealing with different experimental data sets obtained in a single time-resolved experiment, it is often not difficult to correlate events. One plots the development of a SAXS invariant on the same graph as the variations in the spectroscopic absorption band, takes a ruler and determines what comes first. However, when subtle effects are in play and the data are noisy, as one can expect in a time-resolved experiment, the situation might become more complicated. Fortunately, there have been some interesting recent developments. Several years ago, two-dimensional correlation analysis on SAXS–WAXS data sets was attempted in order to determine the exact correlation between parameters obtained from the different data sets (Smirnova 10.We have attempted to give a flavour of what is currently feasible in combining techniques with small- and wide-angle scattering. This is by no means an exhaustive review but rather an attempt to make the case; if one thinks that a combination of techniques might deliver results that would otherwise not be available, one should not be afraid to try. On modern SAXS beamlines many things are possible. It might require some hard work and collaboration with the beamline scientists, but that should also be part of the fun of experimenting!


Correlations between particulate organic carbon (POC) and mineral fluxes in the deep ocean have inspired the inclusion of “ballast effect” parameterizations in carbon cycle models. A recent study demonstrated regional variability in the effect of ballast minerals on the flux of POC in the deep ocean. We have undertaken a similar analysis of shallow export data from the Arctic, Atlantic, and Southern Oceans. Mineral ballasting is of greatest importance in the high-latitude North Atlantic, where 60% of the POC flux is associated with ballast minerals. This fraction drops to around 40% in the Southern Ocean. The remainder of the export flux is not associated with minerals, and this unballasted fraction thus often dominates the export flux. The proportion of mineral-associated POC flux often scales with regional variation in export efficiency (the proportion of primary production that is exported). However, local discrepancies suggest that regional differences in ecology also impact the magnitude of surface export. We propose that POC export will not respond equally across all high-latitude regions to possible future changes in ballast availability. Our previous analysis, based on limited data sets derived from Wilson et al. [Klaas and Archer [Le Moigne et al. [Wilson et al., Wilson et al. [Teff; namely, that Teff is high in calcite-dominated regions and low in opal-dominated regions. While this is the global Teff pattern reported by some studies [François et al., Henson et al., Lam et al., Teff [Buesseler and Boyd, Martin et al., Rynearson et al., Smetacek et al., Further analysis of the global data set of deep particle flux by n et al.  has showd Archer  and Le Me et al.  and othen et al.  hypotheseff). Here we examine the variability in surface mineral ballast Ccs using 234Th-derived estimates of POC and mineral flux in the Arctic, Atlantic, and Southern Oceans (95 data points). We discuss the implications of our results for the role of minerals in the export of POC from the surface ocean.However, it is still unclear how the impact of ballasting on shallow POC export varies regionally and how it is linked to export efficiency  particles collected on Nitex screen using an in situ stand-alone pumping system deployed at a single depth beneath the mixed layer (from 20 to 70 m). Particles were then rinsed off the screen using thorium-free seawater, and the particle suspension split into equal subsamples. Each split was then analyzed for one of the following: 234Th, POC, BSi, and particulate inorganic carbon (PIC) as in Le Moigne et al. [These fluxes were then converted to estimates of downward particle flux using the POC:e et al. .234Th approach has the great advantage of providing a large data set of surface export fluxes, albeit with the caveat that some of the seasonal variability of export flux may be missed. While this is unlikely to be a major issue in low-latitude areas where the seasonal cycle in export is weak [Waniek et al., Waniek et al., Using the Lambert et al. [Kuss and Kremling [Le Moigne et al. [Honda and Watanabe [Planquette et al., −1) for stations in the plumes of chlorophyll a associated with South Georgia and the South Sandwich Islands and the average value from nonplume stations around Crozet [Planquette et al., −1) for our nonplume stations. Table S1 summarizes the values of PAl used for each station. Lithogenic fluxes were estimated as (100/8*Al) [Honda and Watanabe, Gehlen et al., Salter et al., The aluminum concentration in large particles (PAl) was not measured; therefore, literature values were taken from t et al.  and KussKremling  to estime et al.  and HondWatanabe . For theHonda and Watanabe [Le Moigne et al. [total flux) into four portions, three of which are considered to be associated with the mineral phases BSi (POCBSi), calcite (POCpic), and lithogenic material (POClith). The fourth portion is the residual POC flux, which is considered the non-associated fraction (POCres). MLRA takes the following form:We examined the relationship between mineral and POC fluxes using two different techniques. Initially, we used multiple linear regression analysis (MLRA) as in Watanabe  and Le Me et al. . MLRA dia, b, and c are defined as Ccs [Klaas and Archer, d is the nonassociated portion of POC [Armstrong et al., Honda and Watanabe, Le Moigne et al., Wilson et al. [Wilson et al., We then used the geographically weighted regression (GWR) technique. The application of GWR to ocean particle flux data was carefully described and tested by n et al. . We follNakaya et al., http://www.st-andrews.ac.uk/geoinformatics/gwr/gwr-software/). We used an adaptive Gaussian Kernel, which groups the data by the nearest neighbor approach. The Akaike Information Criterion, corrected for small sample bias (AICc) [Akaike, Wilson et al. [We used the GWR 4.0 software [ Akaike, , was usen et al. . We alsoIn the following section, we briefly describe the magnitude of surface POC and mineral export fluxes Figure . The higPIC export Figure b shows sR2 of 0.22 and an AICc of 1139 (Table R2 to 0.52 and reduced the AICc to 1098 (Table p < < 0.001). Formally, we reject the null hypothesis that there is no difference between the GWR and MLRA models. Finally, the test of geographical variability  is high in the SO as previously observed [Le Moigne et al., −2 d−1, with the lowest values seen in the Arctic and the North Atlantic subtropical gyre  and SO  Figures j, and 2mWilson et al., Le Moigne et al., Milliman et al., Ragueneau et al., Wilson et al., To fully understand the interaction between mineral and POC fluxes, one needs to assess the variation of mineral-associated POC fluxes not only at depth [Teff, of flux through the mesopelagic [François et al., eff, we now compare our mineral-associated POC export data to the global pattern of PEeff reported by Siegel et al. [It has been suggested that minerals influence the transfer efficiency, l et al. .Siegel et al. [eff in their data set from the HLNA (0.1–0.25), where our analysis shows the highest proportion PIC- and BSi-associated POC export , where a lower proportion of the total POC export is mineral associated than in the HLNA  [Siegel et al., In the subtropical gyres, where the spatial variability of mineral-associated POC export is reduced and the proportion of the POC flux associated with minerals is low Figures g and 2m,eff. While we can only compare our export data to the climatology of PEeff reported by Siegel et al. [eff at the time of sampling), the comparison suggests the following:Iversen and Ploug, Schmidt et al., Klaas and Archer [Sanders et al. [There are fundamental differences in the ways minerals promote POC export in different oceanic provinces. We know that mineral ballast can have different ballasting properties (particle nucleation and/or added density) depending on mineral type. For example, calcite-ballasted particles generally sink faster than opal-ballasted ones [d Archer  and Sands et al. .eff. For instance, ecosystem functions such as the packaging of slowly sinking phytoplankton cells into large, fast-sinking fecal pellets may increase PEeff regardless of the proportion of POC associated with minerals in the material grazed.Factors other than the magnitude of mineral-associated POC flux are also likely to influence PEThe proportion of POC flux carried by mineral does not so clearly correlate with the PEl et al.  (and noteff in the SO, potentially due to grazing intensity [Maiti et al., Pabortsava, We suggest that these processes are potentially more important in the SO and the subtropical gyres than in the HLNA. This is consistent with observations of an inverse relationship between primary production and PEArmstrong et al., Honda and Watanabe, 2 levels [Kwon et al., Globally, POC not associated with minerals often dominates total POC export out of the surface ocean Figure k. While eff presented by Siegel et al. [eff in the SO and the subtropical gyres. Ecosystem processes like grazing intensity may be more important in these regions.Surface mineral-associated POC fluxes are highly variable Figures i–2k, butl et al. , suggestOur analysis reveals two important messages:eff, Siegel et al. [Teff.Comparing mineral-associated POC export with the global pattern of PEl et al.  has provTeff derived from sea surface temperature are low [Henson et al., Engel et al., Engel et al., Le Moigne et al., Teff in the HLNA is higher than proposed by Henson et al. [Buesseler and Boyd, Martin et al., Rynearson et al., Teff is also estimated to be low [Henson et al., Teff in high-latitude diatom-dominated regions [Assmy et al., Henson et al., Lam et al., Wilson et al., Teff [Smetacek et al., In the HLNA, estimates of n et al. , as was eff/low Teff occurring at high latitudes and low PEeff/high Teff occurring at low latitudes [Henson et al., eff and Teff are likely to be driven in turn by factors beyond just sea surface temperature. Thus, not all high-latitude regions are identical, and they cannot be described by the same “ballast” parameterization in ocean carbon models. For instance, POC export in the HLNA and the SO are unlikely to respond in the same way to any reduction in ballast mineral availability, as might be caused by ocean acidification, increased sea temperature, or deepening of the wind mixed layer [Sarmiento et al., Zondervan, Falkowski et al., 2.Our results suggest that viewing the BCP as a two end-member system with high PE


As such, Safavi and co-workers draw into question this conclusion, based on evidence indicating that neural activity in frontal areas, in particular in the lateral prefrontal cortex, reflects the contents of consciousness in paradigms that do not involve active report.Like any other field, the field of consciousness research benefits from a careful distinction between the concepts involved. An example is the distinction between the state of being conscious  and the contents of consciousness . A similar type of distinction can contribute to the resolution of a debate regarding the role of the frontal cortex in conscious perception. In a recent publication Safavi et al.  respondecontent of consciousness, and neural activity that brings about changes in consciousness during perceptual bistability. Safavi et al. (Here we aim to bring these two positions closer together by making an explicit distinction between neural activity that reflects the i et al.  bring foi et al. . Frässlei et al. . FrässleAs a further point, the “binocular flash suppression” paradigm that provides much of the empirical basis of Safavi and colleagues' argument, manipulates the content of awareness while explicitly avoiding spontaneous switches in perception. This provides a further indication that Frässle et al.'s claims regarding perceptual switches can exist side-by-side with Safavi et al.'s claims regarding stable perception. Indeed, the next paragraph will summarize evidence that frontal involvement in coding the contents of consciousness does not imply frontal involvement in perceptual switches.Whereas the anatomical distribution of activity that reflects the content of consciousness during bi-stable perception depends on the stimulus employed (Logothetis and Schall, As a final remark we would like to stress, as others have (Safavi et al., In conclusion, we agree with the notion, emphasized by Safavi and colleagues, that the field of consciousness research will benefit from an integrated view of evidence from various experimental and neuroscientific paradigms (Aru and Bachmann, 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.


Thirty percent of patients with epilepsy experience seizures despite optimal anticonvulsant drug treatment. Stress is frequently identified by patients with epilepsy as a precipitant of seizures (Spector et al., However, in this commentary we would like to specify that contrary to what it is claimed by the authors, decreasing arousal, using relaxation biofeedback is not the method proposed by Nagai et al. . On the The motivation for believing that EDA relaxation biofeedback may be beneficial for patients with epilepsy comes is understandable given the relationship between stress and increased skin conductance level. It is known that stress and anxiety are commonly reported triggers of seizures Fenwick, . Thus, iDespite strong theoretical underpinnings there is little direct knowledge about the neural mechanisms underlying EDA biofeedback on seizure onset. Scrimali et al.  suggest As evidence of the clinical efficacy of EDA arousal biofeedback in the management of epileptic seizures, Nagai et al.  report aIn conclusion, EDA biofeedback has great potential as a non-pharmacological therapy for the management of refractory epilepsy. The methodology is grounded on theory, and on increasing knowledge regarding the underlying neural mechanisms, and the approach deserves greater clinical attention. There also remains a need for continuing clinical research, such as the case study report by Scrimali et al. . DespiteThe 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.