Timing cues are an essential feature of music. To understand how the brain gives rise to our experience of music we must appreciate how acoustical temporal patterns are integrated over the range of several seconds in order to extract global timing. In music perception, global timing comprises three distinct but often interacting percepts: temporal grouping, beat, and tempo. What directions may we take to further elucidate where and how the global timing of music is processed in the brain? The present perspective addresses this question and describes our current understanding of the neural basis of global timing perception. Rhythm perception is essential to our appreciation of music. Since music is a human construct and rhythm is defined primarily by its use in music, it is unclear whether studies of rhythm in non-human species directly relate to human rhythm perception. In this perspective, we use the term “global timing” as a conceptual framework, to emphasize the temporal computation required to perceive and recognize rhythm that is not unique to musical contexts, but is nonetheless required to perceive and recognize a musical rhythm. With this terminology, we also aim to facilitate the comparison of research findings across different animal models, where perception may take a different behavioral form but neural mechanisms may nevertheless be shared. We discuss the conceptual framework of global timing in light of recent research in both humans and other animal species, examine which brain regions may underlie global timing perception, and speculate on its potential neural mechanisms.local timing of individual events , and analyzes its global features; in human music perception, these include temporal grouping, beat, and tempo . It requires that the listener integrates temporal patterns spanning several seconds. While rhythm is also influenced by other acoustic characteristics such as pitch, intensity, and timber, global timing perception is, by definition, driven by timing. The human auditory system likely processes temporal patterns at two levels—it computes the empo Box . Differeempo Box , or the empo Box . The beaempo Box . In its empo Box . In the empo Box , Deutschempo Box , and Fitempo Box , and theempo Box .Box 1ABox 1BThe schematic depiction indicates the relative tone onsets and duration on time axis (middle), of the perceptual groups based on temporal grouping (top), and the perceived beat (x) where number of stacked x's show the number of overlapping levels in the beat hierarchy  studies in healthy humans have associated a network of cortico-striatal brain areas  with the performance of global timing tasks. These areas show increased activity compared to baseline when participants perform same-different judgments  and electroencephalography (EEG). For example, steady-state evoked potentials have been measured to capture the brains response to beat perception SMA, basal ganglia, and cerebellum (Grahn and Brett, Before global timing information can be processed by multiple higher-order brain structures, the acoustic signal must be encoded in auditory cortex. In cats, inactivation of the anterior, but not the posterior, field of secondary auditory cortex impairs performance on a temporal pattern-matching task (Lomber and Malhotra, In general, the temporal integration window in the auditory pathway varies. Previous reports suggest an integration windows of less than 10 ms in subcortical structures, ~20–25 ms in primary auditory cortex, and more than 100 ms in auditory fields downstream from primary auditory cortex in primates (Lu et al., synfire chain. Neural activity in a synfire chain is sparsely present throughout the song sequence, with each neuron being active at only one single time-point within the sequence and not necessarily correlated with the onset of a syllable (Hahnloser et al., synfire chain (Long et al., synfire chain could be used for computing global timing information in other brain regions.In the zebra finch, area HVC contains neurons that fire during the bird's song production in a mechanism referred to as A different neural mechanism for encoding sequential information has been reported in the hippocampus of rodents. Neurons in the hippocampus are tuned to behaviorally relevant spatial (O'Keefe and Dostrovsky, We suggest global timing as a conceptual framework to investigate the temporal aspects of musical rhythm perception that require listeners to integrate temporal patterns spanning several seconds. Music perception in humans comprises three features of global timing—temporal grouping, beat, and tempo,—which are perceptually distinguishable yet interdependent. Both humans and non-human species make use of temporal grouping and tempo processing in auditory signals. These features are ecologically important in interpreting the vocal calls of many species. In contrast, beat perception—that aspect of global timing that often distinguishes music from other sounds—appears to be limited in most non-human species. Global timing perception relies on neural processes across a wide network of subcortical and cortical brain regions, including the basal ganglia, SMA, and prefrontal cortex, as well as a subset of fields within auditory cortex. The neural mechanisms that underlie global timing perception remain elusive. However, given the limited temporal integration window of neurons, it is likely that sequential activity within a neuronal ensemble is required to encode global timing. Further research into the neural basis of processing outside of auditory cortex may help in understanding how the brain processes global timing. The examination of vocal learners such as parrots or European starlings, that could be capable of beat perception, could be essential toward that goal. In future experiments, designs that allow dissociating temporal grouping, beat and tempo perception processes will be a necessary step toward unraveling neural mechanisms of global timing.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.


Stem Cells 2011;29:169–178Transplantation of bone marrow stem cells into spinal cord lesions enhances axonal regeneration and promotes functional recovery in animal studies. There are two types of adult bone marrow stem cell; hematopoietic stem cells (HSCs), and mesenchymal stem cells (MSCs). The mechanisms by which HSCs and MSCs might promote spinal cord repair following transplantation have been extensively investigated. The objective of this review is to discuss these mechanisms; we briefly consider the controversial topic of HSC and MSC transdifferentiation into central nervous system cells but focus on the neurotrophic, tissue sparing, and reparative action of MSC grafts in the context of the spinal cord injury (SCI) milieu. We then discuss some of the specific issues related to the translation of HSC and MSC therapies for patients with SCI and present a comprehensive critique of the current bone marrow cell clinical trials for the treatment of SCI to date.  When axons in the central nervous system (CNS) are damaged they mount a poor regenerative response due to a combination of inflammation, resulting in extensive neuronal and glial cell death and glial cell activation and hypertrophy, which contributes to the formation of the glial scar. These intrinsic responses to tissue injury both contribute to an environment that is inhibitory to axonal regrowth .Following spinal cord injury (SCI), the blood-brain barrier is disrupted and an influx of inflammatory cells occurs, which is facilitated by their expression of matrix metalloproteinases (MMPs) . MMPs, oSome evidence suggests that inflammation may be a beneficial response to SCI. For example, macrophages phagocytose the myelin debris present in the injured spinal cord, which is known to inhibit axonal regeneration , 7, and Glial scarring involves astrocytes, which are activated in an effort to restore the blood-brain barrier, and oligodendrocytes. The extracellular matrix produced by these scar-associated cells contains a number of molecules that inhibit axonal regrowth  of whichOther inhibitory molecules present within the glial scar include myelin-associated proteins, such as myelin-associated glycoprotein (MAG), Nogo-A, and oligodendrocyte-myelin glycoprotein (OMgp) , 7. MAG There are two types of bone marrow stem cell, hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs), which are known to differentiate into hematopoietic and mesenchymal cell lineages, respectively  , 57, andIt has been proposed that MSCs act as “guiding strands” for regenerating axons across the lesion site in the injured cord and along spinal cord tracts in vivo . TransplA recent study has focused on the ability of MSCs to respond to the environmental stimuli in the injured spinal cord. MSCs that were administered with extracts from injured spinal cord tissue responded by increasing their synthesis of various cytokines, including IL-6, IL-7, and VEGF . The bioA preliminary question for the application of autologous HSCs or MSCs for human SCI cell therapy is whether these cells are available in individuals who have been injured. Early work demonstrated marked and significant changes in the composition of iliac crest tissue in individuals with complete paralysis compared with non-SCI donors . In the In practice, most clinical applications of bone marrow cells for the treatment of SCI have involved the use of whole mononuclear cells preparations (MCPs) –75 and t6 cells are grafted  [Larger patient cohorts would be required to determine the significance of any functional improvements in these patient trials and to assess any associated risks of MCP/MSC or GM-CSF treatments. It is noteworthy that no details of physical rehabilitation were reported in any of these clinical trials other than that “all patients underwent standard physical therapy prior to and after transplantation” . For futrg/iccp) . This inThe potential of bone marrow cell transplantation as a method of repair in the injured CNS may serve a number of different purposes that span various therapeutic targets. Animal studies have demonstrated that transplanted MSCs modify the inflammatory environment in the acute setting and reduce the effects of the inhibitory scar tissue in the subacute/chronic setting to provide a permissive environment for axonal extension. In addition, grafted cells may provide a source of growth factors to enhance axonal elongation across spinal cord lesions. Other studies have suggested that HSCs and MSCs may even transdifferentiate to replace lost or damaged neuronal tissue. Preliminary clinical data indicates that autologous bone marrow cell transplantation and/or GM-CSF administration can be used to treat patients with SCI without any immediate serious complications. These data are promising, but future studies must continue to establish whether bone marrow cell treatments can serve as a safe and functional autologous source for the treatment of the injured CNS.


The current excitement about adult stem and progenitor cells is an opportunity for developmental biologists to shed new light on old problems. Where do differentiated cells in the adult come from, and what else can their progenitor cells do? These questions are especially accessible in the zebrafish, where there is a deep experimental and genetic toolbox, and a developmental pathway with a well-defined metamorphic transition from larvae to adult. One of the premier model systems in this premier-league model organism is pigmentation, since pigment cells in fish—including black melanocytes (known as melanophores in fish), yellow xanthophores, and reflective iridophores—are organized into an array of beautiful, accessible, and evolutionarily diverse patterns that have helped motivate their study PLoS Genetics, Budi et al. In this issue of foxd3 and sox10, markers of both glial and early neural crest cells mitfa::GFP, which in embryos marks pigment cell precursors and differentiating melanocytes mitfa::GFP cells derive from foxd3- and sox10-expressing cells and that some cells expressing these markers become proliferative prior to and during early metamorphosis. All these progenitor cells are strongly associated with peripheral nerves. Crucially, DiI labelling and time-lapse analysis of mitfa::GFP fish demonstrate that a proportion of labelled cells move into the skin, where some differentiate into melanocytes or iridophores.Budi et al. begin their studies by showing that cells that express mitfa::GFP progenitors arrive in the skin in erbb3b and tuba813a mutants, which are known to be required for formation of melanocyte progenitors mitfa::GFP progenitors in mutants for these genes. To what extent these differences can explain the distinctive phenotypes remains to be established, and will likely require detailed mathematical modelling of the patterning process. Nevertheless, this study provides an unprecedented amount of quantitative data on which to base modelling efforts.Further support for the authors' model came from exquisitely detailed quantitative comparisons of progenitor behaviour in key patterning mutants. Many fewer erbb3b  mutant studies. Although this mutation severely reduces the number of pigment cell precursors (as defined by expression of mitfa::GFP) during metamorphosis, the few mutant cells that do survive to reach the hypodermis actually exhibit a greater degree of proliferation and differentiation relative to wild type. This observation seems to demonstrate the often assumed, but rarely shown, regulative behaviour of pigment cells in vivo.An interesting observation emerging from this work comes in particular from the foxd3 or mitfa::GFP-expressing extra-hypodermal cells persisting in adults since this suggests that they may contribute to regeneration of melanocytes. They use an elegant genetic approach to show that adults have limited regenerative capacity for pigment cells, at least in the double-mutant background used. Although not assessed here, it would be interesting to examine whether extra-hypodermal progenitor cell numbers become correspondingly reduced during this experiment.Finally, the fish's ability to regenerate pigment cells is another fascinating aspect of this model system mitfa::GFP transgene as an invaluable marker for the early stages of their development. These breakthroughs inevitably suggest new questions. Are these progenitor cells neural crest-derived? While overwhelmingly likely, this still needs to be formally shown. Do all adult pigment cells arise from these progenitor cells? The authors have not been able to identify the origin of xanthophores, and the possibility that some melanophores and iridophores derive from other cells remains. Where are the pigment cell stem cells? The authors show that progenitors for mitfa::GFP cells express foxd3 and sox10 and reside within the peripheral nervous system. This may suggest a dispersed, axon-associated source of pigment cell progenitors. However, these latter cells themselves may arise from spatially more localized stem cells, perhaps in ganglia or specific niches of the nerves, which simply use the peripheral nerves as convenient tramways for dispersal. Even if the stem cells are dispersed and axon-associated, what do they look like? Do differentiated satellite glia and/or Schwann cells have latent potency to form pigment cells? Or are there distinct, dormant stem cells intermingled with them? For now, a Schwann cell precursor, as in mammals This study provides compelling evidence for a neural origin of adult pigment cells and reveals the foxd3/sox10-expressing cells are tripotent (glia/melanocyte/iridophore), while the mitfa::GFP-expressing cells may be bipotent (melanophore/iridophore), although separate unipotent progenitors for each cell type are not yet ruled out. Detailed lineage studies will address these issues, and may reveal other, unexpected, derivatives.The progenitor populations identified here are abundant during metamorphosis, and it is far from clear that all will form pigment cells. While many may remain as a source of regenerative cells, what cell types can they form? The authors are understandably cautious, but propose that the This landmark study provides compelling evidence for the location and identity of a cellular source of fish adult pigment cells. We can now anticipate comprehensive examination of these cells in the many available mutants, promising mechanistic insight into the enduring problem of adult pigment pattern formation. Beyond this, the identification of the cryptic progenitors of adult pigment cells should make the fish pigmentary system another tractable model in which to explore general principles of adult stem cell biology. For example, issues such as how stem cell potency is controlled and develops, how multiple cell types are generated in appropriate numbers from them, and how these cells are activated during regeneration can all be explored with this system. It is now just a matter of time before fish pigmentation helps colour our understanding of adult stem cell biology!


Alcohol dependence is a chronic relapsing disorder characterized by neuroadaptations that may result in the emergence of negative affective states and stress responses upon discontinuation of alcohol use. Clinical studies have demonstrated that alcohol-dependent people are more sensitive to relapse provoking cues such as alcohol, negative affect, and stress. Moreover, stress relief during protracted abstinence is thought to be a major motivation for excessive alcohol consumption. The relationship between chronic alcohol use, stress, and relapse has implications for the treatment of alcohol dependence. Recent research suggests that neural systems mediating stress responses may offer useful targets for pharmacotherapy of alcoholism. Diagnostic and Statistical Manual of Mental Disorders–Fourth Edition Text Revision (DSM–IV–TR)   characteM–IV–TR) .Several neurochemical systems and brain regions are involved in the development of alcohol dependence , a 41–amino acid neuropeptideSuch alcohol-induced neurobiological changes represent possible molecular targets for pharmacotherapies of alcoholism, which help to facilitate abstinence or greatly reduce alcohol consumption by stabilizing neurobiological systems dysregulated by chronic alcohol use. Medications that normalize the dysregulation or balance of the reward and stress systems may protect against relapse. In fact, evidence shows that pharmacological treatments can support abstinence or decrease the number of heavy drinking days. Three medications are approved for the treatment of alcohol dependence in the United States––disulfiram, naltrexone, and acamprosate. Recent efforts to develop new medications have focused on specific neural responses to factors  that increase risk of relapse to heavy drinking during protracted abstinence. The following sections will describe a series of neuropharmacological agents that alter the stress response and have potential for or have been used in the treatment of alcohol dependence.Recent research has led to the hypothesis that the transition to alcohol dependence involves the dysregulation not only of neural circuits involved in reward but also of circuits that mediate behavioral responses to stressors. Alcohol-induced dysregulation of the brain’s stress and anti-stress systems is hypothesized to contribute to the negative emotional state characteristic of alcohol withdrawal. More specifically, several observations indicate that CRF contributes to the development of alcohol dependence. For example, alcohol is a powerful activator of stress systems involving both the HPA axis and extrahypothalamic CRF systems in the extended amygdala; the latter also become hyperactive during withdrawal, leading to increased CRF levels in certain brain regions  . In animCRHR1 gene and drinking behavior. CRHR1 gene polymorphisms and both binge drinking and lifetime prevalence of alcohol intake in an adolescent sample from the Mannheim Study of Children at RiskCRHR1 gene also were found to moderate the relationship between the number of negative life events and rates of lifetime alcohol use and excessive alcohol use per occasion in the same study sample  seems to be one of the major elements common to depression, anxiety, and addiction. Genetic studies indicate an association between polymorphisms of the y sample , suggestCRHR1 gene that exacerbate a stress-induced susceptibility to alcohol dependence and relapse  have intimate involvement in brain processes relevant to alcohol dependence and contribute to the brain stress activation associated with withdrawal. A study of recently abstinent alcohol-dependent patients revealed elevated plasma levels of norepinephrine and the related neurotransmitter epinephrine , suggestPrazosin, an α1-noradrenergic receptor antagonist, has kindled interest as an effective drug in reducing alcohol use. Pfizer Pharmaceuticals introduced Prazosin in 1973 as an antihypertensive drug. An inexpensive generic drug for many years, prazosin has been used chronically by millions of people for hypertension. It is the most lipid soluble α1-noradrenergic antagonist and the only clinically available α1-noradrenergic antagonist demonstrated to be active at central nervous system sites when administered peripherally . PrazosiA 6-week, double-blind, placebo-controlled pilot study of Prazosin for the treatment of alcohol dependence reported a significant reduction in drinking behavior in actively drinking alcohol dependent patients . Large cTargeting the receptor system for Substance P, which modulates emotional states, has been suggested as a viable therapeutic target for the treatment of alcohol dependence . SubstanA double-blind clinical trial of alcohol dependence found treatment with an NK1 antagonist significantly decreased craving, blunted cortisol responses, and decreased functional magnetic resonance imaging responses to affective stimuli in recently detoxified alcohol-dependent study participants , a 36–amino acid peptide, also is involved in regulating the body’s stress response but with a neural and behavioral profile that in almost every aspect is opposite to that of CRF. For example, NPY has powerful anxiety-reducing effects in animals. It is one of the most abundant neuropeptides in the central nervous system (CNS) and is considered an important regulating factor in emotional behavior. Administration of NPY from an external source  has antianxiety and sedative effects that rely, at least partially, on activation of Yamygdala .Several findings point to a role for NPY produced in the body  in the control of stress- and anxiety-related behaviors, supporting the antistress effects observed following central administration of NPY. In animal models, acute physical restraint, which promotes experimental anxiety, suppresses NPY expression within the amygdala and cortex, an effect that parallels the anxiety-inducing effects of stress. In contrast, repeated exposure to a siren stressor leads to complete behavioral and endocrine habituation, accompanied by an upregulation of amygdalar NPY expression , 2010. TNPY levels are lower in the CeA of alcohol-preferring (P) rats compared to non-P (NP) rats, and NPY infusion in the CeA attenuates the anxiety-like and alcohol drinking behaviors of P rats. Thus, a deficiency in NPY signaling in the CeA may be involved in regulating both anxiety and alcohol-drinking behaviors  and NPY Dynorphins are opioid peptides that derive from the prodynorphin precursor and are the presumed endogenous ligands for the κ opioid receptor . DynorphActivation of the dynorphin/κ receptor system can produce analgesic actions similar to other opioids but also actions that are opposite to those of μ opioid receptors in the motivational domain, where dynorphins produce aversive, dysphoric-like effects in animals and humans . DynorphThe evidence for a role of the dynorphin/κ opioid system in the neuroadaptive actions of ethanol  is based both on biochemical studies and antagonist studies. Chronic self-administration of ethanol in C57BL/6J mice produced increases in dynorphin B in the amygdala and substantia nigra 21 days after cessation of drinking . ChronicStress also increases dynorphin activity , suggestAlcohol has a complex neuropharmacology and can affect many different neurotransmitter systems. Several pharmacological agents that interact with specific neurotransmitter systems affected by alcohol already have shown efficacy in the treatment of alcohol dependence and many exciting experimental agents are on the horizon. Stress relief during protracted abstinence is thought to be a major motivation for excessive alcohol consumption and the present overview outlines several new targets for medications development based on interactions with the brain stress systems. The development of these agents has been based on translational approaches ranging from the use of molecular techniques to understand alcohol neurobiology and identify candidate molecules, to the use of numerous animal models of alcohol-related behaviors to test the effects and mechanisms of action underlying these agents, and finally the use of human clinical trials and laboratory paradigms to evaluate the clinical efficacy of these agents. Future research needs to focus on realizing the therapeutic potential of agents acting on the brain stress systems and examining genetic and patient-specific predictors of treatment response. A better understanding of the mechanisms underlying treatment response could lead to appropriate treatment matching and efficient utilization of such novel medications.


Sepsis is defined as a systemic inflammatory response secondary to a proven or suspected infection. Mechanisms governing this inflammatory response have been shown to be complex and dynamic, involving cross-talk among diverse signaling pathways. However, current knowledge on mechanisms underlying sepsis is far from providing a complete picture of the syndrome, justifying additional efforts that might add to this scenario. Microarray-based expression profiling is a powerful approach for the investigation of complex clinical conditions such as sepsis: the analysis of gene transcription at the genome level potentially avoids results derived from biased assumptions. In this study we investigate whole-genome gene expression profiles of mononuclear cells from survivor and nonsurvivor septic patients.Blood samples were collected at the time of sepsis diagnosis and 7 days later, allowing us to evaluate the role of biological processes or genes possibly involved in patient recovery. Aiming to circumvent, at least partially, the heterogeneity of septic patients, we included only patients admitted with sepsis caused by community-acquired pneumonia. Global gene expression profiling allowed us to characterize early sepsis, as compared with healthy individuals.Our results corroborate literature reports on inflammation response in the early stages of sepsis but highlight great heterogeneity in gene expression during the onset of sepsis. Differences in oxidative stress seem to be associated with clinical outcome, since significant differences in the expression profile of related genes were observed between survivors and nonsurvivors. However, our results substantiate current knowledge supporting that sepsis syndrome development is indeed multifaceted. Although the initial infection of enrolled patients was pneumonia, with no sign of organ failure at the time of diagnosis, 7 days later gene expression profiles seemed to be characteristic for each patient, with no clear pattern of development. This result could be associated with the underlying health status of each one of them, with complications due to sepsis itself as well as with distinct timing for response to treatment.At this point we conclude that studies should focus on studying sepsis at multiple time points, aiming to capture common, although possibly separated in time when patients are considered, biological processes associated with recovery.


The Human Microbiome Project is using next-generation sequencing and metagenomics to characterize microbial communities that inhabit our gastrointestinal tract and other body sites , 2. BaseA recent study by Donohoe et al. now provides considerable mechanistic insight by demonstrating that a fundamental difference in energy metabolism between normal and cancerous colonocytes can explain the butyrate paradox . Normal This study also demonstrated that the role of butyrate in epigenetics is more complicated than previously appreciated. In addition to functioning as an HDAC inhibitor, which was already known, butyrate can also increase histone acetylation by increasing histone acetyltransferase (HAT) activity. When butyrate is metabolized oxidatively, it contributes to acetyl-CoA production not only in the mitochondria but also in the cytosol and nucleus where it serves as an essential co-factor and acetyl-group donor for HATs. This pathway is dependent on the enzyme enzyme ATP citrate lyase (ACL) , and RNAThe study described here primarily involved colorectal cancer cell lines, and it will be important to confirm and extend these findings in gnotobiotic mouse models of colorectal cancer. For example, is it possible to manipulate the microbiota (by colonizing germfree mice with butyrate-producing bacteria) and diet (high fiber) to increase colonic butyrate levels and decrease tumorigenesis? If so, then it might useful to re-evaluate human epidemiologic studies that have had conflicting results regarding fiber consumption and colorectal cancer incidence. For example, might there be an association between participants' microbiota and whether or not a high-fiber diet protects against colorectal cancer? This is an important issue because the incidence of colorectal cancer has increased as we have shifted away from traditional, high-fiber diets towards processed foods that contain less complex carbohydrates and more refined sugars. To counteract this trend, probiotic and prebiotic strategies could possibly be implemented to increase the levels of an endogenous HDAC inhibitor (butyrate) in its normal environment (the colon). This chemoprevention strategy is likely to have fewer adverse effects than the i.v. delivery of synthetic HDAC inhibitors as chemotherapeutic agents .


Nitric oxide (NO) is an important signaling molecule crucial for many physiological processes such as synaptic plasticity, vasomotricity, and inflammation. Neuronal nitric oxide synthase (nNOS) is the enzyme responsible for the synthesis of NO by neurons. In the juvenile and mature hippocampus and neocortex nNOS is primarily expressed by subpopulations of GABAergic interneurons. Over the past two decades, many advances have been achieved in the characterization of neocortical and hippocampal nNOS expressing neurons. In this review, we summarize past and present studies that have characterized the electrophysiological, morphological, molecular, and synaptic properties of these neurons. We also discuss recent studies that have shed light on the developmental origins and specification of GABAergic neurons with specific attention to neocortical and hippocampal nNOS expressing GABAergic neurons. Finally, we summarize the roles of NO and nNOS-expressing inhibitory neurons. Information processing within neocortical and hippocampal circuits relies upon complex interactions between glutamatergic excitatory projection neurons and GABAergic inhibitory neurons. Coordinated cell–cell communication amongst and between these two neuronal populations is essential to maintain a delicate balance between excitatory and inhibitory signaling within the brain and is subject to dynamic regulation by many neuromodulatory substances such as various neuropeptides and nitric oxide (NO)  reactivity and immunostaining with various antibodies against calcium binding proteins, neuropeptides and nNOS, several groups have shown that nNOS GABAergic neurons can be subdivided into several hippocampal and neocortical sub-populations that are summarized in Tables + interneurons may be subdivided based on intensity of nNOS immunoreactivity (see next section), no such distinction exists in the hippocampus. Furthermore, a recent study revealed that interneurons expressing nNOS comprise the most abundant interneuron subpopulation in the hippocampus, in contrast to neocortical observations where parvalbumin (PV) expressing interneurons are considered to be the most abundant interneuron subtypes  and the cornu ammonis (CA). The CA region is classically further divided into CA1–4. In this section of the review, we will mainly focus on results obtained in CA1 where interneuron diversity has been best characterized but will detail other areas when data are available. As in the neocortex, nNOS expressing neurons comprise primarily inhibitory GABAergic neurons although nNOS immunoreactivity is also found in CA1 pyramidal cells. In these glutamatergic excitatory cells staining intensity in mature brain is much weaker than in interneurons and nNOS is observed preferentially in dendritic spines  the neurogliaform cells (NGFC), (2) Ivy cells (IvC), (3) interneurons co-expressing the vasoactive intestinal peptide (VIP) and calretinin (CR), (4) interneurons expressing PV and (5) projection cells. This latter subtype of nNOS the neur+ (SOM+) and PV+ interneurons. NGFCs exhibit a multipolar dendritic network with a high degree of ramification close to the soma without any privileged orientation. The axonal arborization is extremely dense with extensive ramification within the local network and usually radiates beyond the spatial boundaries of the dendritic field  and its border with s. radiatum (sr) of CA1-3, as well as within s. moleculare of the DG  has been found to colocalize with nNOS in both NGFCs and IvCs   and/or the preoptic area (POA)   have revealed that Nkx5.1+ and CR+, but not VIP+, interneurons depend on neocortical activity for their correct migration and positioning  caused an almost complete loss of nNOS+ GABAergic neurons in the hippocampus except for few bipolar interneurons in s.p. reminiscent of the VIP+/CR+ interneurons revealed in the CGE reporter  that project in turn to the stratum lacunosum molecular  to the first postnatal day (P0), a period of intense neuronal migration, nNOS is strikingly expressed by distinct cells types. Indeed, cells migrating in the marginal zone displaying Cajal-Rezius like morphologies express nNOS  and subplate (layers V and VI) regions as small and undifferentiated neurons. Differences in intensity of nNOS immunoreactivity (later mentioned as type I and type II neurons) become evident as early as P7  activities exhibit a remarkable similarity. NADPH-d activity is denser in barrel hollows, regions that receive somatotopic sensory thalamic inputs, and is less active in barrel septa  receptors or other calcium permeable channels , the number of gap-junction coupled neurons decreased  are inhibited by neuromodulatory afferents driving arousal such as acetylcholine, noradrenaline, serotonin, and histamine and that they would be activated when arousal systems are depressed when sleep-promoting substances are released . Once activated nNOS neurons could synchronize EEG activity across neocortical regions through the release of NO, GABA or NPY. Interestingly it has been reported that nNOS knockout mice spend more time than controls in slow wave sleep as monitored by EEG. This suggests that nNOS-type I GABAergic neurons may regulate sleep homeostasis  their laminar position within different circuits; (2) specific combinations of neurochemical markers; (3) their basic morphology; and (4) their electrophysiological features including passive membrane properties, spiking behavior and synaptic connectivity. Various studies including some highlighted above have shown that these criteria are largely dictated by an interneuron's site and time of genesis. However, some studies have also pointed to a role for the cellular environment an interneuron ultimately occupies in refining these properties such as their stratification  and their expression of certain activity regulated markers like NPY. In this respect it should be mentioned that the expression of nNOS appears to be developmentally regulated in various neuronal populations and could be modulated by cellular targets in subpopulations of interneurons . Thus, although challenging, it will be important to determine whether nNOS interneurons are guided to their final location early on, like most interneurons, or are eliminated if inaccurately positioned or if they stop expressing/fail to induce nNOS. An understanding of subtle differences in the genetic makeup/molecular characteristics of divergent nNOS interneuron cohorts may provide insight into these issues. The recent generation and use of Cre reporter animals in association with other techniques have been successfully used to determine the embryonic origin and birthdating of nNOS type I and type II interneurons revealing for the first time their heterogeneity and specificities . The increasing array of transgenic models and genetic tools available  will help advance the pace of this research.The development and plasticity of nNOSInterestingly, the unique features that have been shown to depend on neuronal activity (Verhage 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.


Heightened sensitivity to painful stimulation commonly spreads from the affected limb to the ipsilateral forehead in patients with complex regional pain syndrome (CRPS). In addition, acoustic startle evokes greater auditory discomfort and increases in limb pain when presented on the affected than unaffected side. In contrast, limb pain ordinarily evokes analgesia in the ipsilateral forehead of healthy participants, and acoustic startle suppresses limb pain. Together, these findings suggest that hemilateral and generalized pain control mechanisms are disrupted in CRPS, and that multisensory integrative processes are compromised. Failure to inhibit nociceptive input from the CRPS-affected limb could sensitize spinal and supraspinal neurons that receive convergent nociceptive and auditory information from hemilateral body sites. Somatosensory, auditory, and emotional inputs may then aggravate pain by feeding into this sensitized nociceptive network. In particular, a disturbance in hemilateral pain processing that involves the locus coeruleus could exacerbate the symptoms of CRPS in some patients. Complex regional pain syndrome (CRPS) usually begins after a fracture, contusion or sprain, but can develop after a minor injury that would normally heal quickly. Patients typically describe a burning sensation that is aggravated by movement, the limb being touched, and ambient temperature changes. The pain and sensory disturbances are often so severe that they lead to profound disability   of rats  is common in severely-affected patients with CRPS (de Klaver et al., increased in the forehead of CRPS patients during intensely painful stimulation of the affected limb, suggesting that inhibitory pain modulation processes had failed (Knudsen et al., Limb pain usually suppresses less intense pain elsewhere in the body, including the forehead (Knudsen and Drummond, A shift toward pain facilitation in the rostral projections of the hemilateral pain modulation system might also explain the presence of hyperalgesia in the ipsilateral forehead of CRPS patients (Drummond and Finch, Some observations do not fit with the idea of a central adrenergic influence on pain in CRPS. For example, anecdotal reports suggest that tricyclic antidepressants help some patients (Rowbotham, Similar to most other chronic pain syndromes (Saariaho et al., 1-adrenoceptor blockade for CRPS have been described in case reports (Abram and Lightfoot, 1-adrenoceptors at spinal or supraspinal levels is an intriguing possibility that may have potential treatment implications for certain patients with CRPS or other chronic neuropathic pain syndromes.In terms of treatment, benefits of αThe 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.


This framework describes how each individual's attachment style  may influence the encoding of approach versus aversion tendencies (safety versus threat) in social encounters, implicating the activation of a network of subcortical  and cortical  limbic areas. These basic and automatic affective evaluation mechanisms are in turn modulated by more elaborate and voluntary cognitive control processes, subserving mental state attribution and emotion regulation capacities, implicating a distinct network in medial prefrontal cortex (mPFC), superior temporal sulcus (STS), and temporo-parietal junction (TPJ), among others. Recent neuroimaging data suggest that affective evaluation is decreased in avoidantly but increased in anxiously attached individuals. In turn, although data on cognitive control is still scarce, it points toward a possible enhancement of mental state representations associated with attachment insecurity and particularly anxiety. Emotion regulation strategies such as reappraisal or suppression of social emotions are also differentially modulated by attachment style. This research does not only help better understand the neural underpinnings of human social behavior, but also provides important insights on psychopathological conditions where attachment dysregulation is likely to play an important  role.Since its first description four decades ago, attachment theory (AT) has become one of the principal developmental psychological frameworks for describing the role of individual differences in the establishment and maintenance of social bonds between people. Yet, still little is known about the neurobiological underpinnings of attachment orientations and their well-established impact on a range of social and affective behaviors. In the present review, we summarize data from recent studies using cognitive and imaging approaches to characterize attachment styles and their effect on emotion and social cognition. We propose a functional neuroanatomical framework to integrate the key brain mechanisms involved in the perception and regulation of social emotional information, and their modulation by individual differences in terms of secure  In mammals, including humans, attachment is a major dimension of behavior that can come into play in several domains  Bowlby, , 1982. TAlthough very prominent in developmental psychology  suppression to regulate emotions  positive due to the desire for attention and protection. However, repeated experience of rejection leads to an increased sense of helplessness and vulnerability, paired with doubts about self-worth and -efficacy, leading to a negative internal model of the self and poor self-esteem. Such individuals become highly vigilant to potential threats and rejections. This style is also thought to imply a distinctive emotion regulation strategy, with preferential use of re-appraisal but in the “wrong” direction: instead of decreasing the impact of negative emotions, these subjects actually tend to intensify the impact of negative social signals due to their hypersensitivity to the latter  level and at a more voluntary or explicit (conscious) levels of processing. However, the exact neural mechanisms involved in these effects remain largely unexplored, although general models of social cognition and emotion processing  were observed to show stronger activation, or display increased gray matter volume, in the reward network as well as other interconnected regions such as the hypothalamus or OFC. For instance, the ventral striatum differentially activates in secure mothers seeing images of their own babies with a smiling or a crying facial expression  was inversely correlated with increasing scores on the avoidant attachment dimension  with a distinct social aversion network —schematic drawings of attachment-related scenes depicting either one or two persons—which distinguishes between a resolved In sum, the available data from recent neuroimaging research points to a higher sensitivity to negative social clues, and enhanced recruitment of social aversion or threat systems in relation to anxious attachment, in agreement with previous work suggesting higher vigilance to social emotional cues and hyper-activating of secondary attachment-related strategies in these individuals  information about others , which are also closely related to neurocognitive mechanisms implicated in “emotional contagion”  a wide network of areas in the lateral prefrontal (PFC), OFC, and posterior cingulate cortex (PCC), as well as the precuneus, superior temporal sulcus (STS), and temporo-parietal junction (TPJ), plus specialized sensory regions in superior temporal gyrus, lateral occipital cortex, and fusiform cortex , in relation to others or oneself. However, AT suggests that individual attachment styles also imply distinct modes of emotion regulation strategies , STS, and fusiform gyrus, but they also showed increased BOLD signal change in these areas when hearing baby cries. These results suggest that mothers with secure attachment traits  might more readily engage in complex social behaviors involving mentalization and theory of mind when interacting with children, possibly implying more efficient cognitive processing to represent their mental states in terms of intentions or needs. In turn, this could potentially have beneficial effects on the emerging attachment styles of the child him/herself (see below). Conversely, low scores on the perceived maternal scale reflecting insecure attachment, were associated with increased hippocampus responses to infant cries in the same mothers. As the hippocampus is known to play an important role in stress responses . This somewhat contradicts the hypothesis put forward by the developmental mentalization-based approach of Fonagy and Luyten , or cognitive re-appraisal strategies . Hence, these emotion regulation strategies are generally referred to as distraction, detachment, or reappraisal. By contrast, response-focused emotion regulation involves the (voluntary) inhibition or transformation of the emotional response or behavior after an emotion has ben generated . This strategy is therefore generally referred to as (expressive) suppression. In terms of brain activity patterns, the employment of both emotion regulation strategies has been linked with increased PFC cortical activation related to executive control and/or behavioral inhibition , and a simultaneous modulation  of responses evoked in the emotional evaluation system  during both the “think” and the “don't think” conditions, which was interpreted as a failure of “task-induced deactivation” . This pattern may reflect heightened cognitive and emotional conflict in combination with increased regulatory inhibition during spontaneous viewing of social emotional scenes. Furthermore, during REAP, amygdala activation to negative social images decreased for low, but not high avoidantly attached participants, implying that this emotion regulation strategy was not efficient in reducing affective mentalizing  social behavior have begun to explore the molecular and genetic mechanisms at play in social affective processing, learning, and bonding  receptor-gene polymorphisms, which even correlated with individual measures of emotional reactivity and prosocial temperament in one study  attachment. Such personality traits will produce important and long-lasting influences on social emotional information processing and regulation, associated with differential recruitment of specific functional brain networks for understanding and responding to others in close (or at times less close) relationships.versus threat, or approach versus aversion tendencies in social contexts; and on the other hand, a system for controlled social processing and regulation , operating in a more conscious, voluntary mode, which is involved in representing the mental states of others (theory of mind) and regulating one's own behavior, thoughts, and emotions. These two functional components rely on distinct brain networks , which is primarily involved in encoding basic dimensions of safety  Porges, , essentiavoidant attachment style is characterized by blunted responses in both subparts of the emotional mentalizing systems , reflecting a deactivation of attachment needs. Findings regarding mental state representation specifically related to attachment avoidance are still preliminary and need further confirmation. Yet, avoidant attachment style also appears to imply a distinctive pattern of emotion regulation strategies, with greater reliance on suppression but difficulties in using reappraisal (antecedent-focused regulation) to dampen affective appraisals.According to this model, an anxious attachment style is associated with enhanced responses to social emotional information signaling threat and motivating aversion, mirroring the hyperactivation of attachment needs and subjective lack of control observed behaviorally. This might also be combined with enhanced recruitment of mental state attribution systems in some situations related to fear (trauma in BPD patients). However, emotion regulation appears relatively operational when explicitly required by task instructions, although mingled with greater recruitment of associative memory systems that may promote access to memories of previous attachment contexts.In contrast, an This research highlights the fact that social interactions and emotions therein are susceptible to strong modulations by individual differences, reflecting (among others) the key role of the attachment history of a person as well as possible neurobiological predisposition factors. Such consideration of individual attachment style and history in recent neuroimaging studies appears critical to extend social and affective neuroscience research to a comprehensive and valid framework of socially motivated behaviors, although there still is a lack of experimental investigations of these effects in more complex and “true” social interactions. Future studies should therefore aim at better assessing attachment effects on brain responses during “real” social encounters, or at least in laboratory context resembling the latter as closely as possible, as for example by employing functional near infrared spectroscopy (fNIRS) in two participants at the same time, also referred as to hyperscanning (Cui et al., In addition, despite the fact that most attachment effects described in this review concern healthy non-clinical populations, they also have implications for promoting well-being and reducing social stress, and may in addition provide useful clues regarding attachment system dysregulations in patients with psychopathologies or abnormal social behaviors (Galynker 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.


The cancellation of redundant information is a fundamental feature in many sensory systems. It constitutes, for instance, a key ingredient for the so called "cocktail party problem", in which a relevant signal has to be discerned and separated from other uninteresting stimuli in the auditory system . HoweverOf special interest would be the cancellation mechanisms that are able to perform well under different realistic conditions, such as canceling redundant signals of different intensities. Properly addressing such a contrast-invariant cancellation mechanism could be beneficial to understand sensory processing in the visual system as well, since it is well known that contrast invariance is a prominent feature in neural circuits of primary visual areas .in vivo recordings and computational modeling to study a cerebellar-like circuit in the weakly electric fish which is known to perform cancellation of redundant stimuli. The cancellation is observed in a sub-population of neurons called superficial pyramidal (SP) neurons, which receive input both from electroreceptors encoding sensory input (constituting the feedforward pathway) and from a large population of granule cells  which are ultimately driven by electroreceptors as well (constituting the feedback pathway) [in vivo extracellular recordings, we observe contrast invariance in the cancellation of redundant stimuli in such a system. Our computational model, which improves previous modeling efforts [in vivo observations and provide an explanation for such a contrast-invariant cancellation.In this work, we combine pathway) ,4. Emplo efforts ,6, incorin vivo to date, and (II) provides a strong prediction on the time scale in which potentiation mechanisms in parallel fibers may occur. Our model also predicts that, in order to properly cancel redundant signals at the experimentally observed levels, the average contrast level that must drive the learning lies around contrast levels of 15%. Interestingly, contrast levels around this one are commonly found within the natural environment for communication signals in the weakly electric fish.In addition, the model: (I) gives insight on the activity of granule cells and parallel fibers involved in the feedback pathway, whose activity has not been recorded


Endometrial cancer is the most common gynecologic malignancy in the developed world. Most cases are diagnosed at an early stage and have low-grade histology, portending an overall excellent prognosis. There exists a subgroup of patients with early, high-risk disease, whose management remains controversial, as current data is clouded by inclusion of early stage tumors with different high-risk features for recurrence, unstandardized protocols for surgical staging, and an evolving staging system by which we are grouping these patients. Here, we present preoperative and intraoperative considerations that should be taken into account when planning surgical management for this population of patients. Endometrial cancer is the most common gynecologic malignancy in the developed world. In 2013, it is estimated that over 49,000 new cases will be diagnosed and 4,000 deaths will be reported in the United States [Management of patients with high to intermediate risk endometrial cancer is controversial. This is not surprising as the published literature that we may use to inform intraoperative and adjuvant treatment decisions is difficult to interpret. Studies sometimes include patients that have undergone comprehensive surgical staging that are incompletely staged or that have clinically stage I disease (no lymphadenectomy). Further complicating matters, the International Federation of Gynecology and Obstetrics (FIGO) staging system has changed, thereby muddying the interpretation of results of previous published studies. Finally, the wide-spread adoption of laparoscopy and robotics in the management of this disease has modified the short-term morbidity associated with surgical management. There is a paucity of data, however, on long-term cancer-related outcomes in these patients with early stage, high-intermediate risk disease.It is time to reassess what we know, or think we know, about endometrial cancer. We plan to critically re-review the pertinent data to help address the following questions in this high to intermediate risk cohort: to stage or not to stage? How do we triage whom to stage? Which surgical approach is most appropriate—open, laparoscopic, or robotic? How can we use intraoperative assessments to guide surgical decision making? What do we know about the morbidity of lymphadenectomy? What should we do with unstaged, high/intermediate risk patients? How does this differ if they are comprehensively staged? What is the role of adjuvant radiation and chemotherapy?Endometrial cancer was clinically staged until 1988, when a surgical staging system was adopted by the International Federation of Gynecology and Obstetrics. Endometrial cancer staging includes a total hysterectomy, bilateral salpingo-oophorectomy (BSO), and systematic pelvic and para-aortic lymphadenectomy. The 1988 FIGO staging system is shown in The change in the staging of endometrial cancer is paramount not only in guiding treatment decisions but also impacts how we interpret the literature. Studies published before 2009 include patients that were stratified differently than our current practice. Recognizing this, we need to be aware that patients in these studies may be at a significantly lower risk of recurrence, such as the “old IA” patient compared to “today's IA” patient that may have up to 50% myometrial invasion. In reality, depth of invasion is not a discrete variable, as the staging system artificially creates it, but instead is a continuous variable with risk increasing with each additional millimeter of invasion. We should also keep in mind that, prior to 2009, patients with stage II disease included the cohort of relatively low-risk patients with cervical glandular involvement.The best approach to identifying the high-intermediate risk women pre-or intraoperatively has not been determined and often is surgeon dependent. An argument can be made to perform surgical staging on all women with endometrial cancer regardless of the grade so as to identify all women with advanced disease. Others recommend staging only a selected group of patients to avoid morbidity, citing the apparent lack of survival benefit from comprehensive surgical staging as a rationale for this approach. The decision of who to surgically stage, however, is probably best determined deliberately. Understanding several key factors is critical to making an informed decision regarding the proper surgical procedure: assessing grade, depth of myometrial invasion, tumor size, and lymph node evaluation.It has long been recognized that the tumor grade is an important determinant of lymph node metastases. There are many ways in which tumor grade can be assessed preoperatively, including office endometrial biopsy and dilation and curettage (D&C). The accuracy of each of these methods differs. A retrospective study by Larson et al. examined the use of office pipelle sampling compared with D&C to determine the histologic grade in patients with known endometrial cancer . PipelleAssessing the depth of myometrial invasion is another important aspect of determining the correct surgical procedure for patients with endometrial cancer. Much like grade, depth of invasion can be determined preoperatively, using ultrasound or MRI, but also intraoperatively, with gross visual inspection or frozen section analysis. The diagnostic accuracy for correctly assessing depth of invasion with MRI ranges from 68% to 85% , 11 and Preoperative assessment of lymph node involvement using various radiologic imaging modalities and intraoperative assessment of lymph node involvement by palpation have been proposed by some as useful techniques to assess for metastasis. Unfortunately, for patients undergoing surgery via robotic or laparoscopic approaches, retroperitoneal palpation is impossible. Moreover, the data does not support this approach with regard to diagnostic accuracy . Even whCurrent techniques available for preoperative and intraoperative assessment of tumor characteristics for the purposes of guiding surgical decision making have been shown to have a high degree of accuracy. If preoperative sampling is performed via pipelle endometrial biopsy rather than D&C, there is a higher chance that the tumor will be upgraded on final pathology. Frozen section analysis for grade determination should be strongly considered if this is the case. Gross visual inspection for depth of invasion is fairly reliable. Unfortunately, there are no current methods to determine the presence or absence of lymphovascular space involvement intraoperatively. It should also be recognized that the accuracy of gross inspection falls with increasing the grade of tumor; hence the threshold to perform systemic lymphadenectomy should be lower in this setting.The role of lymphadenectomy in the surgical management of women with early stage endometrial cancer has not been standardized and has led to widely differing practice patterns in the United States. There are still 2 major questions remaining with regards to lymphadenectomy in patients with endometrial cancer (1) Which patients with clinical early stage disease should undergo lymphadenectomy? (2) How extensive of a lymph node dissection should be performed? Two large trials were conducted to help answer these questions.n = 704) or standard surgery plus bilateral pelvic lymphadenectomy (n = 704). The decision to perform para-aortic lymphadenectomy was left to the discretion of the surgeon. The primary outcome of this study was overall survival. The median time of followup was 37 months. Although patients in this study were randomized, baseline characteristics differed significantly between the 2 groups, with more aggressive histologic subtypes and tumors with greater depths of invasion found in the lymphadenectomy arm. These differences were corrected statistically. The hazard ratio for overall survival was 1.04  and for recurrence free survival was 1.25 . Study authors concluded that there were no overall or recurrence-free survival benefits for performing lymphadenectomy and that pelvic lymphadenectomy could not be routinely recommended. Several limitations, however, have been identified with this study, including inadequate pelvic lymph node sampling (median number of resected lymph nodes was 9), inclusion of a large number of patients with low-risk features that may not have ever benefitted from lymphadenectomy, and leaving the decision to perform para-aortic node dissection up to the physician's preference.The Adjuvant External Beam Radiotherapy in the Treatment of Endometrial Cancer (ASTEC) Trial was a multicenter prospective randomized trial conducted from 1998 to 2005 in Europe . Fourteen = 264) or no lymphadenectomy. Similar to the ASTEC trial, performing para-aortic lymph node dissection was left to the discretion of the surgeon. Median followup was 49 months. Disease free survival did not differ significantly between the lymphadenectomy group and the no lymphadenectomy group (81% versus 81.7%). Overall survival also did not differ significantly (85.9% versus 90%). A major criticism of this trial is that the indications for and the extent of lymphadenectomy were not clearly defined.The CONSORT Trial was another Phase III randomized trial that was conducted in Italy to help discern the clinical impact of lymphadenectomy . This stDespite these 2 large prospective trials having been conducted, we still do not have a clear answer about the benefit of lymphadenectomy in early stage patients. Furthermore, it also remains unclear how aggressive of a lymphadenectomy should be performed. Researchers at Mayo designed a prospective trial to help assess lymphatic dissemination patterns . This stAfter critically reviewing these 2 trials, there is still no clear evidence to promote complete surgical staging in women with low-risk stage I endometrial cancer. However, there were not enough early stage high-intermediate risk patients included in these trials to be able to draw separate conclusions about this subset of patients. Complete surgical staging of women with high-intermediate risk features should still be strongly considered.Historically, the surgical approach for the management of all stages of endometrial cancer has been via laparotomy. However, with the increasing incorporation of minimally invasive surgical techniques into gynecology, the use of a laparoscopic approach (both standard laparoscopic and robotic platforms) for treatment of early stage endometrial cancer has become more commonplace. The already established general benefits of minimally invasive surgery, such as decreased blood loss, shorter recovery time, and fewer postoperative complications, become especially attractive when dealing with a population of endometrial cancer patients, who are typically obese with multiple medical comorbidities.A randomized, phase III trial conducted by the Gynecologic Oncology Group (LAP2) was designed with the objective of comparing laparoscopy versus laparotomy for the comprehensive surgical staging of women with clinical stage I to IIA uterine cancers . This trP < 0.0001), fewer perioperative complications , longer operative times , and similar rates of intraoperative complications . Twenty-six percent of patients in the laparoscopy arm, however, required conversion to an open approach. Risk factors for conversion with the strongest correlation included increased body-mass index, older age, and presence of metastatic disease. Removal of pelvic and para-aortic lymph nodes differed significantly between the 2 groups—96% of laparotomy patients and 92% of laparoscopy patients underwent lymph node dissection (P < 0.0001). An equal number of patients in both groups (17%), however, were found to have metastatic disease. These results allowed us to conclude that the laparoscopic approach to staging was safe and did not compromise the ability to perform an adequate staging procedure.Preliminary study outcomes from LAP2 were published in 2009 [Survival data from LAP2 were published in 2012 . During There have been a number of other studies that have also addressed the issue of incorporating minimally invasive approaches into the surgical management of early stage endometrial cancer. A multi-institutional, randomized trial was conducted in the Netherlands from 2007 to 2009, which compared total abdominal hysterectomy versus total laparoscopic hysterectomy for the management of patients with stage I endometrial cancer or complex atypical hyperplasia . The priRobotic surgery has now also become a feasible option for use in the surgical management of early stage endometrial cancers. Multiple retrospective cohort studies have looked at the outcomes and costs of performing staging procedures for endometrial cancer with robotic assistance versus standard laparoscopy and laparotomy –27. AdvaMany factors must be considered when determining the ideal surgical approach for the optimal outcome and the patient benefit. There are advantages and disadvantages to all possible surgical approaches, which makes the decision regarding which approach to pursue a challenging one. At present there will likely always be a place for both laparotomy and laparoscopy. Given the benefits of robotics it would seem unwise for it not to have a place in the armamentarium against early stage endometrial cancer. As we continue to make advances in surgical techniques and technologies, new approaches to surgical management may be implemented and adopted. Continued studies, especially those that include patients with high-intermediate risk features, are necessary to determine the best approach in terms of outcome and cost.Management of women with high-intermediate risk, early stage endometrial cancer remains controversial, and practice patterns vary widely among gynecologic oncologists. While a large amount of time and resources have been invested into the study of women with early stage endometrial cancer, differences in outcomes are difficult to establish since these patients tend to have an overall excellent prognosis. Current data is clouded by inclusion of early stage tumors with different high-risk features for recurrence, unstandardized protocols for surgical staging and an evolving staging system by which we are grouping these patients. Given that a high-intermediate risk population of patients with early stage exists which could benefit from extended surgical staging and adjuvant treatments, we need to invest more into characterizing recurrence patterns and treatment effects in order to adequately treat these patients.POLE ultramutated, (2) microsatelite instability hypermutated, (3) copy-number low, and (4) copy-number high. It has been shown that high grade endometrioid and high grade serous carcinomas can at times be difficult to subtype correctly, with not insignificant rates of intraobserver concordance among pathologists [Preoperative and intraoperative classifications of tumor characteristics through routine and frozen histopathologic assessments can provide us only with information regarding histology, grade, size, and depth of invasion. This information, with the addition of presence or absence of lymphovascular space involvement, can be confirmed by final histopathologic analysis. However, there has now been increased interest in classifying tumor biology based on data obtained from integrated genomic and proteomic analyses of endometrial cancers . Using tologists , 31.Correct classification is of paramount importance, as often Type I endometrial cancers are treated with adjuvant radiation and Type II endometrial cancers are treated with adjuvant chemotherapy-based treatments. As an example, approximately 1/4 of tumors that were classified as high-grade endometrioid on pathologic review actually had similar phenotypes as uterine serous carcinomas when genomic-based analyses were performed. Incorporation of genomic analyses for the purposes of more specific tumor classification into clinical practice may provide us with a way to more appropriately tailor post-operative adjuvant treatments.


Viral encephalomyelitis is an important cause of morbidity and mortality worldwide, and many encephalitic viruses are emerging and re-emerging due to changes in virulence, spread to new geographic regions, and adaptation to new hosts and vectors The primary target cells for most encephalitic viruses are neurons, although a few viruses attack cerebrovascular endothelial cells to cause ischemia and stroke or glial cells to cause demyelination, encephalopathy, or dementia Because infections with encephalitic viruses are initiated outside the CNS , innate and adaptive immune responses are usually mounted rapidly enough to prevent virus entry into the CNS. Therefore, most viruses that can cause encephalitis more often cause asymptomatic infection or a febrile illness without neurologic disease, and encephalomyelitis is an uncommon complication of infection.When a virus does invade the CNS, there are several routes by which infection of neurons can occur. The most common entry point is from the blood, and the level of viremia as a result of virus replication in peripheral organs often correlates with the likelihood of CNS infection. However, the blood–brain barrier (BBB), composed of vascular endothelial cells with tight junctions in contact with the foot processes of astrocytes, inhibits direct access to the brain parenchyma and neurons. Some neurotropic viruses can replicate in cerebrovascular endothelial cells, enter with infected leukocytes, or cross directly into the cerebrospinal fluid (CSF) through the porous capillaries of the choroid plexus. A specialized CNS entry pathway used by several viruses, most notably HSV, varicella zoster, and rabies viruses, is by way of nerve terminals in peripheral organs. These viruses can enter the nerve and then use neural transport mechanisms to transport the infecting virions to the neuronal cell body where replication occurs In tissue culture systems most of these viruses can infect many types of cells, in addition to neurons. Few neuron-specific virus receptors have been identified  but these do not always account for neuronotropism in vivo In addition to fever and headache, signs and symptoms of viral encephalomyelitis typically include evidence of neuronal dysfunction—seizures, cognitive impairment, ataxia, paralysis, etc. Virus replication can damage neurons directly by inducing cell death through apoptotic or necrotic mechanisms  [14]. MaAlthough neuronal virus infection per se is necessary for, and contributes directly to, neuronal dysfunction, the inflammatory response in the CNS is also a major contributor to neuronal damage and can even result in death of nearby uninfected neurons  [19], [2+ T cells) or of MHC class II (CD4+ T cells) and can possess cytotoxic properties. Neurons are long-lived essential cells that cannot be replaced, so a noncytolytic immune mechanism for virus clearance would be advantageous to the host to avoid death of surviving cells. If the immune clearance mechanism is damaging to the infected neuron, then the function of that neuron will be lost and the outcome for the host will be the same as if the virus infection had caused neuronal death.The immune response to infection can contribute to fatal disease, but is also necessary for recovery and virus clearance. Elimination of virus-infected cells from tissue requires elimination of all cells in which the virus is replicating. This can occur either by virus-induced or immune-mediated cytolysis. T cells are ideal for this purpose because they recognize viral antigen as processed antigen only in the context of cell surface–expressed MHC class I (CD8Because mature neurons are relatively resistant to virus-induced cell death Because the noncytolytic process for virus clearance does not completely eliminate viral RNA from neurons, a mechanism for long-term immunologic control of virus replication is needed to prevent virus reactivation or progressive disease Encephalomyelitis resulting from virus infection of neurons is a disease that can be fatal or result in permanent disability due to irreversible damage of infected neurons. The immune response to infection can enhance neuronal damage or can control virus replication by noncytolytic mechanisms and thus determine outcome. However, noncytolytic virus clearance results in persistence of viral nucleic acid in the CNS and thus establishes a need for long-term local immune responses to prevent reactivation of infection and progressive disease. Understanding these mechanisms is necessary for development of strategies for treating and preventing neurologic disease due to viral encephalomyelitis.


The concept of open resource metagenomics includes preparation of physical DNA libraries, preferably in versatile vectors that facilitate screening in a diversity of host organisms, and pooling of clones so that single aliquots containing complete libraries can be easily distributed upon request. Database deposition of associated metadata and sequence data for each library provides researchers with information to select the most appropriate libraries for further research projects. As a starting point, we have established the Canadian MetaMicroBiome Library . The CM Escherichia coli) and stored as a collection of clones.Microbial communities harbor the immense genetic diversity that controls Earth’s biogeochemical cycling. This genetic diversity has a concomitantly immense potential for applications in bio-product synthesis, green chemistry and pharmaceutical and bio-energy sectors. Metagenomic libraries provide a window into this largely untapped reservoir of nucleic acid diversity. Individual metagenomic libraries have been generated from a variety of terrestrial and aquatic environments, and these have been prepared either as sequence-based libraries for analysis and submission to public databases, or as DNA libraries captured in a host organism  has recognized the drawbacks of the lack of sample metadata submission, or the submission of metadata structured at the discretion of individual researchers ,3; the rWhile the scientific community recognizes the importance of appropriate metadata collection for genomics and metagenomics research, the accompanying physical metagenomic libraries are not commonly generated and maintained as shared resources. Since the publication of the first metagenomic libraries from marine water samples ,6, nucleE. coli as the host. Another example of multiple applications for metagenomic libraries includes cosmid libraries generated from activated sludge and soils. These originally underwent selection for clones conferring the ability to utilize D-3-hydroxybutyrate as sole carbon source in both E. coli and Ensifer meliloti (Sinorhizobium meliloti) surrogate hosts [E. meliloti or Agrobacterium tumefaciens as a surrogate host. These examples illustrate the sustained value of metagenomic libraries for use in a range of hosts for uncovering a wide variety of different genes, enzymes and functions related to small-molecule metabolites in the environment.Given that the success of a phenotypic screen or selection hinges on the screening strategy, expression host and the function being sought, the libraries are able to yield nearly limitless value for additional combinations and comparisons in later studies. Several examples already exist of individual laboratories mining the same metagenomic libraries for myriad functions across multiple studies. One of the earliest metagenomic studies involved capturing soil DNA in large-insert BAC libraries, followed by screens to identify clones that coded for activities including DNase, antibacterial, lipase, amylase, cellulase, chitinase, esterase, keratinase and protease . The samte hosts . Subsequte hosts , phosphate hosts , and polte hosts , all in International Journal of Systematic and Evolutionary Microbiology, “characterization of prokaryote strains must include all relevant metadata ” [Molecular Microbiology journal stipulates that “authors will distribute freely any strains, clones or antibodies described therein for use in academic research”, which is similar to the recommendation for authors from the Journal of Bacteriology that mutant strains be “available from a national collection or will be made available in a timely fashion, at reasonable cost, and in limited quantities to members of the scientific community for noncommercial purposes”.Unlike the examples above, most metagenomic libraries are prepared for individual projects and are not necessarily maintained in a manner that facilitates convenient and low-cost distribution. This situation hinders both repetition of the work done previously and exploration of the captured DNA for additional enzymatic functions. These libraries, similar to cultured isolates or mutant strains, could have value extending far beyond the initial publication. According to the iables)” . For mutA tradition of sharing has been integral to the development of the science of microbiology. From the earliest days, sharing of pure cultures has been essential to experimental replication and validation, and for the definition of bacterial types. The validity of patent claims requires access to cultures, and such access is legally mandated. Central culture collections such as the American Type Culture Collection (ATCC), Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) and hundreds of others around the world  have ensOne reason for the lack of storage and distribution standards for physical metagenomic libraries may be that the customary formats for storage are 96- or 384-well plates. Multi-well plates require extensive storage space. For example, a single 100,000-clone metagenomic library from a soil sample would be represented by approximately ~1000 96-well plates or ~250 384-well plates, which could fill an entire ultrafreezer with only 96-well plates. Due to the obvious challenges of permanent storage of large numbers of plate-arrayed metagenomic libraries, we argue that libraries should also be preserved in a format that is amenable to low-cost shipping and storage so these valuable resources remain available for long-term distribution subsequent to publication.We propose an open resource model for archiving published libraries using an alternative clone pooling strategy for storage and distribution. The features of this model include (a) cloning of large-size inserts into versatile vectors for downstream screening or selection in a variety of hosts, (b) pooling of clones for facilitating convenient library distribution in standard microcentrifuge tubes, and (c) extensive metadata and associated sequence-based characterization of libraries.E. coli). Indeed, screening in multiple hosts can reveal target clones that would not have been evident otherwise [en masse transfer of cloned genomic DNA from the library vector backbone to host-specific destination vectors, resulting in expression clones appropriate for the desired surrogate host.Cloning of inserts into broad host-range vectors helps circumvent one of the main limitations of metagenomic library functional screens: the inability to express many genes in a single host  could be approached to offer permanent storage for these genetic resources. Ultimately, the scientific value of environmental metagenomic libraries is analogous to the agricultural value of seeds from Earth’s plants, which are protected within a distributed system of Seedbanks.Altogether, these approaches are affordable and would result in libraries being made readily available to all researchers, and allow future grant proposals to leverage existing libraries from international research colleagues. Each functional study using the library could be related to the sequence data, as well as other functional studies using the same library, and would ensure that the original creators of the library are appropriately cited. Open resource metagenomics helps circumvent the traditional cycle of requiring new samples to be collected for each subsequent study. With such an approach, metagenomic libraries could be considered akin to isolates and mutant strains that are housed in culture collections. Unlike established culture collections, our recommendation would be that open resource metagenomics be established conceptually as a federated system in which individual laboratories maintain control and maintenance of pooled metagenomic libraries, ready for distribution upon request. As the Of course, as with isolates and mutant strains, there remain outstanding challenges that could prevent universal adoption of open resource metagenomics. For example, many funding agencies require and encourage the involvement of for-profit industry, which typically includes agreements with regards to intellectual property. Under these circumstances, the reality may be that unique arrangements will be required to blend open resource metagenomics with industrial collaborations. These arrangements may include delayed library release or the maintenance of particular libraries outside of the research literature. In addition, some laboratories may require a specific security level for labs handling and receiving metagenomic material and others may require signed agreements for transfers of research materials. Of course another important barrier to a central repository would be financial implications. As with culture collections, sustained funding is essential to maintain resources for sample submission, cataloguing and distribution. Although we are seeking appropriate funding towards establishing such a centralized system, the methodological approaches outlined above will enable the immediate adoption of a distributed open resource system for researchers preparing new metagenomic libraries. These are issues that we are working to resolve and we urge the research community to join this effort in moving toward a system of open resource metagenomics for greatest international collaborative benefit.2BL [2BL was initiated with the construction of cosmid libraries cloning DNA isolated from soil samples collected from across Canada spanning multiple biomes and ecozones , a publecozones . These s2BL libraries are prepared using derivatives of the IncP cosmid pRK7813 [in vivo [in vitro [E. coli and as extracted cosmid clone DNA. For each sample prepared thus far, libraries contain pools of between 10,000 and 250,000 cosmids. Copies of these libraries are stored as permanent frozen archives and purified cosmid DNA, in duplicate freezers with CO2 backup systems to ensure security of the resource. If required, additional library expansions can be generated for distribution to companies or academic institutions at minimal cost per library. Distribution of libraries is accompanied by materials transfer agreements to ensure consistency with requirements of the UN Convention on Biodiversity [The CM pRK7813  that hav[in vivo  or in viin vitro  reaction2BL resource was initiated by preparing a cornfield soil metagenomic library using the versatile IncP cosmid vector pJC8 . A total of 79,058 clones with ~33 kb random inserts have been generated. The resulting soil DNA library was calculated to contain 2,640 Mb of metagenomic DNA, which represents approximately 561 genomes, assuming average bacterial genome size of 4.7 Mb in the soil sample [The CMl sample .2BL to include samples from additional environments and have partnered with international research initiatives such as the Earth Microbiome Project to provide sequence-based context for libraries intended for functional screens and selections. Ultimately, open resource metagenomics, and initiatives such as CM2BL, will provide the international scientific community with cost-recovery access to a collection of thoroughly characterized genetic materials for the phenotypic screening of enzymes and other gene products with truly novel functions independent of sequence-based surveys, available for distribution upon request.Considering that methods for preparing metagenomic libraries have largely remained constant since their conception in the 1990s, these proposed standards for metadata, storage and distribution of metagenomic libraries are anticipated to remain relevant for decades. We are expanding CM