We used SILAC (stable isotope labelling of amino acids in cell culture) and high-throughput quantitative MS mass spectrometry to analyse the protein composition of highly purified WT wild type adenoviruses, mutant adenoviruses lacking an internal protein component (protein V) and recombinant adenoviruses of the type commonly used in gene therapy, including one virus that had been used in a clinical trial. We found that the viral protein abundance and composition were consistent across all types of virus examined except for the virus lacking protein V, which also had reduced amounts of another viral core protein, protein VII. In all the samples analysed we found no evidence of consistent packaging or contamination with cellular proteins. We believe this technique is a powerful method to analyse the protein composition of this important gene therapy vector and genetically engineered or synthetic virus-like particles. The raw data have been deposited at proteomexchange, identifer PXD001120. Adenoviruses are non-enveloped, icosahedral and enclose a dsDNA genome of ~36 000 bp . The majProteomic analysis of adenovirus particles by tandem MS (MS/MS) has identified peptides for most of the adenovirus structural proteins along with some phosphorylation sites  composition and even biosynthetic nanoparticle composition. For example, adenoviruses have been modified structurally to make therapeutic virus or useful VLPs to deliver therapeutic agents , as described previously . Some adenovirus structural proteins are made as precursor proteins that are proteolytically matured by the virus-coded 23K proteinase. Thus, protein entries corresponding to the mature, processed viral proteins need to be included in the protein database.We compared batches prepared at the University of Bristol and at Washington University in St Louis, and one sample of recombinant adenovirus Ag85A used recently in a clinical trial (We detected a mean of 556 peptides in each mixture of viruses  and idenWe compared our data on the mature particles with that of previous studies (Turning to the analysis of immature particles, they show greatly reduced amounts of proteins V, VII and TP compared with mature particles . This isAs expected, protein V was present in similar amounts in mature WT Ad5 and both recombinant adenoviruses. Moreover, proteins pre-VII and mature VII were present at a notably reduced amount in mature dV/TSB as compared with the mature WT Ad5 and both recombinant adenoviruses . This apWe found no evidence that WT Ad5, the recombinant adenoviruses and dV/TSB have cellular proteins consistently associated with or packaged into virus particles. As we reliably detect and quantify viral proteins with a copy number of just two, our data make it less likely that there are cellular proteins deliberately packaged or associated with the virus particles. What is important to bear in mind, however, is that other cellular proteins or post-translationally modified proteins may not be detected by our analysis. In addition, it is also possible that specific combinations of cell line and preparation method may allow consistent association of a protein with the virus in that context only.We have analysed the composition of WT, recombinant and dV/TSB particles using SILAC-based quantitative proteomics. To the best of our knowledge, this kind of analysis has only been used before to examine pseudorabies (


Many defining human characteristics including theory of mind, culture and language relate to our sociality, and facilitate the formation and maintenance of cooperative relationships. Therefore, deciphering the context in which our sociality evolved is invaluable in understanding what makes us unique as a species. Much work has emphasised group-level competition, such as warfare, in moulding human cooperation and sociality. However, competition and cooperation also occur within groups; and inter-individual differences in sociality have reported fitness implications in numerous non-human taxa. Here we investigate whether differential access to cooperation  is likely to lead to variation in fitness at the individual level among BaYaka hunter-gatherers. Using economic gift games we find that relational wealth: a) displays individual-level variation; b) provides advantages in buffering food risk, and is positively associated with body mass index (BMI) and female fertility; c) is partially heritable. These results highlight that individual-level processes may have been fundamental in the extension of human cooperation beyond small units of related individuals, and in shaping our sociality. Additionally, the findings offer insight in to trends related to human sociality found from research in other fields such as psychology and epidemiology. Many unique aspects of human sociality such as language, theory of mind and cultural norms have been proposed to provide the framework for human cooperative behaviour1236Inter-individual differences in sociality have been reported to have fitness implications in numerous taxa. For instance, in various non-human primates, greater social integration or social capital has been associated with increased longevity, offspring survival or mating access89Substantial research into the evolution of human sociality and cooperation has focused on group-level explanations. Such explanations emphasise the importance of cooperation between unrelated individuals in large scale warfare and resource competition between groupsWe attempt to identify the role of inter-individual variation in relational wealth in the dynamics of within-group competition among BaYaka hunter-gatherers. The BaYaka are simple and mobile hunter-gatherers - they consume food they forage soon after acquisition and lack storage mechanisms, and are also politically egalitarian; such societies are the best extant approximation of the ecological conditions under which our species evolved. Therefore, although the BaYaka are an extant population of cognitively modern humans, their forager lifestyle offers a valuable opportunity for inference regarding human evolutionary history.et al. hypothesise that since activities such as food sharing are often not uniform in hunter-gatherer groups, markets for cooperative partners emerge and social capital is likely to become relevant for consumption patterns and fitnessWhereas in other subsistence modes food storage is an option, simple hunter-gatherers rely profoundly on food sharing to mitigate risks associated with the unpredictability of their foraging niche141522Here we use economic gift games to construct and compare individual cooperative networks in three BaYaka camps. Our results demonstrate the presence of individual variation in relational wealth, which is particularly striking among men. We show that those with more relational wealth receive food transfers from a larger number of individuals than their peers, and this is reflected in their significantly higher BMI; women with more relational wealth also have significantly higher age-specific fertility. The data also suggest there is a heritable component to relational wealth, and that cooperative alliances may be transmitted inter-generationally. While cooperation may have been important for increasing group resilience in warfare and resource competition, our findings indicate that individual level competition for cooperation within the group may also have been fundamental in shaping human sociality.2 = 0.036) (see We constructed an adult-to-adult gift network by playing a honey stick gift game (HSGG)036) see . The ext036) see . The lacThis trend may reflect the fact that usually male hunting production is both more variable and shared more widely than female gathered foods in hunter-gatherer socities25Using multiple regression we found a significant association between our measure of relational wealth (HSGG nominations) and the number of camp members from whom an individual receives food in real world transfers   in which nuclear families reside; camp size tends to vary from 10–60 individuals, and genetic relatedness within camps is lowOur study uses data from the Mbendjele BaYaka, a subgroup of the BaYaka who speak Mbendjele language and whose residence spans across the forests of Congo and Central African Republic. BaYaka subsistence techniques include hunting, trapping, fishing, gathering and honey collecting; as well as some trade with neighbouring farmer groups. Food sharing is an integral component of BaYaka subsistence and culture. The BaYaka live in et al. Participants were asked to accompany the researcher and translator to a private area.Participants were shown three honey sticks, and told that real honey was within each batton.Participants were told they must decide to whom they would like us (the researchers) to give the honey sticks.Participants were told they could give freely i.e. all three sticks to one individual or one stick to three different individuals etc.Participants were told they could nominate any adult in their camp other than themselves.After the games had been completed with all adults in camp, the honey sticks were distributed according to the results.This game was played with all willing members of a camp and was completed as quickly as possible, usually within 2–3 days in each camp. All instructions were spoken in French by the researcher, and then immediately repeated in Mbendjele by the translator. The game was based on the procedure of Apicella Households were observed by JT over a series of two to four hour time blocks, with households observed for a total of 24 or 36 hours depending on the camp. Observations were evenly distributed between 6 am and 6 pm and spread over several days. During observation periods, a record was made of all food produced by a focal household. If division of resource packages occurred, all recipient households were identified. For all food cooked and consumed by the household, the type and amount of food were recorded and all those who ate the food were identified.We measured height and weight of all willing and non-pregnant adults in each camp in order to calculate BMI. Height was measured to the nearest mm using a Harpenden anthropometer, and weight using a Philipps mechanical scale.In all analyses, relational wealth is calculated as the number of nominations received in the HSGG standardised by camp and sex.We use multiple regression to analyse the relationship between relational wealth and number of food sharing donors. The response variable is the number of different camp members observed to share food with ego during food transfer observations. The predictor is ego’s relational wealth, and controls are ego’s sex, age /over 45 since there is a significant decline in BMI for these age-groups in our sample. We also control for camp membership .2 as controls to account for the quadratic relationship between age and fertility. To check whether reverse causality may explain the significant association found, we conduct a gamma correlation between female relational wealth and the number of dependent offspring in their household. The gamma correlation is conducted using the rococo package in R; and is selected as it is appropriate for variables which contain many ties, such as number of dependent offspring.We use multiple regression to analyse the relationship between relational wealth and female fertility. We use age and ageFor correlation analyses of ego’s and ego’s parents’ relational wealth we also use gamma correlations because they are appropriate for small sample sizes and data with ties.How to cite this article: Chaudhary, N. et al. Competition for Cooperation: variability, benefits and heritability of relational wealth in hunter-gatherers. Sci. Rep.6, 29120; doi: 10.1038/srep29120 (2016).


No such relationships were found in control subjects. These data suggest that symptoms of fatigue in CFS subjects were associated with reduced responsivity of the basal ganglia, possibly involving the disruption of projections from the globus pallidus to thalamic and cortical networks.Reduced basal ganglia function has been associated with fatigue in neurologic disorders, as well as in patients exposed to chronic immune stimulation. Patients with chronic fatigue syndrome (CFS) have been shown to exhibit symptoms suggestive of decreased basal ganglia function including psychomotor slowing, which in turn was correlated with fatigue. In addition, CFS patients have been found to exhibit increased markers of immune activation. In order to directly test the hypothesis of decreased basal ganglia function in CFS, we used functional magnetic resonance imaging to examine neural activation in the basal ganglia to a reward-processing (monetary gambling) task in a community sample of 59 male and female subjects, including 18 patients diagnosed with CFS according to 1994 CDC criteria and 41 non-fatigued healthy controls. For each subject, the average effect of winning vs. losing during the gambling task in regions of interest (ROI) corresponding to the caudate nucleus, putamen, and globus pallidus was extracted for group comparisons and correlational analyses. Compared to non-fatigued controls, patients with CFS exhibited significantly decreased activation in the right caudate (p = 0.01) and right globus pallidus (p = 0.02). Decreased activation in the right globus pallidus was significantly correlated with increased mental fatigue (r There has been increasing interest in the neural correlates of fatigue and other symptoms that afflict individuals with chronic fatigue syndrome (CFS). CFS has devastating effects on social and occupational function, leading to significant personal and public health costs In order to further explore basal ganglia function in patients with CFS and its relationship with CFS-related symptoms, we conducted a fMRI study in CFS patients to examine the basal ganglia response to hedonic reward during a gambling task previously shown to activate relevant basal ganglia nuclei a priori by the Emory and Centers for Disease Control and Prevention Institutional Review Boards. All subjects provided written informed consent prior to study participation.The study was approved Participants in this study represent a subgroup of persons enrolled in the baseline and follow-up survey of CFS in Georgia Exclusion criteria for all subjects included: left-handedness (to minimize the variance associated with the effect of handedness on brain structure and function); presence of metal in the body; claustrophobia; and a reading level below grade 7 . Subjects with a score >60 on the 20-item, self report Zung Self Rating Depression Scale (Zung SDS)(indicating more than mild depressive symptoms) were also excluded A previously published gambling task, proven effective in eliciting specific activation of basal ganglia structures, was adapted for the study Three T2*-weighted functional runs  and a high-resolution T1-weighted anatomical scan  were collected on a Siemens Trio 3T MRI scanner. All fMRI processing was performed using the Analysis of Functional NeuroImage (AFNI) software package To focus the analysis on the specific basal ganglia regions activated by the task, the following procedure was employed: (1) a whole-brain group analysis revealing the general activation pattern for the Win-Lose contrast across all subjects was performed; (2) the resultant statistical parametric brain map thresholded at p<0.05, corrected for multiple comparisons, was intersected with basal ganglia regions of interest , obtained from a probabilistic cytoarchitectonic brain atlas included in the AFNI distribution; (3) for each subject, the average value of Win-Lose activation contrast in each ROI was extracted for group comparisons and correlational analyses.To determine the status of participant's illness at the time of the scan, questionnaires were administered the evening before the scan. Fatigue was assessed using the 20-item Multidimensional Fatigue Inventory (MFI) 20, a self-rating scale yielding specific subscales for general fatigue, physical fatigue, reduced activity, reduced motivation and mental fatigue, with the summed score for each subscale ranging from 4 (best) to 20 (worst)Differences between groups in sociodemographic and clinical variables were assessed using t-tests for continuous measures and chi-square tests (or Fisher's Exact Test where appropriate) for categorical variables. In cases of unequal variances, Welch's t-test was employed. Evoked activity in right, left and bilateral caudate, putamen and globus pallidus was determined by averaging the estimated percent blood oxygen level-dependent (BOLD) signal change corresponding to the Win-Lose contrast across all ROI voxels as described above. Differences in overall neural activity between groups across ROIs in the three right and left basal ganglia nuclei  were analyzed using a multivariate analysis of covariance (MANCOVA) controlling for age sex, race and body mass index (BMI) as well as the presence or absence of an anxiety disorder or the use of medications categorized by class see . The impEighteen CFS patients and 41 healthy controls qualified for the study and completed the fMRI procedure. Subjects were well-matched on age, sex, race, education, BMI and medication status . HoweverAs shown in 2 = 0.491, p = 0.001), general fatigue  and reduced activity  subscales of the MFI  but not for the reduced motivation subscale of the MFI . No significant correlations were found between symptoms of fatigue and the right caudate nucleus in CFS subjects. In addition, there were no significant correlations in control subjects between basal ganglia activation in any of the identified ROIs and fatigue symptoms , along with the substantia nigra pars reticulata, are main basal ganglia outputs to the thalamus and receive significant dopaminergic projections from the substantia nigra as well as inhibitory GABA input from the striatum One mechanism that may contribute to alterations in function and neurotransmission in the basal ganglia in CFS is inflammation There are several strengths and weaknesses of this study which should be considered in the interpretation of the data. First, the sample size of CFS subjects was relatively small. Although there were many more CFS subjects identified by our initial sampling strategy, we adhered to strict inclusion and exclusion criteria to limit the potential influence of factors such as psychotropic medications and psychiatric conditions like depression, both of which can have a profound effect on neuroimaging data. Moreover, we used validated assessment tools to quantify the degree of pathology in each of the domains relevant to the diagnosis of CFS. This attempt to provide a relatively homogenous sample of CFS patients also runs the risk of producing results that are not generalizable to the “typical” CFS patient. In addition, these data represent averaged results from each of the samples, and observation of the data indicates there was significant overlap between the groups. Thus, not all persons in our study exhibited reduced neural activation in the basal ganglia, although all CFS subjects exhibited significant fatigue. Therefore, although alterations in basal ganglia function may characterize a subgroup of CFS patients, such alterations do not typify all CFS subjects and do not account for all instances of fatigue. In this study non-fatigued controls were used as the comparison group as a starting place in investigating biologic associations with CFS. These non-fatigued controls exhibited some low level of minor medical problems including allergies, muscle aches and pains, gastrointestinal distress, high blood pressure and hypercholesterolemia for which they were taking medications, but not at rates that differed from our CFS sample. Nevertheless, in order to evaluate whether the findings are specific to CFS, additional studies comparing persons with CFS to persons with more significant illnesses or chronic fatigue would be useful to determine if our findings are typical of CFS patients, or patients with fatigue or illness in general. Finally, it should be noted that the CFS patients in this study were not drawn from a sample of subjects seeking care at an academic or private practice specialty clinic providing tertiary care. Rather the subjects studied herein were a community-based sample, and despite the rigorous inclusion and exclusion criteria, might more likely reflect findings in the average patient.CFS subjects free of psychotropic medications and without significant psychiatric disease were found to exhibit reduced activation of basal ganglia structures that correlated with symptoms of fatigue. The findings are consistent with diseases known to affect the basal ganglia and are consistent with basal ganglia changes seen after administration of a variety of stimuli that induce an inflammatory response.


Here, we present a label-free high resolution optical coherence tomography (OCT) based lymphangiography (OLAG) within human skin in vivo. OLAG enables rapid (~seconds) mapping of lymphatic networks, along with blood vessel networks, over 8 mm x 8 mm of human skin and 5 mm x 5 mm of human areola. Moreover, lymphatic system’s response to inflammation within human skin is monitored throughout an acne lesion development over 7 days. The demonstrated results promise OLAG as a revolutionary tool in the clinical research and treatment of patients with pathologic conditions such as cancer, diabetes, and autoimmune diseases.Due to the limitations of current imaging techniques, visualization of lymphatic capillaries within tissue  Lymphatic system drains lymph fluid from the extracellular space into larger ducts through unidirectional, thin-walled capillaries and collecting vessel network. To transport the fluid, lymphangions, the vascular smooth muscle cells, rhythmically constrict and relax, and valve leaflets open and close in an orchestrated manner to mediate unidirectional flowMost of the current and emerging lymphatic imaging approaches require the administration of exogenous contrast agents either directly into lymphatics via the cannulation of a lymphatic vessel or indirectly into lymphatic plexus via intradermal injection. Unfortunately, most of the existing contrast agents can be toxic and induce side effects to patientsin vivo animal models of disease. So far, none of these techniques has shown its potential in clinical investigations. Optical coherence tomography (OCT) is an emerging tool for non-invasive, label-free in vivo imaging of tissuein vivo blood perfusion map in tissue beds down to capillary level using Fourier domain OCT141619Despite its prominent role in healthy and pathological processes, there are only few methods reported in the literature1011in vivo mouse ear model using OCT111211in vivo mouse ear models, however, they have not been able to apply to human skin due to obvious challenges facing optical imaging, i.e., the thicker and higher scattering nature of human skin compared to mouse ear skin. The contrast of the dark areas to the surrounding tissue becomes worse and segmentation algorithms fail to provide a clear separation of these areas from the surrounding tissue in OCT images of human skin.Previously, label-free imaging of lymphatic vessels has been demonstrated on an in vivo. Moreover, lymphatic system’s response to inflammation is monitored on an acne lesion within human skin for 7 days. Results showed that OLAG is promising to provide a remarkable alternative tool in the investigation and treatment of pathologic mechanisms involving lymphatic system without the use of exogenous contrast agents on patients.We developed a method to contrast lymphatic vessels within highly scattering tissue by reducing light attenuation effect on OCT cross-sectional images and extracting lymphatic vessels through re-arranged enface OCT images . With thUsing OCT, we imaged the skin in the ventral side of the human lower arm, and presented the results in The areolar dermis consists of highly active lymphatic plexusDuring inflammation, the lymphatic drainage is activated along with the vascular remodelling. Therefore, the monitoring of a simple inflammation phenomenon could be a good surrogate for demonstrating the usefulness of the proposed OLAG technique. Here, human skin’s inflammatory state and its recovery are monitored for 7 days within an acne lesion using OMAG and OLAG and shown in The volumetric depth-resolved MIP of OMAG images clearly show the changes in microvascular activity within the region of interest. In the first 3 days, inflammation causes edema in the tissue, which is circled in Breakage of dermal balance and inflammatory response leads to dense microvasculature in Lymphangiogenesis is critical to maintain internal stability of the immune system by reducing edema and inflammation in the tissue25in vivo status of tumor induced lymphangiogenesis and to monitor the treatment modalities on lymphatics. Currently, subdermal injection of contrast agents is required for imaging lymphatic vessels in clinics, but they can create complications in patients so it is considered as significantly invasive in the clinical care as well as in the clinical researchThere are no truly non-invasive ways to assess the in vivo, in the absence of exogenous contrast agents. We also observed the variations in the dynamics of lymphatic vessels in the case of inflammation that has been developed by acne vulgaris. To the best of our knowledge, this is the first study imaging the lymphatic system within human skin without any interventional radiology procedures. Our technique offers a unique ability to quickly visualize the microvascular and lymphatic response to stimulation, non-invasively. It may be used to image functional and constitutional changes through lymphangiogenesis and angiogenesis in the patients with cancer in response to tumor growth. OLAG results from subareolar lymphatic plexus  system operating at 1310 nm central wavelength to implement our methods to image blood and lymphatic microcirculation within man skin . The sysin vivo human skin.In this study, we elected the use of SS-OCT over SD-OCT system for the following reasons: its longer imaging range, less depth-dependent signal roll-off, and less motion-induced signal loss due to fringe washoutThe optical power of incident light upon the sample was ~5.2 mW, well below the American National Standards Institute (ANSI) standards (Z136.1) for the safe use of near infrared light at 1310 nm on skinTo visualize the volumetric microvasculature up to capillary level, an OMAG based OCT angiography technique was utilizedAcquired 3D OMAG datasets were visualized through volume rendering and a Gaussian filter of 3 × 3 kernel size was applied for noise reduction. Maximum intensity projection (MIP) of the OMAG blood flow image was produced by mapping the maximal value in each A-line on to an enface 2D plane which was used to visualize the blood vessel networks.Accurate detection of lymphatic vessels within human skin requires OCT images with good contrast. Non-uniform intensity distribution of the OCT images due to light attenuation and heterogeneous tissue properties make it difficult to extract lymphatic vessels within highly scattering tissue. In order to mitigate this problem, OCT structural images were processed in axial and lateral directions and the contrast was enhanced using a processing protocol described below.The contrast of the original OCT structural images were first improved using attenuation compensation algorithmHow to cite this article: Baran, U. et al. OCT-based label-free in vivo lymphangiography within human skin and areola. Sci. Rep.6, 21122; doi: 10.1038/srep21122 (2016).


Attention-deficit/hyperactivity disorder (ADHD) is commonly associated with disordered or disturbed sleep. The relationships of ADHD with sleep problems, psychiatric comorbidities and medications are complex and multidirectional. Evidence from published studies comparing sleep in individuals with ADHD with typically developing controls is most concordant for associations of ADHD with: hypopnea/apnea and peripheral limb movements in sleep or nocturnal motricity in polysomnographic studies; increased sleep onset latency and shorter sleep time in actigraphic studies; and bedtime resistance, difficulty with morning awakenings, sleep onset difficulties, sleep-disordered breathing, night awakenings and daytime sleepiness in subjective studies. ADHD is also frequently coincident with sleep disorders . Psychostimulant medications are associated with disrupted or disturbed sleep, but also ‘paradoxically’ calm some patients with ADHD for sleep by alleviating their symptoms. Long-acting formulations may have insufficient duration of action, leading to symptom rebound at bedtime. Current guidelines recommend assessment of sleep disturbance during evaluation of ADHD, and before initiation of pharmacotherapy, with healthy sleep practices the first-line option for addressing sleep problems. This review aims to provide a comprehensive overview of the relationships between ADHD and sleep, and presents a conceptual model of the modes of interaction: ADHD may cause sleep problems as an intrinsic feature of the disorder; sleep problems may cause or mimic ADHD; ADHD and sleep problems may interact, with reciprocal causation and possible involvement of comorbidity; and ADHD and sleep problems may share a common underlying neurological etiology. Diagnostic and Statistical Manual of Mental Disorders (DSM), Fifth Edition   is a common neurodevelopmental disorder that has been estimated to affect approximately 5.3 % of children and adolescents worldwide  and subjective measures  are used to assess sleep in patients with ADHD.Polysomnography involves simultaneous and continuous measurement of multiple physiological parameters, and is usually conducted in a sleep laboratory. A combination of electroencephalography, electrocardiography, electromyography, electrooculography, pneumography and pulse oximetry is typical, sometimes together with audiovisual recordings.Polysomnography is considered the ‘gold standard’ for the objective measurement of sleep  sleep in children and adolescents with ADHD , and there is a need for larger-scale, multicenter studies , naturalistic studies are subject to uncontrolled variables such as parents’ work schedules, parenting style, family structure, child habits and employment in teenagers Beebe . SeveralInternational Classification of Sleep Disorders  describes a spectrum of conditions ranging from obstructive sleep apnea (OSA) to primary snoring O’Brien . SDB hasSurgical treatment of children with OSA via adenotonsillectomy in prospective, interventional studies has been reported to be associated with improvements in neuropsychological behavior Beebe , academiRestless legs syndrome (RLS) is a neurological disorder characterized by an irresistible urge to move the legs to relieve uncomfortable sensations at rest . In a 2009 systematic review, the frequency of somnolence reported as a TEAE in placebo-controlled clinical trials of atomoxetine was reported to range from 15 to 17 %   is also approved in North America with a similar indication  soon before bedtime; sleeping only in bed and using the bed only for sleeping; and attention to environmental factors such as bedroom furniture, lighting and temperature  and bedtime fading, which involves identifying a bedtime at which the child falls asleep within about 15 min, and gradually setting bedtime earlier until the desired bedtime is achieved, while keeping wake time fixed and disallowing sleep at other times   should be ruled out before diagnosing or treating ADHD. Obesity and psychiatric comorbidities  can also lead to sleep problems, and need to be identified and treated appropriately. The multifaceted effects of stimulant pharmacotherapy on sleep in patients with ADHD are particularly important for clinicians to understand when evaluating treatment options for patients. Stimulant medications may disrupt or improve sleep in different patients, depending not only on the nature of the patient’s illness, but also on the drug dose, class, formulation and duration of efficacy. Effective management of sleep problems associated with ADHD and its treatment may not only alleviate sleep-related symptoms, but also improve quality of life in parents or carers of children with disruptive bedtime behavior or insomnia.In the near term, new understanding of how sleep interacts with ADHD and how this affects treatment choices is likely to come from clinical studies. Polysomnographic studies need to be larger and better controlled than previous studies, and should not overlook subtle polysomnographic signals such as microarousals and the time structure of PLMS. There is also a need to follow up the recent data indicating a link between ADHD, obesity, daytime sleepiness and circadian-rhythm alterations. Conspicuously absent from the current literature is any convincing demonstration that pharmacological or non-pharmacological intervention to improve sleep actually leads to improved ADHD symptoms or reduced functional impairment associated with the disorder. Similarly, as children become adolescents, they may experience sleep loss, but do any adolescents benefit from more or better sleep in terms of preventing worsening of ADHD? Crucial to both these questions is the possibility that specific ADHD phenotypes are associated with, or characterized by, particular types of sleep-related problems. In addition to helping clinicians make treatment decisions, the ability to subcategorize patients with ADHD based on their sleep phenotype may help shed light on areas where current data are conflicting. Furthermore, such phenotypic classification is probably essential for increased sensitivity in genomic screens for ADHD-associated polymorphisms. The overlapping neurochemical and neuroanatomical systems involved in regulating sleep, attention, arousal and circadian rhythms are the subjects of current basic research (Owens et al.


However, this kind of therapy is far from ideal as physiological demand for steroids varies considerably throughout the day and increases during periods of stress. The development of alternative curative strategies is therefore needed. In this review, we describe the latest technologies aimed at either isolating or generating  The adrenal cortex is the primary site of steroid synthesis, producing glucocorticoids under the control of the hypothalamic–pituitary–adrenal (HPA) axis and mineralocorticoids under the control of the renin–angiotensin system. Glucocorticoids affect carbohydrate metabolism and mediate the mammalian stress response, while mineralocorticoids control blood volume and salt homeostasis. For this reason, the adrenal cortex is essential for life.in vivo  . At arouin vivo . Other tin vivo , 7; pre-in vivo , 9; dosain vivo , whose min vivo , such asin vivo –15]. The tracing , 17. Not tracing ].The adult gland is divided into at least three distinct zones arranged concentrically around the medulla; the zona glomerulosa , which lies just underneath the capsule, the zona fasciculata , and then the zona reticularis . Zonation occurs around birth .Primary adrenal insufficiency is due to a number of adrenal disorders, and management of these patients can be challenging . CurrentAdrenal autotransplantation in humans is poorly characterized. A few surgeons tested the potential benefits of adrenal autotransplantation in patients with Cushing’s disease since the early 1960s. At that time, replacement therapy with synthetic steroids was becoming the gold standard for patients with Cushing’s disease but they hypothesized that autotransplantation would be an effective alternative to lifelong therapy –26. AdreThe refinement of allotransplantation and xenotransplantation has been the focus of more recent research, where a variety of animal models have been employed. Hornsby and colleagues performed initial work, demonstrating that transplantation of human adrenocortical cells into adrenalectomized severe combined immunodeficiency (scid) mice could be an effective technique for the treatment of adrenal insufficiency . They alThrough their work, they also showed that transplanted adrenocortical cells (of either human or bovine origin) could survive and form a functional, vascularized tissue that was able to replace the host animal’s organ. The newly formed tissue was able to produce and secrete the necessary amounts of cortisol, albeit aldosterone was only produced by primary bovine adrenocortical cells, but not from clonal cells . One posin vitro leading to improved efficacy  have been shown to be very effective in xenotransplantation procedures. This technique aids the expansion of adrenocortical cells efficacy , 41. Altefficacy .Other groups looked into harnessing the properties of the adrenal cortex extracellular matrix (ECM). It is well established that the ECM plays a significant role in organ growth and development, as well as function, by providing structural support and by regulating cell signaling and cell-to-cell interactions . The ECMFurther work to assess the function of implanted cells seeded on a collagen matrix, using a mouse model with adrenal insufficiency, determined that treatment involving staged bilateral adrenalectomy of the animals and implantation of collagen sponges each time was the most beneficial. This protocol lead to a 100% survival rate  and reversal of adrenal insufficiency, supported by restoration of corticosterone levels and the expression of adrenal markers by the transplanted cells .In vitro studies, using porcine decellularized adrenal ECM as a scaffold for human fetal adrenal cells, resulted in successful cell attachment and a good cell function, the latter assessed by cell proliferation and cortisol secretion . Other clinical trials are in the pipeline, a culmination of decades of intense in vitro and animal studies carried out by dozens of laboratories  by expressing genes encoding four transcription factors . IPSCs cCyp11a1 together with an induction of steroidogenesis. However, these cells were incapable of producing different steroid hormones de novo, since only progesterone was detected after treatment with 20α-hydroxycholesterol, a freely diffusible form of cholesterol  to differentiate into a steroidogenic lineage using adenoviral-mediated SF1 overexpression . In thisThe same group later obtained steroidogenic cells from mouse adipose tissue-derived mesenchymal cells (AMCs), using a similar strategy . Remarka+ BMCs injected into rat testes showed upregulation of steroidogenic enzymes in the engrafted cells  stably expressing SF1  were able to upregulate steroidogenic enzymes and produce both adrenal and gonadal steroids after treatment with cAMP. Immunohistochemical analysis of rat GFPed cells . Taken tin vivo, since LRH-1 is barely detectable in the adrenal cortex but highly expressed in the gonads, while SF1 is abundantly expressed in both adrenals and gonads   and umbiRecently, several groups have changed their focus onto ESCs, as the first attempts to reprogram these cells were only partially successful . Yazawa Sonoyama et al. demonstrated the capacity of human ES cells (hESCs) to become steroidogenic . Using ain vivo models of adrenal insufficiency , they provide strong evidence that cells can be reprogramed to a steroidogenic phenotype through overexpression of SF1 and cAMP treatment. The choice of the most appropriate source of cells as substrates for reprograming is still debated and might differ depending on downstream applications. Differences in species, cell/tissue source, cellular development stage, epigenetic landscape, SF1/transcription factors dosage  the conditions to obtain cells with restricted adrenocortical-specific gene expression and hormone production are far from optimized (see above). (2) In most of the protocols, upon overexpression of SF1, cells terminally differentiate with a consequent growth arrest. This makes it difficult to prepare the large amount of cells needed for cell therapy. The use of inducible vectors should allow the expression of SF1 (and/or other factors) at a desired culture time-point when there are enough cells for clinical purposes. (3) The methodologies used until now are all based on the overexpression of SF1 exogenously, either episomally or virally. Optimization of the culture conditions in a gene delivery-free model of reprograming might help to avoid the safety concerns about these cells. The use of excisable vectors together with the new CRISPR-dCasVP64 technology to activate specific transgenes without the use of transcription factors  might reA major factor limiting the application of stem-cell therapy in patients transplanted with ECs-derived adrenocortical cells or even autologous IPSCs-derived adrenocortical cells  is the recipient’s need to adhere to lifelong immunosuppression. An effort going back a couple of decades in the field of biomaterials is providing us with the technology, which can make an impact in the clinical setting today; in fact, several encapsulation devices (endowed with excellent biocompatibility) have been developed and are undergoing clinical testing in patients with type-I diabetes. Animal studies  and publIn recent times, a fast-paced development of gene-editing technologies using different approaches has made it possible to correct known disease-causing mutations without leaving any footprint . This tePotential future strategies, as well as pioneering past and present attempts to cure adrenal insufficiency are outlined in Figure In conclusion, while autotransplantation has long been discarded, allotransplantation is currently being considered only in specific clinical settings and xenotransplantation has not reached the bedside, stem-cell biology has granted great promise for tissue engineering and regenerative medicine. Treatments employing stem cells offer the potential to successfully treat patients with new modalities in the future, by essentially regenerating and replacing non-functional tissues or organs. A permanent collaborative and multidisciplinary effort carried out by translational scientists with expertise in stem cells, bioengineering, and material science is undeniably necessary to fully develop alternative treatments for adrenal insufficiency that have true clinical relevance.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.


There is an increased need for additional care and support services for the elderly population. It is important to identify what support older people need once they are discharged from hospital and to ensure continuity of care. There is a large evidence base focusing on enhanced discharge services and their impact on patients. The services show some potential benefits, but there are inconsistent findings across reviews. Furthermore, it is unclear what elements of enhanced discharge interventions could be most beneficial to older people. This meta-review aims to identify existing systematic reviews of discharge interventions for older people, identify potentially effective elements of enhanced discharge services for this patient group and identify areas where further work may still be needed.The search will aim to identify English language systematic reviews that have assessed the effectiveness of discharge interventions for older people. The following databases will be searched: Medline, Embase, PsycINFO, HMIC, Social Policy and Practice, CINAHL, the Cochrane Library, ASSIA, Social Science Citation Index and the Grey Literature Report. The search strategy will comprise the keywords ‘systematic reviews’, ‘older people’ and ‘discharge’. Discharge interventions must aim to support older patients before, during and/or after discharge from hospital. Outcomes of interest will include mortality, readmissions, length of hospital stay, patient health status, patient and carer satisfaction and staff views. Abstract, title and full text screening will be conducted independently by two reviewers. Data extracted from reviews will include review characteristics, patient population, review quality score, outcome measures and review findings, and a narrative synthesis will be conducted.This review will identify existing reviews of discharge interventions and appraise how these interventions can impact outcomes in older people such as readmissions, health status, length of hospital stay and mortality. The review could inform practice and will help identify where further research is needed.CRD42015025737.PROSPERO The online version of this article (doi:10.1186/s13643-016-0222-8) contains supplementary material, which is available to authorized users. There has been a recent movement in healthcare policy and practice towards providing additional patient-centred care , to support the needs of the growing population of older patients (people aged over 60) , 2. SuboThe older populations are more likely than younger individuals to suffer from multiple morbidities and problems such as frailty , 6. CondPrimary research on discharge interventions has led to a substantial number of systematic reviews being conducted in this area, but they have varied in their focus. Some reviews have not focused on a specific population –12 whilsThere is evidence to suggest that in some contexts, discharge interventions can reduce length of hospital stay , 12; redIt would be useful to identify what discharge interventions there are specifically for the older population in systematic review evidence, as there are numerous discharge interventions available for this group. These may include assessing the rehabilitation needs of patients, planning and delivering specific support in particular settings to patients after they are discharged  , 12, 21.Meta-reviews are useful when systematic reviews have already been conducted in the research area of interest, as they can provide new insights into the existing evidence base . A meta-Identify reviews of different discharge interventions which aim to support older patients.Identify the characteristics of the different subgroups of older patients included in these reviews.Patient mortality, health status, quality of life, dependency (using activities of daily living scores) and satisfaction.Patient hospital length of stay and readmission rates.Carer outcomes including acceptability of intervention, satisfaction, burden and quality of life.Staff outcomes including views on service and workload.Costs and resource use.Identify any interventions or specific elements of interventions which have been effective in achieving clinical, patient, staff and carer outcomes:Identify which patient-reported outcome measures have been used to measure outcomes in older patients.Identify areas where further research is needed.The protocol for this meta-review has been written in accordance with the PRISMA-P  guidelines  and has A clearly stated set of objectives with pre-defined eligibility criteria for studies.An explicit, reproducible methodology.A systematic search that attempts to identify all studies that would meet the eligibility criteria.An assessment of the validity of the findings of the included studies, for example through the assessment of risk of bias.A systematic presentation, and synthesis, of the characteristics and findings of the included studies.Only systematic reviews, meta-analysis or other systematic meta-reviews will be included in this meta-review. Eligible reviews must have the following key characteristics as defined by Higgins and Green :A clearlThe population of interest for this review is older patients (i.e. those over 60 years of age) . Where oDischarge planning schemes: primarily interventions that utilise comprehensive discharge planning protocols.Discharge support schemes: a variety of models in which new and existing services are targeted at recently discharged patients, including schemes with early discharge from inpatient hospital care.Geriatric assessment programmes: assessment services focused on hospital inpatients and patients recently discharged from hospital.Educational programmes: a fairly distinct group of studies with objectives of educating patients in aspects of management of their illness.Systematic reviews that focus on interventions that provide additional support or adapt the process around the time of discharge for older patients who are a hospital inpatient. This could be an intervention before, during and/or after discharge from hospital. A previous systematic review  developeThese categories are not mutually exclusive, but they will provide structure to the review and analysis . Any posDue to hospitals offering different types of discharge services and support, comparators of any type of standard care or alternative interventions will be included in this review. Where possible, the services used as comparators will be summarised and then grouped to allow comparison.MortalityReadmissions Length of hospital stayPatient health status Patient satisfactionAdmission to institutional careCarer outcomes Staff outcomes (including staff views and workload)Care process measures (e.g. number of complications)Costs and resource use Outcome measures of interest will include (but are not restricted to) the following:Interventions that focus solely on populations that cannot be classified as older people and where there is no stand-alone data that can be extracted on the population of interest, e.g. a review focusing on post-pregnancy discharge services.Interventions which do not aim to support patients around the time of discharge from hospital.Specialist mental health discharge services except discharge services for delirium or dementia. These services will be included as these conditions are common in the older population .Reviews where there is no full paper and/or results such as abstract only citations from conference proceedings or published protocols of systematic reviews.Reviews written in languages other than English (there is no restriction regarding the language of source data included in the reviews).Reviews focusing solely on less economically developed countries (LEDCs). Interventions in LEDCs will not be comparable due to different standards of living, quality of medical care, life expectancies and quality of life. If there is uncertainty regarding whether a country is a more or less economically developed country, the United Nations country classification document will be used .Interventions evaluating the effectiveness of drugs administered to patients.Reviews scoring less than 5 on the AMSTAR (a measurement tool to assess systematic reviews) checklist see ‘’ sectionThis meta-review will have no restriction on review publication date. Due to pragmatic reasons, the search has been limited to reviews written in English, which could introduce a language bias .Medline (using Ovid)Embase (using Ovid)PsycINFO (using Ovid)Health Management Information Consortium (HMIC) (using Ovid)Social Policy and Practice (using Ovid) Cumulative Index to Nursing and Allied Health Literature (CINAHL)Cochrane Library (includes Database of Abstracts of Reviews of Effects (DARE))Applied Social Sciences Index and Abstracts (ASSIA)Social Science Citation Indexwww.greylit.org)Grey Literature Report  and filters for ‘systematic reviews’, ‘older people’ and ‘discharge’.Data are available on the effectiveness of different filters to identify systematic reviews, and previous studies have conducted reviews of the best search strategies for identifying systematic reviews in databases  , 31. WheThe preliminary search strategy for Medline is provided in Additional file The study is considered eligible by both of the reviewers.The reviewer is unsure of a review’s eligibility and the full text needs to be reviewed.There is disagreement between the two reviewers as to whether the study is relevant.First, all of the citations will be transferred to Refworks and duplicates will be removed. Citations will then be exported into Microsoft Excel to check for any remaining duplicates. The titles and abstracts of all citations will be independently screened for eligibility against the inclusion and exclusion criteria by two reviewers. A review will be taken forward to the full text screening stage if . If the two reviewers do not agree about inclusion of a review, they will meet to discuss it. If agreement regarding inclusion/exclusion cannot be reached, the review will be discussed by the wider research team who will then make a decision. Results of the search and screening process will be reported in a PRISMA flow diagram. The above screening process will be piloted on the first 100 results of the Ovid Medline search, and this will allow the process to be refined if necessary before completing the whole review. Cohen’s kappa index of inter-rater reliability will be used to assess agreement between the two reviewers. A score between 0.61 and 1.0 will be considered acceptable as anything above 0.61 is defined as substantial or almost perfect agreement .Reference ID numberCitationCountry of publicationIdentifying featuresType of review (for example systematic review or meta-review)Discharge planning schemesDischarge support schemesGeriatric assessment programmesEducational programmesWhich category does the review fit into as defined by Parker et al.?Databases searched Additional sources (for example reference lists or contacting experts in field)Years searchedGeographical scope Language restrictions of included studies (for example English)Healthcare settings included in review Overall aim of the reviewReview research questions/objectivesStudy designs included , controlled before and after studies or cohort studies)Number of studies included in reviewNumber of published articles included in reviewData synthesis AMSTAR score Review characteristicsPatient population included in reviewPatient age rangeTotal number of participants includedReview participantsGeneral description of intervention(s)General description of comparator(s)Evidence of theories or conceptual frameworks informing the interventionDid intervention involve different care provided by different staff, in different settings or at different time points? Where was the intervention(s) administered? Staff involved in intervention(s) Timescales over which interventions were administeredDo interventions have a ‘handover’ back into routine care or other service? InterventionsAll primary outcomeAll secondary outcome(s)Outcomes measuredResults of outcomes of interest (review outcomes of interest listed)Intervention settingIntervention componentsIntervention comparatorIntervention populationIntervention providersIntervention resourcesWe will collect data on each outcome regarding whether any of the following has had an impact on each outcome:Review conclusion(s)Patient reported outcome measures that are identified in this review.Review authors’ assessment of overall quality of the evidence reported in the reviewResults of any relevant subgroup analysisDetailed in results sectionDetailed in discussion sectionAny barriers and/or facilitators for implementing the interventionsReview conclusion(s)If none, consider the Population, Intervention, Comparison, Outcomes and Setting (PICOS) detailed in the systematic review question and whether these are represented in the results .Further research/work or gaps identified in reviewFindingsData extraction will be completed independently by the two reviewers. Any disagreements will be discussed, and if agreement cannot be reached, they will be discussed by the wider research team. Extracted data will be collated in word documents, and possible answers with tick boxes will be used where possible. Please see below details of what the data extraction form will include and brief examples of tick boxes we will have:AMSTAR will be used to assess the quality of the systematic reviews, and the tool consists of an 11-item checklist . SystemaThe search strategy for this meta-review has been designed to be comprehensive and includes sources of grey literature. It is accepted that there may be some publication bias in this meta-review due to only including reviews published in English.The meta-review will include moderate to high quality systematic reviews, and the AMSTAR checklist includes questions on publication bias assessment, whether the literature search was comprehensive and whether status of publication was used as an inclusion criteria. The scores of the AMSTAR checklist will be reviewed to assess if included reviews have scored well regarding publication bias, and this data will be reported.A number of previous systematic reviews have faced challenges when comparing study results such as missing information , 16, difDue to the expected heterogeneity in intervention designs and characteristics, a more narrative synthesis will be performed. Included reviews will be categorised, analysed and presented according to the Parker et al.  categoriIn addition to discussing the study protocol with the study team, the National Institute for Health Research (NIHR) Collaboration for Leadership in Applied Health Research and Care (CLAHRC) West Midlands Chronic Disease theme patient group have been consulted to discuss the review. The patient group will also be consulted to discuss the interpretation of the findings and draft paper and to incorporate what they consider relevant to patient and public audiences . FurtherThere are not considered to be any ethical concerns regarding undertaking this meta-review. Study findings will be disseminated and published. The findings from the work will inform ongoing and future work which is being undertaken by the NIHR CLAHRC West Midlands Chronic Disease Theme. As well as academic publications, the findings of the review will be disseminated to hospital teams in the West Midlands where discharge interventions for older people are part of hospital services or are being planned.The large number of existing reviews on this subject has made the evidence base challenging to interpret, and it would benefit from being systematically appraised. Systematic reviews of RCTs are considered the highest level of evidence available . This re


Age-related osteoporotic fractures are major health care problem worldwide and are the result of impaired bone formation, decreased bone mass and bone fragility. Bone formation is accomplished by skeletal stem cells (SSC) that are recruited to bone surfaces from bone marrow microenvironment. This review discusses targeting SSC to enhance bone formation and to abolish age-related bone fragility in the context of using stem cells for treatment of age-related disorders. Recent studies are presented that have demonstrated that SSC exhibit impaired functions during aging due to intrinsic senescence-related changes as well as the presence of senescent microenvironment. Also, a number of approaches aiming at increasing bone formation through targeting SSC and that include systemic SSC transplantation, systemic SSC targeting using aptamers or antibodies, use of therapeutic screteome and tissue engineering approaches will be presented and discussed. Aging is the most important risk factor for fragility fracture leading to the highly prevalent disease osteoporosis. Osteoporosis is defined as a disease of low bone mass and bone architectural deterioration that lead to bone fragility  to bone formation surfaces. Genetic studies of SSC and osteoblast ablation in mice demonstrated a significant decrease in bone formation . In some of the current trials, SSC are loaded on biomaterial matrices (scaffold). The biomaterial available for bone tissue regeneration can be classified as either biologically-derived polymers isolated from extracellular matrix, plants or seaweeds e.g. collagen, fibronectin, alginate or synthetic material e.g. hydroxyapatite, tricalcium phosphate ceramics, polylactide and polyglycolide or a combination of these. It is also possible to “functionalize” the scaffold adding a biological material e.g. siRNA, miRNA or a small molecule that can direct the differentiation of stem cells or SSCs into bone lineage (Andersen et al. SSC-based therapeutics has been employed for tissue regeneration and repair of both skeletal and non-skeletal tissues. For skeletal tissue regeneration, the concept is that SSC will home to bone and participate in bone regeneration (Shen et al. As an alternative approach for transplantation of in vitro expanded and differentiated SSC, is to target the resident SSC. This approach is clinically relevant for treatment of age-related impaired bone formation. As mentioned above, in the elderly and osteoporotic patients, SSC can respond to biological stimuli. In order to target SSC specifically, a large number of studies have tried to identify molecules that are enriched in SSC populations using global proteome analysis (Kristensen et al. The contribution of senescent SSC to skeletal aging is increasingly recognized, and supported by a large number of in vitro and in vivo studies. Recent studies suggest that extrinsic factors present in the aging microenvironment play a dominant role in impairing SSC functions during aging. These studies also show that SSC obtained from elderly persons and patients with osteoporosis maintain responses to extrinsic stimuli. Thus, “rejuvenation” of SSC is possible treatment option for age-related skeletal diseases. Advances in identification of rejuvenating molecules, molecular targets and in vivo systemic delivery systems targeting SSC, will enable the use of these novel therapies in clinical practice.


Reperfusion of ischemic cardiac tissue is the standard treatment for improving clinical outcome following myocardial infarction but is inevitably associated with ischemia-reperfusion injury (IRI). Ischemic myocardial injury can be alleviated by exposing the heart to brief episodes of sublethal ischemia-reperfusion prior to the ischemic insult, a phenomenon that has been termed ischemic preconditioning (IPC). Similarly, remote IPC (RIPC) is defined as transient episodes of ischemia at a distant site before a subsequent prolonged injury of the target organ. In this setting, adaptive responses to hypoxia/ischemia in peripheral tissues include the release of soluble factors that have the potential to protect cardiomyocytes remotely. Oxygen fluctuations is a hallmark of insufficient tissue perfusion and ischemic episodes. Emerging evidence indicates that prolyl hydroxylase oxygen sensors (PHDs) and hypoxia-inducible transcription factors (HIFs) are critical regulators of IPC and RIPC. In this review, we discuss recent findings concerning the role of the PHD-HIF axis in IPC and RIPC-mediated cardioprotection and examine molecular pathways and cell types that might be involved. We also appraise the therapeutic value of targeting the PHD-HIF axis to enhance cardiac tolerance against IRI. Ischemic heart disease represents a heterogeneous group of pathological conditions characterized by insufficient perfusion of the heart. Coronary artery revascularization and reperfusion of ischemic cardiac tissue is the standard treatment in patients with myocardial infarction. However, reperfusion by itself causes additional tissue injury  and is associated with a burst of reactive oxygen species (ROS) production and intracellular calcium overload  and the oxygen sensors  are key players in the cellular response to hypoxia. Insufficient oxygen supply to tissues rapidly increases the protein levels of HIFα subunits, which heterodimerize with their HIFß partner to activate transcription of a large number of target genes involved in the adaptive cellular and systemic response to oxygen deprivation Semenza, . HIFα suIPC is an effective strategy to protect the heart from IRI. This approach requires one or more short sublethal cycles of IR by coronary occlusion to protect the heart against a subsequent longer period of ischemia followed by reperfusion. Emerging evidence suggests that this phenomenon is mediated by the PHD-HIF oxygen-sensing pathway and has been reviewed elsewhere in detail , with partial reduction of PHD2, display a modest stabilization of HIF factors and a subsequent increase in some, but not all, metabolic genes previously identified as HIF-1α target genes  -1, -3, and -4 , the archetype HIF-responsive gene, which we recently found elevated in vhl-deficient hearts  activated by receptor-dependent and-independent mechanisms, which attenuate mPTP opening and promote cardioprotection during the reperfusion stage of IR  and, importantly, iNOS deficiency abrogates the late phase of IPC  are protective against future ischemic insults to organs and tissues distant from the regional area of initial ischemia, with similar efficacy as local IPC. This unique and innovative treatment approach has been named remote ischemic preconditioning or RIPC  could directly impact endothelial cells to mediate heart protection by regulating the coronary diameter, endothelial permeability or vascular tone. In this respect, the presence of Epo receptors on endothelial cells and the activation of the PI3K/Akt pathway in response to Epo have been previously evaluated  as well as systemic protective mechanisms initiated by extracellular factors , which contribute to myocardium salvage during reperfusion. The extracellular component of cardioprotection is strongly supported by the benefits of RIPC that rely on soluble factors released by peripheral ischemic tissues , which subsequently activate cardioprotective RISK pathways in cardiac tissue Figure  and furtThe 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.


Although alcohol consumption is a leading cause of preventable death and social problems worldwide, many studies have found an association between low-dose consumption and reduced risk of cardiovascular (CVD) disease. Despite important limitations in the underlying research, this idea has been promoted extensively, used to argue against the adoption of effective alcohol policies and led some doctors to advise patients to drink for better health.There are a number of grounds for scepticism about these claims. There have been no randomised studies of low-dose alcohol consumption with disease or death outcomes to confirm findings from non-randomised studies. There are many methodological problems with observational studies, most importantly confounding and misclassification. In contrast to observational studies, a recent large Mendelian randomisation study found that having a genetic disposition that causes less drinking was associated with a significantly reduced risk of coronary disease, even among those who consume modest amounts of alcohol. Recent research has also challenged some of the purported mechanisms for the protective effect of moderate drinking, eg the effect on and impact of blood lipids.This paper will provide methodological critique of the scientific basis for the claim that low-dose alcohol confers health benefits. It is concluded that the protective effects of moderate drinking may be spurious. Governments should strengthen effective alcohol control policies to reduce alcohol-related deaths, social problems and economic costs. Physician advice to patients should focus on reducing consumption among current drinkers, and should discourage drinking initiation or increased consumption on the basis of health-related considerations.


Menstrual-derived stem cells (MenSCs) are a new source of mesenchymal stem cells isolated from the menstrual fluid. Currently, there is a growing interest in their clinical potential due to fact that they are multipotent, highly proliferative, and easy to obtain in a non-invasive manner. Sampling can be repeated periodically in a simplified and reproducible manner devoid of complications that no existing cell source can match. MenSCs are also free of ethical dilemmas, and display novel properties with regard to presently known adult derived stem cells. This review details their distinctive biological properties regarding immunophenotype and function, proliferation rate, differentiation potential, and paracrine effects mediated by secreted factors. Their possible role in antenatal diagnosis is also discussed. While more insight on their immunomodulatory and diagnostic properties is needed, the impact of clinical and epidemiological factors, such as age, use of contraceptives, or hormonal status still requires further investigations to properly assess their current and future use in clinical application and diagnosis. Therefore, many studies have focused on the search for novel stem cells that can be effectively used for therapeutic purposes without these limitations. While each clinical application will have its own selection criteria for choosing the most appropriate MSCs source, a representation of a decision tree based on six sources of MSCs and five different criteria related to their availability, isolation procedure, and different properties is presented in Figure Mesenchymal stem cells (MSCs) are pluripotent progenitor cells with self-renewing capacity and potential ability of differentiating into various specialized cell types under specific conditions. Adult stem cells are derived from different sources, such as bone marrow, adipose tissue (AD), or post-natal tissues such as umbilical cords and placenta. MSC have recently received a great deal of attention because of their therapeutic potential for treating immune mediated or neoplasic human diseases. However, the difficulty of isolating adult stem cells from diverse tissues due to the invasiveness of the extraction methods and the need for A study published in 2007 identified and characterized a new source of stem cells within the menstrual fluid. They showed that menstrual-derived stem cells (MenSCs) are a highly proliferative stem cell population that is able to differentiate under standard laboratory conditions into specific-tissue cells of three germ layers . These cA detailed characterization of the MenSCs is a pre-requisite for a head-to-head comparison with related cells from other sources. This will pave the way for evaluating possible advantages of MenSCs and also their safety/efficacy profile for clinical applications.Meng et al. showed that MenSCs from the menstrual fluid of young healthy women grew at a rate of one doubling every 19.4 h, which is twice faster than bone marrow-derived MSCs (BM-MSCs), estimated at 40–45 h in early passages . The uppin vitro without any mutation or visible abnormality at the chromosomal level reported so far. They maintained a telomerase activity greater than 50% even at passage (P) 12 compared with human embryonic stem cells  on MenSCs . They alIn addition, these multipotent cells had the ability to differentiate into respiratory epithelial cells, neurocytes, myocytes, endothelial cells, pancreatic cells, hepatocytes, adipocytes, and osteocytes . A recenin vitro, and that differentiation status was determined only phenotypically using specific antibodies , promote regulatory T cell generation (Tregs)  , and to  (Tregs) . While ein vitro  have been identified as constituents of the in vitro cultured MSC secretome. Additionally, several reports also showed that MSCs are able to secrete large amounts of micro and nanovesicles such as exosomes (The potency in tissue restoration mediated by paracrine factors of a broad range of bioactive molecules (secretome) produced by MSCs has raised interest in further exploring this aspect for potential therapeutic applications. This mechanism includes various main actions: immunomodulation, anti-apoptosis, angiogenesis , and supexosomes . The exoexosomes , 41. Exoexosomes , AD 43)ex vivo eexosomes . Severalexosomes . Since MEscherichia coli inhibiting bacterial growth  determined that OGD-exposed primary rat neurons that were co-cultured with MenSCs or exposed to MenSCs-conditioned medium (MenSCs-CM) exhibited a significant decrease in cell death , cytokine growth factors  and angiogenic factors  ll death . It has ll death .+CD133+ HSCs expansion although it was lower than that of the direct cell interactions with the stromal cells. These results suggest that MenSCs might display a quantitative and/or qualitatively distinct “secretome,” or panel of surface molecules capable of exerting distinct contact and paracrine effects on their targets. Furthermore, their protein expression profile can also be modified through the overexpression of factors of interest as they were shown to be permissive for retroviral transduction  cultures, than BM-MSCs under cell-to-cell contact conditions (submitted data). We also showed that the non-contact condition (transwell) resulted in the CD34sduction .Taken together, these studies suggest that MenSCs share some properties with other MSCs but might functionally produce factors that are specific to them. This can be investigated through a comparative analysis of their secretome under different stimulation conditions, including a profiling of their exosome content.6 MenSCs were injected subcutaneously in both male and female immunocompromised NOD/SCID il2rγ null mice. No sign of tumor development or toxicity was detected after a 16 weeks follow-up (unpublished data). In a different experimental setting  conditions with proper product release criteria. Another concern is the development of endometriosis and the possibility of activation or progression of dormant tumors. To address this aspect, we performed a chronic tumorigenicity and toxicity studies, where progressive doses from 1 to 10 setting , injecteude mice . Accordiude mice .The first report of clinical usage of MenSCs involved the allogenic injection of four patients with Multiple sclerosis, with a total dose of 16–30 million cells. Treated patients showed no apparent physical or serological abnormalities at follow-up . More reIn all the MenSCs studies mentioned in this review, cells were isolated from healthy donors. There are no published reports yet characterizing the property changes of MenSCs isolated from epidemiologically different background donors. Thus the effects of age, hormonal status , or prior contraceptive usage remain unexplored. Since stem cells are sensitive to environmental changes and stress conditions, one can only speculate if these variations might affect their function and properties. While it is known that proliferation and therapeutic potential are greatly impacted by the pathological conditions of the donors, little is known on the extent of the effect of these physiological changes on MenSCs. An epidemiological study comparing the secretome, phenotype, and immunomodulatory among other properties would present a valuable guide for the formulation of inclusion and exclusion criteria of donors for a stem cell-based therapy.As MSCs properties are modulated by environment factors, it also becomes important to analyze the role of these changes in pathological conditions.Of the 130 million newborns each year, 8 million die before their first birthday. A contributing factor in many of these deaths is poor pregnancy outcome as a result of a complication of pregnancy, including hypertensive syndromes ; poor fetal growth ; gestational diabetes and preterm birth. Each occurs with an incidence of 5–10% and are responsible for the majority of obstetric and pediatric morbidity and mortality and can permanently impact on lifelong health. As an example, PE has become one of the main causes of maternal and fetal morbidity and mortality in the world, and has also been strongly associated with an increased risk of later-life death due to cardiovascular disease, independent from other risk factors –53. On tIt is now clear that the physiopathological process of many pregnancy diseases begins with an inadequate trophoblast invasion early in pregnancy . SeveralRecent data, suggest that microvesicles (MV) are released from the placenta and their concentration in maternal plasma increases during normal pregnancy , 60. TheAlthough MenSCs have been tested only in very limited disease models, these cells have been shown to possess various regenerative properties under physiological and pathological conditions. From a translational point of view, MenSCs appear to have practical and also biological advantages over other stem cell sources. While some clinical research group and companies launched clinical trials using these cell, these fast developments in the face of lacking data, underscore the need to characterize the differentiation potential and immunological properties of well defined populations of MenSCs. The need for this type of information is decisive with respect to the development of safe and effective cell therapies for clinical application in human diseases.The other to be investigated property of MenSCs is their potential as biomarkers that could be highly informative of the risk of asymptomatic early pregnant women subsequently developing complications of pregnancy. Such tests will offer valuable clinical information that will provide an opportunity for timely and appropriate intervention.Future research and new evidence would greatly contribute to propulse MenSCs to the top list of best proven source of MSCs for new therapies and novel diagnostic tools.Maroun Khoury is the CSO of Cells for Cells and REGENERO, Francisca Alcayaga-Miranda received research funds from Cells for Cells. The other authors declare no conflicts of interest.


Over the past two decades it has become evident that more than 50 % of all heart failure patients suffer of heart failure with preserved left ventricular (LV) ejection fraction (HFpEF), previously called diastolic heart failure because of important involvement of diastolic LV dysfunction. The advances and current status of HFpEF research are described in this special focus issue of the Netherlands Heart Journal. HFpEF is the most common heart failure phenotype in ageing societies, highly prevalent in elderly women and frequently accompanied by comorbidities which result from a detrimental lifestyle, such as obesity, metabolic syndrome, type 2 diabetes mellitus and salt-sensitive arterial hypertension . DespiteA targeted strategy is needed to prevent the progression from obesity and type 2 diabetes towards diastolic LV dysfunction and HFpEF. Available heart failure therapies might exert beneficial effects at an early stage of HFpEF, which is, however, poorly recognised with many patients initially presenting with an advanced form of heart failure. Recognition of an early stage of diastolic dysfunction and initiation of heart failure therapy should be done in primary care, and novel staged therapeutic strategies should be tested to prevent or retard the progression from diastolic LV dysfunction to end-stage heart failure. To battle this life-threatening chronic disease, specialists from different areas of expertise need to join forces.Based on recent findings in cardiac tissue samples from HFpEF patients, a novel paradigm on HFpEF pathogenesis was formulated . This pa


Genomics promises exciting advances towards the important conservation goal of maximizing evolutionary potential, notwithstanding associated challenges. Here, we explore some of the complexity of adaptation genetics and discuss the strengths and limitations of genomics as a tool for characterizing evolutionary potential in the context of conservation management. Many traits are polygenic and can be strongly influenced by minor differences in regulatory networks and by epigenetic variation not visible in DNA sequence. Much of this critical complexity is difficult to detect using methods commonly used to identify adaptive variation, and this needs appropriate consideration when planning genomic screens, and when basing management decisions on genomic data. When the genomic basis of adaptation and future threats are well understood, it may be appropriate to focus management on particular adaptive traits. For more typical conservations scenarios, we argue that screening genome-wide variation should be a sensible approach that may provide a generalized measure of evolutionary potential that accounts for the contributions of small-effect loci and cryptic variation and is robust to uncertainty about future change and required adaptive response(s). The best conservation outcomes should be achieved when genomic estimates of evolutionary potential are used within an adaptive management framework. The rapid pace of human-driven global environmental change is a well-recognized threat to global biodiversity  to track suitable environments. However, whilst tracking environments spatiotemporally can be an important response to environmental change, concomitant factors (e.g. habitat loss and fragmentation) can limit the effectiveness of the response  or over multiple generations through evolutionary adaptation. There is increasing recognition of, and experimental evidence for, the importance of plastic responses to climate change  have not proven good predictors of genome-wide diversity and/or evolutionary potential  outline challenges associated with estimating evolutionary potential in wild populations, (ii) summarize knowledge of the genetic and epigenetic components of evolutionary potential, (iii) review current methods and techniques for characterizing adaptively important variation, (iv) develop recommendations for estimating evolutionary potential in a management context, and (v) outline future research directions. Whilst our primary focus is restricted to genomics as a tool for estimating evolutionary potential in wild populations, we would also direct readers to exciting advances in the parallel field of phenomics which, through use of high-dimensional phenotype data, offers an alternative, nongenetic approach to estimating evolutionary potential  and environmental components. To allow comparison across traits and populations, measures of additive genetic variance must be standardized Houle . In quanEven with recent genomic advances , a quantitative genetics approach to measuring evolutionary potential in natural populations has some limitations   Hill , genomicow) Hill . There aow) Hill .The eukaryote genome is broadly comprised of exons , introns (noncoding sequence in a gene) and intergenic regions (noncoding sequence between genes). In the human genome, only ∼1.5% of sequence codes for amino acids, but around 10–15% of sequence is estimated to be functionally constrained, highlighting the evolutionary significance of noncoding sequence (Ponting and Hardison cis-regulatory elements in the upstream 5' untranslated region (UTR), the downstream 3'UTR, the promoter region and within the coding sequence), they can also occur at greater distances (e.g. trans-regulatory elements); enhancers often lie 10–100 kb from their target genes , populations containing large amounts of cryptic variation adapted six times faster to a new chemical environment than did populations with little cryptic variation , releases a variety of different phenotypes that are not expressed under benign conditions (Chen and Wagner Drosophila melanogaster (Takahashi A large amount of genetic variation that is present but invisible at the phenotypic level (i.e. cryptic variation) enables populations to adapt faster to environmental change  is a contagious cancer that has devastated Tasmanian devil Arabidopsis: Atwell et al. Drosophila: Burke et al. It is now understood that many – if not most – complex phenotypic traits are polygenic: they are controlled by a large number of interacting alleles of individual small phenotypic effect  have revealed many novel variants associated with complex traits, but have not proven a panacea; significantly associated variants are collectively still unable to explain a substantial part of heritable variation in complex traits  in humans  could not be attributed to individual SNPs, but could be explained by positive covariance in allelic effects. Similarly, for another well-studied tree species, the loblolly pine Pinus taeda, Eckert et al.  , gene expression studies were unleashed for use in nonmodel organisms by the advent of high-throughput sequencing of RNA (RNA-seq). Many such studies begin with a Expression of many genes is highly tissue-specific , or not currently adaptive, may still contribute to a population's evolutionary potential. Much of this complexity is difficult to detect using standard population genomic methods, and limitations of current methods need to be considered when designing the screening approach, and when basing management decisions on genomic data (e.g. by screening coding and noncoding parts of the genome and not routinely biasing management decisions to large-effect loci). Embracing recent advances in other fields  and factoring in the contributions of polygenic adaptation, as well as cryptic and epigenetic variation, should result in better-informed applications of conservation genomics and better estimates of evolutionary potential than have been previously possible for most wild organisms.sensu Frankham et al. As we are unlikely to know in advance the best course of conservation action, a risk-management approach is appropriate for making initial decisions . There are well-known examples of traits known to be controlled by a small number of large-effect QTLs, including armour plating associated with marine-freshwater divergence in sticklebacks, , anonymous large-effect loci associated with the adaptive divergence can be detected using standard genome-scan methods  that provides resistance to disease epizootics in the wild could be used to inform selection of resistant or immune individuals for captive breeding, reintroductions and other forms of population genetic augmentation. Key examples might include resistance to chytrid fungus in frogs (Savage and Zamudio Bufo marinus), which cause almost certain, rapid death in susceptible individuals, leading to massive population declines and local extinctions  that will be required. An extreme illustration is provided by toxic invasive cane toads variances, genetic correlations and genetic architecture) will be limited, and/or the predictability of environmental change and the required adaptive response(s) will be low. Under this scenario, screening genome-wide variation should be a sensible approach that may provide a generalized measure of evolutionary potential that accounts for the contributions of small-effect loci and cryptic variation and is robust to uncertainty about future change and required adaptive response(s). We use the term ‘genome-wide variation’ in a broad sense to reflect variation sampled representatively across the genome, which could encompass a range of metrics, including allelic diversity, heterozygosity and weighted measures that place greater importance on specific loci of interest within the context of genome-wide variation.FST-outlier loci. Second, and more significantly, for highly polygenic traits, genome-wide variation currently provides the best predictions of phenotypes  were derived using the relatedness matrix and then validated using extensive simulations and known pedigree information.In some conservation situations , which may often be challenging in wild populations , additive genetic (co)variances and genetic correlations could be estimated in natural populations using large-scale genomic data without the need for detailed pedigrees is an exciting one and would open up a raft of possibilities for exploring trait architecture and evolutionary potential in nonmodel systems . The authors were able to identify major QTL influencing the thermal sensitivity of flowering time and to build a genetic model that was able to successfully predict flowering time of given genotypes in their future climate scenarios. Approaches that link genotypes to their environments within an experimental evolution framework can identify the loci that may promote adaptive responses to environmental change. Although there are obvious limitations to applying these experimental approaches directly to many wild populations, there is good scope for application to commercially valuable crop and animal species and potential for some information to transfer to related wild taxa  under future environments  have looked for correlations between traditional measures of standing genetic diversity (e.g. based on a handful of microsatellite markers) and either fitness and/or evolutionary potential in response to environmental stressors (Reed and Frankham A feasible approach might be to calibrate genomic estimates of evolutionary potential in nonmodel organisms with estimates from model populations where evolution has been more rigorously measured. Past empirical studies of model organisms (e.g. Here, we have outlined some of the complexity regarding adaptation genetics and explored limitations and recent advances in the application of genomic tools to conservation. Recent advances in the fields of quantitative and human genetics have revealed that adaptively important variation is dispersed throughout the genome, with regulatory variants and epigenetic mechanisms playing an important role in shaping gene expression and rapid adaptive evolution. Polygenic adaptation is widespread and even cryptic genetic variation that is hidden at the phenotypic level may make an important contribution to evolutionary potential. The complexity of adaptation genetics needs to be appropriately considered when planning genomic screens and subsequently when basing management decisions on genomic data. When the genomic basis of adaptation and future threats are well understood, it may be appropriate to focus management on particular adaptive traits that are likely to be of importance. For more typical conservation situations, we argue that screening genome-wide variation may provide bet-hedging estimates of evolutionary potential that account for small-effect and cryptic variants and are relatively robust to uncertainty about future environments and required adaptive change. Mining such data to understand adaptation within an adaptive management framework of conservation actions, integrating genomic advances into a quantitative genetics framework, empirically testing the effectiveness of genomic measures of evolutionary potential relevant to population persistence, and exploration of the relative contributions of epigenetic and genetic mechanisms to evolutionary potential are exciting fields in the development of conservation genomics.


Irritable bowel syndrome (IBS) is a common functional gastrointestinal disorder. While abdominal pain is a dominant symptom of IBS, many sufferers also report widespread hypersensitivity and present with other chronic pain conditions. The presence of widespread hypersensitivity and extra-intestinal pain conditions suggests central nervous dysfunction. While central nervous system dysfunction may involve the spinal cord  and brain, this review will focus on one brain mechanism, descending pain modulation.We will conduct a comprehensive search for the articles indexed in the databases Ovid MEDLINE, Ovid Embase, Ovid PsycINFO and Cochrane Central Register of Controlled Trial (CENTRAL) from their inception to August 2015, that report on any aspect of descending pain modulation in irritable bowel syndrome. Two independent reviewers will screen studies for eligibility, assess risk of bias and extract relevant data. Results will be tabulated and, if possible, a meta-analysis will be carried out.The systematic review outlined in this protocol aims to summarise current knowledge regarding descending pain modulation in IBS.CRD42015024284PROSPERO  Irritable bowel syndrome (IBS) is a chronic gastrointestinal disorder characterised by abdominal pain, discomfort and altered bowel habit .A significant amount of research suggests that the abdominal pain associated with IBS is due to peripheral changes occurring in the gut  –4. HowevWhile peripheral mechanisms remain important in IBS, when widespread hypersensitivity and extra-intestinal pain syndromes occur, this suggests a central nervous system involvement –11. CentThe term descending pain modulation (DPM) is used to describe the process mediated by pathways descending from the brain stem that modulate spinal cord ascending nociceptive transmission. Descending neural modulatory circuits from the brain are able to inhibit or facilitate ascending nociceptive transmission and are also known to be influenced by cognitive processes and mood. Ultimately, changes in DPM are felt to profoundly influence pain perception.We will undertake a systematic review of the research examining descending pain modulation in IBS. As part of this review, we will examine the influence of cognitive processes and mood on descending modulation .This systematic review will summarise current knowledge and identify unresolved questions regarding descending pain modulation in IBS.This systematic review aims to answer the following research questions: (1) Do people with IBS when compared with healthy controls demonstrate a consistent difference in descending pain modulation as assessed by neurophysiological testing  and (2) If descending pain modulation is altered in people with IBS compared with healthy controls, what is the magnitude of that difference?The protocol of this systematic review has been registered on PROSPERO 2015   from their inception to August 2015. A comprehensive search strategy has been designed with the assistance of an experienced research librarian and adjusted to account for differences in indexing across databases. Articles identified through reference lists of included studies and relevant systematic reviews will be considered for inclusion based on their title. In addition, cross-checking of references, citations in review papers, and communication with scientists who have been working in the field will also be carried out. Non-English language studies will be included, where a translation can be made available. See We will include studies of adults (18 years or older) who have been diagnosed with IBS where the diagnosis has been made according to the ROME criteria .Studies of adults with comorbid intestinal disorders  and/or extra-intestinal disorders  alongside IBS will also be included.The outcome of interest is descending pain modulation function in people with IBS as assessed by neurophysiological testing i.e. conditioned pain modulation (CPM) or offset analgesia (OA). The neurophysiological protocol has to use a painful test stimulus assessed both prior to and during and/or after the presentation of a conditioning stimulus. Descending pain modulation in the control group would be expected to be quantified as normally responsive. Those studies where descending pain modulation in control participants are not quantified as normally responsive, will be examined as a potential source of variability and will be considered as a candidate for a stratified meta-analysis.The study designs considered relevant to this review include cross-sectional studies, case-control studies, cohort studies and randomised controlled trials. Qualitative studies will be excluded. We will exclude intervention studies if assessment of descending pain modulation is only reported after treatment . Reference lists of relevant systematic reviews will be checked in order to identify relevant primary studies, but systematic reviews will otherwise be excluded.Following the initial database search, duplicate papers will be removed and two reviewers will independently screen titles and abstracts of selected studies to identify those, which are potentially suitable and meet the inclusion criteria.These studies will then be reviewed in full text by two reviewers to determine their eligibility using a standardised eligibility criteria sheet. Reasons for exclusion of papers will be recorded when screening full papers. Any discrepancies in this final list will be discussed between the reviewers, with an independent reviewer consulted if a consensus cannot be attained.Data from the included studies will be extracted using a standardised eligibility criteria sheet. Any discrepancies in this final list will be discussed between the reviewers, with an independent reviewer consulted if a consensus cannot be attained. In addition, authors of papers will be contacted where there is incomplete data or data that needs further clarification.The following data will be collected from the included studies. General study information: authors, year of publication, sample size, language; study design: cross-sectional, case-control, observational study or clinical trial; clinical setting: primary care, specialist clinic, hospital outpatient department; participant characteristics: age, gender, classification or diagnostic criteria used, duration of pain, severity of pain; measure of descending pain modulation: conditioning stimulus, test stimulus, outcome measure and time points outcome measure were recorded; measure of cognitive processes and mood: assessment type used.Several risks of bias tools including the non-randomised studies of interventions (ACROBAT-NRSI)  and the The Quality in Prognostic Studies (QUIPS) tool developed by Hayden, van der Windt, Cartwright, Cote´, Bombardier  was alsoWith these limitations taken together, a modified version of the QUIPS risk of bias tool was employed. The risk of bias tool developed for the current review consists of six categories including: 1) The criteria outline by the ROME foundation  to asses The critResults for descending pain modulation function within a study will be tabulated according to the type of measure used for assessment . The effect size for descending pain modulation function within a study will then be calculated using means and standard deviations at baseline (prior to testing) and during or following testing in the patient and control populations. When a variety of effect size measures are reported, they will be converted to a common measure when that is possible, such as to Cohen’s d  or corre2 values [For each measure available, the random effects model of DerSimonian and Laird  will be 2 values  to quant2 values . Where sIn addition, if there are adequate data in the included studies, a separate analysis will also be carried out to investigate the influence that cognitive processes  and mood (e.g. depression and anxiety) have on neurophysiological measures of descending modulation.Lastly, if the heterogeneity of the data does not allow pooling and quantitative statistics, a narrative synthesis of the findings will be undertaken and the implications critically discussed.This systematic review will consolidate our current knowledge of descending pain modulation in IBS.