There is not too much success in the antioxidant treatment of heart deceases in humans. However a new approach is now developed that suggests that depending on their structures and concentrations antioxidants can exhibit much more complicated functions in many pathological disorders. It is now well established that physiological free radicals superoxide and nitric oxide together with their derivatives hydrogen peroxide and peroxynitrite  and reactive nitrogen species (RNS)) play a more important role in heart diseases through their signaling functions. Correspondingly this work is dedicated to the consideration of damaging signaling by ROS and RNS in various heart and vascular disorders: heart failure (congestive heart failure or CHF), left ventricular hypertrophy (LVH), coronary heart disease, cardiac arrhythmias, and so forth. It will be demonstrated that ROS overproduction (oxidative stress) is a main origin of the transformation of normal physiological signaling processes into the damaging ones. Furthermore the favorable effects of low/moderate oxidative stress through preconditioning mechanisms in ischemia/reperfusion will be considered. And in the last part we will discuss the possibility of efficient application of antioxidants and enzyme/gene inhibitors for the regulation of damaging ROS signaling in heart disorders. Heart disease (cardiopathy) and cardiovascular diseases are a group of numerous pathological disorders such as heart failure (congestive heart failure or CHF), left ventricular hypertrophy (LVH), coronary heart disease, cardiac arrhythmias, and so forth, in which signaling processes of reactive oxygen and reactive nitrogen species (ROS and RNS) play an important  role. Contemporary studies identified major sources of ROS and RNS productions: NADPH oxidases (Nox), xanthine oxidase, mitochondria, and nitric oxide synthases (NOS). As a rule, heart and cardiovascular diseases are characterized by ROS overproduction whereas the formation of major RNSs nitric oxide  and peroxynitrite (diamagnetic molecule) can decrease or increase depending on the nature of heart injury. Free radicals are usually considered to be the damaging factors in various pathologies, but on the other hand ROS and RNS are important signaling species in many physiological and pathophysiological processes. For example the critical role of these species has been shown in preconditioning and other survival processes (see below). A major aim of this work is to consider the role of ROS and RNS signaling in various heart and cardiovascular diseases.NADPH oxidases generate superoxide by the one-electron reduction of dioxygen:2O2+NADPHphox) was an important factor of the development of Ang II-induced cardiac hypertrophy independently of the change in blood pressure in mice. Similar effect of NADPH oxidase-derived superoxide was demonstrated by Nakagami et al. [Phagocyte NADPH oxidase Nox2 plays important role in heart injury. Bendall et al.  found thi et al.  Li et ali et al.  showed ti et al. . Similari et al.  demonstri et al. . β) induced NADPH oxidase activation and ROS overproduction that accelerated atherosclerosis, hypertension, and myocardial remodeling in apolipoprotein E-deficient (apoE(−/−) mice. It was found that Rac1 initiated hypertrophic response in the heart dependent on NADPH oxidase-generated ROS . Hingtgephox  on internal membranes in epithelial cells. It was also found that in contrast to the other NADPH oxidase isoforms Nox4 produced mainly hydrogen peroxide and the very small amounts of superoxide. Cytosolic oxidase proteins or the GTPase Rac are not required for the activity of this enzyme. Serrander et al. [NADPH oxidase Nox4 is a NADPH oxidase isoenzyme which plays an important role in heart and vascular diseases. Martyn et al.  suggester et al.  also fou There is a lot of uncertainty in the studies of Nox4-dependent ROS production. Conclusion that Nox4 produces mainly hydrogen peroxide and not superoxide contradicts a majority of the other experimental data. It is possible that unreliable methods such as nitroblue tetrazolium (NBT) reduction were applied for superoxide detection in the above works –16. On t Nox4 is apparently located in different way in cardiac cells comparing to other NADPH oxidases. Thus Kuroda et al.  found th Surprisingly, Zhang et al.  found th Obviously these findings contradict those obtained by Kuroda et al. . In bothZhang et al.  suggestePrincipal reaction catalyzed by xanthine oxidase (XO) is the oxidation of xanthine into uric acid: ablished  and Chamablished  suggesteablished . Althougablished , 27.  However, it is now generally agreed that there is a significant increase in cellular XO level and activity in the cardiovascular system under pathological conditions even though these changes may not be easily detected under physiological conditions. For example Thompson-Gorman and Zweier measured xanthine oxidase-mediated free radical generation in isolated rat heart . They fo Similarly to NADPH oxidases xanthine oxidase stimulated many ROS-dependent heart disorders. Landmesser et al.  showed tα) and induced the activation of xanthine oxidase and superoxide generation leading to coronary endothelial dysfunction in a murine model. Yamamoto et al. [ Duncan et al.  found tho et al.  showed to et al.  demonstrMitochondria are an essential ROS producer in heart and vascular diseases although its significance as a ROS source comparing to NADPH oxidases and xanthine oxidase remains a subject for discussion. It is now well established that superoxide is generated by mitochondria due to electron leak from the two electron carriers of respiratory chain; these sources of superoxide are Complex I and Complex III . Many auα-induced mitochondrial superoxide production impaired respiratory complex I activity and led to mitochondrial damage in the left ventricle (LV) in rats. Mitochondrial depolarization and enhanced ROS production mediated by lipoxygenase and arachidonic acid were probably responsible for arrhythmias following ischemia-reperfusion injury [ Redout et al.  have stun injury . Howevern injury  found thNitric oxide synthases (NOS) catalyze conversion of L-arginine to L-citrulline and nitric oxide but under uncoupling conditions these enzymes also produce superoxide:oxidants . Thus Lioxidants  showed toxidants  found th As NOSs are able to produce both RNS and ROS, the effects of these enzymes on cardiovascular system can be very complicated—they can enhance or diminish heart damage. Nitric oxide is the endothelium-derived relaxing factor (EDRF); therefore its function must be mainly favorable at the heart. However, the diffusion-controlled reaction of nitric oxide with superoxide produces the highly reactive peroxynitrite, a really harmful agent. Khadour et al.  have stuRegulation of superoxide/nitric oxide balance is important for the prevention of heart damage. This balance is partly achieved by the interaction of principal enzymatic ROS producers during pathological changes in heart. Thus Saavedra et al.  showed t Saraiva et al.  suggesteA brief episode of myocardial ischemia makes the heart remarkably resistant to a subsequent ischemia, the phenomenon named ischemic preconditioning. It has now been shown that ROS and RNS signaling play an important role in ischemic preconditioning and cardioprotection. For example it was found that 30 min of ischemia triggered by acetylcholine and an opioid receptor in isolated rabbit hearts stimulated preconditioning which included the activation of ROS- and RNS-dependent cascade of the epidermal growth factor (EGF) receptor, phosphatidylinositol 3-kinase (PI3-K), protein kinase B (Akt), nitric oxide synthase (NOS), and ROS-dependent opening of mitochondrial (mito)K(ATP) channels . This ca Kimura et al.  found th It is usually proposed that both K(ATP) channel opening and ischemic preconditioning protect the ischemic heart by acting at K(ATP) channels in the inner mitochondrial membrane. However Brennan et al.  found th Van-Cuong et al.  studied δ, PI3K, and ERK kinases. It is thought that these processes stimulate the inhibition of mitochondrial permeability transition pore formation and trigger the entrance into the preconditioned state. In recent work Vigneron et al. [β (GSK-3β), the opening of mitoK(ATP), and ROS generation activating the target of rapamycin (mTOR) pathway and induced cardioprotection. These findings suggest that cardioprotection involved a prosurvival mTOR pathway. Cohen et al.  have revn et al.  showed t Overproduction of ROS and deregulation of RNS production are important factors of the development of heart and cardiovascular diseases. Mechanisms of ROS and RNS generation by major producers, NADPH oxidases, xanthine oxidase, mitochondria, and nitric oxide synthases in these diseases as well as in preconditioning were discussed above and presented in κB and cardiomyocyte hypertrophy in mice. As it has been already noted, Akt also participates in preconditioning [Among various enzymes, protein kinases B and C and mitogen-activated protein kinases (MAPK) play a very important role in ROS and RNS-dependent enzymatic cascades responsible for heart damage. One of these kinases is the serine/threonine protein kinase B (Akt). It has already been noted that Akt participates in Nox2-initiated Ang II-dependent cardiomyocyte hypertrophy . Recentlitioning , 65. Theitioning  found thitioning  proposedδ, and PKCε in heart damaging processes has been also demonstrated [δ or selective activation of PKCε reduced oxidative damage in the heart following myocardial infarction. cGMP-dependent protein kinase (PKG) showed protective activity in the heart [ Participation of protein kinases PKC, PKCnstrated , 71, 87.he heart , 89.Gaitanaki et al. [β-Estradiol (E2) improved congestive heart failure (CHF) in rats by antioxidative mechanism that involved thioredoxin (Trx) upregulation, the inhibition of Rac1 mediated NADPH oxidase activity, and the apoptosis signal-regulating kinase 1(ASK-1)/JNK/p38-mediated apoptosis. Cai et al. [α inhibited endothelium-dependent NO-mediated dilation of coronary arterioles from porcine heart by the ceramide-induced activation of JNK and subsequent production of superoxide by xanthine oxidase. Widder et al.  found thi et al.  showed ti et al.  demonstri et al.  showed ti et al.  studied i et al.  showed tvia a novel zinc-finger protein, MCPIP (MCP-1-induced protein). MCPIP stimulated enzymatic cascade through the activation of MAP kinases JNK and p38. These findings suggested that MCPIP induced ROS/RNS production that stimulated ER stress, autophagy and apoptosis. Hikoso et al. [κB in cardiomyocytes in response to pressure overload. They demonstrated that IκB kinase (IKK)-(NF-κB) signaling cascade was protective in cardiomyocytes due to the attenuation of oxidative stress and JNK activation. Younce and Kolattukudy  studied o et al.  investigα) is upregulated in a number of cardiomyopathies inducing adverse cardiac remodeling and dilation due to the degradation of the extracellular matrix by matrix metalloproteinases (MMPs). Awad et al. [α (rTNF) induced stronger superoxide production and increased expression of several MMPs in mouse neonatal cardiomyocytes comparing to cardiofibroblasts. Phosphatidylinositol 3-kinase (PI3K-γ) mediated TNF-dependent superoxide production and MMP expression. Lu et al. [ It has been shown that tumor necrosis factor  proteins depend on ROS levels and can stimulate or decrease longevity of experimental animals. Klotho gene regulates cell senescence and aging. Mechanisms of ROS regulation in such gene/enzymatic processes were recently discussed [ It is known that several genes  regulate ROS formation in pathological states. Thus it has been shown that the suppression of iscussed .  These genes can also participate in ROS-dependent heart damage. Alcendor et al.  showed tα1-adrenergtic receptor (α1-AR) pathway together with protein kinases PKCε and PKCδ and induced Akt-FOXO3a phosphorylation in rat cardiomyocytes. Sengupta et al. [ Guo et al.  showed ta et al.  showed ta et al. .Heart Diseases and Antioxidants), therefore we need to choose some principal studies. Antioxidants and free radical scavengers can suppress free radical-dependent heart disorders by direct reactions with reactive hydroxyl and peroxy free radicals or through the regulation of ROS signaling in gene- and enzyme-catalytic cascades. We will consider some important examples. For a long time antioxidants were considered important pharmacological agents for the treatment of heart diseases. It is impossible to discuss even a small part of works published on this problem  against ischemia-reperfusion injury in the heart. It has been proposed that Sildenafil, a selective inhibitor of phosphodiesterase type 5 induced powerful protection against myocardial I/R injury through the activation of cGMP-dependent protein kinase G (PKG). Das et al.  suggeste Thus antioxidants of various classes might be considered to be of potential use for the prevention and maybe treatment of heart and cardiovascular diseases.The data discussed in this work show that practically all pathological disorders in heart and cardiovascular system are associated with damaging ROS signaling. These damaging processes can be initiated by ROS overproduced by various sources such as NADPH oxidases, mitochondria, xanthine oxidase, and NO synthases , or normα, or monocyte chemotactic protein-1 (MCP-1) are responsible for ROS overproduction. ROS overproduction might be also initiated by cellular disorders for example initiated by hyperthermia  increased the expression of thioredoxin (Trx) and the suppression of NADPH oxidase/ASK-1/c-Jun/p38 cascade causing a decrease in apoptosis and the stimulation of CHF recovery [ On the other hand ROS signaling in enzymatic/gene cascades might also decrease disorders in heart and cardiovascular system. As is seen from farction . 17β-estrecovery . Decreasrecovery , 78.  It is of interest that the participation of certain genes in signaling pathways frequently led to diminishing heart disorders. As is seen from  As ROS overproduction is one of the most important stimulators of heart disorders, it is logically to suggest that the application of antioxidants should be useful for fighting the heart and cardiovascular diseases. It has been shown above that the effects of antioxidants are very complicated and changed from direct interactions of antioxidants with reactive free radicals to the influence of enzymatic/gene pathways. For example protein kinases and the antioxidant/prooxidant genes can enhance or diminish ROS formation stimulating surviving and death of cardiomyocytes.  Another source of difficulty is dependence of the effects of antioxidants or prooxidants and various ROS inhibitors dependent on the levels of oxidative stress. As it was pointed out, low levels of ROS (low oxidative stress) might have prosurvival effects while high ROS levels (severe oxidative stress) will damage biomolecules. These effects can in turn be complicated by the survival effects of prooxidants-induced preconditioning and the induction of antioxidant enzymes (MnSOD and CuZnSOD).  Therefore a major question is how we might take into account all miscellaneous effects of antioxidants, prooxidants, and gene/enzymatic inhibitors for the selection of the right compounds and methods in the antioxidant (or prooxidant) treatment of heart diseases? Of course there is no simple answer. However, it is probably useful before beginning the treatment of patients to consider possible effects of the selected compound in major ROS and RNS signaling processes.


Vitamin C and the Common Cold” Linus Pauling included an interesting chapter on “human biochemical individuality” that defined some important parameters on individual human genotypic versus phenotypic variation, based in part on studies from hemoglobin genetics . If we assume that there are about 26,600 human genes available to be expressed in each cell and that each gene is responsible for at least one inherited trait or genetic function (a number that is probably vastly underestimated), in a global human population exceeding 7 billion it then becomes exceedingly difficult to define “human genetic normalcy.” These ideas form the basis for the evolving concept of “human genetic individuality” and our ongoing efforts to better understand the genotypic basis of human phenotypic diversity in both health and disease  and miRNA abundance, speciation and complexity in defined brain anatomical regions from different human samples. These studies have been very valuable since the profiling of mRNA and/or miRNA can provide a powerful “snapshot” into the physiological status of a human cell or tissue in health and disease, and may even be predictive for the prognosis and/or diagnosis for the future outcomes of other AD patients. Steady-state mRNA and miRNA levels from different individuals clearly indicates that the abundance and speciation of these RNAs within clearly defined anatomical regions can significantly differ between samples analyzed, suggesting that genetic variation and extraneous effects, including age, gender, body mass index (BMI), apolipoprotein E (ApoE), beta-amyloid cleavage enzyme (BACE) and other AD-relevant allele status, life-style and intrinsic population effects can influence the profile of mRNA or miRNA abundance and complexity (Colangelo et al., APOE genotype (Tang et al., To illustrate one important example is the miRNA abundance and speciation of a small family of inducible, NF-kB-sensitive miRNAs in two different American populations—Caucasian Americans and African Americans afflicted with AD Figure . A pathoVariation in miRNA patterns lends further strength to the idea that AD is not a single, definable neurological disease entity such as sickle-cell anemia Pauling, , but rat“human genetic individuality.” If molecular-genetic and epigenetic profiles of AD brain samples are any indication of AD phenotypic variation then there may be real and significant inter-ethnic differences in AD epidemiology, incidence, disease course and progression. This further suggests that an equally wide variety of diagnostic and individualistic prevention and treatment strategies will be required to more effectively address such progressive, age-related neurological disorders of the human CNS, including the implementation of novel combinatorial therapeutic strategies such as anti-NF-kB and anti-miRNA approaches that have not yet been considered (Lukiw, Lastly, much independently derived data comparable to that shown in Figure d Lukiw, .


Dear Editor,I read with great interest the work published by Mogadam et al., entitled "Comparison of Analgesic Effect between Gabapentin and Diclofenac on Post-Operative Pain in Patients Undergoing Tonsillectomy" . I found


Synovial sarcomas commonly occur in the soft tissue of the extremities, while a primary occurrence in the mediastinum is quite rare. The current study reports the case of an 11-year-old male who presented with a neck mass, which computed tomography showed was due to a giant mediastinal mass involving the thyroid gland. The tumor was resected by thoracotomy and diagnosed as monophasic synovial sarcoma by histopathology. The patient received adjuvant combination chemotherapy and radiation therapy following surgery. At the 3-month follow-up, no local tumor recurrence was found. The present case report highlights the significance of recognizing the unusual presentation and clinical manifestation of synovial sarcoma to aid clinical management. Written informed consent was obtained from the patient’s family. Synovial sarcoma is a type of mesenchymal tissue cell tumor that exhibits epithelial differentiation, which most frequently arises in the extremities, while a primary occurrence in the mediastinum is quite rare ,2. Prima2 in the left thyroid gland, however nothing of significance was revealed in the chest or abdominal regions. Computed tomography (CT) scans  scans  and 2 shmination  showed mmination –7, confimination  showed amination  was perfet al in 2002 as a type of mesenchymal tissue cell tumor that exhibits epithelial differentiation , which met al, of the Primary mediastinal synovial sarcoma is a type of malignant tumor with no specific differences from other mediastinal tumors with regard to clinical manifestation, imaging or histology, therefore, it is difficult to diagnose. With regard to clinical manifestations, mediastinal synovial sarcomas reveal various initial symptoms due to their different scopes of infringement. The common symptoms include chest pain –7, shortet al Although mediastinal synovial sarcomas have limited clinical data and no standardized treatments, complete surgical excision remains the cornerstone of therapy ,6,8,9. Aet al retrospe al(et al, adjuvan al(et al also fou al(et al. In the  al(et al, the 5-y al(et al recently al(et al,9,17. Th al(et al and 9 sh al(et al, it was  al(et al must be A previous study  showed tMediastinal synovial sarcomas are malignant tumors with a low incidence, no specific clinical manifestations and a lack of unified and effective treatments. These factors highlight the challenges preventing its diagnosis and treatment in clinics. The existing data supports surgery as the preferred treatment for mediastinal synovial sarcoma, but appropriate auxiliary treatments, including radiotherapy and chemotherapy, must be taken into consideration according to the factors affecting prognosis. In addition, the SYT-SSX fusion gene in synovial sarcoma must be further investigated in hope of identifying novel targeted therapies.


Clustering and classification of large-scale chemical data are essential for navigation, analysis and knowledge discovery in a wide variety of chemical application domains. The maximum common structure (MCS) for a group of compounds is an important element of such classification, providing insight into activity patterns and enabling scaffold alignment for a more consistent 2D depiction. Most modern, exact MCS implementations use back-tracking  or cliqu


A goal of the COG Ewing Sarcoma (ES) Biology Committee is enabling identification of reliable biomarkers that can predict treatment response and outcome through the use of prospectively collected tissues and correlative studies in concert with COG therapeutic studies. In this report, we aim to provide a concise review of the most well-characterized prognostic biomarkers in ES, and to provide recommendations concerning design and implementation of future biomarker studies. Of particular interest and potentially high clinical relevance are studies of cell-cycle proteins, sub-clinical disease, and copy number alterations. We discuss findings of particular interest from recent biomarker studies and examine factors important to the success of identifying and validating clinically relevant biomarkers in ES. A number of promising biomarkers have demonstrated prognostic significance in numerous retrospective studies and now need to be validated prospectively in larger cohorts of equivalently treated patients. The eventual goal of refining the discovery and use of clinically relevant biomarkers is the development of patient specific ES therapeutic modalities. First described by James Ewing as an endothelioma of bone Ewing, , Ewing sIt was the advent of consistency in diagnosis that enabled cooperative groups worldwide to develop multi-center ES clinical trials. Over the past three decades these trials have systematically evaluated and optimized local and systemic treatment protocols for patients with ES , and increasing patient age have all been implicated as negative prognostic features. None of these are as significant as the presence of metastatic disease and studies have demonstrated variability in these individual features , and sub-clinical disease measurement. Fusion type will be discussed to demonstrate the importance of prospective evaluation and validation of biomarkers in the context of evolving therapy. The remaining categories were selected for in depth discussion after consideration of REMARK criteria. In the following sections we will highlight the features of each of these putative biomarkers that lead us to propose that their parallel evaluation and validation in the next series of prospective therapeutic trials is warranted. Several of these, including CNAs and cell-cycle proteins were recently discussed at a European Network for Cancer Research in Children and Adolescents (ENCCA) summit of 35 international experts   and RB1. Kovar et al. (CDKN2A deletions in 30% of tumors (N = 8/27) and 52% of ES cell lines (N = 12/23) and several retrospective studies have demonstrated an association between CDKN2A alterations and clinical outcome in ES patients. Wei et al. , while Tsuchiya et al. (CDKN2A deletions in 17% of tumor samples (N = 4/24). Patients in both studies were found to have worse disease-specific survival in univariate and multivariate analyses. Maitra et al. (CDKN2A downregulation by immunohistochemistry in 20% of patients (N = 4/20), and this correlated with metastatic disease at presentation and trended toward shortened survival. A meta-analysis examining the prognostic significance of CDKN2A alterations in ES based on six separate studies (N = 188) concluded that the estimated pooled risk ratio (RR) for worse outcome with CDKN2A alterations was 2.17  and the estimated pooled RR of metastasis at diagnosis was 2.60   and 10% of primary tumors (N = 4/42)  and this over-expression was associated with advanced disease at diagnosis, poorer treatment response, and a worse overall survival. Significantly, this effect was independent of site, local treatment, or tumor necrosis. Similarly, a study by de Alava et al. (N = 6/55) and increased p53 protein expression was found to be the strongest prognostic factor that was associated with worse overall survival. Huang et al. (TP53 mutations in 13.3% of patient samples (N = 8/60), as well as CDKN2A homozygous deletions in another 13.3% of samples (N = 8/60). TP53 mutations and/or CDKN2A deletions were significantly associated with a poor response to chemotherapy (P < 0.0001) and, in a multivariate analysis, TP53 and/or CDKN2A alteration status as a single combined variable was identified as the most significant prognostic factor (P < 0.001). Finally, using immunohistochemistry and fluorescent in situ hybridization (FISH), Lopez-Guerrero et al. (P = 0.025), and worse progression-free survival (P = 0.012) and disease-specific survival (P = 0.006) in patients with localized disease.The potential of u et al.  detecteda et al.  identifig et al.  reportedo et al.  analyzedTP53 and CDKN2A as negative prognostic biomarkers in ES. Currently, COG and the ES Biology Committee are performing a large-scale analysis of TP53 and CDKN2A status in over 150 prospectively collected tumors from patients treated on the most recent AEWS0031 therapeutic study. Should this study confirm prior observations, analysis of these cell-cycle regulatory proteins will become a strong candidate for inclusion as a prognostic biomarker that can inform treatment decisions in future clinical trials.In summary, compelling data from several retrospective studies implicates alterations of Genomic instability with subsequent CNAs have been well-documented in ES and these alterations have been recently reviewed by Jahromi et al. . The recIn summary, independent studies of both small and large tumor cohorts have identified individual and global patterns of CNAs as putative prognostic biomarkers in ES. We anticipate that the continued improvement in next generation sequencing platforms will allow for greater characterization of structural variations in tumors, and will generate even more data to test associations between CNAs and clinical outcome. It is the recommendation of this committee that tumor and germline DNA be collected from all patients registered on future therapeutic studies of ES in order that CNAs and other genetic mutations can be evaluated as prognostic and predictive biomarkers in homogeneously treated patients. To that end, COG has discussed the prospective incorporation of CNA and genomic analysis in their upcoming ES trial for relapsed/refractory patients.Assessment of minimal residual disease (MRD) has been established as a critical part of therapeutic decision making in childhood acute lymphoblastic leukemia  protocols and therefore received similar therapy , and 19% of patients (N = 18/92) with non-metastatic disease at presentation. Circulating transcripts were identified in 20% of patients (N = 29/144) at diagnosis, and were more frequently observed in patients with large tumor burdens. In patients with localized disease, RT-PCR positivity in bone marrow and peripheral blood correlated with significantly poorer outcomes. In contrast, a study of peripheral blood samples from 26 children was unable to identify a significant progression-free survival difference in patients with detectable fusion transcript at diagnosis  with localized disease, 50% of patients (N = 3/6) with isolated pulmonary metastases, and 100% of patients (N = 6/6) with bone metastases. However, the study did not establish a correlation between marrow positivity for ES transcript and progression-free disease. Results of other smaller studies have been recently summarized by Wagner et al.  for molecular diagnosis, isolation of quality RNA has become less practical. Feasibility will only diminish as additional rare non-EWSR1 translocations are identified.To rigorously address whether the detection of circulating tumor transcript is of prognostic significance, the multi-center European EURO-E.W.I.N.G. 99 trial prospectively collected bone marrow samples for over 10 years. As the first large prospective trial examining sub-clinical disease via RT-PCR in ES patients, the findings of this study will be critical to evaluate the feasibility and usefulness of this modality as a biomarker for ES. Based on our own experience with a much smaller cohort of patients in COG we, as a committee, are skeptical that RT-PCR-based assays will be clinically optimal for prognostication and treatment stratification. We base this assertion on our combined observations regarding issues of technical reproducibility of the assay between individual laboratories, and the technical expertise required to consistently obtain sufficient quality RNA for valid and reliable RT-PCR analysis. Although these issues could be addressed with the establishment of a central College of American Pathologists (CAP)-Clinical Laboratory Improvement Act (CLIA)-certified reference laboratory, the issue of RNA degradation in sample shipments would remain. In addition, RT-PCR-based analysis requires knowledge of the precise breakpoint. With the increasing use at many COG institutions of closed needle biopsy for diagnostic tissue collection and fluorescence In summary, although of potential prognostic significance, technical and logistic realities regarding tissue collection and RNA-based studies of blood and bone marrow specimens significantly diminish this committee’s enthusiasm for RT-PCR analysis of sub-clinical disease in routine clinical practice. Should the aforementioned Euro-Ewing study validate RT-PCR of bone marrow as a significant prognostic variable, this issue will need to be re-addressed. At such time, consideration would need to be given to optimizing collection and submission of quality RNA and to creation of a CAP-CLIA certified COG reference laboratory.5 peripheral blood mononuclear cells. Ash et al. -directed therapy. Activating mutations in the EGFR gene are detectable in only a small minority of NSCLC patients but it is these patients who selectively respond to EGFR-directed therapy  patients who will respond to Epidermal Growth Factor Receptor (in vitro and in vivo models of ES are highly sensitive to the PARP1 inhibitor Olaparib alone and in combination with the drug temozolomide. Moreover, in a drug screening of several hundred cancer cell lines a marked and selective susceptibility of ES cell lines to Olaparib was also discovered (Garnett et al., The findings that only a small subset of patients with relapsed ES respond to IGF-1R targeted monotherapy serve as a sobering example of the critical need for predictive biomarkers in this disease. As trials investigating novel agents move forward, it is paramount that strategies that will permit evaluation of predictive biomarkers be simultaneously implemented. This will enable identification of patients who may preferentially benefit from such interventions in the future and allow for more selective inclusion and exclusion of patients in a manner that will lead to improved response rates. One potential treatment modality to emerge from recent pre-clinical investigations is PARP1 inhibition. PARP1 is a key enzyme involved in single-strand repair of DNA (Wang et al., Numerous prognostic biomarker studies for ES have been published in recent years. Of particular interest and potentially high clinical relevance are studies of cell-cycle proteins, sub-clinical disease, and CNAs. All of these have demonstrated prognostic significance in numerous retrospective studies and now need to be validated prospectively in larger cohorts of equivalently treated patients. The challenges in identifying and validating clinically relevant biomarkers in ES highlight a significant hurdle for the individualization of therapy in any rare cancer. Prospective therapeutic trials with standardized treatments remain the optimum source of biologic material and clinical correlative information to drive successful biomarker identification. Since these trials can take years to complete it is essential that biomarker studies be meticulously designed and incorporated up front in therapeutic studies. It is imperative that these studies are designed vigilantly to maximize levels of evidence and ensure adherence to REMARK guidelines. In addition, biomarkers that can be tested and validated on blood or fixed tumor specimens will have the best chance of translation into routine clinical practice. As new agents are developed, predictive biomarkers will need to be developed to assess the benefit of these therapies and rationally design treatment stratification based on likelihood of response. The choice of technical platforms must also be carefully considered in trials involving rare diseases. Although characteristics such as sensitivity are important when choosing a methodology, issues such as availability, cost-effectiveness, and sample requirements are equally important. Rare cancers require the participation of multiple institutions, and it is imperative that samples from each site are similarly collected and processed. Cooperative groups can play a critical role to ensure that biomarker studies are carefully selected, rigorously designed and, whenever possible, incorporated into therapeutic studies.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.


Graft versus host disease (GVHD) is the major complication of allogeneic hematopoietic stem cell transplantation. GVHD is characterized by an imbalance between the effector and regulatory arms of the immune system which results in the over production of inflammatory cytokines. Moreover, there is a persistent reduction in the number of regulatory T (Treg) cells which limits the ability of the immune system to re-calibrate this proinflammatory environment. Treg cells are comprised of both natural and induced populations which have unique ontological and developmental characteristics that impact how they function within the context of immune regulation. In this review, we summarize pre-clinical data derived from experimental murine models that have examined the role of both natural and induced Treg cells in the biology of GVHD. We also review the clinical studies which have begun to employ Treg cells as a form of adoptive cellular therapy for the prevention of GVHD in human transplant recipients. Although hematopoietic stem cell transplantation (HSCT) has been a successful therapeutic strategy for treating hematological malignancies for several decades, its broad application is limited by the high incidence of graft versus host disease (GVHD). GVHD is primarily a donor T cell-mediated syndrome whereby T cells in the graft elicit an immune response, resulting in host tissue damage . These cells, termed Treg cells, express the forkhead box transcription factor Foxp3, which is both necessary and sufficient for the suppressive ability of Treg cells  cells comprise 5–10% of the CD4+ T cell compartment and develop in the thymus  which are the sites of GVHD-associated tissue damage  which is a ligand for the CD1d molecule has been shown to expand donor-derived Treg cells in a dose-dependent manner and reduce GVHD-associated mortality (Duramad et al., in vivo expansion of nTreg cells and in vivo conversion and/or expansion of iTreg cells. Thus, it is difficult to exclude that these approaches may also result in the expansion of iTreg cell populations as well.Pharmacological strategies have also been tested in murine GVHD models to determine whether Treg cell numbers can be augmented after allogeneic HSCT. To that end, Shin et al.  demonstrde novo iTreg cell generation in recipient mice is negligible during GVHD (Chen et al., in vitro induction/expansion of this population followed by adoptive transfer into recipient animals. In initial studies, iTreg cells were stimulated with allogeneic dendritic cells or treatment with anti-CD3/anti-CD28 antibodies in the presence of TGF-β and IL-2 to induce Foxp3 expression. Administration of in vitro-differentiated iTreg cells along with BM grafts containing alloreactive donor T cells did not result in any significant protection from lethal aGVHD (Koenecke et al., in vivo survival of these cells which was accompanied by instability of Foxp3 expression, resulting in a loss of suppressive function early post transplantation (Koenecke et al., While the majority of rodent models of GVHD have focused on the biology of nTreg cells, there has been much less attention devoted to the role of iTreg cells in GVHD biology. This has been due, in part, to the fact that there are no proven cell surface markers that distinguish nTreg cells from iTreg cells. Consequently, isolation of a pure iTreg cell population from donor animals for selective adoptive transfer studies is not currently feasible. Furthermore, in vivo is not clear, but one potential explanation is that the proinflammatory cytokine milieu that occurs during GVHD may also render iTreg cells more unstable. Supporting this premise are data demonstrating that in vivo-derived iTreg cell conversion is significantly enhanced when mice are treated with monoclonal antibodies that block signaling through IL-6 or IL-21 which serves to reduce inflammatory cytokine production (Bucher et al., in vivo, an alternative approach has involved the culture of CD4+CD25− T cells with the hypomethylating agent 5-azacytidine. Choi et al. (+ T cells both in vitro and in vivo, and that transplantation of these cells ameliorated GVHD severity.The reason that iTreg cells are unstable i et al.  reported+ Treg cell populations.We would note that instability of Foxp3 expression has also been noted to occur in nTreg cells in non-transplant models (Zhou et al., + Treg cells are classically defined as being a subset of the CD4+ T cell compartment. However, a CD8+ Foxp3+ Treg population has been described and found to be capable of suppressing T cell responses in animal models of autoimmunity and allergen exposure (Hahn et al., + Foxp3+ T cells have also been documented in the tumor microenvironment of patients with colon and prostate cancer, suggesting that they may be a mechanism by which tumors escape immune surveillance (Kiniwa et al., + Foxp3+ iTreg cells are induced early during GVHD (Beres et al., + counterparts, these cells were found to be dependent on TGF-β and IL-2 for induction (Sawamukai et al., + or CD8+ iTreg cell population was required to prevent increased GVHD-associated mortality (Beres et al., + iTreg cells could be expanded in GVHD recipients using IL-2 antibody complexes in conjunction with Rapamycin as has been previously described with CD4+ Treg cells (Shin et al., + Foxp3+ T cells have been induced in vitro and found to suppress GVHD in a humanized mouse model (Zheng et al., in vitro or in vivo methodological approaches for translational application.Foxp3+ Treg cells to CD8+ T cells was significantly decreased at the mucosal interface of GVHD patients as compared to patients with intestinal inflammation unrelated to GVHD (Rieger et al., The approach that has been employed to address whether Treg cells may serve to modulate the severity of GVHD in man has been to correlate the absolute number and/or frequency of Tregs with the subsequent incidence and severity of aGVHD and cGVHD. Several reports have demonstrated a decreased frequency of Treg cells in the peripheral blood of patients with high clinical grades of aGVHD as compared to patients with lower grade aGVHD or no GVHD (Li et al., +CD25hi Treg cells as compared to individuals without GVHD. This was supported by a more recent study that reported increased peripheral Treg cell numbers in transplant recipients that developed cGVHD with no prior aGVHD diagnosis (Ukena et al., It is important to note, however, that not all studies have demonstrated a correlation between reduced Treg frequency and GVHD severity. Clark et al.  observedThe reason for the differences observed in these studies is not entirely clear. For the most part, however, studies that have failed to demonstrate that a reduction in Treg cell frequency and/or absolute numbers is associated with increased GVHD severity have relied on CD25 expression to delineate Treg cell populations, whereas those that have reported a positive correlation have tended to employ Foxp3 expression as a readout for this Treg cell population. Thus, it is possible that the reliance on different phenotypic markers may result in somewhat different populations being examined and be a potential explanation for these discordant results.+Foxp3+ Treg cells in the peripheral blood of the donor negatively correlated with the incidence of GVHD in the graft recipient. Several subsequent studies confirmed this correlation in recipients of HLA-identical sibling and unrelated donor stem cell grafts (Pabst et al., +CD25+CD127lo Treg cells as compared to bone marrow grafts, the frequency of peripheral blood Treg cells is reversed post transplantation. This was presumed to be due to the fact that PBSC Treg cells tended to be CD62Llo as a consequence of both granulocyte colony stimulating factor treatment for mobilization and the subsequent leukapheresis process (Blache et al., + Treg cells in the graft have been found to correlate with reduced GVHD incidence (Lu et al., + Treg cell population is more potent at suppressing GVHD than the corresponding CD62Llo population (Taylor et al., An alternative approach to examine the effect of Treg cells on GVHD severity in human allogeneic HSCT has been to assess the number of donor-derived Treg cells within the graft prior to transplantation. In this regard, Rezvani et al.  determin+CD25hi CD127− or CD4+CD25hiICOS+ Treg populations were likely to be most suitable for human adoptive transfer studies. Many groups have also identified in vitro expansion protocols that yield high number of Treg cells for adoptive transfer (Karakhanova et al., ex vivo-expanded cells also ameliorated disease in a xenograft model of GVHD (Chakraborty et al., Less than two decades after their discovery, Treg cells are now entering into clinical trials in allogeneic HSCT recipients. Pre-clinical murine models of GVHD have provided much insight into Treg cell-based therapy, but most mouse studies have been performed using Foxp3-GFP reporter mice where Foxp3-expressing Treg cells can be definitively isolated for adoptive transfer studies. This is not a luxury that is available in human studies where CD25 expression necessarily serves as a surrogate for Foxp3. However, since CD25 is upregulated on all activated T cells, further phenotypic characterization of these cells has been generally thought to be necessary for their use in man. To that end, Ukena et al.  comparedOne of the first reported clinical studies was conducted by Brunstein et al. who performed a phase I clinical prophylaxis trial with cord blood-derived Treg cells. The rationale for the use of cord blood-derived Treg cells was based, in part, on earlier studies, that had shown that they express similar levels of CTLA-4, Foxp3, GITR, and CD25 as adult peripheral blood Treg cells, and when stimulated by alloantigen, were potent suppressors of T cell expansion (Chang et al., A second clinical trial was performed by Di Ianni et al. , in whicThere has been one small study involving two patients in which expanded Treg cells were administered to patients with documented GVHD (Trzonkowski et al., + donor leukocyte infusions, used to treat relapsed hematologic malignancies post HSCT, resulted in Treg cell expansion in vivo. This same group then utilized this strategy to treat glucocorticoid refractory cGVHD patients. The administration of low dose IL-2 was associated with an amelioration of disease severity and this correlated with an increase in the number of Treg cells (Koreth et al., in vivo expansion of Treg cells may be a more clinically feasible strategy to enhance Treg reconstitution post transplantation, as compared to more costly expansion strategies.An alternative approach to harness the potential for Treg cell therapy in humans is based on the requirement of these cells for IL-2. Specifically, Zorn et al.  found thCollectively, these studies are exciting evidence that Treg cell therapy has now entered into the clinic. Going forward, well-designed trials will be necessary to determine whether these cells are indeed capable of preventing and/or treating patients with established GVHD.+ nTreg cells, CD4+ iTreg cells, and CD8+ iTreg cells in GVHD biology remain unclear. Elucidating the mechanisms by which the respective cell subsets function may provide insight for developing better therapeutic strategies. Furthermore, additional studies are required to ascertain whether CD4+ and CD8+ Treg cell populations function cooperatively or whether they have overlapping redundant roles in GVHD biology.Despite the significant progress that has been made in understanding the role of Treg cells in GVHD biology, a number of questions remain. First of all, the relative roles of CD4in vitro, have unstable Foxp3 expression. Since Foxp3 expression is necessary for suppressive function, further inquiry is needed to determine whether Foxp3 expression can be stabilized especially under pro inflammatory conditions which characterizes the GVHD milieu (Koenecke et al., Secondly, accumulating evidence indicates that Treg populations, particularly those that are expanded ex vivo expansion remain unresolved. Moreover, it is possible that alloantigen-specific Treg cells may be more potent in suppressing GVHD, and should be studied further (Albert et al., Finally, current Treg cell-based immunotherapy approaches rely on the expansion of polyclonal populations of Treg cells. The best source of Treg cells and the optimal culture conditions for 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.


Neuropeptide W (NPW), which was first isolated from the porcine hypothalamus, exists in two forms, consisting of 23 (NPW23) or 30 (NPW30) amino acids. These neuropeptides bind to one of two NPW receptors, either NPBWR1 (otherwise known as GPR7) or NPBWR2 (GPR8), which belong to the G protein-coupled receptor family. GPR7 is expressed in the brain and peripheral organs of both humans and rodents, whereas GPR8 is not found in rodents. GPR7 mRNA in rodents is widely expressed in several hypothalamic regions, including the paraventricular, supraoptic, ventromedial, dorsomedial, suprachiasmatic, and arcuate nuclei. These observations suggest that GPR7 plays a crucial role in the modulation of neuroendocrine function. The intracerebroventricular infusion of NPW has been shown to suppress food intake and body weight and to increase both heat production and body temperature, suggesting that NPW functions as an endogenous catabolic signaling molecule. Here we summarize our current understanding of the distribution and function of NPW in the brain. In 1995, O'Dowd et al. . In 2002in situ hybridization histochemistry has revealed that NPW mRNA is expressed in a few restricted brain regions, including the rat periaqueductal gray (PAG), Edinger-Westphal nucleus (EW), and dorsal raphe nucleus , PAG, superficial gray layer of the superior colliculus, and subfornical organ. In general, NPBWR1 is most commonly expressed at high levels in the amygdala (Singh et al., In humans, RT-PCR analysis has demonstrated that NPBWR1 mRNA is highly expressed in the amygdala, hippocampus, neocortex, and hypothalamus (Lee et al., i et al.  reportedi et al.  used [12NPBWR1 knockout mice are hyperphagic and show decreased energy expenditure, suggesting that NPW may act as a modulator of feeding. icv infusion of NPW in male rats has been shown to increase food intake during the first 2 h in the light phase (Shimomura et al., ob/ob and db/db mice. Therefore, NPW may play important roles in feeding and energy metabolism, functioning as a substitute for leptin (Date et al., We have carried out a series of neuroanatomical studies to examine the neural relationship between NPW and other neuropeptides involved in the regulation of feeding. Very close neuronal interactions were observed between NPW-containing nerve fibers and orexin- or melanin-concentrating hormone-containing neuronal cell bodies and nerve fibers in the rat brain (Takenoya et al., 2 consumption and increased CO2 production, as well as an increase in body temperature (Mondal et al., Very recently, Skrzypski et al.  have demImmunohistochemical studies have reported that NPBWR1 is expressed in the PVN, pituitary gland, and adrenal medulla in the human, mouse and rat (O'Dowd et al., in vitro study has reported that log molar NPW23 concentrations, significantly alter prolactin, growth hormone and ACTH release from dispersed rat anterior pituitary cells (Baker et al., in vivo studies have revealed that icv infusion of NPW23 stimulates an increase in plasma prolactin levels (Baker et al., On the other hand, icv infusion of NPW23 stimulates prolactin release in the rat (Shimomura et al., Niimi and Murao  have repThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


In western industrialized nations approximately 25% of all deaths are caused directly or indirectly by the consumption of psychotropic substances. Substance-related addictions therefore constitute the most frequently occurring psychiatric disease category (McGinnis and Foege, In their recently published manuscript Pascoli et al.  elucidatSince the first description of a patient, whose alcohol addictive behavior was positively influenced by high-frequency accumbal DBS (Kuhn et al., Although the mechanisms of deep brain stimulation are, despite its decade-long application, not yet fully understood it has been argued that the modulation of dopaminergic transmission – especially in the context of addictive disorders – underlies the therapeutic effect. While this assumption could only be demonstrated for the rat nucleus accumbens (Sesia et al., The influence of impaired dopaminergic functioning on cognition and neuronal plasticity has been much debated. It is assumed that an activation of accumbal D1 receptors by phasic dopamine release facilitates limbic afferents and promotes cortical synaptic plasticity. At the same time, D2 receptor activation might promote the reinforcement of prefrontal influences, which is especially relevant in the context of addiction given the reduced density of both striatal and accumbal D2 receptors in addicted patients (Lee et al., The results of Pascoli et al.  help expThe short-term application of psychotropic substances, in this case cocaine, apparently triggers a fast modification of D1-receptor expression in terms of focal synaptic plasticity and could, hence, bring forth a lasting shift in the D1/D2 ratio, which eventually might lead to a fortification of limbic and drug-associated afferents.Current pharmacologic approaches to ameliorate the dysregulated dopaminergic system are not sufficiently effective (Spanagel and Vengeline,


Cells with damaged and unrepaired genomes represent a potential threat for the organism and therefore their elimination is beneficial. The biological safeguard barriers responsible for elimination of such hazardous cells rely on two principles: intrinsic - achieved through cellular senescence/apoptosis, and extrinsic - performed by the immune system. The concept of immune system-mediated clearance of senescent cells is well established , howeverResearch performed in the last two decades shows that cellular senescence as an essentially irreversible block of cell proliferation can be triggered or bypassed via manipulation of expression levels of several dozens of genes indicating the complexity and redundancy of regulatory machineries controlling this antitumor barrier. While the regulatory circuits are not understood in much detail, the unifying feature of cellular senescence is the activation of cell cycle checkpoints that block cell-cycle progression at G1/S or G2/M boundary. Checkpoint activation reflects suprathreshold long-term expression, commonly triggered by persistent DNA damage response (DDR) signaling, of protein inhibitors of cyclin-dependent kinases (CDKs), the key drivers of cell-cycle progression. The multiple pathways leading to the induction of individual  inhibitors of CDKs (CDKi) form the basis for redundancy of mechanisms for induction and maintenance of senescence. The robustness of response, manifested as the possibility to impose senescence even in tumor cells lacking two pivotal senescence mediators p53 and Rb, stems from the fact that the regulatory circuits are interconnected by numerous cross-talks of signaling pathways and several feedback mechanisms [et al. reported recently [per se . Production of ROS in cells during inflammation contributes to aging and development of age-related diseases. Nox4 is a member of the NADPH oxidase family known to regulate production of ROS, especially superoxide forms to induce DNA damage and premature senescence. Weyemi et al. showed that knock-down of NOX4 decreased RAS-induced DDR [et al. reported that Ras-induced senescence is mediated via Nox1 and Nox4, and overexpression of both genes is sufficient to induce senescence via activated DDR [Cytokines secreted by senescent cells play here the important role not only in shaping the senescent phenotype by autocrine and paracrine signaling and reinforcing the cell-cycle block by secondary induction of diverse CDKi, but also, as Hubackova recently , by caus se Fig. . In thisated DDR .et al. did not explore the presence of cytokine-induced secondary bystander senescence in vivo, this can be anticipated based on the recent study of Braumuller et al. [et al. show that T-helper-1 cell cytokines TNFα and IFNγ cooperatively induce senescence in mouse beta-cell tumors, both in vivo and in vitro, indicating a reciprocal relationship between the immune system and cellular senescence. Importantly, the concerted action of these two T-cell produced cytokines induced senescence in cancer cells indicating the role of immune surveillance in control of tumor cell proliferation by cytokine-induced senescence. Although not assessed in their study, it can be predicted based on previous reports that the observed senescence-inducing effect in response to both IFNγ and TNFα is also triggered by DNA damage. TNFα and IFNγ were also found to increase both intracellular and extracellular ROS production. Importantly, binding of NFκB, a crucial mediator of cytokine effects, on NOX4 promoter was observed [Even though Hubackova r et al. . The cytobserved . Thus, NIt is becoming widely accepted that secretome of senescent cells can also modulate the microenvironment, in both normal or tumor tissues. However, due to the overall complexity and variability of the secreted cytokine species, dependent on specific cell types, (patho)physiological context and senescence-inducing stimulus, it is currently hard to predict all outcomes of senescence-associated cytokine signals on tissue homeostasis. Nevertheless, what can be conceived now, is that the genotoxic effects of several cytokine species produced by senescent cells can spread damage in tissues manifested as secondary (and tertiary) senescence and thus to contribute to aging and pathogenesis of aging-associated diseases. Therefore, the elimination of senescent cells from the organism may provide a rejuvenation effect, as has already been documented in progeroid mice .


IFT122, and demonstrated impaired ciliogenesis in patient fibroblasts. This report on IFT122 broadens the phenotype of CED and expands its allelic heterogeneity.Cranioectodermal dysplasia (CED) is a very rare autosomal recessive disorder characterized by a recognizable craniofacial profile in addition to ectodermal manifestations involving the skin, hair, and teeth. Four genes are known to be mutated in this disorder, all involved in the ciliary intraflagellar transport confirming that CED is a ciliopathy. In a multiplex consanguineous family with typical CED features in addition to intellectual disability and severe cutis laxa, we used autozygosity-guided candidate gene analysis to identify a novel homozygous mutation in  IFT122,WDR35,C14ORF179, and WDR19)  is a skeletal dysplasia characterized by typical craniofacial features in the form of dolichocephaly, sagittal craniosynostosis, and facial dysmorphism , and skeletal anomalies in the form of narrow thorax and short extremities, in addition to ectodermal dysplastic features in the form of thin sparse scalp hair and micro/hypodontia   and was absent from 374 Saudi control chromosomes.In this report, we describe a multiplex family containing three affected siblings born to healthy first cousin Saudi parents Fig. A. In addIFT122 as a novel CED disease gene (Walczak-Sztulpa et al. IFT122 in CED (Tsurusaki et al. In order to confirm the pathogenicity of this mutation, we cultured fibroblasts from the foreskin of the younger brother following circumcision and proceeded with stress-induced ciliogenesis assay, essentially as described before (Shaheen et al.


Recent progress in the field of epigenetics has provided a new study angle for our research efforts on reproductive medicine and gynecologic malignancies. We have acquired valuable insight into the regulatory mechanism and biological effects of DNA methylation and histone modification, the two major epigenetic pathways. The newly acquired knowledge effectively complements that gained from the genetic standpoint and holds great potential for the prevention, diagnosis, risk assessment, and treatment of these diseases. Specifically, the DNA methylation and imprinting mechanisms are implicated in fertilization, early embryonic development, placental function, and pathogenesis of preeclampsia and intrauterine growth retardation. Aberrant DNA methylation and chromatin modification lead to gene-specific silencing of numerous tumor suppressor genes, DNA repair genes, and steroid hormone receptors. This special issue presents a collection of peer-reviewed papers focusing on these areas. While the issue is not intended as an exhaustive representation of all of the potential topics, they nevertheless provide insightful and multifaceted information that we consider a pleasure to share with the readers.This special issue includes 9 articles: three of which are related to the IGF-II imprinting in placenta and the effects of reproduction procedures on imprinting; two describe epigenetic mechanisms and genetic test for infertility; another paper documents the effects of in vitro maturation on histone acetylation in oocytes and early cleavage embryos; two address DNA methylation changes in cancers; one paper discusses rational design of primer for methylation assays.Effects of in vitro maturation on histone acetylation in metaphase II oocytes and early cleavage embryos,” Wang et al. document a reduced expression of histone acetyltransferase GCN5 (GCN5) and histone deacetylase 1 (HDAC1) in two-cell embryos but a normal level of these enzymes after the two-cell stage. The results indicate that in vitro maturation could affect protein and gene expression related to histone acetylation in oocytes and early cleavage embryos. However, by function of selection, parts of the changes could be recovered in late embryo development. In the first paper entitled “Imprinting and promoter usage of insulin-like growth factor II in twin discordant placenta,” Luo et al. analyze the imprinting and promoter usage of IGF-II in placenta of normal twins and twins with weight or phenotype discordance and conclude that promoter 3 specific LOI of the IGF-II gene may be closely related to phenotype discordance, but not to weight discordance. In the second paper entitled “Oxidative stress and DNA methylation in prostate cancer,” Donkena et al. present a comprehensive review on the effects of oxidative stress on DNA methylation and cancer progression, life style and diet as factors involved in ontogenesis and epigenetic interference for cancer prevention, and DNA methylation as a biomarker for cancer detection. Updates on the application of DNMT inhibitors to chemotherapy are also provided. In the third paper entitled “Preimplantation genetic screening: an effective testing for infertile and repeated miscarriage patients?,” Wang et al. compare results from different laboratories on preimplantation screening of aneuploidy and assess the efficacy, risks, and benefits of the procedure. They conclude that the use of preimplantation genetic screening should be reconsidered.In the fourth paper entitled “Study on the imprinting status of insulin-like growth factor II (IGF-II) gene in villus during 6–10 gestational weeks,” Chen et al. compared the rate of loss of GF-II imprinting in the placental villous tissues between normal and abnormal embryo development and observed a significantly increased loss of imprinting in the abnormal group, suggesting that the imprinting status of IGF-II may be functionally related to embryo development.In the fifth paper entitled “Effects of assisted reproduction technology on placental imprinted gene expression,” Katagiri et al. investigate the impact of assisted reproduction techniques (ART) on imprinted gene expression in human placenta. Different changes in the mRNA levels of imprinted genes are observed in the ART group compared with the spontaneous conception group, suggesting that ART may modify epigenetic status.In the sixth paper entitled “Specificity of methylation assays in cancer research: a guideline for designing primers and probes,” Barekati et al. discuss the critical parameters to be considered for a rational design of PCR primers used for the detection of methylated DNA. The authors also provided applicable tools/algorithms and useful websites.In the seventh paper entitled “Epigenetic regulatory mechanisms associated with infertility,” Minocherhomji et al. review the epigenetic mechanisms involved in spermatogenesis and infertility. Topics discussed in detail include the regulation and potential role of epigenetics in infertility by high-order chromatin organization, epigenetic control of genes associated with pericentromeric regions of chromosome 9 and Y, and noncoding RNAs. In the eighth paper entitled “In the ninth paper entitled “Hypermethylation of SOX2 promoter in endometrial carcinogenesis”, Wong et al. report their studies on the methylation profiles of SOX2, a gene encoding the stem cell-related transcription factor SOX2 in endometrial carcinomas. Compared to normal control tissues, cancer tissues show hypermethylation and decreased expression of SOX2. The authors conclude that epigenetic silencing mechanisms may play a crucial role in transcriptional regulation of SOX2 and loss of SOX2 expression.Shi-Wen JiangShi-Wen JiangBrian BrostBrian BrostSean DowdySean DowdyXing XieXing XieFan JinFan Jin


Physiotherapists play an inherent role in the multidisciplinary palliative care team. Existing knowledge, attitudes, beliefs and experiences influence their team participation in palliative care.The objective of this study was to assess the changes in knowledge, attitudes, beliefs and experiences among student physiotherapists who attended a palliative care training program.Preliminary quasi-experimental study design, conducted at an academic institution.Fifty-two student physiotherapists of either gender  of age (20.51±1.78 years) who attended a palliative care training program which comprised lectures and case examples of six-hours duration participated in this study. The study was performed after getting institutional approval and obtaining participants’ written informed consent. The lecture content comprised WHO definition of palliative care, spiritual aspects of life, death and healing, principles, levels and models of palliative care, and role of physiotherapists in a palliative care team. The physical therapy in palliative care-knowledge, attitudes, beliefs and experiences scale - modified from palliative care attitudes scale were used for assessing the participants before and after the program.t-test and Wilcoxon signed rank test at 95% confidence interval using SPSS 11.5 for Windows.Paired P<0.05) were noted for all four subscales- knowledge (7.84±4.61 points), attitudes (9.46±8.06 points), beliefs (4.88±3.29 points) and experiences (15.8±11.28 points) out of a total score of 104 points.Statistically significant differences (The focus-group training program produced a significant positive change about palliative care in knowledge, attitudes, beliefs and experiences among student physiotherapists. However, communication and coordination within the palliative care team has to be improved to minimize the negative impact of symptom distress on patient well-being and quality of life. Also, physical therapists must develop strategies for patient empowerment and methods for assessing and evaluating qualitative aspects of physical therapy in palliative cancer care.et al.,[According to Glazer-Waldman et al., the attiet al., if patieet al.[et al.[Previous studies that evaluated effects of training in palliative care utilized the educational program as part of a curriculum development among other healthcare professionals like critical care medicine trainees, medical et al. which wal.[et al.et al.[Effects of education and training programs on knowledge, attitudes, beliefs and practice were studied by other authors in the fields of nursing and child health.14 In phyet al. studied et al. who founExisting knowledge, attitudes, beliefs and experiences influence their team participation in palliative care. The objective of this study has been to assess if any change that occurred in knowledge, attitudes, beliefs and experiences among physical therapy students who attended a 6-hour educational training program on palliative care.The study ethical conduct was approved by the Institution and written informed consent was obtained from all participants prior to commencement of the study.Study design: This study used a pre-post quasi-experimental design, in which data were collected from a convenience sample of physical therapy students who attended a formal training program on palliative care and role of physical therapy.Participants: Final year physical therapy students of the institution attended upon free registration. The physical therapy curriculum had four years of academic training followed by six months of internship in undergraduate degree program. The students had previous experience of evaluating and treating pain in their third and early fourth year of academics during their clinical postings in multispeciality government hospital focused in treatment of cancer patients. Participants were given an initial description and instructions on filling the questionnaires by a qualified physical therapist.Training program: The training program was an elective educational intervention which comprised a lecture, case examples and active demonstration which consisted of WHO definition of palliative care, spiritual aspects of life, death and healing, principles, levels and models of palliative care, and role of physiotherapists in a palliative care team- for a total duration of six hours, where direct contact training of physiotherapists in palliative care emphasizing on introduction to palliative care, principles and models of assessment and care of persons at hospice, supportive, palliative, end-of-life and bereavement perspectives, goal planning and strategic implementation of physical therapy treatment methods in palliative care was done. Interaction duration of 15 min was allowed both between participants and also with the invited speaker (first author), who was a qualified physical therapist, experienced in pain relief methods for seven years.The training program required free registration of the participants and was conducted annually on a not-for-profit basis by the institution. The training program is one of its kinds listed in International Association for Hospice and Palliative Care website.et al.[Outcome measurement: The data was collected pre and post-training through a self-administered questionnaire- physical therapy in palliative care-knowledge, attitudes, beliefs and experiences scale . The scale is a 37-item self-report measure consisting of both qualitative and quantitative data. The full version of the questionnaire is given in et al. on critiThe PTiPC-KABE scale was was pilot-tested for test-retest reliability and was then used for assessing the participants before and after the program as a primary outcome measure. To avoid bias and to maintain participant anonymity, coding and decoding during data mining and analysis was done by another independent blinded physical therapist.Evaluation of test-retest reliability of PTiPC-KABE scale: For this purpose, we randomly selected 24 physical therapy students by Lots method from the same sample and we administered the questionnaire twice with a time-gap of 1 h, 2 h before the commencement of the actual study.Participants who could not understand English, who refused consent or who returned incomplete questionnaires were excluded from analysis. Completed questionnaires were collected by another qualified physical therapist, who was blinded to the study. The data were initially mined and entered for analysis by another blinded physical therapist.t-test and Wilcoxon signed rank test. Test for normality was done using Kolmogorov-Smirnof test. All analyses were done using SPSS 11.5 for windows at 95% confidence interval.Evaluation of test-retest reliability was done using intra-class correlation coefficient (ICC) for the total scores of two trials of the PTiPC-KABE scale. A mixed-methods model of primary analysis was done to compare pre-post scores using paired Of the total 82 physical therapy students- who attended the training program, 76 provided consent and were willing to participate in this study. 24 filled for the initial test-retest reliability of the scale (ICC=.80-.90) for the total score and the rest 52 were considered for this pre-post study. A total of 52 participants- 12 male (23.1%) and 40 female (76.9%) with age 20.51±1.78 years- filled the questionnaire twice- before the commencement of training program and after the completion of the training program.P<.05) changes post-program was observed for all items of the PTiPC-KABE scale after we adjusted for acquiescence bias of Likert scaling. We combined the moderately agree and strongly agree into “agree” and same for disagree option. No response was given a score of “zero”, disagree was given “one”, neutral/ unsure was given “two” and agree was given “three”. Such a scoring helped us to have a continuous data for computation purposes. We also inverted the scores for the items with negative impact on palliative care such as items 10, 17, 24, 27-33. Thus a total score can thus be derived after adding all the raw scores.Statistically significant , attitudes (9.46±8.06 points), beliefs (4.88±3.29 points) and experiences (15.8±11.28 points) subscales of PTiPC-KABE scale when analyzed using paired t-test .t-test [There was an overall statistically significant change in total score of 36.61±14.35 points out of a total of 104 points (35.2% change) when analyzed using paired t-test .Our study is the first of its kind reported on physiotherapist population and effects of a palliative care training program. One of the positive consequences of such training programs is the development of curricular content to suit student needs. Such educational interventions pave way to develop better curricular models and to integrate into regular professional and post-professional education.et al.[Marion et al. describeGoals of care should guide use of technology; and decision-making should be patient- centered.Know how to use prognostic scoring systems and recognize their limitations; be familiar with the ethical principles and guidelines for forgoing life-sustaining treatments; know how to withdraw life-sustaining treatments in ways that avoid distress; know how to monitor, identify and treat symptoms of distress, discomfort, anxiety or pain; and, know how to use opiates and sedatives to titrate to effect.Respect patients as individuals with diverse preferences; value the role of families in shared decision-making; respect ethnic and religious differences; and, be aware of their own values regarding death.Listen and communicate effectively and empathically; form trusting relationships with patients and families; explain details of illness, treatment strategy and goals of care simply; collaborate with the patient and family in balancing benefits and burdens; make recommendations and decisions in the face of uncertainty; give advice based on the patient’s preferences; talk comfortably about death, dying and loss; support families or other intimates during their bereavement; and, work effectively in collaboration with multidisciplinary teams.et al,[We see these- knowledge and attitudes among physical therapists largely shown to influence their practice patterns and levels of care. Earlier reports by Weed and Zimny, Zimny anet al, commenteet al.[et al.[Population-based interventions are much better as educational interventions aimed at team training in palliative care. Sato et al. found pol.[et al. showed pet al.[The study had few limitations- the lack of established validity of the questionnaire used and lack of long-term follow-up of participants to observe knowledge translation into action process. We understand that measurement error of the said questionnaire would be nullified in pre-post designs if the scale has appreciable test-retest reliability. Accordingly the test-retest reliability was excellent. Further research on validation is in progress. Long-term carry-over effect of gained knowledge and its implication into actual real-life situation in palliative care is also under scrutiny currently. Long-lasting improvements in knowledge was found to be absent among medical and nursing students education previously by Velayuthan et al. who alsoet al.[The implementation of the above suggested findings, however, face their own barriers. Lai et al. listed tet al.,[There are already well-established multidisciplinary palliative care training centers for healthcare professionals in operation in Kerala as described by Seamark et al., to name According to Fins and Nilson, as acadeet al.[et al,[et al.[Considering that physical therapists have largely favorable attitudes toward evidence-based practice as reported by Jette et al. and a tret al. and a wil.[et al, there isl,[et al. This knoPhysical therapists, let’s now move on, from ignorance to knowledge.The focused training program on palliative care had positively influenced a group of physiotherapy students in this study by bringing about a significant change in their knowledge, attitudes, beliefs and experiences.


Further expanding the major findings of the landmark study by Harrison and colleagues showing that late-life pharmacological inhibition of the enzyme mammalian Target Of Rapamycin (mTOR) -a conserved integrator of nutrient and growth factor signaling- is sufficient to significantly extend lifespan in mice on rapamycin feeding , Selman If activation of AMPK (and/or inhibition of mTOR/S6K1) constitutes the best metabolic response to avoid that a normal growth rate would compromise somatic integrity in progeroid animals , 5, it iEven if findings from Colman and colleagues demonstrating significant lifespan extension in rhesus monkeys can definitely substantiate CR as an effective pro-longevity strategy for humans , many peMetformin treatment has been recently found to selectively target tumor-initiating cancer stem cells (CSCs) in breast cancer cell cultures -31, likeTranslation of in vitro results into in vivo applications may significantly alter, however, the assumed CRM nature of the biguanide metformin. A recent study by Anisimov and colleagues published in a previous issue of Aging (Albany NY) may certainly establish that metformin should be defined as geroprotective or gerosuppressant rather than bona fide CRM. Long-living female mice from the outbred SHR strain were fed metformin in drinking water beginning at 3, 9 or 15 months of age and they were then analyzed for reproductive aging, mean & maximal lifespan and incidence of malignant tumors . MetformEarlier studies from the same group showed that metformin treatment increased mean lifespan of SHR mice by >37% and increased maximum lifespan by >10 months . MetformIf metformin both prevents cancer and extends lifespan in cancer-prone (e.g. neu-N transgenic mouse model of mammary cancer) strains of rodents, metformin's ability to prolong lifespan without affecting cancer in non-cancer-prone strain of rodents (e. g. SHR) may suggest that metformin can prolong life (and delay aging) by mechanisms unrelated to its ability to suppress cancer. It may not, however, if this discrepancy simply relies with the cellular response that oncogenes and tumor suppressors have on the phenotype of energetic metabolism. Derived from the FVB/N strain, neu-N transgenic mice express the nontransforming rat neu cDNA under the control of a mammary-specific promoter . As a coWhen considering recent studies describing that pre-malignant intraepithelial neoplasias such as ductal carcinoma in situ (DCIS) of the breast do contain pre-existing carcinoma precursor cells , 64, it It remains to be elucidated how metformin treatment, if started early and/or late in life, might significantly impact on late-onset diseases, like Alzheimer dementia, coronary artery diseases or, perhaps more importantly, certain familial forms of cancer. In this regard, an ideal scenario relates to genetic susceptibility to breast and ovarian cancer in women arising from a mutation in the BRCA1 gene, which is one of the most widespread genetic diseases , 77. WomAnisimov and colleagues studied markers of cellular senescence in fibroblasts from skin of SHR mice treated with metformin since the 3rd and the 9th months of life . AlthougAs mentioned above, one of the hallmarks of senescent cells is that the senescence growth arrest is essentially permanent and cannot be reversed by known physiological stimuli . TherefoPioneeringly revealed by current Anisimov's studies, a previously unrecognized ability of metformin to regulate the senescent phenotype in vivo may provide crucial insights to definitely distinguish a/the molecular mechanisms underlying metformin-promoted anti-aging and/or anti-cancer effects. On the one hand, studies of human tissues and cancer-prone mice argue strongly that cellular senescence suppresses cancer in vivo -114. SenAn unexplored scenario relates to metformin's ability to induce and/or enhance senescence responses in tumor cells themselves. Senescence-inducing stressors such as oxidative damage, DNA damage and/or oncogenes normally reach sufficient intensity to trigger senescence at the pre-malignant tumor stage. In agreement with a role of senescence in cancer prevention, the subsequent invasive progression of pre-malignant lesions almost inevitably involves one or more events that inhibit or impair the senescence pathway . Pret al [et al [et al [Landmark studies of Lin et al  and Campl [et al  have recl [et al . Senescel [et al ? Althougl [et al ; b.) phal [et al  to promol [et al , 129. Inl [et al , Skp2 inl [et al -132 gettl [et al , 134, itl [et al . Becausel [et al ), a molel [et al -139. Finl [et al -142, it l [et al  and to pl [et al .Cellular senescence can drive are-related pathologies other than cancer. If metformin reduced abundance of senescent cells is central to metformin's ability to actively regulate lifespan extension, it would be interesting to test if metformin treatment may decelerate rapid accumulation of senescent cells and/or the rapid development of age-related phenotypes including growth retardation, loss of weight, lipodystrophy (i.e. loss of adiposity), hair loss and bone density defects. A mouse model of Hutchinson-Gilford progeria syndrome  can be extremely informative because it develops phenotypes that overlap with those of HGPS children and do not include cancer even when causing a significant alteration in the number and proliferative capacity of epidermal stem cells , 146. AdThe key question that remains to be answered in Anisimov's studies relates to the molecular mechanism(s) through which metformin treatment increased lifespan and delayed tumor formation in female SHR mice. An ever-growing experimental body of evidence strongly suggests that metformin, at the cellular level, works as an efficient deactivator of the mTOR/S6K1gerogenetic pathway owing its ability to activate the metabolic rheostat AMPK . Althoug


This strategy might yield a novel class of more efficacious anti-epileptics with fewer side effects by specifically addressing disease pathophysiology. Moreover, this strategy may have ramifications for other adult seizure syndromes in which GABA receptor-mediated depolarizations play a pathogenic role, such as temporal lobe epilepsy.Neonatal intensive care has advanced rapidly in the last 40 years, with dramatic decreases in mortality and morbidity; however, for neonatal seizures, neither therapies nor outcomes have changed significantly. Basic and clinical studies indicate that seizures in neonates have long-term neurodevelopmental and psychiatric consequences, highlighting the need for novel pharmacotherapeutics. First-line treatments targeting GABAA receptors, like barbiturates and benzodiazepines, are limited in their efficacy and carry significant risks to the developing brain. Here, we review the use of current GABA agonist therapies for neonatal seizures and suggest other treatment strategies given recent developments in the understanding of disease pathogenesis. One promising avenue is the indirect manipulation of the GABAergic system, via the modulation of neuronal Cl Seizures are the most common neurological emergency in the neonate, with an estimated prevalence of 1.8–5 seizures/1000 live births in the US Jensen, . In factA receptor, or by antagonism of NMDA receptors. Although these drugs are effective in adults, they do not control neonatal seizures well. When given as monotherapy, phenobarbital controls seizures in less than half of newborns  often secondary to an increased neuronal accumulation of Cl−, though − homeostasis. Among these are the bumetanide-sensitive Na+-K+-Cl−co-transporter NKCC1, which transports Cl−into the cell, and KCC2, which normally extrudes it  is actively enrolling patients in a randomized, double-blind, controlled dose-escalation study of bumetanide as an add-on therapy to treat refractory seizures caused by hypoxic-ischemic encephalopathy. The estimated study completion date is December 2015. A second trial (NCT01434225) is being performed by a large, multi-center European group in an “open-label,” dose-escalation fashion to assess the effect of bumetanide in addition to phenobarbital for the treatment of neonatal seizures caused by hypoxic-ischemic encephalopathy. Data from these pilot studies will be utilized to guide the design of larger Phase III trials that will determine the efficacy of bumetanide in the treatment of neonatal seizures.GABA via NKCC1-knockdown or KCC2-overexpression may result in cortical neurons with fewer and shorter dendrites (Cancedda et al., − gradients in GABA transmission. Nevertheless, in the context of our enhanced understanding of the mechanisms of GABA signaling in the neonatal brain, novel therapies that target Cl− homeostasis are promising new targets for the treatment of neonatal seizures (Loscher et al., − homeostasis should also be considered. One unexplored mechanism of Cl− modulation is the manipulation of neuronal Cl− gradients via targeting of the serine-threonine regulatory kinases of NKCC1 and KCC2 (Kahle et al., in vitro, and does so via changes in transporter phosphorylation at critical regulatory residues (Figure − homeostasis, such as temporal lobe epilepsy, in which abnormal expression of Cl− channels also renders GABA excitatory (Palma et al., There are some concerns about the modulation of depolarizing GABA responses based on its importance during neurodevelopment. Studies suggest the “premature” shift of Es Figure  (Kahle es Figure . Given tThe 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.


Dear Editor:i.e.: ‘every cell has a sex’ In 2001, The United States Institute of Medicine (IOM) Committee on Understanding the Biology of Sex and Gender Differences concluded that ‘Sex…should be considered when designing and analysing studies in all areas and at all levels of biomedical and health-related research…’ and stated an apparent paradox It is unchallenged that there are health differences between males and females and that social and cultural factors could contribute to the observed differences. Anyway, the sex-dependent differences also have a biological base which sometimes has not been deeply investigated. Scientists studying health differences between male and female aim to both considering social/cultural environment and investigating biological/molecular mechanisms different between sexes. Some experimental studies have elucidated important differences in cell death control et al., via an apoptosis-inducing factor-dependent pathway (a caspase-independent pathway) in male neurons while proceeds via a cytochrome C-dependent pathway (a process mediated by caspase activation) in female neurons in vivo studies. In particular, it has been demonstrated that microRNA-23a, by binding the mRNA of the caspase inhibitor named XIAP, induces its translational repression in females, leading to enhanced caspase signalling in the ischaemic female brain. This effect has been shown to be independent of circulating oestrogen levels , contributing to female cellular higher resistance after ischaemic brain injury. Gupta NC et al. in vitro study. In addition, at least to some extent, sex-matched or sex-unmatched cell controls may be necessary in many experimental settings. In conclusion, sex-related differences in cell death mechanism may have strong implications for experimental studies and sexual dimorphism dependent on chromosomal rather than hormonal differences have important implications for planning preclinical studies and clinical interventions.However, it is currently emerging that some cell death programs are differentially controlled by sex-related hormone-independent cellular genetics. Differences in cell death sensitivity in male and female may then occur in the absence of an hormonal context. This is not an immediately obvious finding; Penaloza C


Frontiers in Neurology, on the treatment of West syndrome (WS) by using valproate as monotherapy, prompted us to rethink about the past and present treatment strategies and the outcome in this severe epileptic syndrome.The recent article by Chandra et al. , publishEven though the study by Chandra et al.  leaves sHistorically, some previous studies in children with WS, which employed valproate as monotherapy, proved effective in controlling either hypsarrhythmia and/or the epileptic spasms –6. BesidOverall, the treatment strategies in WS [either the first-line treatments  or the more classical non-golden treatments ] are based on the assumption that an early initiation of therapy coupled with a rapid control of seizures in these patients may prevent the arrest or the decline in cognitive development.clinical spasms with onset in infancy is wider than previously thought and is currently comprised under the umbrella term of Infantile Spasms syndrome (ISs), which defines an epileptic syndrome (occurring in children younger than 1 year – rarely older than 2 years), with clinical  spasms usually occurring in clusters whose most characteristic EEG finding is hypsarrhythmia . WS refers to a form (a subset) of ISs, characterized by the combination of clustered spasms and hypsarrhythmia on an EEG. Additional (less common) phenotypes falling within the ISs include the so-called infantile spasms single-spasm variant (ISSV: in which spasms may occur singly rather than in clusters), hypsarrhythmia without infantile spasms [HWIS: in which hypsarrhythmia can be  recorded without any evidence of clinical spasms], and infantile spasms without hypsarrhythmia  . There  Osborne . Notably Osborne . In thisFor all the above reasons, the results of Chandra et al.  highligh


Atrial fibrillation (AF), as a sustained arrhythmia, is featured by uncoordinated atrial activation with the consequent deterioration of mechanical function in the atrium (Mestroni, Three classical models have been proposed for the genesis of AF, including focal activity, single-circuit reentry, and multiple-circuit reentry model (Fatkin et al., Although AF is a most prevalent type of arrhythmia, its genetic etiology remains unclear (Franco et al., Pitx2a, Pitx2b, and Pitx2c (Schweickert et al., Pitx2d is also expressed in human, functioning as a dominant negative protein (Cox et al., The Pitx2 gene mainly encodes three distinct isoforms including 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.


Rhizobia are also controlled by systemically regulated or autoregulated processes on top of the basic innate immunity response. The induction of systemic immunity responses like ISR (induced systemic resistance) by some beneficial rhizosphere bacteria or the SAR (systemic acquired resistance) response provoked by pathogens are results of multiple response cascades employed by the plant host to respond to microbial and other environmental interactions. However, the entire response network is by far not yet revealed. For example, bacteria-induced plant responses resulting in improved resistance towards pathogens can also be due to the perception of secondary metabolites, like the surfactin lipopeptide, produced by certain biocontrol Bacilli  such as chitin or lipochitooligosaccharides, peptidoglycan, lipopolysaccharides or flagellum structures, and the initiation of efficient plant defence reactions  type is tightly regulated in response to cell density or the cell “quorum”  or pathogenic (Pseudomonas aeruginosa) bacteria provoked at concentrations as low as nano- to micromolar significant changes in the accumulation of over 150 proteins. Auxin-responsive and flavonoid synthesis proteins were induced and also a secretion of plant metabolites that mimic QS compounds were found, which may have the potential to disrupt QS signaling by associated bacteria. In tomato plants, a specific induction of systemic resistance proteins after inoculation of the roots with C4- and C6-side chain AHL-producing Serratia liquefaciens MG1 was discovered independently  in MG1-inoculated plants. Furthermore, Serratia plymuthica HRO-C48, producing C4-/C6- and OHC4-/OHC6-homoserine lactones, is able to induce ISR-like systemic protection of bean and tomato plants against the fungal leaf pathogen Botrytis cinnera; this response was greatly reduced with mutants impaired in AHL-production  N-acyl AHL-compounds in a different manner: C4- and C6- homoserine lactones alter the expression of selected hormonal regulated genes which results in changes of the plant's hormone content, in particular an increased auxin/cytokinin ratio   Urban) (Götz et al., Lotus corniculatus produce lactonases which degrade AHLs and prevent their uptake and transport. In barley, it could further be demonstrated that C8- and C10-AHLs are taken up in a cell energy dependent manner by ABC-transporters into the root and transported via the central cylinder into the shoot (Sieper et al., The first demonstration of specific responses of a plant to bacterial AHLs was demonstrated for the legumes o et al.  found thu et al.  and Jin u et al.  demonstrRhizobia (Pérez-Montano et al., Gypsophila was shown to influence QS, type III secretion system and gall formation activity by Pantoea plantarum (Chalupowicz et al., Serratia marcescens 90–166 (Ryu et al., Interestingly, several plants have been demonstrated to produce AHL-mimic substances or to develop other activities influencing QS of plant associated bacteria (Gao et al., Pseudomonas aeruginosa, was shown to selectively impair the regulation of the nuclear transcription factor NF-κ B which controls innate immune responses in mammalian cells (Kravchenko et al., N-acylhomoserine lactones as modifying bacterial effector molecules.Uptake of AHL-compounds and specific perception of AHLs in animal cells were also studied intensively in recent years (Teplitski et al., The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


We discuss the cellular machinery required for CIL and the molecular signals that regulate it. We focus on our recent finding that in prostate cancer cells, Contact inhibition of locomotion is regulated by a balance between EphA and EphB receptor signalling. We show that, as recently described for chick heart fibroblasts, microtubule dynamics are required for Contact inhibition of locomotion in prostate cancer cells and we propose that stabilization of microtubules could account for defective Contact inhibition of locomotion between cancer cells and noncancer cells.Contact inhibition of locomotion (CIL) occurs when a cell stops migrating in a particular direction upon contact with another cell. Many cancer cells show Contact inhibition of locomotion when contacting one another but display contact-unimpeded migration following collision with noncancer cells. Here we review current understanding of Contact inhibition of locomotion, from Abercrombie's historical studies of cells in tissue culture to more recent analyses of Contact inhibition of locomotion  This indicates that CIL has occurred. However, when PC-3 cells are treated with EphA2/EphA4 siRNA, the difference between free migration and migration following contact was significantly reduced, indicating that these cells do not display CIL . There is no significant difference between the free and contact Cx values of taxol-treated cells indicating that taxol treatment leads to failure of CIL  and suggests that defective CIL in heterotypic collisions between PC-3 cells and fibroblasts is mediated by EphB3 and EphB4 signalling. DU-145 cells may not display defective CIL because they do not have increased expression of EphB receptors and so EphA signalling predominates and CIL occurs in heterotypic collisions between DU-145 cells and fibroblasts (Astin et al., Reverse-transcription PCR profiling of the Eph receptor and ephrin expression in PC-3 and DU-145 cells indicated that PC-3 cells have increased expression of EphB3 and EphB4 compared to DU-145 cells (Astin Here we show that CIL between prostate cancer cells is regulated by EphA receptors, specifically EphA2 and EphA4. These receptors appear to act together to coordinate CIL. By contrast, PC-3 cells display contact-unimpeded migration following collisions with fibroblasts. We find that this is due to their increased expression levels of EphB3 and EphB4, which engage ephrin-B2 expressed on fibroblasts. Knockdown of these two EphB receptors can restore CIL between PC-3 cells and fibroblasts. We propose that during cell–cell collisions, cell migratory responses are regulated by a balance between repulsive EphA versus attractive EphB signalling.et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., et al., Recently, we have shown that Cdc42 is activated downstream of EphB receptors while RhoA is activated following EphA receptor activation (Astin et al., et al., et al., et al., et al., Our finding that EphB4 is increased in the more metastatic cell line, PC-3, compared to the less metastatic prostate cancer cell line, DU-145, is consistent with previous reports (Xia in vitro and in vivo (Abercrombie & Heaysman, et al., et al., et al., Since Abercrombie's early observations on the social behaviour of migrating cells in tissue culture, several studies have shown that CIL is an important process in defining the migratory behaviour of cells