Coronary artery disease is the leading cause of death worldwide. Many trials to date have investigated the diagnostic accuracy of coronary computed tomography angiography (CCTA) when compared to the gold standard diagnostic test, invasive coronary angiography. However, whether the use of a non-invasive anatomical test, such as CCTA, can translate into improved patient risk stratification, management and outcome has yet to be established. The Scottish COmputed Tomography of the HEART (SCOT-HEART) trial sought to address these questions and determined whether CCTA, when used in addition to standard care, could aid the diagnosis, further investigation and treatment of patients referred to the cardiology clinic with suspected angina due to coronary heart disease. In this trial, CCTA clarified the diagnosis of angina due to coronary heart disease in a quarter of patients and this led to major alterations in treatment and management that appeared to reduce the risk of subsequent coronary heart disease death or non-fatal myocardial infarction. The SCOT-Heart trial has established that CCTA is a valuable diagnostic test in patients with suspected angina pectoris due to coronary heart disease and leads to greater clarity, more focused appropriate treatments and better coronary heart disease outcomes. Despite significant advances in detection and management, atherosclerotic coronary artery disease remains a leading cause of ill health and mortality throughout the world . InvasivThe evolution of coronary computed tomography angiography (CCTA) has seen improved temporal and spatial resolution and has raised the promise of providing such an accurate non-invasive anatomical evaluation of the coronary arteries. More recently, questions have focused, not only on the diagnostic accuracy of CCTA, but also its clinical application and value in improving patient outcomes Table .Table 1SThe Scottish COmputed Tomography of the HEART (SCOT-HEART) trial was designed as a multicenter randomised controlled trial to assess systematically the role of CCTA in the diagnosis, management and prognosis of patients referred to the cardiology clinic with recent onset stable chest pain. The main questions addressed by the SCOT-HEART trial included whether CCTA could be of value when used alongside standard care in improving not only diagnosis but also further management and patient outcomes ••.Chest pain remains an extremely frequent presentation and clinical history alone is often inadequate at accurately diagnosing and risk stratifying patients with suspected angina secondary to coronary heart disease. A report by Sekhri et al. investigated the outcome of patients referred with chest pain to cardiology clinics and found that, whilst those patients diagnosed with angina had a greater risk of cardiovascular events, patients diagnosed with non-cardiac chest pain accounted for almost one third of cardiovascular-related deaths over a median follow-up period of 2.5 years . PatientThe diagnostic accuracy of CCTA has been well demonstrated in previous studies . InitialFrom the existing evidence, it is discernible that CCTA holds greatest value in the reliable exclusion of coronary artery disease, particularly in low- or intermediate-risk patients. Additionally, its use aids the diagnosis of coronary heart disease by reliably detecting the presence of coronary atherosclerosis. However, its reliability can falter in the accurate grading of stenosis and has been especially impeded by the over estimation of stenotic plaques, particularly in the presence of significant coronary calcification Fig. . This haThe primary objective of the SCOTHEART trial was to investigate the role of CCTA in the clinical diagnosis of angina, in order to test the generalisability of this non-invasive test in the real-world setting. The trial was intentionally designed to include a broad population of patients, truly reflective of those presenting to the cardiology clinic with stable chest pain of recent onset. Almost half of all eligible patients were recruited to the study, including patients with high calcium scores, high body mass index and atrial fibrillation. A total number of 4146 patients were randomised (1:1) to standard care alone (including cardiovascular risk assessment) or to standard care plus coronary calcium score and CCTA. Randomisation used minimisation to match for age, sex, BMI, diabetes, history of coronary heart disease, atrial fibrillation and baseline diagnosis of angina due to coronary heart disease. At baseline, cardiovascular risk was calculated using the ASSIGN score . CliniciAs opposed to comparing CCTA directly with functional testing, the SCOT-HEART trial was designed to assess the role of CCTA in addition to usual standard care, to investigate the benefit of this anatomical investigation in clarifying the diagnosis of angina secondary to coronary heart disease. In fact, the majority of patients (85 %) underwent exercise ECG prior to CCTA. The exercise ECG was abnormal in 15 % of patients ••. When For a diagnostic test to be useful, it must increase the clinician’s certainty of detecting or excluding a specific condition. At 6 weeks follow–up, the diagnosis of coronary heart disease, as defined by the attending clinician’s report, was reclassified in 27 % of patients assigned to CCTA in addition to standard care, compared with only 1 % of patients assigned to standard care alone. Similarly, the diagnosis of angina secondary to coronary heart disease changed in 23 % of patients assigned to CCTA compared to 1 % assigned to standard care. The use of CCTA doubled the certainty of diagnosing both coronary heart disease and angina secondary to coronary heart disease ••.In summary, the results of the SCOT-HEART trial demonstrate CCTA provides a non-invasive test that can identify or exclude coronary atherosclerosis and increase the certainty of the clinical diagnosis of angina secondary to coronary heart disease in patients presenting with recent onset stable chest pain.Following previous evidence highlighting the reduced positive predictive value in assessing the severity of coronary stenosis, there is a concern that CCTA will increase the number of invasive angiograms showing non-obstructive disease. In the SCOT-HEART trial, whilst the use of CCTA was associated with a small increase in early referrals, there was no overall difference in the rates of invasive coronary angiography. In patients where CCTA triggered a new request for invasive coronary angiography, the majority (73 %) had evidence of obstructive disease and over half (58 %) underwent coronary revascularisation ••. Furthp 0.02) [Another large randomised trial assessing the clinical use of CCTA, the Prospective Multicenter Imaging Study for Evaluation of Chest Pain (PROMISE) study, compared the use of a diagnostic testing strategy using CCTA with functional testing  ••. The rp 0.02) ••. OveraHistorically, the use of CCTA has been associated with radiation doses of up to 20 mSv and this has led to concern regarding its widespread adoption as a first line investigation . HoweverIn the SCOTHEART and PROMISE trials, the number of adverse reactions secondary to CCTA was very low 2 %), and all of these were recorded as mild and self-limiting  %, and a••.As a diagnostic tool, CCTA is therefore a safe, reliable and reproducible test which can add valuable anatomical information about the coronary arterial circulation. The clinical implementation of this tool in addition to exercise electrocardiography provides a means whereby an accessible and cost-effective functional assessment can be coupled with an accurate anatomical test, clarifying the diagnosis of myocardial ischaemia secondary to coronary heart disease and reducing the need for further stress testing. By clarifying the diagnosis of angina secondary to coronary heart disease, this allows appropriate tailoring of subsequent management, including focusing invasive angiography to those in whom this is necessary. Similarly, such diagnostic clarification avoids the labelling and lifelong adherence to unnecessary medication in those patients in whom coronary heart disease is excluded. Evidence has concluded that a normal CCTA confers an excellent prognosis and extends a “warranty period” of at least 7 years , 25.Following the acquisition of evidence supporting the diagnostic accuracy and clinical use of CCTA in the detection of coronary artery disease, and since the rapid development of cardiovascular imaging, there has been a drive to examine the impact of diagnostic imaging on both risk stratification and clinical outcomes.Early studies examining the role of CCTA in accurate risk stratification and prognosis were largely limited to single centres and small patient cohorts . In ordeIn contrast, for symptomatic patients with suspected coronary heart disease, the use of CCTA measures of coronary heart disease improved discrimination of patients at risk of death or myocardial infarction when added sequentially to traditional risk models, clinical variables and coronary artery calcium score .p = 0.0527) of coronary heart disease death and non-fatal MI over a median follow up period of 1.7 years. This appears to be attributable to changes in preventative therapies and coronary revascularisation [p = 0.049) [In order for a diagnostic test to alter outcomes, the results must be interpreted and clinical knowledge used to translate this information into further treatment decisions, such as the initiation and adherence to evidence-based medication. Therefore, it is challenging for a diagnostic test to have a direct effect on clinical outcomes. However, we have learned from the SCOTHEART trial that the use of CCTA leads to large changes in treatment decisions and these changes appeared to reduce the subsequent risk  ••. HowevIn both the SCOTHEART and PROMISE trials, the overall absolute event rates were low, reflective of the fact that the majority of patients had either normal coronary arteries or mild coronary artery disease , 11••. FIn the SCOTHEART trial, patients’ symptoms were reassessed at 6 weeks and there was no significant change in symptom frequency or severity between the CCTA and standard care groups. However, this was often the point at which the clinician was fully informed regarding the outcome of CCTA in this arm of the trial, and so could be considered too early to account for subsequent alterations in diagnosis or management. Longer follow-up is thus required to fully ascertain the effect of CCTA on patient-centred outcomes ••.In addition to the accurate diagnosis of coronary atherosclerosis, the evolution of CCTA has enabled the non-invasive characterization of plaque morphology, which raises promise for its future in accurate risk stratification and its potential to reduce future major cardiac events. An advantage of CCTA over invasive coronary angiography is the ability to visualise the vessel wall, providing the potential to identify high-risk features of coronary plaque despite a preserved vessel lumen . A meta-CT) have been highlighted in three large multicenter studies  which have compared the accuracy of FFRCT with invasive FFR measurements [CT has a pooled sensitivity similar to CCTA  but improves specificity in both a per-vessel and per-patient analysis  [CT has raised promise for its potential to exclude ischaemia caused by intermediate grade lesions, potentially avoiding unnecessary invasive angiography [CT: Outcome and Resource Impacts (PLATFORM) study has investigated the clinical use of FFRCT and the results revealed that CCTA with FFRCT did in fact lead to a marked reduction in the number of invasive angiography showing no obstructive coronary artery disease [In addition to the detection of high-risk plaque features on CCTA, the evolution of approaches to combine this anatomical information with physiological measures of coronary blood flow and perfusion holds promise for the future of CCTA. Recent developments in the calculation of fractional flow reserve noninvasively  . The higiography . Recentl disease . This waAnother technique which holds promise for the future potential of CT to evaluate cardiac function and physiology is the assessment of myocardial perfusion by CT. Studies comparing CT myocardial perfusion with alternative functional imaging techniques including MRI and radionucleotide perfusion imaging have demonstrated sensitivities of 83–91 % and specificities of 72–98 % for CT myocardial perfusion imaging . The mulCCTA continues to evolve as a rapidly developing imaging modality and has the ability to clarify the diagnosis of angina secondary to coronary heart disease. Results from the SCOTHEART trial have highlighted that routine application of CCTA, in addition to other clinical tools including exercise ECG, helps guide patient management, select appropriate treatments and appears to improve clinical outcomes including myocardial infarction. The potential ability of CCTA to evaluate and provide in depth information about plaque morphology could lead to the identification of specific high-risk characteristics within the vulnerable plaque, allowing sophisticated and accurate risk stratification. Furthermore, the evolution of these methods to combine physiological evaluation with CCTA raise the promise for the adoption of a single imaging platform to provide accurate diagnostic information, guide management and ultimately improve cardiovascular outcomes.


Advances in Nuclear Medicine physics enabled the specialty of Nuclear Medicine and directed research in other aspects of radiotracer imaging, ultimately leading to Nuclear Medicine’s emergence as an important component of current medical practice.in vivo biology without perturbing it will assure its ongoing role in a practice of medicine increasingly driven by molecular biology. However, in the future, it is likely that advances in molecular biology and radiopharmaceutical chemistry will increasingly direct future developments in Nuclear Medicine physics, rather than relying on physics as the primary driver of advances in Nuclear Medicine.Nuclear Medicine’s unique ability to characterize in vivo biology, and by combining the strengths of radiotracer imaging with other imaging modalities in hybrid devices, with the overall goal to enhance Nuclear Medicine’s ability to characterize regional in vivo biology.Working hand-in-hand with clinicians, chemists, and biologists, Nuclear Medicine physicists can greatly enhance the specialty by creating more sensitive and robust imaging devices, by enabling more facile and sophisticated image analysis to yield quantitative measures of regional  Physics advances helped create Nuclear Medicine and brought the field to the forefront of medicine as a specialty driven by physical science and technical innovation.From a historical perspective, it can be said that Nuclear Medicine was the direct result of advances in nuclear physics and their translation to medicine. In many ways, physics drove the creation of the field, including our abilities to produce radioisotopes suitable for human use, to capture quantitative data and/or images made possible by these radioisotopes, and to estimate the quantity of radioactive material that could be safely and effectively administered to patients for diagnosis and treatment. Advances in the physics of radioisotope production and radiation detection drove many other aspects of Nuclear Medicine research and development - including the type of radiopharmaceuticals used , predict response to specific therapies (prediction), and to measure the efficacy of treatment (response). Thus, the focus of radiopharmaceutical imaging will increasingly move away from detection and diagnosis towards characterization. Biomarkers, namely biologic measures that characterize disease status or predict disease behavior , 5, playmedicine , 7. Thismedicine .There is increasing concern about the risk of low-level radiation exposure arising from diagnostic imaging . This isRadionuclide therapy already plays an important role in the treatment of several diseases, including hyperthyroidism and thyroid cancer. The unqualified success of Ra-223 dichloride as a therapeutic radionuclide capable of palliating bone pain and extending survival in castrate-resistant prostate cancer  suggestsThe need to reduce radiation exposure, to accurately quantify regional imaging probe biodistribution, and to maximize the molecular information derived from imaging will require improvements in device sensitivity. Most current imaging research has been focused on detectability, i.e., simply visualizing a finding above background. The evolution towards accurate quantification of dynamic processes requires the consideration of not only sensitivity but also temporal resolution. An example is tumor receptor imaging, where target expression may be limited, and the quantity of tracer bound to target will be less than tracers like FDG that are enzymatically trapped . ImproveThe need for analytic validity of quantitative nuclear imaging measurements , 22 - naAdvances in computing and the science of molecular imaging analysis will lead to images more relevant to the needs of molecular medicine. Images of regional radiotracer uptake should be accompanied by images of key quantitative molecular parameters, made possible by powerful and user-friendly image analysis software . This soin vivo biology. For example, the combination of hyperpolarized 13C probes [The introduction of hybrid imaging in the form for PET/CT and SPECT/CT has changed the way Nuclear Medicine is practiced by offering the ability to directly relate regional radiotracer uptake to structural anatomy . More reC probes  and nuclC probes . Other min vivo molecular biology assure Nuclear Medicine an ongoing and significant role in medical science and the practice of medicine. Whereas advances in physics drove the early creation and evolution of the field of Nuclear Medicine, future developments will be increasingly guided by advances in molecular biology and molecular imaging probe chemistry. In turn, advances in the specialty supported by Nuclear Medicine physics will lead to advances in Nuclear Medicine’s ability to measure regional in vivo biology as an essential component of current and future medical care.The increasing importance of molecular medicine and Nuclear Medicine’s unique ability to quantify regional


CSMD1) gene harbors signals provided by clusters of nearby SNPs with 10-2 > p > 10-8 associations in genome wide association (GWAS) studies of addiction-related phenotypes. A CSMD1 intron 3 SNP displays p < 10-8 association with schizophrenia and more modest associations with individual differences in performance on tests of cognitive abilities. CSDM1 encodes a cell adhesion molecule likely to influence development, connections and plasticity of brain circuits in which it is expressed. We tested association between CSMD1 genotypes and expression of its mRNA in postmortem human brains (n = 181). Expression of CSMD1 mRNA in human postmortem cerebral cortical samples differs 15–25%, in individuals with different alleles of simple sequence length and SNP polymorphisms located in the gene’s third/fifth introns, providing nominal though not Bonferroni-corrected significance. These data support mice with altered CSMD1 expression as models for common human CSMD1 allelic variation. We tested baseline and/or cocaine-evoked addiction, emotion, motor and memory-related behaviors in +/- and -/- csmd1 knockout mice on mixed and on C57-backcrossed genetic backgrounds. Initial csmd1 knockout mice on mixed genetic backgrounds displayed a variety of coat colors and sizable individual differences in responses during behavioral testing. Backcrossed mice displayed uniform black coat colors. Cocaine conditioned place preference testing revealed significant influences of genotype (p = 0.02). Homozygote knockouts displayed poorer performance on aspects of the Morris water maze task. They displayed increased locomotion in some, though not all, environments. The combined data thus support roles for common level-of-expression CSMD1 variation in a drug reward phenotype relevant to addiction and in cognitive differences that might be relevant to schizophrenia. Mouse model results can complement data from human association findings of modest magnitude that identify likely polygenic influences.The CUB and sushi multiple domains 1 ( CSMD1) has been identified in case vs control genome-wide association (GWAS) samples for addiction- related phenotypes that include extensive use of an addictive substance, vulnerability to develop dependence on an addictive substance, ability to quit smoking and time from initiation of tobacco use to development of nicotine dependence. The modest association signals identified in these studies come from clusters of nearby single nucleotide polymorphisms (SNPs) with 10−2 > p > 10−8 association that often do not survive conservative Bonferroni corrections . Th. Th33]. Locomotion and locomotor habituation measured in 42 x 42 cm dark and sound attenuated boxes to which the mice had not been previously exposed for 120 min trials did not identify significant influences of genotype . Scheffe’s post-hoc testing showed striking significance for the differences between homozygotes and their wild-type siblings (p = 3.7 x 10−12) and a moderate difference between wild type and heterozygous mice (p = 0.045). There was a significant effect of gender (p = 1.0 x 10−11) but no significant genotype*sex interaction (p = 0.800).Locomotion during the first 20 min conditioning session was assessed when mice were confined to 20 x 20 cm portions of the conditioned place preference boxes after receiving their first saline or cocaine injections. Saline-injected csmd1 knockouts displayed significantly increased locomotion . There was a trend towards genotype*dose interaction (p = 0.054). Cocaine injections increased locomotion in wild type and heterozygous mice , but did not significantly increase locomotion in the homozygotes (p = 0.078).Locomotor sensitization was sought by comparing locomotion during the second vs first cocaine conditioning sessions. Mice of different genotypes failed to display significant overall differences between the quantities of locomotion that they displayed during these two sessions (repeated measures ANCOVA effect of genotype (Table J in −26), significant genotype*dose interactions (p = 0.007) and significant dose*session interactions (p = 2.0 x 10−13).Memory: Morris water maze: Mice of all csmd1 genotypes swam through the Morris water maze apparatus with similar velocities  between these markers in the individuals whose brains we sampled here. Indeed, there is substantial variation in the patterns and extents of linkage disequilibrium across the 5’ aspects of the CSMD1 gene in samples from different populations (http://hapmap.ncbi.nlm.nih.gov/cgi-perl/gbrowse/hapmap3r2_B36/#search), supporting the idea that use of the markers described here will provide differing patterns of information about nearby variations in samples from distinct populations.The intron 5 SNP rs10503223 and the intron 3 trinucleotide repeat annotated as rs71534387 are each associated with modest-to-moderate individual differences in levels of expression of CSMD1 mRNA that provide results at the margins of Bonferroni-corrected statistical significance. Variants in 5’ aspects of many genes are associated with their levels of expression . The relcsmd1 knockouts [csmd1 mice, we have found only modest (25–30%) increases in cerebral cortical levels of expression of mRNAs for the two CSMD1 family members, CSMD2 and CSMD3 , providing additional evidence for the modest magnitude of the adaptive brain changes caused by lifelong reductions in CSMD1 expression .The differences in expression associated with these markers’ alleles suggest that heterozygous and/or homozygous knockout mice with reduced expression provide valid models for common variation in CSMD1 in ways that accord nicely with their manifestation of phenotypes relevant to addiction and cognitive abilities. Based on the human associations with addiction phenotypes, it is thus of interest both that there are significant differences in cocaine conditioned place preference in the heterozygous knockout mice and that these differences display some evidence for specificity. Heterozygous CSMD1 knockout mice fail to display reductions in tests of locomotion or swimming speed that would provide confounding explanations for altered performance in tests of preference for places paired with cocaine or the Morris water maze. There is no evidence that heterozygous knockout alters the conditioned locomotion that can be exerted by cocaine during conditioned place preference testing. There is no enhanced lethality or other profound physiological alteration noted with the lifelong reductions in CSMD1 expression found in heterozygous knockouts. Others have reported only modest changes in expression of genes other than CSMD1 in initial nockouts . In csmdet al, submitted) and receptors , providenotypes .S1 Table(XLSX)Click here for additional data file.S2 TableTable A: Original spreadsheet and statistical analysis of the 60 min locomotion in the original CSMD1 line. Mice were injected with saline or 10 mg/kg s.c. cocaine, as indicated and immediately placed into dark, sound-attenuating locomotor boxes (42 x 42 cm). Total distance traveled was calculated from infrared beam breaks by an Optovarimax ATS System. Table B: Original spreadsheet and statistical analysis of the Rotarod test in the original CSMD1 line. Latencies (s) to fall off on each day are presented. Table C: Original spreadsheet and statistical analysis of the weights of the back-crossed CSMD1 mice. Table D: Original spreadsheet and statistical analysis of the hanging wire test of the back-crossed CSMD1 mice. Table E: Original spreadsheet and statistical analysis of the Rotarod test of the back-crossed CSMD1 mice. Latencies (s) to fall off on each day are presented. Table F: Original spreadsheet and statistical analysis of the Dark box emergence and the open field tests of the back-crossed CSMD1 mice. For the Dark box emergence test, the testing cage (18 x 36 cm) consisted of two compartments: a dark chamber (18 x 18 cm) with black walls and a small opening (5 cm) leading to a Plexiglas compartment. An animal was placed into the dark chamber. The latency to emerge and the time spent outside the dark chamber during the 10 min trial were measured using the Optovarimax system . For the Open Field test, mice were placed singly in an open field (42 x 42 cm) for ten minutes. The time each animal spent in the central quadrant was recorded. Table G: Original spreadsheet and statistical analysis of the Conditioned Place preference (CPP) test of the back-crossed CSMD1 mice. Times spent on the cocaine-paired side during two pre-tests  and post-test (Wire-T3) and the difference (Ave ∆T) between the post-test and average of the two pretests (Ave T 1–2) are presented. Table H: Original spreadsheet and statistical analysis of the locomotion recorded during the CPP pre-test (T1- Loco) of the back-crossed CSMD1 mice. Table I: Original spreadsheet and statistical analysis of the locomotion recorded during the first CPP conditioning session (1st COC Loco) with the back-crossed CSMD1 mice. Table J: Original spreadsheet and statistical analysis of the locomotor sensitiziation of the back-crossed CSMD1 mice that occurred during the two CPP conditioning sessions . Table K: Original spreadsheet and statistical analysis of the Morris water maze (MWM) learning curves of the back-crossed CSMD1 mice. Values for each day are averages of four trials performed on that day. Table L: Original spreadsheet and statistical analysis of the Morris water maze (MWM) Latencies to reach the platform during each trial on day 1 are presented. Table M: Original spreadsheet and statistical analysis of the results from the (MWM) probe trial.(XLSX)Click here for additional data file.


Exenatide is a GLP-1 (glucagon-like peptide-1) agonist that has been approved in the UK for use in the management of Type 2 Diabetes Mellitus (T2DM) since 2006. It acts by increasing glucose-induced insulin release and by reducing glucagon secretion postprandially. It therefore increases insulin secretion and reduces glucose levels, especially postprandially. It also reduces gastric emptying and acts centrally to promote satiety. In clinical practice it reduces HbA1c , fasting and postprandial blood glucose levels and is the only antidiabetic agent  to promote weight loss . It can be used as monotherapy or in combination with metformin and/or sulphonylureas (SU) and/or thiazolinediones (TZD). When compared with insulin it causes similar reductions in HbA1c and glucose levels, but unlike insulin it has the advantage of inducing weight loss. Its main side effect is gastrointestinal (GI) disturbances; nausea is the commonest GI adverse effect, albeit usually mild and transient. Hypoglycaemia is uncommon, especially when used as monotherapy or in combination with metformin. In this review article we scrutinize the currently available evidence for use of exenatide in the management of T2DM. Gut hormones are well recognized to modulate glucose homeostasis. They are normally secreted from the gut after glucose ingestion, and help regulate the secretion of insulin and glucagon (from the pancreatic islet cells), which in turn regulate hepatic glucose metabolism. These gut hormones were first hypothesized to exist when it was noted that ingested glucose elicits a larger and longer-lasting insulin response compared with intravenous glucose, suggesting that a mechanism existed within the gut that enhances insulin release in response to a meal or oral glucose . This isGLP-1 exerts wide array of actions involving an effect on pancreatic islets, as well as extra-pancreatic effects involving actions on hepatocytes, adipocytes, skeletal muscles and central nervous system. In the pancreas it modulates glucose dependent insulin secretion and somatostatin secretion. It also leads to suppression of inappropriate glucagon secretion in the postprandial state. Peripherally, it stimulates glucose uptake and disposal by skeletal and adipose tissues. It also delays gastric emptying and stimulates the satiety centre. GIP-1 also modulates glucose dependent insulin secretion, stimulates glucose uptake and enhances growth, differentiation and neogensis of the beta cells of the pancreas . MicroscThe incretin effect is well documented to be impaired in patients with T2DM with the primary defect lying at the level of beta cells , but impInitial evidence for the efficacy of supplementing incretin activity in patients with T2DM came from short-term experiments in which exogenous, native GLP-1 was administered by continuous intravenous (IV) or subcutaneous (SC) infusion. In these experiments, GLP-1 was shown to reduce blood glucose levels to near normal, restore post-meal insulin secretion and improve beta cell responsiveness to oral glucose . Two facHeloderma suspectum). This synthetic GLP-1 agonist was developed and marketed by Eli Lilly, as exendine-4 ; it shares 53% homology with the GLP-1 amino acid sequence, and binds avidly to the GLP-1 receptors, but is resistant to the action of DPP-4. As a result of this, exanatide has a greatly extended duration of insulinotrophic activity compared to endogenous GLP-1, with a half-life of 2.4 hours versus 2–3 minutes, respectively. Through its amino acid sequence homology with GLP-1, it is able to interact with GLP-1 receptors and to mimic all aspects of anti-diabetic activities of GLP-1. Thus, and as a result of stimulation of GLP-1 receptors, exenatide has been found to result in stimulation of the glucose-dependent insulin secretion, suppression of postprandial glucagon secretion, slowing of gastric emptying and reduction of food intake via promotion of satiety . Although data are limited, current evidence indicates that exenatide LAR is an effective and safe antidiabetic agent ,34. A 15vs. 2.87 kg, respectively). Liraglutide is however more expensive than exenatide and exenatide remains the first line incretin-mimetic treatment at present, given the large volume of positive evidence attached to it. Nevertheless, it is worth noting that liraglutide has the advantage of once daily subcutaneous injections which can be appealing to some of our needlephobic patients.At present, liraglutide is the only other GLP-1 agonist that is approved for use in the management of T2DM. Exenatide has been compared with liraglutide in the 26-week LEAD 6 trial, involving 464 patients with T2DM treated with maximally toletated dose of metformin and/or SU . Patientet al. [et al., exenatide use in type 2 diabetics, naïve to treatment with other antidiabetic medicines, was associated with improvements in HbA1c, fasting and postprandial glucose, weight and beta-cell function [There is some preliminary evidence to suggest that monotherapy with exenatide may be as effective as a combination of exenatide with oral antidiabetic agents. In a study by Nelson et al.  who condfunction . Indeed, since November 2009 exenatide has received a monotherapy indication from the US Food and Drug Administration (FDA), indicating that it can be used safely and effectively as a single diabetes agent in conjunction with dietary measures and exercise. The most common adverse effects observed with exenatide therapy ,13,14,15The incidence of mild to moderate hypoglycemia was less common when exenatide was used as monotherapy (5%); however, when used in combination with other anti-diabetic agents the incidence of mild to moderate hypoglycaemia was 5% with metformin and up to 36% with a SU. Thus, when combining exenatide with a SU, a reduction in the SU dose should be considered.The effect of renal impairment on the pharmacokinetics of exenatide was also studied in 31 patients, of whom only one had T2DM . ExenatiThere is no evidence that exenatide adversely affects any of the other diabetic microvascular complications, although there is a case report were the introduction of exenatide caused a dramatic drop in HbA1c and a subsequent deterioration of underlying diabetic retinopathy . Beyond their effects on glycaemic control and body weight exenatide has demonstrated additional novel features including positive effects on beta cells, as well as an improvement in lipid profile and blood pressure. The current evidence demonstrated that incretin mimetics improve some parameters of beta cell function during treatment. Although beta cell function cannot be measured directly in human subjects, indices of beta cell mass and function can be estimated indirectly by assessing some markers such as Homeostasis Model of Assessment: Beta-cell function (HOMA-B), the proinsulin to insulin ratio and the arginine stimulated C-peptide. Exenatide treatment has been shown to improve beta cell function; the HOMA-B value increased from baseline by 32% in exenatide treated patients following 24 weeks treatment compared to only 6% for placebo (p = 0.02) . FollowiStudies have suggested cardio-protective activities of GLP-1 agonists, although long-term studies to prove cardiovascular benefit are necessary in order to make firm conclusions on that. Current evidence has shown positive effects of exenatide on surrogate cardiovascular parameters such as systolic blood pressure, triglycerides, endothelial function and brain natriuretic peptide. Exenatide studies have demonstrated a marked improvement of lipid profile, with substantial reduction in apolipoprotein B (ApoB) (-5.2 mg/dL), triglycerides (-7.3 mg/dL), and an increase in high density lipoprotein cholesterol (+4.5 mg/dL), together with small reductions in total cholesterol and low density lipoprotein cholesterol . In addiAlthough this review article mainly relates to the use of exenatide in T2DM, we feel it is imperative to mention three more potential thrilling avenues for use of GLP-1 agonists:(1). Pre-diabetes state; the DECODE study showed an increased cardiovascular disease (CVD)-related morbidity and mortality in the pre-diabetes period ; by the (2). ‘Stress-diabetes’ in the peri-operative and hospital inpatient setting; GLP-1 agonists can be tried in these situations as they counteract the effect of ‘stress hormones’ (steroids and glucagon) and in view of the low incidence of hypoglycaemia. They are especially beneficial in those with pre-diabetes or in T2DM treated with oral antidiabetic medications . However(3). Steroid-induced diabetes; GLP-1 agonists counteract the inhibitory effects of steroids (such as dexamethasone) on the beta cell secretion of insulin. This effect may relate partly on the action of GLP-1 agonists on a transcription factor, called PDX-1  . Exenatide appears to be a promising agent in the multitude of drugs used in the management of T2DM. Perhaps the most significant beneficial effect of exenatide is the glucose dependent insulin secretion, which probably mimics the physiological post-prandial insulin secretion. It is therefore associated with low rates of hypoglycaemia. In addition to this, and probably more significant to most of our type 2 diabetic patients exenatide will provoke significant weight loss. Its main disadvantage is that it cannot be taken orally. However; it holds great potential to tackle the long lasting issue of secondary failure to oral hypoglycaemic agents. Its effects on some of the core pathophysiological defects, its positive effect on beta cell mass and function and the parameters of the metabolic syndrome holds great promise; long-term cardiovascular outcome studies are desperately awaited in that respect. Exenatide appears to be an attractive combination with most of the existing oral agents (metformin and/or SU and/or TZD); its use with TZD and possibly with  insulin may hold specific advantages.


Kaposi's sarcoma (KS) is a tumor derived from the endothelial cell lineage caused by Kaposi sarcoma-associated virus (KSHV), also known as human herpes virus 8 (HHV-8). Four subtypes of KS have been described: classical KS, African endemic KS, immunosuppression-associated KS and AIDS-associated KS. Over 95% of the lesions have been found to be infected with HHV-8, regardless of the clinical subtype. Classical KS usually occurs in elderly men from the Mediterranean region. It is a chronic, slowly progressing disorder, usually confined to the skin, which only rarely affects other organs.We report the case of a 56-year-old male patient who addressed to our clinic presenting extensive violaceous plaques comprising both feet and lower half of the calves, the right hand, the dorsum of the left hand, as well as smaller lesions located on the lower and upper extremities and the torso. Several violaceous firm nodules of various diameters were scattered on the surface of the plaques. The lesions were associated with extensive oedema and therefore the functionality of the right hand was severely impaired and the patient had walking difficulties. The patient asserts that the lesions had first occurred two years prior to the presentation on the lower extremities and rapidly enlarged. He was diagnosed with lichen planus and was treated with topical and systemic glucocorticoids, which helped reduce the oedema but had little effect on the cutaneous lesions. When the systemic glucocorticoid treatment was ceased, the oedema rapidly recurred.The HIV testing turned out negative. Laboratory findings were within normal range. A biopsy was taken from one of the lesions and the clinical suspicion of Kaposi's sarcoma was confirmed. The patient was treated with radiotherapy and the oedema rapidly reduced, the patient regaining the functionality of his hands and feet.The classical form of Kaposi's sarcoma is usually a chronic, indolent disorder, slowly progressing over a period of several years or decades. The particularity of the case is the rapid progression in an otherwise healthy patient.Written informed consent was obtained from the patient for publication of this Case report and any accompanying images. A copy of the written consent is available for review by the Editor of this journal.


The molecular hallmarks of ATL comprise outstanding deregulations of signaling pathways that control cell cycle, apoptosis resistance, and proliferation of leukemic cells. Using integrative analyses of primary ATL cells, we discovered unique molecular characteristics of ATL . Genetic and epigenetic disruption leads to numerous gene expression alterations that dominate disorders of homeostasis and characteristics of ATL. In particular, a novel tumor suppressor miR-31 is completely lost in all ATL cases, leading to constitutive NF-kB activation via NIK overexpression. Polycomb family directly involves in the silencing of miR-31, providing a novel interconnection between epigenetic reprogramming and NF-kB pathway. In addition, we recently unveiled molecular mechanisms how Polycomb-dependent epigenetic perturbation is abnormally sustained in HTLV-1 infected and leukemic cells. We discuss the recent discovery of molecular hallmarks of potential generality, an abnormal miRNA pattern and epigenetic reprogramming, which strongly involve the imbalance of the molecular network of lymphocytes. Because epigenetic marks are potentially reversible, development of genuine epigenetic-targeted therapy drugs holds great promise in HTLV-1-related diseases. Furthermore, we also introduce additional signaling pathways affecting leukemic cell fate. Pathway analysis based on our comprehensive dataset and biological studies suggest breaking down of the essential signaling pathways such as Hedgehog and p38 pathways, which support the biological properties of ATL. Because these organized principles may be directly associated with the clinical traits of ATL, targeting the one outstanding hallmark or co-targeting of multiple molecular hallmarks in mechanism-guided combinations will result in more effective and durable therapies for aggressive ATL.


Acute pancreatitis (AP) is a frequent gastrointestinal disorder that causes significant morbidity, and its incidence has been progressively increasing. AP starts as a local inflammation in the pancreas that often leads to systemic inflammatory response and complications. Soluble epoxide hydrolase (sEH) is a cytosolic enzyme whose inhibition in murine models has beneficial effects in inflammatory diseases, but its significance in AP remains unexplored.Ephx2 knockout (KO) mice to determine the effects of sEH deficiency on cerulein- and arginine-induced AP. sEH expression increased at the protein and messenger RNA levels, as well as enzymatic activity in the early phase of cerulein- and arginine-induced AP in mice. In addition, amylase and lipase levels were lower in cerulein-treated Ephx2 KO mice compared with controls. Moreover, pancreatic mRNA and serum concentrations of the inflammatory cytokines IL-1B and IL-6 were lower in cerulein-treated Ephx2 KO mice compared with controls. Further, Ephx2 KO mice exhibited decreased cerulein- and arginine-induced NF-κB inflammatory response, MAPKs activation and decreased cell death. Conclusions -These findings demonstrate a novel role for sEH in the progression of cerulein- and arginine-induced AP.To investigate whether sEH may have a causal role in AP we utilized  Acute pancreatitis (AP) is a frequent gastrointestinal disorder that causes significant morbidity Several animal models have been utilized to study the pathogenesis of AP; one of the most common is cerulein-induced pancreatitis Ephx2 deficiency and the development of selective sEH pharmacological inhibitors (sEHI) Soluble epoxide hydrolase (sEH) is a ubiquitously expressed predominantly cytosolic enzyme with C-terminal epoxide hydrolase and N-terminal lipid phosphatase activities Ephx2 knockout (KO) mice, and the underlying molecular mechanism investigated.The role of sEH in AP has heretofore remained unexplored but a growing body of evidence implicates sEH in pancreatic endocrine function. sEH deficiency and pharmacological inhibition promote insulin secretion and reduce islet apoptosis in a type 1 diabetes model Ephx2 gene Ephx2 KO and wild type (WT) male mice were fed standard laboratory chow  at weaning. Acute pancreatitis was induced in 8–12 week old male WT and KO mice using cerulein or arginine. For cerulein-induced AP, mice were fasted overnight then injected intraperiotoneally with cerulein (50 µg/kg body weight) 12 consecutive times, at 1 h intervals. The control group was administered DMSO as a vehicle control for cerulein. Animals were sacrificed 2 h after the last injection  and blood was collected to determine serum lipase and amylase using commercial kits (Sigma) according to the manufacturer’s instructions. Levels of serum cytokines were measured using a Multiplex kit  according to the manufacturer’s protocol. Another group of animals was used for arginine-induced AP as previously described, with modifications Mice with targeted disruption in exon 1 of the S178/180/pIKKα) for each animal and for non-phosphorylated proteins as total protein expression normalized to Tubulin (such as sEH/Tubulin) for each animal.Pancreata were lysed using radio-immunoprecipitation assay (RIPA) buffer . Lysates were clarified by centrifugation at 13,000 rpm for 10 min, and protein concentrations were determined using a bicinchoninic acid protein assay kit  according to the manufacturer’s instructions. Proteins were resolved by SDS-PAGE and transferred to PVDF membranes. Immunoblotting of lysates and immunoprecipitates was performed with antibodies for sEH (generated by the Hammock laboratory), cleaved Caspases 8, 9 and 3, PARP, SHP1 and Tubulin , pp38 (Thr180/Tyr182), p38, pJNK (Thr183/Tyr185), JNK, pIKKα/β (Ser178/180), IKKα/β, pIκBα (Ser32), IκBα, pNF-κBp65 (Ser536), NF-κBp65 and NF-κBp50 . After incubation with the appropriate secondary antibodies, proteins were visualized using enhanced chemiluminescence . Pixel intensities of immunoreactive bands were quantified using ImageQuant 5.0 software (Molecular Dynamics). For phosphorylated proteins data are presented as phosphorylation level normalized to total protein expression . Relative gene expression was quantitated using the ΔCT method with appropriate primers (Tata-box binding protein (Tbp). Briefly, the threshold cycle (Ct) was determined and relative gene expression was calculated as follows: fold change = 2-Δ(ΔCt), where ΔCt = Ct target gene-Ct TBP (cycle difference) and Δ(ΔCt) = Ct (treated mice)−/Ct (control mice).Total RNA was extracted from pancreata using TRIzol reagent (Invitrogen). cDNA was generated using high-capacity cDNA synthesis Kit (Applied Biosystems).  primers  and normEphx2 KO male mice were injected intraperiotoneally with cerulein or DMSO (50 µg/kg body weight) 12 consecutive times, at 1 h intervals then sacrificed 48 h after the first injection. A portion of the pancreas was fixed in 4% paraformaldehyde overnight, embedded in paraffin and 5 µm sections were stained with hematoxylin and eosin (H&E) to observe morphological changes. Histological analysis was initially performed in a blinded fashion. Histological scoring of pancreatic sections was performed to grade the extent of pancreatic parenchyma edema , cell vacuolation , inflammation , and acinar cell necrosis  as previously described WT and Pancreata were homogenized as described in a previous publication Data are expressed as means±standard error of the mean (SEM). Data comparisons were performed using Tukey’s-Kramer honest significant difference analyses using the JMP program (SAS Institute). Differences were considered significant at P≤0.05 and highly significant at P≤0.01.Expression of pancreatic sEH was determined in wild type mice without and with cerulein-induced pancreatitis. AP was induced in mice with repetitive intraperitoneal injections of cerulein as detailed in Ephx2 whole-body KO mice. Immunoblot analysis of total pancreas lysates demonstrated ablation of sEH expression in KO mice compared with controls  polymerase (PARP) (Mitogen-activated protein kinases (MAPKs) including p38, ERK1/2 and JNK1/2 are induced rapidly and transiently during experimental AP in rodents  KO mice . Similar KO mice . After ee (PARP) . sEH defe (PARP) . In addie (PARP) . CollectEphx2 KO mice. Together, these findings demonstrate a novel role for sEH in the pancreas and suggest that sEH pharmacological inhibition may be of therapeutic value in AP.The development of AP involves a complex cascade of events that are triggered by acinar cells, but the underlying mechanisms regulating the initiation and severity of the disease are not well understood. In the current study, we investigated the role of sEH in AP using two rodent models cerulein- and arginine-induced AP. We report increased sEH expression and activity during the early phase of AP. Importantly, sEH deficiency mitigated the effects of cerulein- and arginine-induced AP in mice. This was associated with decreased cerulein- and arginine-induced NF-κB inflammatory response and decreased cell death in Alterations in gene and protein expression during the initiation phase of AP play a significant role in the progression and severity of the disease Il-1b, Il-6 and Tnfa expression and decreased serum levels of IL-1B, IL-6. Pro-inflammatory cytokines play a pivotal role in the progression and severity of pancreatitis Ephx2 KO mice may be associated with alterations in expression of anti-inflammatory cytokines. Nevertheless, it is reasonable to stipulate that the protective effects of sEH deficiency could be mediated, at least in part, through the attenuation of the inflammatory response. It is important to note that since Ephx2 KO mice exhibit global sEH deficiency the inflammatory response is likely regulated by the systemic effects of sEH deletion. Accordingly, additional studies are warranted to determine the effects of specific pancreatic sEH deficiency on cytokine expression and development of AP.Using a genetic approach, we demonstrated that sEH deficiency ameliorated the course of AP as evidenced by pancreas histology, reduced amylase and lipase, decreased pancreatic sEH deficiency modulated cerulein- and arginine-induced NF-κB inflammatory response and MAPKs signaling. NF-κB inflammatory response is activated early in AP and plays an important role in disease pathogenesis The current studies suggest that sEH inhibition in the pancreas may represent a potential approach for treating acute pancreatitis; however it is important to note that the effects of pancreas-specific sEH deficiency on AP remain to be determined. Further, the therapeutic effects of sEH pharmacological inhibition after the development of AP need to be evaluated. Nevertheless, the findings presented herein uncover a novel role for sEH in AP and suggest that interventions designed to inhibit pancreatic sEH may be of value in combating this disease.


EETs are rapidly metabolized to dihydroxyeicosatrienoic acids (DHETs) by the soluble epoxide hydrolase (sEH). We hypothesized that sEH gene (EPHX2) deletion would increase endogenous EET levels and thereby protect against I/R-induced acute kidney injury (AKI).Kidney damage was evaluated in male wildtype (WT) and sEH-knockout (KO)-mice that underwent 22-min renal ischemia followed by two days of reperfusion. CYP-eicosanoids were analyzed by liquid chromatography tandem mass spectrometry.Contrary to our initial hypothesis, renal function declined more severely in sEH-KO mice as indicated by higher serum creatinine and urea levels. The sEH-KO-mice also featured stronger tubular lesion scores, tubular apoptosis, and inflammatory cell infiltration. Plasma and renal EET/DHET-ratios were higher in sEH-KO than WT mice, thus confirming the expected metabolic consequences of sEH deficiency. However, CYP-eicosanoid profiling also revealed that renal, but not plasma and hepatic, 20-HETE levels were significantly increased in sEH-KO compared to WT mice. In line with this finding, renal expression of Cyp4a12a, the murine 20-HETE-generating CYP-enzyme, was up-regulated both at the mRNA and protein level, and Cyp4a12a immunostaining was more intense in the renal arterioles of sEH-KO compared with WT mice.These results indicate that the potential beneficial effects of reducing EET degradation were obliterated by a thus far unknown mechanism leading to kidney-specific up-regulation of 20-HETE formation in sEH-KO-mice. Renal ischemia-reperfusion (I/R) is one of the major causes of acute kidney injury (AKI) . IschemiRecent preclinical studies indicate that arachidonic acid (AA) metabolites generated by cytochrome P450 (CYP) enzymes play an important role in the development of I/R-injury in the kidney –12, hearEETs share the capacity of 20-HETE to inhibit tubular sodium transport but show a profile of vascular activities that opposes that of 20-HETE ,24. EETsIn the present study, we used sEH-knockout and corresponding wildtype mice to test the hypothesis that sEH gene deficiency protects against renal I/R-injury. Unexpectedly, we observed that EPHX2 deletion aggravated the disease process. Searching for the potential mechanisms, we found that the sEH-KO mice displayed kidney-specific upregulation of 20-HETE formation.The sEH-KO mice were originally established by Boehringer Ingelheim Pharmaceuticals, Inc  and were5′-CCACCTACCTTGTGCTTGCC-3′, SEPOH-RP, 5′-GGGAAGAGGGGAAGGATTGT-3′, and LTR2, 5′-AAATGGCGTTACTTAAGCTAGCTTGC-3′. The PCR products were separated on 2% agarose gels and visualized under UV light after staining with GelRedTM . The genotype-specific PCR products had a size of 308 (homozygous WT) and 230 bp (homozygous sEH-KO mice).Genomic DNA was extracted from mouse tail biopsies for polymerase chain reactions (PCR)-based genotyping. The primers  were as follows: SEPOH-FP, Renal and hepatic cytosolic fractions were prepared as described previously . The assMale mice were used at the age of 10–13 weeks, 25–30 g in weight. Animals were anesthetized with isoflurane  and placed on a temperature-controlled heating table, maintaining the body temperature in the range of 36.5–37.5°C. After removal of the right kidney, ischemia was induced in the remaining kidney by applying a non-traumatic vascular clamp  to the left renal pedicle for 22 min. Renal-reflow was confirmed after releasing the clamp by visual inspection. Before wound closure, 1 ml of pre-warmed (37°C) saline was placed in the abdominal cavity to prevent dehydration. For analgesia, mice received a single injection of buprenorphine followed by tramadol in drinking water during the next two days. Sham (control) groups went through the identical procedures including uninephrectomy, however, without clamping of renal pedicle. One day after surgery, the animals were individually placed in metabolic cages for urine collection over a period of 24 h. Blood, urine, and kidneys tissue samples were collected 48 h after I/R. The kidneys were cut in half through the long axis. One half of the kidney was fixed in 4% paraformaldehyde for paraffin embedding, while the second half was snap frozen in liquid nitrogen and stored at −80°C for subsequent mRNA, protein, or CYP-eicosanoid analysis. Before analysis, the stored samples were homogenized in liquid nitrogen using a Biopulverizer . Completely untreated sEH-KO and WT mice were used for evaluating the oxylipin profiles, sEH activities and Cyp4a12a expression under baseline conditions.Creatinine and urea nitrogen in serum and urine were measured by an automated chemistry analyzer. Histomorphologic analysis of Hematoxylin and Eosin (HE) and Periodic Acid-Schiff (PAS) stained renal paraffin embedded sections (2 μm) were used to determine acute tubular necrosis (ATN) score as described previously . TUNEL sTotal RNA was extracted with the Qiazol RNeasy Micro kit including DNase digestion  and then reverse-transcribed into cDNA using a high-capacity cDNA reverse-transcription kit . Subsequent TaqMan analysis of Cyp4a12a and sEH mRNA expression was conducted as described previously . The relAliquots (30 μg of protein per lane) of kidney or liver homogenates were separated by 10% SDS-PAGE and transferred onto PVDF membranes . Recombinant Cyp4a12a protein was included as positive control. The primary antibodies used were raised in rabbits against mouse Cyp4a12a , sEH  served as internal standards. Subsequent analysis of the endogenous eicosanoid profiles was performed by liquid chromatography tandem mass spectrometry  as established previously [Plasma and homogenized tissue (kidney and liver) samples were subjected to alkaline hydrolysis and solid-phase extraction was performed as described previously . 10 ng oeviously . ResultsStatistical analysis was performed by using GraphPad Prism 5 software . All results were tested for normal distribution and expressed as mean ± standard error of mean (SEM). Two-tailed t-test was used for comparing the difference in terms of mean values between two different groups. The significance of variability among multi groups was evaluated by one-way ANOVA with a Bonferroni multiple comparison post-test. P<0.05 (*), <0.01 (**) and <0.001 (***) were considered as statistically significant.Evaluation of renal and hepatic sEH activities was performed using 14,15-EET as natural substrate and analyzing its conversion to 14,15-DHET by RP-HPLC. The cytosolic fractions prepared from the organs of WT mice metabolized 14,15-EET with hydrolase activities of about 20 kidney,  and 60 nP<0.001; urea: 561.6 ± 29.63 vs. 404.4 ± 13.93 mg/dl, P<0.001; Serum creatinine and urea levels were determined two days after reperfusion in order to evaluate the extent of I/R-induced impairment of renal function. In WT mice, I/R resulted in a 3.5-fold increase of serum creatinine  and a 5.P<0.001; P<0.001; P<0.001; In line with the differences observed in renal functional impairment, sEH-KO mice showed higher tubular necrosis scores , strongeSearching for potential mechanisms predisposing the sEH-KO mice to increased I/R-induced renal damage, we compared the oxylipin profiles of WT and sEH-KO mice under baseline conditions. Our LC-MS/MS analysis included several of the linoleic acid- and AA-derived endogenous substrates and products of sEH-mediated hydrolysis in order to evaluate the direct metabolic consequences of sEH deficiency. Moreover, we determined the levels of various AA-derived monohydroxy-metabolites (HETEs) to gain insight into sEH deficiency-associated alterations in other branches of AA metabolism. The sEH product/substrate-ratios were reduced in sEH-KO compared to WT mice, as congruently demonstrated by the oxylipin profiles of plasma, kidney, and liver samples Figs . In partP<0.05; P<0.05; Searching for the origin of increased renal 20-HETE levels in sEH-KO mice, we analyzed untreated kidneys from sEH-KO and WT mice for the expression of Cyp4a12a, the major murine 20-HETE generating enzyme . As showTo visualize the intrarenal localization of Cyp4a12a, kidney sections were incubated with a peptide-specific Cyp4a12a antibody followed by a fluorescent labeled secondary antibody. Immunostaining occurred in renal vascular and tubular structures and could be blocked at both sites by pre-saturating the Cyp4a12a antibody with the corresponding synthetic peptide . TubularIn this study, we investigated the effect of global sEH gene disruption on the development of ischemic AKI in mice. Similar to our previous experiments in rat , we usedIn apparent contrast to our findings with the sEH-KO mice, pharmacological inhibition of sEH activity was recently shown to protect against renal I/R injury in mice . MoreoveMoreover, both sEH-deletion and sEH-inhibition may cause the development of compensatory mechanisms in response to either increased levels of epoxy-metabolites or reduced levels of the corresponding hydrolysis products. In line with this hypothesis, sEH-inhibition shifted the renal AA metabolism towards the lipoxygenase pathway and failed to elicit renoprotective effects in the 5/6-nephrectomy mouse model . A shiftEETs function as vasodilators in a variety of vascular beds raising the possibility that systemic hemodynamic effects might impair renal blood flow and thus contribute to the increased renal I/R injury observed in sEH-KO mice. However, like sEH gene deletion also pharmacological sEH inhibition increases endogenous EET levels, but protects against renal I/R injury in mice . MoreoveThe mechanisms are currently unclear that lead to the observed upregulation of renal 20-HETE formation in sEH-KO mice. Renal Cyp4a12a expression is largely male-specific and can be further increased by treating C57Bl/6 mice with androgens  resultinTaken together, our findings suggest that increased Cyp4a12a-mediated 20-HETE formation in renal vessels might be responsible for the increased susceptibility of male sEH-KO mice to renal I/R-injury. Supporting this notion, vascular overproduction of 20-HETE has the potential of mediating sustained vasoconstriction  and to pS1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file.


One day, in 1987, Ron Duman and I were setting up our new laboratories in CMHC, the Connecticut Mental Health Center, a state mental hospital jointly run by the State of Connecticut and Yale University. Since the 1960’s, CMHC was unique as a state mental hospital in that it included a basic neuroscience research group embedded within inpatient and outpatient units that care for the area’s mentally ill. Duman and I were newly hired assistant professors and our appointments at Yale were made possible by the conversion of a basement corridor, which had heretofore been used to teach psychiatry residents group and family therapy, into biochemical laboratories. As Duman and I were setting up our new labs, we decided to name our corridor “The Laboratory of Molecular Psychiatry,” and we had the gumption to post a huge sign with this new name, which we had made at a local photocopying store, without any permission from our higher-ups. Thus was born the field of Molecular Psychiatry!The name Molecular Psychiatry was particularly provocative in 1987. One of our fellow faculty researchers, a behavioral neuroscientist, posted a sign on his laboratory, “Laboratory of Molecular Fear.” Another faculty member in the department labeled his office, “Laboratory of Molecular Psychoanalysis.” I’m not sure what was most discomfiting about Molecular Psychiatry. The name made eminent sense to Duman and me, and our primary mentors at Yale, George Heninger and George Aghajanian. The 1980’s marked the dawn of the molecular revolution in medicine and we all believed that brain diseases that manifest themselves in behavioral abnormalities would prove to be no different from diseases of all other organ systems, namely, that psychiatric disorders have fundamental molecular lesions that need to be identified and understood and, with that knowledge, reversed or prevented.Duman and I, and our many colleagues at Yale and across the country and world, set out to establish the molecular foundations of psychiatry. This molecular approach to psychiatry depended on the tremendous advances in biological psychiatry and neuropharmacology that had taken place the previous several decades. These advances included the development of animal models of particular mental disorders as well as an increasingly sophisticated understanding of the brain’s neurotransmitter systems and their regulation by all major classes of psychotropic drugs. The goals of Molecular Psychiatry, as we first formulated them, are several fold. First, the field aims to identify abnormalities in particular neuronal cell types which account for the behavioral pathology seen in the animal models of psychiatric disorders A second goal is to understand, at the molecular and cellular levels, how psychotropic drugs produce their clinically relevant actions and reverse the behavioral pathology in the animal models. A third goal is to understand how the genetic underpinnings of an individual animal combine with a host of environmental factors to determine the individual’s inherent vulnerability for developing that behavioral pathology. The fourth and ultimate goal is to translate this information to clinical populations: to identify the genetic variants that help determine individual risk for mental illness, to study the molecular pathophysiology of mental disorders in humans, and to use the increasing knowledge of this pathophysiology to develop improved treatments, even preventive measures, for these conditions.The Molecular Foundations of Psychiatry, with Steve Hyman, then at Harvard and one of our fellow pioneering molecular psychiatrists. Hyman and I also organized a lead symposium at the Society for Neuroscience meeting in 1995 on Molecular Psychiatry which was widely attended. Duman, Hyman, and I, alone and in various combinations, along with many colleagues in the field, also published several articles in the mid-1990’s that defined this burgeoning new field and laid out its promise for the future. By the late 1990’s the field had matured to a point where a new journal, Molecular Psychiatry, was formed by the Nature publishing group. Molecular Psychiatry made it!Although first greeted with disbelief and even some snickers, we achieved several milestones in those early years that helped establish Molecular Psychiatry as a bona fide field. Duman and I organized several symposia for the American College of Neuropsychopharmacology and the American Psychiatric Association in the early 1990’s. In 1993, I published a book, bona fide genetic variations that confer risk for psychiatric disease in people, and currently we have discovered well less than 10% of the genetic causes of these complex syndromes. Nevertheless, we remain optimistic that molecular approaches will lead to revolutionary advances in psychiatry—the brain and its disorders are simply far more complicated than we ever imagined. An impressive array of new and extremely powerful experimental approaches, in genetics, epigenetics, optogenetics, multi-unit recording from awake, behaving animals, and brain imaging, among others, should set the pace for accelerating advances over the next decade. We hope and expect that we will at long last witness the molecular revolution fully transforming the field of psychiatry into a modern medical subspecialty.Despite this promise for the field, the past two decades have been very frustrating. We have learned a great deal about the molecular actions of psychotropic drugs and stress in animal models, but we have not yet been able to use this information to develop new diagnostic tests (biomarkers) or improved treatments for mental illness. Furthermore, only in the past 2–3 years have we begun to identify In this context, I greatly welcome the foundation of the Journal of Molecular Psychiatry (JMP), because its modern open-access format will further disseminate and strengthen of the idea of molecular psychiatry.


The recent advances in gene expression analysis as well as protein and metabolite quantification enable genome-scale capturing of complex biological processes at the molecular level in crop field trials. This opens up new possibilities for understanding the molecular and environmental complexity of field-based systems and thus shedding light on the black box between genotype and environment, which in agriculture always is influenced by a multi-stress environment and includes management interventions. Nevertheless, combining different types of data obtained from the field and making biological sense out of large datasets remain challenging. Here we highlight the need to create a cross-disciplinary platform for innovative experimental design, sampling and subsequent analysis of large-scale molecular data obtained in field trials. For these reasons we put forward the term field-omics: “Field-omics strives to couple information from genomes, transcriptomes, proteomes, metabolomes and metagenomes to the long-established practice in crop science of conducting field trials as well as to adapt current strategies for recording and analysing field data to facilitate integration with ‘-omics’ data.” In the nineteenth century agricultural field experiments became widely practiced and have since been a central part of crop science. These trials have on several occasions driven scientific discovery forward and given valuable insights on, for example, the effects of fertilization and crop-rotation on yield or the transmission of plant pathogens. The results have been applied both in extension services and environmental regulations Alm, . They haThe recent flurry of technological advancements by, e.g., Next-Generation sequencing (NGS) and mass spectrometry (MS), have enabled genome-scale capturing of biological processes at the molecular level  metabolomics revealed a large variability in phenotypic plasticity of individual plant species  remain underused and the practice needs to increase  have been effective in analysing clinical data  of biological processes or states, which are robust enough to function in the various environments experienced in field conditions, need to be identified. It is possible that more relevant ones or signatures of several (“-omics” profiles; Figure Besides the potential use in breeding, “-omics” profiles could be important in precision agriculture and influence crop systems and farming practices. Several qPCR-based projects and patents use the expression of certain genes as biomarkers, e.g., to screen for good elicitors of plant defense in both laboratory and field (Brisset 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.


Hyaluronan is a negatively charged polydisperse polysaccharide where both its size and tissue concentration play an important role in many physiological and pathological processes. The various functions of hyaluronan depend on its molecular size. Up to now, it has been difficult to study the role of hyaluronan in diseases with pathological changes in the extracellular matrix where availability is low or tissue samples are small. Difficulty to obtain large enough biopsies from human diseased tissue or tissue from animal models has also restricted the study of hyaluronan. In this paper, we demonstrate that gas-phase electrophoretic molecular mobility analyzer (GEMMA) can be used to estimate the distribution of hyaluronan molecular sizes in biological samples with a limited amount of hyaluronan. The low detection level of the GEMMA method allows for estimation of hyaluronan molecular sizes from different parts of small organs. Hence, the GEMMA method opens opportunity to attain a profile over the distribution of hyaluronan molecular sizes and estimate changes caused by disease or experimental conditions that has not been possible to obtain before. N-acetyl-D-glucosamine and D-glucuronic acid [7 Da as a single molecule [Hyaluronan (HA) is a negatively charged polysaccharide consisting of an unbranched repetitive dimer ofnic acid . It is umolecule .In vertebrates, so far, three HA synthases are described, HA synthases 1, 2, and 3 (HAS 1–3)  and apprHA is present in various forms in the tissues of the body: as a freely circulating molecule bound to hyaluronan-binding proteins, loosely associated with tissue or anchored to the cell membrane via receptors. A complex of HA bound to proteins has different properties and functions depending on its molecular weight, concentration, and the protein –8. HA caThe various functions of HA depend on its molecular weight –16. HighTo understand the role of HA in pathological conditions as well as in developmental processes, it is important to analyze its molecular size distribution with a reliable method suitable also for small amounts of tissues and fluids.μL [Gas-phase electrophoretic molecular mobility analyzer (GEMMA)  was originally established for globular proteins and relatively weak protein complexes , 20 and μL . The lowμL .Due to the physical properties of HA and the shape dependence of the GEMMA method, a higher resolution is achieved in the LMW range than in the HMW range of HA . The GEMThe low detection level of the GEMMA method opens new opportunities to study HA molecular size distributions from tissues in diseases that have not been possible to study before. In this paper, we demonstrate that the GEMMA method can be used for estimation of HA molecular weight distributions in biological samples with a limited amount of HA, such as from small biopsies with only few mg of available tissue.Adult white Leghorn chicken controls from a commercial population were bred and maintained at the Animal Facilities at the National Veterinary Institute in Uppsala, Sweden. A sagittal sample was collected from the middle part of the comb. These samples were frozen in −80°C. Approval from the Ethical Committee for animal experiments in Uppsala was obtained for all experiments involving the chickens (C307/11).A heart from a wild moose was a kind gift from a local moose hunter. The heart was kept in −20°C and myocardial septum tissue samples were taken for analysis.Human myocardial septum tissue was obtained from Oslo, Norway, from a cardiac donor who was rejected for surgical reasons. The myocardium was kept on ice for 1–4 hours before tissue sampling with subsequent freezing in −80°C. Approval for the study was obtained from the Regional Ethical Committee  in Norway.Kidney tissue was excised from adult Sprague-Dawley rats and samples from the inner medulla (papilla) were frozen in −80°C. Approval from the Ethical Committee for animal experiments in Uppsala was obtained for all experiments involving the rats (C169/11).Except the wild moose, all animals in this project were handled by trained personnel and reared according to the guidelines from the Swedish Board of Agriculture.To isolate HA from tissue samples a slightly modified protocol from Tolg et al.  was usedA brief summary of the protocol follows.To digest proteins in the homogenized tissues proteinase K (Sigma-Aldrich) was used in a deferoxamine mesylate containing buffer to prevent HA degradation. HA was extracted in chloroform by liquid-liquid extraction and precipitated by ethanol (EtOH) 99%.To digest nucleic acids, the HA containing pellets were dissolved in a buffer containing benzonase (Sigma-Aldrich) followed by digestion of chondroitin by chondroitinase ABC (Sigma-Aldrich). To avoid digestion of HA the chondroitin digestions were allowed to carry on for exactly 10 min at 37°C . ChlorofThe crude HA-samples were purified on an anion exchange minispin column (Thermo Scientific). To remove the salt, the eluted HA fractions were dialyzed thoroughly against 20 mM of ammonium acetate at pH 8.0. The final aliquots were ready for GEMMA analysis and were subsequently diluted according to μL before the start of the measurements, and each GEMMA run used between 125 and 300 nL of sample. All GEMMA measurements were performed as described in Malm et al. [All HA molecular weight analyses were performed using GEMMA. Depending on the tissue origin, sample size, and initial concentration the HA samples were diluted between 1 and 100 times prior to GEMMA analysis. The final volume for all GEMMA measurements was adjusted to ca 20 m et al.  with theTo show the potential of GEMMA analysis in HA research we show examples of HA size distribution in four different tissues and species, heart tissue from human and wild moose, chicken comb, and rat kidney.Weight needed for analysis was about 50 mg wet weight for heart and kidney and ca 20 mg for chicken comb samples. Purified HA extract of the heart and kidney were not necessary to dilute whereas the chicken comb HA required a 100-fold dilution prior to analysis to prevent overload of the instrument . DiffereIn this paper, some examples are given that demonstrate the ability of GEMMA to separate LMW from HMW HA in small biological samples. This opens up for the possibility to study the involvement of LMW HA in pathological conditions which have not been possible before due to lack of material.The strength of the GEMMA compared to other methods for HA molecular size determination is the low detection level. SEC with multiangle light scattering (MALS), sedimentation techniques, flow field-flow fractionation, or gel electrophoresis usually has a detection limit of >0.01 mg/mL –28. SincAs expected, moose cardiac tissue and adult chicken comb showed HMW HA content around 5000 kDA. In contrast, besides the native HMW HA seen in the inner medulla of rat kidney, a large fraction of LMW HA around 30 kDa was also found. Both overexpression of hyaluronidase 2 or hyaluronidase 1 deficiency could potentially lead to accumulation of fragmented LMW HA as seen for this sample. However, in the kidney the amount of HA changes rapidly according to body hydration, and an extensive turnover of HA in a dehydrated kidney is to be expected . Thus, sAnalysis of HA standards and ladders from Hyalose L.L.C. showed that also the biggest molecular weight (6 MDa) did not have an EMD over 9 nm (data not shown). The GEMMA spectrum of the human heart sample showed a peak center at EMD ca 9 nm, indicating abnormally large HA. This could possibly be cross-linked HA molecules or HA cross-linked with proteins. The latter case would indicate a protection of the crosslinking protein from degradation during the extraction process by HA shielding it.We have seen this very high molecular weight HA in samples from very young chicken combs (data not shown). Thus, it is likely that this very high molecular weight HA is extracted only from certain types of tissues, for example, human heart and young chicken comb tissues which excludes the extraction process as source of the crosslinking. The very high molecular weight HA in the young chicken comb is degraded by hyaluronidases but possible trace amount of some other molecules forcing crosslinking could hide in the baseline noise. This highlights the need to develop a reliable HA specific affinity extraction which would be useful for all variants of HA molecular size determination methods.Up to now, it has been very difficult to study diseases with pathological changes in the extracellular matrix where availability is low or tissue samples are small in size. Difficulty to obtain large enough biopsies for HA analysis from tissues of human disease or animal disease models has in many cases restricted the study of HA in such diseases. The low detection level of the GEMMA method now allows for studies of diseases where HA is involved. In addition, the method allows for estimation of the distribution of HA molecular sizes from different parts of small organs, for example, rat kidney . Hence,


Doubly curved nanofiber-reinforced optically transparent composites with low thermal expansion of 15 ppm/k are prepared by hot pressing vacuum-filtered Pickering emulsions of hydrophobic acrylic resin monomer, hydrophilic chitin nanofibers and water. The coalescence of acrylic monomer droplets in the emulsion is prevented by the chitin nanofibers network. This transparent composite has 3D shape moldability, making it attractive for optical precision parts. Glass is the most common transparent material, and is used extensively in different industries, particularly automobiles and electronics, because of its low thermal expansion, hardness, and abrasion resistance. However, because of its high density and difficulty in forming complex shapes, there is a strong demand to find alternatives to glass with lower weight, low thermal expansion and high mechanical strength. Plastic is an attractive candidate because it can possess reasonable moldability and optical properties. However, the high coefficient of thermal expansion and low rigidity and strength of plastics hinder their use in certain applications. One efficient route to improve the thermal expansion and mechanical properties of plastics while minimizing loss of transparency is to use fillers with a diameter much smaller than the wavelength of visible light2As next-generation environmentally friendly materials, bio-based nanofillers, especially cellulose and chitin nanofibers with a width of 10–20 nm, have been attracting considerable interest because of the development of new efficient isolation processes4568111213The major obstacle in reinforcing optically transparent matrices with cellulose and chitin nanofibers has been the difficulty in attaining well-dispersed nanofibers in a hydrophobic matrix. When cellulose nanofibers are directly mixed with a hydrophobic matrix resin, satisfactory nanofibers dispersion is seldom achieved because the nanofibers tend to aggregate. Various processing strategies have encountered challenges caused by nanofiber aggregation. For example, cellulose nanofibers hydrophobized by acetylation were mixed carefully in acetone with amorphous polylactic acid, but the film obtained was opaqueTherefore, to prepare optically transparent nanocomposites, cellulose or chitin nanofibers are first dispersed in water and then filtered to form nanofiber sheets. The sheets are then impregnated with a curable resin such as epoxy or acrylic resin, which replace the air interstices and yield transparent nanocomposites with low thermal expansion111213An emulsion is a system of dispersed droplets of one immiscible liquid in another stabilized by emulsifiers, while Pickering emulsions are solely stabilized by small particles19212223242526Commercial α-chitin powder from crab shells was used as a starting material. Dry chitin powder was dispersed in water at a concentration of 0.4 wt% to form slurry. Several drops of acetic acid were added to the chitin slurry to adjust the pH to 3–4 and facilitate fibrillation. The slurry was crushed roughly with a domestic blender, stirred for 1 h to remove air bubbles and then passed through a high-pressure homogenizer  30 times.An aqueous suspension containing a mixture of acrylic resin and chitin nanofibers (3 wt%) was prepared by mixing a suspension of chitin nanofibers (0.5 g) in water (166 g) with acrylic resin  for 60 min with vigorous stirring. The suspension was then subjected to mechanical agitation  for 15 min. The resulting emulsion was diluted to a concentration of approximately 0.1% w/w using distilled water and observed using an optical microscope . The slurry was also freeze-dried at −40 °C, coated with Pt (thickness of ca.2 nm) in an ion-sputter coater , and observed by FE-SEM (JSM-6700F JEOL).To avoid multiple scattering effects, the emulsion samples were diluted to a concentration of approximately 0.1% w/w with distilled water and stirred gently with a spatula until completely dispersed. Samples were analyzed by a laser scattering particle size distribution analyzer .The emulsion was vacuum filtered through a metal mesh (300 mesh) and polytetrafluoroethylene membrane filter  to produce a wet thin sheet with a diameter of 40 mm. The sheets were dried at 50 °C in a convection oven for 4 h. The fracture surface of the sheet was observed by FE-SEM. Transmission electron microscopy (TEM) observation was carried out using a JEOL JEM1230 instrument at an accelerating voltage of 120 kV.2, F300S UV lamp system and LC6 benchtop conveyor, Fusion UV Systems, Inc.) to obtain the composite sheets. The fiber content of the composites was calculated based on the original weight of fibers and the final oven-dried weight of the composites. The fiber content of the composites was 14%.Acrylic resin-chitin nanofiber sheets were placed between two glass plates and mechanically pressed at 110 °C for 10 min under different pressures. The samples were cooled to ambient temperature under pressure and then immediately cured using UV curing equipment  with an integrating sphere with a diameter of 60 mm. Regular transmittance was measured by placing the sample 25 cm from the entrance port of the integrating sphere. CTE values were measured at tensile mode by a thermomechanical analyzer . Samples had a length of 25 mm, width of 3 mm, thickness of 150–160 μm and span of 20 mm via a vertically adjustable quartz glass probe. The specimen is held between two clamps and then suspended in the measuring device. To prevent the specimen from twisting or curling, it is loaded with a slight tensile force. Measurements were carried out three times during elongation at a heating rate of 5 °C/min in a nitrogen atmosphere at a load of 3 g. CTE were determined as mean values at 20–150 °C in the second run.Because the resin matrix contained a uniform distribution of nanofibers, the freeze-dried mat was compressed at 15 MPa and 110 °C to remove trapped air. However, this did not result in a transparent composite. Although the resin-nanofiber mat was deformed under loading, the trapped air was not fully removed because of the rigid 3D nanofiber structure. Therefore, we used filtration to form composite sheets. When the mixture was slowly filtered through a 0.1-μm filter membrane to obtain a wet sheet, a mixture of acrylic monomer and chitin nanofibers stayed on the filter. Because the nanofibers were concentrated during filtration to form a dense network, a large amount of hydrophobic acrylic resin was trapped in the hydrophilic nanofiber network and retained in the sheet. The fiber content of the dried composites was 14%. The success of this process was confirmed by regular light transmittance measurements. The regular transmittances of the sheets before and after pressing at different pressures are compared in Another notable feature of the molded transparent films is their low coefficient of thermal expansion (CTE), as shown in The mechanism responsible for the high level of transparency obtained by using a Pickering emulsion to form these nanocomposites can be explained as follows. Once the water was removed by vacuum filtration, the resin droplets that were covered with chitin nanofibers formed a concentrated network that trapped the resin droplets. During drying at 50 °C, the resin softened and droplets flattened, which is consistent with the observed lamellar structure . From thThe resin-nanofiber sheet was very soft and could be enlarged during hot pressing because of the plasticity of the resin. This allowed us to prepare a 3D die specifically for molding and hot pressing the sheet, as shown in In summary, a new process to fabricate doubly curved optically transparent nanocomposites with low thermal expansion has been established. Emulsions of chitin nanofibers and acrylic resin are stabilized by the hydrophilic and high-specific-surface-area chitin nanofiber networks preventing the coalescence of tiny emulsion resin droplets. The chitin nanofibers and hot pressing at high pressure increase the transparency and decrease the CTE of the composites. This composite will be useful in 3D optical precision parts such as optical lenses, as flexible optically transparent substrates for display and solar cells indented with tiny, precise lenses on their surface, and transparent 3D engineering parts.How to cite this article: Shams, M. I. and Yano, H. Doubly curved nanofiber-reinforced optically transparent composites. Sci. Rep.5, 16421; doi: 10.1038/srep16421 (2015).


Electrospun fibrous membranes find place in diverse applications like sensors, filters, fuel cell membranes, scaffolds for tissue engineering, organic electronics etc. The objectives of present work are to electrospun polyacrylonitrile (PAN) nanofibers and PAN–CNT nanocomposite nanofibers and convert into carbon nanofiber and carbon-CNT composite nanofiber. The work was divided into two parts, development of nanofibers and composite nanofiber. The PAN nanofibers were produced from 9 wt% PAN solution by electrospinning technique. In another case PAN–CNT composite nanofibers were developed from different concentrations of MWCNTs (1–3 wt%) in 9 wt% PAN solution by electrospinning. Both types of nanofibers were undergone through oxidation, stabilization, carbonization and graphitization. At each stage of processing of carbon and carbon-CNT composite nanofibers were characterized by SEM, AFM, TGA and XRD. It was observed that diameter of nanofiber varies with processing parameters such as applied voltage tip to collector distance, flow rate of solution and polymer concentrations etc. while in case of PAN–CNT composite nanofiber diameter decreases with increasing concentration of CNT in PAN solution. Also with stabilization, carbonization and graphitization diameter of nanofiber decreases. SEM images shows that the minimum fiber diameter in case of 3 wt% of CNT solution because as viscosity increases it reduces the phase separation of PAN and solvent and as a consequence increases in the fiber diameter. AFM images shows that surface of film is irregular which give idea about mat type orientation of fibers. XRD results show that degree of graphitization increases on increasing CNT concentration because of additional stresses exerting on the nanofiber surface in the immediate vicinity of CNTs. TGA results shows wt loss decreases as CNT concentration increases in fibers. Electrospinning is a most efficient technique to generate fibers with submicron diameters. Electrospun nanofibers mats are more promising because of high surface area and porosity, which have applications in air filtration, tissue engineering, drug delivery, and energy storage materials  of 99 % purity (B. Pt. = 157 °C) and also Functionalized Carbon Nanotubes (CNT) are used as fillers  of 99 % purity (B. Pt. = 157 °C) from Fisher Scientific. Functionalized Carbon Nanotubes (CNT) from Nanocyl.The 9 wt% PAN copolymer solution was prepared by taking 0.9 g chopped PAN microfibers was socked in 9.1 g of DMF and mix with glass rod. The mix was sonicated in ultrasonicator till it gives a clear solution and stirred continuously on magnetic stirrer for 5 h to mix well the contents.N,N-dimethylformamide. The mixture was kept in sonicator for 5–8 h to break bundles of CNTs and magnetic stirring to disperse well CNTs in DMF. A good dispersion was achieved in 7 h. Calculated quantity of PAN copolymer chopped microfibers were added in dispersion solution and kept on magnetic stirrer for overnight to mix well the polymer in CNT dispersion. Solution was sonicated for 1 h and then electrospun with ESPIN instrument at voltage 15 kV, flow rate 0.2 ml/h, drum speed 2000 rpm, distance 15 cm to produce PAN–CNT composite nanofibers. It was difficult to prepare solution using higher concentration of CNT. The problem faced was that, in high quantity of CNTs polymer not get dispersed well and form a solid mass. So both CNTs and polymer were dispersed in DMF separately. CNTs and DMF mixture was sonicated for 1 h till all bundles breakup and then kept on magnetic stirrer, good dispersion observed in sun light. Polymer was soaked in DMF well and sonicated for 1 h. Then it was kept on magnetic stirrer till a clear solution was obtained. Same procedure was carried out for PAN–CNT concentrations (2–3 wt%).PAN–CNT (1–3 wt%) 9 % solutions were prepared in following steps: at first 0.01 g functionalized CNTs were dispersed in Both types of nanofibers were undergone through oxidation, stabilization, carbonization and graphitization. At each stage of processing of carbon and carbon-CNT composite nanofibers were characterized by SEM, AFM, XRD and TGA.The concentration, viscosity, and conductivity of the solution as well as the applied voltage and distance between the charged electrode and the grounded target were adjusted in order to obtain stable electrospinning jet Drozin . Figure 2 and from bar analysis of surface of film, nanofibers have diameter in the range 75–100 nm & depth of film is 273 nm /(0.3440–0.3354)) × 100 and d200 = nλ/2sinθ where λ is wavelength, θ is diffraction angle and 0.3440 is interlayer spacing of fully non graphitized carbon (nm). Degree of graphitization for PAN nanofibers is −20 % and for PAN–CNT nanofibers is −41.86. Degree of graphitization increases due to presence of CNT in fibers.Figures Mettle Toledo TGA star system was used for TGA analysis up to 1000 °C temp and rate of 10°/c in the nitrogen atmosphere at flow rat 10 ml/min. Figure Figure In the present study we have successfully used the electrospinning for the synthesis of PAN–CNT carbon nanofibers. SEM images shows that the fiber diameter varies from 75 to 1500 nm on increasing concentration of CNT. The presence of black color in fiber, it indicates the presence of CNTs in nanofibers. AFM images show that the surface morphology of the composite nanofibers is smooth at lower concentration of MWCNT but rough at high concentration of MWNTs. Since the MWNTs possess a high electron density compared with the PAN polymer matrix, the nanotubes appear as darker tubular structures embedded in the PAN nanofibers. It is also seen that on Stabilization fibers diameter decreases due to shrinkage in fibers. Degree of graphitization for PAN nanofibers is −20 % and for PAN–CNT nanofibers is −41.86. Degree of graphitization increases due to presence of CNT in fibers. TGA results shows that wt loss is small in case of PAN nanofibers because of high surface area of nanofibers while in case of carbonized PAN–MWCNT carbon nanofibers wt loss is small as comparison to PAN–CNT stabilized because of increase in stability in structure, and removal of nitrogen, water and hydrogen. HCN and another gases and also carbon content increases on carbonization.