Endogenous retroviruses (ERVs) have much lower mutation rates than exogenous retroviruses (XRVs), therefore they are an important tool in analysing the long-term evolutionary history of retroviruses. Their transmission patterns also act as useful markers when studying host phylogenetics.[Oryctolagus cuniculus) [Lepus europeaus) [Microcebus murinus) [Cheirogaelus medius) [gamma- and beta- retroviruses, which are present in many vertebrate species The aim of this project is to look for and characterise further examples of primate ERVs and therefore gain insight into the evolutionary history of retroviruses and their primate hosts. This will involve data mining of primate genomes for previously unknown ERVs as well as PCR based screening of genera for which genome sequences are not available. Preliminary work suggests that prosimian primate genomes contain previously unclassified endogenous lentiviruses, so PCR-based screening will be used with DNA samples from lemurs, bushbabies and lorises to sequence and characterise these viruses.Recently, the first examples of endogenous lentivirus were characterised. The earliest was rabbit endogenous lentivirus type K in the European rabbit pol), group-specific antigen (gag) and envelope (env) genes of representative XRVs from all known retroviral genera were used to identify regions of the host genome which are highly similar to retroviral genes. We have validated this method by successfully searching the human genome for known ERV insertions. Regions of interest are then extracted and analysed in detail.The bioinformatics analysis component of this project used Exonerate  to perfopol gene  on sampM. murinus genome sequence identified 3542 ERV-like regions. This included 256 regions of less than 10,000 base pairs which incorporated gag, pol and env-like sequences in the correct orientation. The subset of pol genes which were within these 256 proviruses appear to be closely related to known gamma- and beta-retroviruses from other species.Bioinformatics analysis of the low-coverage Varecia variegata, Eulemur rufus and Mirza coquereli; the loris Perodictus potto and the bushbaby Galago moholi, although these insertions are yet to be further characterised.Preliminary laboratory work has shown previously uncharacterised lentiviral insertions in several species of lemur - murinus genome. Preliminary laboratory results suggest that there are undescribed endogenous lentiviruses in further prosimian primate species. Methodology has been established which can be used to screen further species for proviral insertions and to explore these in detail.Multiple potential ERV sequences have been found in the M.


Entropy is an elusive and somehow non-intuitive concept. Nevertheless, entropy governs spontaneous thermodynamic processes as important contribution to Gibbs Free Energy. Information theory defines Shannon entropy as a measure for uncertainty. In the context of protein binding the inherent link between flexibility, thus conformational entropy, and substrate specificity is discussed. Substrate promiscuity of proteases is quantified as cleavage entropy correlating local binding site flexibility directly with substrate readout. Caspases are examined as example protease family, where active site dynamics play a major role in mediating substrate specificity. Direct comparison of entropy in substrate data allows highlighting previously unexpected similarities in substrate recognition in proteases. Promiscuous binding to several protease targets demonstrates the emerging importance of quantitative studies on binding specificity. Shannon entropy applied to probability densities is used to rationalize ordering or disordering by binding processes. We have developed a data-driven method to reconstruct probability densities from discrete sampling by computer simulations. Application to solvent degrees of freedom leads to excellent correlation with experimental data.


Bisphenol A (BPA) is a chemical widely used to make polycarbonate plastics and epoxy resins lining food and beverage containers. A number of in vitro and in vivo studies have demonstrated that BPA has an estrogenic effect by binding to the nuclear estrogen receptor, and early BPA exposure could in induce early puberty. However, effects of human exposure to BPA on pubertal onset and the association of gonadotropin levels have not been fully evaluated. We aimed to study whether serum bisphenol A levels are associated with central precocious puberty (CPP) in Korean children.A total of 103 girls  were enrolled. Pubertal staging, anthropometry, bone maturation were assessed. Gonadotropin releasing hormone-stimulation test were conducted to determine the basal and peak levels of luteinizing hormone (LH). Serum bisphenol A levels were analysed by gas chromatography/mass spectrometry method. Geometric mean serum BPA levels were higher in CPP girls than in controls . In partial correlation analysis controlling for age and body mass index, serum BPA level showed significant positive correlation with bone age , fat mass , waist circumference , basal LH levels , and peak LH levels . Bone age, height, basal/peak LH levels and prevalence of CPP increased significantly with increasing tertile of serum BPA. Increased risk of CPP [Odds ratio] was observed across increasing serum BPA tertile  after adjusting for age and BMI.Serum BPA level was higher in CPP girls compared with controls, and higher serum BPA level was associated with increased risk of CPP. Prospective studies are needed to determine potential causal links between BPA exposure and CPP.


We present a case of myocardial infarction in a 19 year old female with cystic fibrosis who had a heart and lung transplant performed at the age of four years old. She presented atypically with a one day history of severe, intermittent, central, sharp chest pain, radiating to her back and down her left arm. A coronary angiogram showed proximal stenosis of the left anterior descending artery and right coronary artery. She was treated with percutaneous coronary intervention, involving drug eluting stents to the left anterior descending artery (LAD) and the right coronary artery (RCA). In this study we discuss the pathophysiology, investigations and treatment of cardiac transplant vasculopathy. Although complete reversal of LAD and RCA stenosis was achieved, routine follow-up with coronary angiography and careful control of cardiac risk factors will be important to identify and reduce future restenosis and adverse cardiac events. Heart and lung transplantation in children with cystic fibrosis (CF) and end-stage lung disease are now rarely performed, with bilateral sequential lung transplantation (BSLT) being the preferred procedure . AlthougA 19 year old female with CF who had a heart and lung transplantation in 1997 presented to the Emergency Department (ED) with a one day history of severe, intermittent, central, sharp chest pain, radiating to her back and down her left arm. This was associated with intermittent paraesthesia, pallor and coldness in her left hand. The chest pain was worse on deep inspiration but was not positional or exacerbated by movement. Over the previous four weeks she had been having a productive cough with a reduced exercise tolerance but no chest pain.9/L), creatinine (138 μmol/L) and urea (12.8 μmol/L).On examination there was no swelling, pallor or erythema of her left arm/hand and no chest wall tenderness. She was well perfused with a regular pulse and a stable blood pressure. No murmur or added heart sound was present and vesicular breath sounds were heard on auscultation. A chest radiograph showed no acute changes and an initial electrocardiogram (ECG) taken in the ED showed a sinus tachycardia with no ST or T wave changes. Initial blood tests were normal apart from an elevated white cell count  was 10,801. A computed tomography pulmonary angiogram (CTPA) the following day showed no pulmonary embolus (PE) and no vascular abnormalities. A coronary multi-slice computed tomography (MSCT) was not performed as it is not used as an investigation of acute MI in inpatients at St James’s University Hospital.Later that day the patient was reviewed by a cardiology registrar who performed a bedside echocardiogram. This demonstrated moderate left ventricular impairment due to extensive apical akinesis and anterior/antero-septal hypokinesis with an ejection fraction of 42%. No valvular dysfunction was identified. That evening the patient received percutaneous coronary intervention (PCI) which illustrated a 95% proximal stenosis of the left anterior descending artery (LAD), normal left circumflex artery (LCx) and 80-95% proximal stenosis of the right coronary artery (RCA) Figure . Drug elCardiac transplant vasculopathy (CTV) is a rapidly progressive form of atherosclerosis involving thickening of the intima, leading to stenosis of coronary artery grafts and subsequent myocardial ischaemia . It is gThe development of CTV is thought to be due to a combination of an immune-mediated process and non-immunologic risk factors resulting in endothelial injury and subsequent fibroelastic proliferation of the intima . Risk faThe diagnosis of CTV is particularly challenging as it rarely presents with typical angina, despite 10-30% of patients with heart transplants having partial reinnervation . HoweverIn many transplant centres, patients with heart transplants receive annual coronary angiography. However, due to the diffuse nature of CTV, angiography may underestimate the severity of disease or miss it completely . The patThe gold standard investigation of CTV is intravascular ultrasound (IVUS) which has the ability to assess the amount of intimal thickness and identify vessel wall morphology . A studyA number of non-invasive radiological modalities (e.g. dobutamine stress echocardiography (DSE), myocardial perfusion imaging (MPI) and multislice computed tomography (CT) coronary angiography) have also been utilized to assist in the diagnosis of CTV, with variable degrees of sensitivity and specificity . A pilotThe most effective treatment of CTV is prevention and modification of risk factors using antihypertensive agents , statins (e.g. pravastatin) and strict diabetic control. Coronary artery bypass grafting is rarely an option in CTV due to the diffuse nature of the disease process and the limited availability of donor hearts makes re-transplantation unlikely. In patients with heart transplants presenting with chest pain  or features of congestive heart failure, if the ECG or echocardiogram show ischaemic changes or wall motion abnormalities, acute coronary syndrome should be suspected and patients should be treated accordingly with antiplatelet agents and PCI .A systematic review of six small, retrospective, nonrandomised studies comparing the use of bare-metal stents (BMS) with drug-eluting stents (DES) in the treatment of CTV (328 lesions treated with BMS and 287 lesions treated with DES) showed that DES were associated with a lower restenosis rate at 6–12 months, but in only half of the six studies were the differences statistically significant . There wA recent retrospective nonrandomised study involving 34 patients receiving a total of 46 stents (27 DES vs 19 BMS) is the first to suggest a clinical benefit  with DES compared to BMS in the treatment of CTV . RandomiMaintenance immunosuppression following heart transplantation usually consists of a triple therapy of corticosteroids , a calcineurin inhibitor (CNI) (cyclosporine or tacrolimus) and an antiproliferative agent . AzathioTacrolimus and cyclosporine have failed to demonstrate prevention of CTV after heart transplantation and there appears to be little difference between these CNIs in terms of long-term development of CTV . HoweverIn terms of CTV prevention, the optimum maintenance immunosupression regime is unclear and a long-term randomised trial is required to determine this. The maintenance immunosupression regime following heart and lung transplantation for the patient in this case consisted of tacrolimus and prednisolone without the use of an antiproliferative agent. There was no change in the maintenance immunosupression regime post MI.MI must be considered in young patients with CF and a heart and lung transplant with atypical chest pain. Although complete reversal of LAD and RCA stenosis was achieved in this case with the use of PCI and DES, routine follow-up with coronary angiography and careful control of cardiac risk factors will be important to identify and reduce future restenosis and adverse cardiac events.Written informed consent was obtained from the patient for publication of this case report and any accompanying images.BMS: Bare-metal stents; BSLT: Bilateral sequential lung transplantation; CF: Cystic fibrosis; CFRD: Cystic fibrosis related diabetes; CMV: Cytomegalovirus; CNI: Calcineurin inhibitor; CTV: Cardiac transplant vasculopathy; DES: Drug eluting stents; DSE: Dobutamine stress echocardiography; ECG: Electrocardiogram; ED: Emergency Department; CTPA: Computed tomography pulmonary angiogram; IVUS: Intravascular ultrasound; LAD: Left anterior descending artery; LCx: Left circumflex artery; MI: Myocardial infarction; PCI: Percutaneous coronary intervention; PE: Pulmonary embolus; RCA: Right coronary artery; MPI: Myocardial perfusion imaging; CT: Computed tomography.The authors declare that they have no competing interests.JE and DP were both involved in the management of the case. JE wrote the case report with assistance from DP. Both authors read and approved the final manuscript.


The patient with a history of bone pain and muscle weakness, was thought to have oncogenic osteomalacia as a result of biochemical investigations and directed to Nuclear Medicine Department for a whole-body bone scintigraphy and 111In-octreotide scintigraphy. There was no focal pathologic tracer uptake, but generalized marked increase in skeletal uptake on bone scintigraphy. Octreotide scintigraphy showed accumulation of octreotide in the region of the left lobe of the thyroid gland in the neck. Thereafter, parathyroid scintigraphy was performed with technetium-99m labeled metroxy-isobutyl-isonitryl (99mTc-MIB) and MIBI scan demonstrated radiotracer uptake at the same location with octreotide scintigraphy. The patient underwent left inferior parathyroidectomy and histopathology confirmed a parathyroid adenoma. Somatostatin receptor positive parathyroid adenoma may show octreotide uptake. Octreotide scintigraphy may be promising and indicate a possibility of using somatostatin analogues for the medical treatment of somatostatin receptor positiveConflict of interest:None declared. HowevA 48-year-old woman was presented to our department with history of bone pain, muscle weakness. Biochemical investigations showed hypophosphatemia ; increased alkaline phosphatase ; normocalcaemia  supporting hypocalcaemic onkogenic osteomalacia.Whole body bone scintigraphy was performed after intravenous administration of 20 mCi (740 MBq) of methyleno-diphosphonate labeled with technetium-99m (Tc- 99m MDP). Bone scan showed metabolic bone disorder featured by the following characteristics: Prominent tracer uptake in the calvarium, mandible, spine, rib, and bilateral femora; intense uptake at the costochondral junctures and faint kidney visualization . BesidesParathyroid adenoma is part of a spectrum of parathyroid proliferative disorder that includes parathyroid hyperplasia, parathyroid adenoma and parathyroid carcinoma . Eighty Patients with primary hyperparathyroidism may present clinical evidence of elevated serum calcium levels which include non-specific symptoms such as fatigue, pain and weakness as well as polydipsia, polyuria, and nephrolithiasis . SonograSome studies also have suggested a role for somatostatin analogues in the medical treatment of hyperparathyroidism ,12,13. FIn conclusion, when a focal uptake was observed in the region of the thyroid gland on octreotide scintigraphy, the presence of parathyroid adenoma should also be considered. Parathyroid tumors expressing somatostatin receptors may show octreotide uptake and octreotide scan may be promising and indicate a possibility of using somatostatin analogues for medical treatment of somatostatin receptor positive parathyroid tumors in some situations which surgery is not possible and parathyroid tumor recurrence.


Dear Editor,Atrial fibrillation (AF) is one of the most frequent complications after cardiac surgery. It occurs in approximately 20% to 35% of patients after coronary artery bypass graft (CABG) surgery and in more than 50% of patients after valve surgery . AF afteAlthough many of potential risk factors for development of post-operative AF have previously been demonstrated in cardiac surgery patients ; the resRegarding the relatively high prevalence of opium abuse among patients undergoing cardiac surgery and considering its potential role in prediction of AF after cardiac surgery, anesthesiologists and cardiac surgeons should provide better preventive measures when planning surgery in opium addict patients. Since most of the studies performed in this field were retrospective, with inherent limitations, future prospective studies are needed to confirm this association.


In the traditional view on brain activity dynamics, the cognitive flow of information wanders through multiple stable states driven by task-dependent inputs -3. This It has been recently proposed that such transient states are basically shaped by anatomical connectivity and transitions between them occur even in the absence of external stimuli : Noise eA different metaphor of transient brain dynamics was proposed by Rabinovich and colleagues . In suchIn this work we develop an empirical criterion to discern whether observable neural ensemble activity can be originated by non-autonomous yet deterministic dynamical systems or rather by stochastic fluctuations between temporally attracting states. Towards this goal, we used in vivo multiple single-unit recording in rodent frontal cortex during a decision making task. Effective dynamics of neural activity is first empirically reconstructed in nonlinear state spaces . Then, tUnderlying dynamics of recorded ensemble activity is probably driven by a slowly drifting, non-autonomous dynamical system containing low-order nonlinear interactions.


Colorectal cancers are the third most common type of cancers globally, affecting both sexes . As of nNine normal and 25 tumoral colon tissue sections (3mm diameter x 10 µm thick) embedded in the form of paraffinized tissue microarray, stabilized in an agarose matrix were analyzed directly by IR imaging. To avoid chemical deparaffinization, a modified Extended Multiplicative Signal Correction (EMSC) method[EMSC permitted mathematical correction of the spectral interferences originating from paraffin and agarose. K-means classification allowed to identify and to distinguish important histological features of the colonic tissues such as crypts, lamina propria, tumor, etc. When compared to HPS stained images, after whole slide image analyzed with crop and score Calopix module from TRIBVN or through pathologist control, color-coded spectral images not only reveal features representative of the biochemical make up of the tissues, but also highlight additional features like intra-tumoral heterogeneity and tumor-associated stroma, which are difficult to discern by conventional histopathology. The LDA prediction model was promising since an average sensitivity of 91 % was achieved in the identification and prediction of tumoral tissues.IR imaging allowed differentiating and detecting normal and tumoral colon tissue features based on their intrinsic biochemical information. This chemical-free approach on paraffinized tissue biopsies combined with multivariate statistical image analysis opens a new avenue for numerical spectral histopathology and appears as a promising tool for colon cancer diagnosis. Further work to improve the model and to predict tumors in blind samples is ongoing.


The regulation of gene expression plays a pivotal role in complex phenotypes, and epigenetic mechanisms such as DNA methylation are essential to this process. The availability of next-generation sequencing technologies allows us to study epigenetic variation at an unprecedented level of resolution. Even so, our understanding of the underlying sources of epigenetic variability remains limited. Twin studies have played an essential role in estimating phenotypic heritability, and these now offer an opportunity to study epigenetic variation as a dynamic quantitative trait. High monozygotic twin discordance rates for common diseases suggest that unexplained environmental or epigenetic factors could be involved. Recent genome-wide epigenetic studies in disease-discordant monozygotic twins emphasize the power of this design to successfully identify epigenetic changes associated with complex traits. We describe how large-scale epigenetic studies of twins can improve our understanding of how genetic, environmental and stochastic factors impact upon epigenetics, and how such studies can provide a comprehensive understanding of how epigenetic variation affects complex traits. The term epigenetics was originally introduced to describe how interactions between genetics and environment can give rise to phenotypes during development de novo methylation and the maintenance of methylation patterns during replication Epigenetic mechanisms are present in many taxa, but DNA methylation has been most extensively studied in mammals where it appears to be restricted to the cytosine base, and especially in the context of CpG dinucleotides. CpG dinucleotides are cytosine–phosphate–guanine sequences that typically cluster in genomic regions referred to as CpG islands (CGI), which are often located in gene promoters and exhibit low levels of DNA methylation. However, DNA methylation in mammals can also occur outside the CpG context, and this has been reported for example in embryonic stem cells H19 locus control imprinting and gene expression at the maternally imprinted and transcriptionally-silenced insulin-like growth factor II (IGF2) locus and at the paternally imprinted and silenced H19 region de novo DNA methylation has also been suggested  twins are traditionally regarded as genetically identical, therefore any phenotypic differences within MZ twin pairs are classically attributed to environmental factors. However, epigenetic variants can also associate with phenotypic differences, and the identification and interpretation of such associations is currently an important area of research. Epigenetic studies of disease-discordant MZ twins, who are completely matched for genetics, age, sex, cohort effects, maternal influences and common environment, and are closely matched for other environmental factors, should be considerably more powerful in detecting disease-related epigenetic differences than epigenetic studies of unrelated disease cases and controls with different life-histories. In the following sections we consider the value of twin studies in epigenetics and discuss recent findings highlighting the possibility that epigenetic variation can be transmitted through generations and impact upon common diseases.Heritability is the proportion of the phenotypic variance in the population that is attributed to genetic variation. Heritability estimates are traditionally obtained by comparing the extent of similarity between relatives in classical twin studies, twin-adoption studies, sib/half-sib studies, and transgenerational family studies. Each has weaknesses, but for most traits twin studies are generally regarded as the most reliable because they are unbiased by age effects and offer the ability to separate common environment from genetic effects post hoc in twins who differed most in lifestyle. However, the study was cross-sectional instead of longitudinal, potentially obscuring comparison of individual variability; furthermore, comparing epigenetic patterns in fast-growing children and adults might not be generalizable to the aging process. To this end, a recent longitudinal study of twin epigenetic heritability assayed DNA methylation patterns in the promoters of three genes in 46 MZ and 45 DZ twin pairs sampled at 5 and 10 years of age Several studies have examined DNA methylation patterns in twins. Early work focused on X-chromosome inactivation patterns in females – where one X chromosome is inactivated at random and the silent state of the inactive X-chromosome is maintained by DNA methylation IGF2) gene and the paternally imprinted H19 region, estimating high epigenetic heritability in those regions using multiple tissues from 182 newborn MZ and DZ twins Substantive evidence for epigenetic heritability has been obtained from further studies of age-matched twins using larger samples Overall, these findings confirm that DNA methylation is a heritable trait on a genome-wide basis, but also highlight the importance of taking age into account when studying epigenetic processes. Furthermore, the results are consistent with recent population-based findings of quantitative trait loci (QTL) for DNA methylation DNA methylation patterns can be affected by genetic variation, environmental changes, heritable and non-heritable changes in other epigenetic processes , and stochastic changes over time. All these factors can contribute to DNA methylation heritability estimates, which are therefore time-, tissue-, locus- and population-specific. There are three further important aspects of epigenetic heritability. First, does twin epigenetic heritability reflect stability in methylation transmission during mitosis and meiosis? Second, does epigenetic variation contribute to phenotype heritability? Finally, how do epigenetic heritability findings relate to time-dependent methylation changes?In mammals, maintenance DNA methyltransferases and histone methyltransferases ensure propagation of epigenetic marks through mitotic cell divisions with high fidelity  in two discordant or concordant pairs of schizophrenic twins; this study found greater methylation differences between discordant MZ twins than in unrelated cases Recently, several epigenetic studies of MZ discordant twins have examined differences in DNA methylation profiles, aiming to identify differentially methylated regions (DMRs) in human disease The power of discordant MZ twin studies to detect DMRs will depend on a number of factors, including the effect size of the epigenetic change on the phenotype, the similarity of methylation profiles between MZ twins, sample size, and the sensitivity and coverage of the methylation assay. An estimate of the power of the discordant MZ twin design for a specific microarray methylation assay There are several aspects of epigenetic studies where twins present novel opportunities to understand the biology and the mechanisms underlying complex traits. We suggest a few examples.http://www.muther.ac.uk/), which aims to assay gene expression variation in multiple tissues in twins and to identify regulatory genetic variants, can be linked with epigenetic data to explore the tissue specificity and functional consequences of epigenetic variation in twins. This project is being extended to RNA sequencing (via the EUroBATS project), and this will also allow differential allelic expression to be explored. Allele-specific expression (ASE) patterns are relatively common and are under strict genetic control Twin resources such as the MuTHER (multiple tissue human expression resource) project  affect epigenetics. ARTs include IVF and related technologies http://www.twinsuk.ac.uk/) aims to discover methylated genes responsible for discordance of ten common traits and diseases. The study is using MeDIP-seq on blood samples to assay epigenomic differences in 5000 adult UK twins aged 16–85, discordant and concordant for a wide variety of diseases and environments. Next-generation sequencing, although currently at significant cost, has the potential to prove powerful in detecting disease-related methylation differences at a high level of resolution in a sample of this size. Another ongoing large-scale prospective study consists of a cohort of Australian newborn twins http://www.euengage.org/), where MeCAP-seq is being performed by sample pooling across multiple traits in discordant twins.A recent large-scale study (EpiTwin – Two further areas that can benefit from the epigenetic differences observed in MZ twins are forensic science and medical transplantation. Although twins are no more likely to be criminals than the general population, one in 250 people are MZ twins and legal cases involving MZ twins are high-profile. For example, the genetic identity of MZ twins can allow twins to provide each other with alibis in criminal cases. Differences in epigenetic profiles between MZ twins, if consistently replicated, could in future lead to closing this loophole. In transplantation there are reports of occasional graft failures in identical twins. Studying subtle differences in twin epigenetic profiles could improve transplantation outcomes, where small epigenetic changes of immune-related genes in the host or in transplanted organs could affect transplant success The study of epigenetic profiles in twins offers an excellent opportunity to understand the causes and consequences of epigenetic variation. Twin epigenetic heritability estimates tell us about the genetic control of DNA methylation variability and the stability of methylation patterns during cell division. The contribution of epigenetic variants to complex phenotypes can be assessed using disease-discordant MZ twins who are otherwise matched for genetics, age, sex, cohort effects, maternal effects and a common environment. These twin designs are considerably more powerful discovery tools than studies on singletons. In the near future, large-scale epigenetic studies in twins across different ages, tissues, and diseases will improve our understanding of the etiology and mechanisms of a wide range of common complex traits and diseases.


Vaccination strategies capable of eliciting neutralizing antibody responses to HIV remain elusive despite extensive efforts. Alternative antibody functions offer opportunities for protection without necessarily achieving broad neutralization breadth. Viral immune exclusion through aggregation has been proposed as an alternative protection pathway, but mechanisms for studying this phenomenon at the scale necessary for clinical trials have not been explored.Concentrated fluorescent virions of two colors, suspended in hydroxyethylcellulose (HEC) gel, which has been formulated to simulate the diffusion characteristics of cervical mucus, are imaged over time. Mean squared displacement and incidence of colocalized viral particles are determined. The addition of monoclonal antibodies of various specificities and isotypes affects these parameters is explored. Immunoglobulin isolated from HIV-1 positive individuals was examined as well. Correlative scanning electron micrographs of the same preparations were performed to confirm the nature of suspected aggregates.Multimeric antibodies, rather than monomeric isoforms, selectively hinder the diffusion characteristics of colocalized virions, more so than non-aggregated virions. The incidence of colocalized virions is also increased in a concentration dependent manner. Excessive antibody or virus concentration is seen to obviate aggregate formation. These results are seen in both monoclonal antibody experiments alongside polyclonal patient IgA isolated from breast milk and IgM from serum.Virus aggregation has been demonstrated as a feasible effector function of HIV-1 specific antibody preparations. This assay platform is amenable to adaptation for medium to high throughput sample screening and low sample size. Monoclonal antibodies as well as patient samples are able to induce similar behaviors. Measurement of viral aggregation as a corollary of vaccine efficacy is deserving of further exploration in a clinical setting.


The resulting adducts were fully characterized by spectroscopic and analytical methods; they constitute interesting substrates for further organic transformations.The chemical behaviour of various alkyl-substituted, acyclic conjugated bisallenes in reactions involving polar intermediates and/or transition states has been investigated on a broad scale for the first time. The reactions studied include lithiation, reaction of the thus formed organolithium salts with various electrophiles , oxidation to cyclopentenones and epoxides, hydrohalogenation , halogenation (Br


X-linked hypohidrotic ectodermal dyplasia (XLHED) is the most common form of ectodermal dysplasia. It often presents with ocular symptoms, already in early childhood. Multiple ophthalmological tests are available, but in early childhood only tests of lower invasiveness can be applied. We have evaluated tear film tests and ocular surface staining as screening methods in pediatric patients with signs of ectodermal dysplasia. These tests may also be used in adults with confirmed XLHED to determine the severity of ocular surface disease.Twelve children and 14 adults with XLHED were subjected to a panel of tests including the ocular surface disease index (OSDI), non-invasive measurement of tear film break-up time (NIBUT), osmolarity, Schirmer test, lissamine green staining, fluorescein staining, meibography and infrared thermography. Sensitivity and specificity were determined for single tests and selected test combinations. For adults with XLHED, OSDI, NIBUT and osmolarity were the best single routine tests . Their combination increased the sensitivity to 92.8%. Meibography yielded optimal results (100% sensitivity and specificity). Infrared thermography revealed a typical pattern for XLHED. In children with XLHED, NIBUT or OSDI were the most convincing single tests , combination of which increased the sensitivity to 100%. More invasive tests such as meibography and infrared thermography led to good results if they were tolerated.Tear film tests can help to establish an early diagnosis in individuals with suspected XLHED, even before genetic test results are available. Meibomian gland disorder and resulting hyperevaporative dry eye are typical features of XLHED. Once the diagnosis is made, tear film tests are an important instrument to establish an effective therapy of dry eye disease.


Spinal cord injury (SCI) causes chronic peripheral sensitization of nociceptors and persistent generation of spontaneous action potentials (SA) in peripheral branches and the somata of hyperexcitable nociceptors within dorsal root ganglia (DRG). Here it is proposed that SCI triggers in numerous nociceptors a persistent hyperfunctional state  that originally evolved as an adaptive response to compensate for loss of sensory terminals after severe but survivable peripheral injury. In this hypothesis, nociceptor somata monitor the status of their own receptive field and the rest of the body by integrating signals received by their peripheral and central branches and the soma itself. A nociceptor switches into a potentially permanent hyperfunctional state when central neural, glial, and inflammatory signal combinations are detected that indicate extensive peripheral injury. Similar signal combinations are produced by SCI and disseminated widely to uninjured as well as injured nociceptors. This paper focuses on the uninjured nociceptors that are altered by SCI. Enhanced activity generated in below-level nociceptors promotes below-level central sensitization, somatic and autonomic hyperreflexia, and visceral dysfunction. If sufficient ascending fibers survive, enhanced activity in below-level nociceptors contributes to below-level pain. Nociceptor activity generated above the injury level contributes to at- and above-level sensitization and pain (evoked and spontaneous). Thus, SCI triggers a potent nociceptor state that may have been adaptive (from an evolutionary perspective) after severe peripheral injury but is maladaptive after SCI. Evidence that hyperfunctional nociceptors make large contributions to behavioral hypersensitivity after SCI suggests that nociceptor-specific ion channels required for nociceptor SA and hypersensitivity offer promising targets for treating chronic pain and hyperreflexia after SCI. My focus is on the many uninjured nociceptors that are chronically altered by SCI and positioned to help drive below-level and at-level pain.Neuropathic pain in general and spinal cord injury (SCI) pain in particular are usually viewed as maladaptive consequences of neural injury , sprout new branches within the dorsal horn after SCI, which potentially could lead to more extensive nociceptive input to dorsal horn neurons  3 days to 8 months after SCI compared to what has been reported (0–20%) for dissociated small DRG neurons sampled from rats with peripheral neuropathy   under these quite different conditions  can be triggered in nociceptors by a combination of events after SCI that mimic patterns of signals used by nociceptors to recognize particularly severe e Figure . I then Long-lasting changes in intrinsic functional properties of primary sensory neurons are likely to depend upon changes in gene expression in these neurons  to the nociceptor's own branches. This includes immediate injury discharge and both positive injury signals , and negative injury signals  transported from damaged axons to the nucleus , which are unlikely to have axons projecting close enough to the lesion to experience axotomy or other forms of direct damage     are conveyed directly to cells within the DRG after peripheral injury/inflammation and after SCI Figure . Periphe2)  that avoids apparent tissue inflammation produce behavioral hypersensitivity for at least 1 month afterwards, which is accompanied by Nav1.8 upregulation and is dependent upon Nav1.8 expression, suggesting a key role for intrinsically altered nociceptors  continuing release of extrinsic signals, such as inflammatory mediators, that continuously refresh the hyperfunctional state, (2) positive feedback loops between nociceptor activity and inflammatory and retrograde synaptic effects, and (3) switching of the nociceptor into a potentially permanent intrinsic hyperfunctional state that remains after the extrinsic induction signals fade (“nociceptor memory”). First, widespread inflammation may persist chronically after SCI . Somata of nociceptors identified thus far in vertebrates, molluscs, and annelids are often located centrally, distant from their peripheral receptive fields, which ensures that these cells can survive sublethal injury that destroys their peripheral branches. In the mollusc, Why haven't chronic, somally expressed HSA states in nociceptors been reported previously in mammalian models of long-lasting pain? Few chronic studies have tested somal excitability in nociceptors, or recorded under conditions where nociceptor SA generated in the DRG could be distinguished from SA generated peripherally. Moreover, most studies of dissociated nociceptors have examined ionic currents under voltage clamp, without testing for somal hyperexcitability or SA. In addition, common models of long-lasting pain, including models based on cutaneous injection of inflammogens, surgical incision models (which produce relatively little inflammation or nerve damage), and various nerve injury models may not mimic adequately severe peripheral injuries that involve extensive amounts of tissue destruction  combined with prolonged inflammation—i.e., the traumatic conditions most likely to result in chronic pain in humans. Thus, while standard pain models produce many sensitizing effects on nociceptors  and other hyperfunctional alterations in nociceptors contribute significantly to human at- and below level pain, as well as to hyperreflexia and visceral problems after SCI, pharmacological agents that selectively target nociceptor hyperexcitability should be clinically useful. A number of genes important for excitability are preferentially expressed in nociceptors, raising the possibility that blocking their function could ameliorate some of the suffering caused by SCI while producing minimal side effects. Indeed, preliminary results show that SA in dissociated nociceptors depends upon nociceptor-specific Nav1.8 channels, and indicate that knocking down the expression of these channels can attenuate behavioral hypersensitivity after SCI (Yang et al., The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Daphnia magna) – bacterium (Pasteuria ramosa) system typifies such specificity and high virulence. We studied the North American host Daphnia dentifera and its natural parasite Pasteuria ramosa, and also found strong genetic specificity for infection success and high virulence. These results suggest that Pasteuria could promote Red Queen dynamics with D. dentifera populations as well. However, the Red Queen might be undermined in this system by selection from a more common yeast parasite (Metschnikowia bicuspidata). Resistance to the yeast did not correlate with resistance to Pasteuria among host genotypes, suggesting that selection by Metschnikowia should proceed relatively independently of selection by Pasteuria.The Red Queen hypothesis can explain the maintenance of host and parasite diversity. However, the Red Queen requires genetic specificity for infection risk  and strongly virulent effects of infection on host fitness. A European crustacean ( R0 when all else is equal, Genetic specificity between hosts and their parasites shapes the ecology and evolution of infectious disease Daphnia magna and its sterilizing bacterial parasite Pasteuria ramosa exemplify genetic specificity Pasteuria – the physical attachment of spores to the host oesophageal wall Pasteuria inflicts high virulence on infected hosts through sterilization Pasteuria epidemics can become large D. magna-Pasteuria system The European freshwater crustacean Pasteuria could maintain genetic diversity in other susceptible host species. Therefore, Pasteuria could act as a general catalyst of Red Queen-driven coevolution in freshwater systems. To test this possibility, we looked for genetic specificity for infection with Pasteuria in a natural North American host, Daphnia dentifera, using an established experimental design Pasteuria castrates its European D. magna hosts: Pasteuria's specificity and virulence effects are general, the European Daphnia – Pasteuria story may apply more broadly.These results suggest that D. dentifera populations, epidemics of the yeast Metschnikowia bicuspidata are usually larger and more frequent than Pasteuria epidemics Metschnikowia does not, however, exhibit genetic specificity with D. dentiferaPasteuria may not necessarily resist Metschnikowia. Such parasite-specific resistance  and five Pasteuria isolates  all originated from Midland Lake, Greene County, Indiana, USA. The five Pasteuria isolates were taken from five different infected hosts in 2010  and were propagated by feeding homogenized, infected hosts to a single host genotype (H119). These isolates likely consist of a mix of Pasteuria clones and represent ecologically relevant sub-populations of Pasteuria . We used only this lab-based culture because Metschnikowia isolated from different lakes and in different years show no genetic variation in infectivity or virulence on D. dentiferaD. dentifera (which is not a protected or endangered species), but permission was obtained for access to lakes.The experiments used host and parasite cultures and previously established laboratory protocols. The six clonal genotypes of Pasteuria isolates, one Metschnikowia isolate, and a sham-exposure control), replicated 15 times, for a total of 630 experimental units. Four of the Pasteuria replicates were lost due to accidental death after treatment exposure. Maternal lines consisted of individual Daphnia kept under favourable conditions (20°C and 16∶8 hour light/dark) in 40 mL of medium . Experimental animals consisted of second clutch offspring of third-generation maternal lines and were also kept in 40 mL of medium and fed ample food. In the parasite exposure treatments, each neonate (<24 hour old) received a high dose of spores  produced by gently crushing and diluting infected hosts. These doses aimed to yield similar infection levels among parasite species (based on pilot data). Sham controls received 100 μL of a Daphnia solution (50 uninfected hosts ground in 10 ml nanopure water). Replicates were fed lower food  during exposure to increase spore uptake by hosts 6Ankistrodesmus algal cells) again. Hosts were checked daily for reproduction and mortality for 25 days. Medium was refreshed every 2–3 days.The experimental design involved a factorial manipulation of these hosts and parasites. We crossed the six host genotypes with seven parasite treatments (five http://www.r-project.org). We analysed infection risk  by testing the fixed effects of host genotype and parasite species using data from only parasite-exposed hosts (not sham-exposed hosts). We tested for genetic specificity of Pasteuria infection using the fixed effects of host genotype and parasite isolate. Both Generalized Linear Models (GLM) were estimated with a binomial error distribution, and significance of each treatment was evaluated with deviance tests . However, in previous Daphnia-parasite studies, genotype is fitted as a fixed effect, a random effect or both (e.g. Metschnikowia and Pasteuria (averaged over the five isolates) for each host genotype using a Spearman rank correlation.All analyses were performed using the statistical package R . Fecundity was analysed by testing the fixed effects of infection status (infected or not), host genotype, and parasite species. Both host survival and r were analysed by testing the fixed effects of host infection status (infected or not) and parasite species. All of these tests used data from parasite-exposed hosts only; for r, the means of data from each infection category and parasite species for each host genotype were used.We examined the fitness consequences of infection by the two parasites using three metrics: production of host offspring (fecundity), host survival, and the instantaneous rate of host population growth, Metschnikowia's capacity to disrupt Red Queen dynamics between D. dentifera and Pasteuria should be reduced if the host pays an activation cost of resistance to Metschnikowia  We looked for evidence of an activation cost of resistance to Metschnikowia by comparing host fecundity, survival, and r in controls (sham-exposures) with hosts that were exposed to parasites but did not suffer infection .Fecundity across treatments was examined using a GLM fit with a quasipoisson error distribution (used due to over-dispersion of the fecundity data), and survival analyses used Cox's proportional hazards. Measures of Pasteuria) and between parasite species. Infection risk from Pasteuria depended on the specific combination of host and parasite genotypes . As expected based on prior studies, host genotype strongly influenced infection risk from Metschnikowia . There was, however, no correlation between a genotype's risk of infection by Metschnikowia and its overall risk of infection by Pasteuria  depended on both infection status and parasite species: infection with either parasite reduced r, but Pasteuria infections reduced r more than Metschnikowia infections. Thus, Pasteuria was more virulent . Thus, there was no evidence for a fitness cost of mobilising resistance mechanisms against Metschnikowia  to a lesser extent than Pasteuria  should reduce host population densities, even during small epidemics Due to differences in specificity and virulence, these two parasites may exert different ecological impacts on host populations. Theory suggests that specificity can reduce invasion success of parasites; i.e., they will have a lower reproductive ratio Pasteuria and Metschnikowia could either amplify or dampen Red Queen oscillations. A negative correlation would indicate antagonistic pleiotropy, where resistance to Pasteuria would come at a cost to resistance to MetschnikowiaMetschnikowia and Pasteuria suggests evolution of D. dentifera in response to the two parasites should proceed relatively independently. Thus, we do not expect Metschnikowia epidemics to amplify or dampen Red Queen dynamics between D. dentifera and Pasteuria.Correlations between host resistance to Pasteuria could promote genetic diversity in multiple host species through host-parasite coevolution. Pasteuria exhibits strong genetic specificity and high virulence with its North American host, D. dentifera, just as it does with its European host, D. magna. Can Pasteuria then promote host diversity in North American lakes? We cannot yet fully answer that question with these or other data, but the D. dentifera-Pasteuria system in isolation certainly contains several of the essential ingredients of the Red Queen. Still, it seems likely that the larger epidemics of the more common Metschnikowia yeast may mean that parasite-mediated selection in D. dentifera is driven primarily by Metschnikowia, making Red Queen dynamics relatively rare. This possibility further highlights the need to consider other co-occurring parasites when exploring host-parasite coevolution.In conclusion,


Cardiogenic shock is very uncommon in healthy people. The differential diagnosis for patients with acute heart failure in previously healthy hearts includes acute myocardial infarction and myocarditis. However, many drugs can also depress myocardial function. Propofol and fentanyl are frequently used during different medical procedures. The cardiovascular depressive effect of both drugs has been well established, but the development of cardiogenic shock is very rare when these agents are used.After a minor surgical intervention, a 32-year-old Caucasian woman with no significant medical history went into sudden hemodynamic deterioration due to acute heart failure. An urgent echocardiogram showed severe biventricular dysfunction and an estimated left ventricular ejection fraction of 20%. Extracorporeal life support and mechanical ventilation were required. Five days later her ventricular function had fully recovered, which allowed the progressive withdrawal of medical treatment. Prior to her hospital discharge, cardiac MRI showed neither edema nor pathological deposits on the delayed contrast enhancement sequences. At her six-month follow-up examination, the patient was asymptomatic and did not require treatment.Although there are many causes of cardiogenic shock, the presence of abrupt hemodynamic deterioration and the absence of a clear cause could be related to the use of propofol and fentanyl. Cardiogenic shock is the most serious form of presentation of heart failure (HF). The anticipation of hemodynamic deterioration and multiple organ failure in these patients is very important in terms of survival. The outcome for patients with refractory acute cardiogenic shock remains disproportionately poor. Technological advances in recent years have enabled the development of new treatments, such as extracorporeal life support (ELS). ELS is a variation of cardiopulmonary bypass which could improve the outcomes of patients in cardiogenic shock . AlthougWe report the case of a 32-year-old Caucasian woman who experienced sudden, severe hemodynamic deterioration after undergoing a minor surgical procedure. Her medical history was unremarkable except for a vaginal delivery two years before. She underwent surgery to remove a Bartholin cyst, and no infection in the gland was found. The operation was performed while the patient was under sedation and being given an analgesic. Spontaneous breathing was maintained by infusing a propofol bolus 1 mg/kg) and fentanyl 100 μg intravenously. During surgery, the patient remained hemodynamically stable. She has nausea and vomiting in the early post-operative period, which were treated with intravenous ondansetron 4 mg). A few minutes later the patient went into sudden hemodynamic deterioration, with sinus tachycardia (113 regular beats/minute) and hypotension (50/30 mmHg). Pulse oximetry showed that her oxygen saturation level had decreased to 80% despite oxygen supplementation through a face mask (fraction of inspired oxygen 40%). In this clinical situation, we treated her with intravenous dopamine and dobutamine, as well as with mechanical ventilation because of global respiratory failure . The electrocardiogram showed sinus tachycardia. Signs of HF were found on her chest X-ray, and urgent transthoracic echocardiography (TTE) revealed severe biventricular dysfunction with global hypokinesia and a LVEF estimated to be 35%. Coronary angiography showed no coronary lesions, and an intra-aortic balloon pump was inserted for counterpulsation. Repeat TTE revealed a LVEF of 20% with a dilated left ventricle  showing severe left ventricular dysfunction during the acute phase.Click here for fileShort-axis view showing severe left ventricular dysfunction. Transthoracic echocardiography  showing severe left ventricular dysfunction during the acute phase.Click here for fileApical four-chamber view after total recovery of left ventricular function. Transthoracic echocardiography  showing total recovery of left ventricular function before discharge.Click here for fileShort-axis view after total recovery of left ventricular function. Transthoracic echocardiography  showing total recovery of left ventricular function before discharge.Click here for file


Cellular and molecular regulation of the immune system is exquisitely controlled in the brain and is disrupted in neoplasia. Despite accumulation of immune cells in the tumor microenvironment, glioblastoma (GBM) growth persists while the mechanisms suppressing immune function remain largely unknown. Myeloid derived suppressor cells (MDSCs) are a heterogeneous class of immune cells responsible for immunomodulation through the suppression of cytotoxic T-cells. These populations are elevated in the peripheral blood of GBM patients, but their roles within the GBM microenvironment are uncharacterized. Through immunofluorescence analysis, we identified MDSCs within human GBM specimens, suggesting an immunosuppressive phenotype marked by arginase 1 (Arg1) staining. We have also observed co-localization between MDSCs and GBM cancer stem cells (CSCs) in both human tissues and mouse xenografts, leading to the hypothesis that CSCs recruit MDSCs to the tumor microenvironment, promote their survival, and that MDSCs are responsible for the immune-evasive properties of CSCs Fig. . Intracr


Hypertrophic cardiomyopathy (HCM) is most common in patients under 40 years and is associated with early sudden death, but HCM is increasingly recognized in elderly patients. We sought to characterize and identify features unique to elderly HCM patients when compared with youthful patients using CMR.Cine CMR and late gadolinium enhancement (LGE) imaging were performed in 902 HCM patients, including 202 young , and 143 elderly patients .Elderly HCM patients had a lower proportion of males . They had significantly lower left ventricular (LV) mass  and lower maximal LV wall thickness . The most common maximal thickness segment in the elderly patients was in the basal anterior septum . Elderly patients were also more likely to have the thickest segment located within the septum . Elderly patients were far less likely to have massive hypertrophy  .Elderly HCM patients had a smaller LV cavity size, with lower LV end-diastolic volume  and smaller LV end-diastolic dimension . The elderly patients had similar LV ejection fraction (LVEF)  and similar rates of depressed (<60%) LVEF .The prevalence of LGE (a marker of fibrosis) was similar in both groups . When present, both LGE mass  and percentage of LV with LGE  were similar.Substantial amounts of late gadolinium enhancement are compatible with normal longevity in many patients with hypertrophic cardiomyopathy. Presence of LGE may thus have different prognostic significance across the vast spectrum of HCM. This has potential implications for using LGE as a risk stratification tool for sudden death in HCM.Nil.


Successful scientific investigations at CAM institutions are often hampered by limited research infrastructure. Enhancing the clinical trial capabilities of our chiropractic research center was one goal of 2 NIH/NCCAM developmental center grants.Web applications were enhanced and customized based on individual project needs and trial team requests. Templates were built to support quick development and standard functionality of project web applications. We also developed standardized processes to effectively provide project-specific participant management, data collection, staff training, and quality control.Web applications included: the Centralized Participant Database System and Project/Users Permissions System to securely capture participant contact information and control personnel access to web modules; a real-time participant tracking report to monitor recruitment and participant flow; a role-specific Reminder System to track outstanding activities requiring personnel follow-up; and double key-entry verification on web data used for an adaptive treatment allocation algorithm. Templates of these tools can be incorporated into any new trial’s web application. Study protocol templates facilitate timely, accurate translation of research proposals into detailed protocols for IRB applications, DSMC review and training. A revised training plan included role-specific training logs for tracking training objective completion and a thorough certification process for key protocols to ensure accurate and consistent performance. Quality control templates support the informed consent process and fidelity of trial interventions and outcomes assessment. We implemented these enhanced resources in 5 clinical trials. To date, our team has completed 3378 phone screens, conducted 1439 first and 473 second in-person screening visits, enrolled 394 participants, and followed 315 through trial end. The study protocol templates have been used in two additional clinical trials currently undergoing IRB review.We expanded our capacity to support multiple, diverse trials with complex methodologies. This is a successful example of a CAM institution successfully leveraging research grant funding to enhance research infrastructure.


Synapses in neuron networks filter incoming spikes with a wide variety of time constants, affecting the stability of various collective dynamics , the selHere, we addressed this issue working out a unified framework in which from small to large synaptic time scales the same approximated theoretical description holds for the firing rate dynamics of spiking neuron networks. Starting from single-neuron stochastic dynamics, we derived a set of ordinary differential equations for the population emission rate relying on the spectral expansion of the associated Fokker-Planck equation, eventually extending to the colored noise case a previous derivation obtained under white noise hypothesis . The resMoreover, such perturbative approach allowed to avoid the analytical difficulties of dealing with multi-dimensional Fokker-Planck equations with discontinuous boundary conditions. A simplification which helped in highlighting as main result that networks of neurons with non-instantaneous synapses can, under fairly broad assumptions, be described by equivalent networks with suitable distributions of transmission delays. A formal analogy valid from simple VIF models to more realistic point-like simplifications like LIF or EIF neurons.


Late gadolinium enhancement (LGE) is excellent for identifying focal lesions in the myocardium , but diffuse homogeneous abnormalities are not readily detectable. Quantitative measurement of the myocardial extracellular volume (ECV) fraction using MRI has recently been validated histologically, and implemented for parametric imaging in patients. We hypothesized that quantification of ECV using ECV imaging could detect focal lesions detected by LGE, and diffuse homogenous disturbances in the myocardial extracellular volume fraction which are not detectable by LGE. The objective of the study was to examine the diagnostic utility of ECV MRI for detecting myocardial tissue abnormalities compared to LGE.Consecutive patients referred for clinical cardiac MRI evaluation of known or suspected heart disease were prospectively enrolled . MRI was undertaken at 1.5T using a Modified Look-Locker Inversion recovery (MOLLI) sequence before and 15 minutes after an intravenous bolus of a gadolinium-based extracellular contrast agent . Maps of the extracellular volume fraction (ECV images) were generated using MOLLI-derived T1-mapping before and after contrast, calibrated to hematocrit from a venous blood sample. Motion correction and co-registration was performed offline using an automated dedicated algorithm. ECV images were interpreted quantitatively and the results were compared to clinical assessment of LGE images. Increased myocardial ECV was determined by quantitative comparison to established normal values where the 95% confidence limits are 20-32%.Out of 32 patients, 10 had focal lesions . All lesions identified by increased ECV were also identified by blinded clinical read of LGE images (100% agreement). Among patients with normal LGE findings (n=22), two patients (9%) had diffusely increased myocardial ECV . The figure shows examples of LGE and ECV images in a normal finding, a focal lesion and a case with diffusely increased ECV.Our initial findings suggest that ECV and LGE imaging have excellent diagnostic agreement for identifying focal myocardial findings such as those seen in myocardial infarction and non-ischemic heart disease. The ability of ECV imaging to quantify and detect diffuse abnormalities in myocardial ECV increased the number of diagnostic findings compared to LGE in 9% of cases with otherwise normal LGE. Further studies in larger populations are warranted to explore the diagnostic and prognostic benefits of ECV imaging in assessment of diffuse pathologies of the myocardial extracellular space.The study has been funded in part by a grant (PI: Dr. Ugander) from the Swedish Society for Medical Research.


Central statistical monitoring (CSM) can identify trial misconduct, and help to prioritise on-site visits and additional training. Key risk indicators (KRIs) focus CSM on variables most likely to affect study reliability or patient safety. We developed the use of robust minimum covariance determinant (MCD) distances to detect outlying centres in the context of KRI analyses.Initially, a summary statistic (e.g. mean) describing the KRI is calculated for each centre and robust MCD-based distances are calculated by the FAST-MCD algorithm . For each KRI, robust estimates of the multivariate location Robust MCD distances offer a flexible approach for analysing both univariate and multivariate KRIs and can be implemented in standard statistical software.


Major trauma is the leading causing of death in young adults across the globe. Themortality from traumatic cardiac arrest remains high but survival with good neurologicaloutcome from cardiopulmonary arrest following major trauma has now been reported. Rapid,effective intervention is required to address potential reversible causes of traumaticcardiac arrest if the victim is to survive. There is no standard treatment algorithm fortraumatic cardiac arrest. We present a simple algorithm to manage the major traumapatient in actual or near cardiac arrest.We reviewed current pre-hospital clinical practice and the published literature on majortrauma management. An algorithm was developed and used regularly by London’s AirAmbulance pre-hospital physician/paramedic trauma team.The algorithm addresses the need treat potential reversible causes of traumatic cardiacarrest. This includes immediate resuscitative thoracotomy in cases of penetrating chestor abdominal trauma resulting in cardiac arrest, airway management, optimisingoxygenation, reversal of hypovolaemia using intravenous/intraosseous fluid replacementand chest decompression to exclude tension pneumothorax.A standard approach to traumatic cardiac arrest is feasible. Use of a treatmentalgorithm can rapidly, simultaneously address reversible causes of traumatic cardiacarrest and has the potential to save lives.


MDLcompress. We apply this tool to explore the relationship between miRNAs, single nucleotide polymorphisms (SNPs), and breast cancer. Our new algorithm outperforms other grammar-based coding methods, such as DNA Sequitur, while retaining a two-part code that highlights biologically significant phrases. The deep recursion of MDLcompress, together with its explicit two-part coding, enables it to identify biologically meaningful sequence without needlessly restrictive priors. The ability to quantify cost in bits for phrases in the MDL model allows prediction of regions where SNPs may have the most impact on biological activity. MDLcompress improves on our previous algorithm in execution time through an innovative data structure, and in specificity of motif detection (compression) through improved heuristics. An MDLcompress analysis of 144 over expressed genes from the breast cancer cell line BT474 has identified novel motifs, including potential microRNA (miRNA) binding sites that are candidates for experimental validation.We describe initial results of miRNA sequence analysis with the optimal symbol compression ratio (OSCR) algorithm and recast this grammar inference algorithm as an improved minimum description length (MDL) learning tool:


Optical coherence tomography (OCT) has enhanced our understanding of changes in different ocular layers when axial myopia progresses and the globe is stretched. These findings consist of dehiscence of retinal layers known as retinoschisis, paravascular inner retinal cleavage, cysts and lamellar holes, peripapillary intrachoroidal cavitation, tractional internal limiting membrane detachment, macular holes (lamellar and full thickness), posterior retinal detachment, and choroidal neovascular membranes. In this review, recent observations regarding retinal changes in highly myopic eyes explored by OCT are described to highlight structural findings that cannot be diagnosed by simple ophthalmoscopy. Pathologic or high myopia is defined as a refractive error of −6.00 diopters (D) or greater and an axial length exceeding 26 mm.2Optical coherence tomography (OCT) has recently made it possible to explore changes in ocular layers as axial myopia progresses and the globe is stretched. These findings consist of dehiscence of retinal layers known as retinoschisis, paravascular inner retinal cysts and lamellar holes, peripapillary intrachoroidal cavitation , tractional internal limiting membrane detachment, macular holes (lamellar and full thickness), posterior retinal detachment and choroidal neovascular membranes.In this review, we describe retinal changes in highly myopic eyes according to findings observed by OCT.The early stages of myopic retinal changes can easily be underestimated by biomicroscopy, angiography or ultrasonography, and therefore remain undiagnosed. Recently, OCT with its fine cross-sectional imagery of retinal structures has greatly facilitated the evaluation of posterior vitreoretinal anatomy in eyes with high myopia. It can also reveal otherwise undetectable retinal changes in asymptomatic patients. The OCT ophthalmoscope produces longitudinal retinal B-scans and coronal C-scans.B-scan produces cross-sectional images of retinal morphology which bears strong resemblance to histology. OCT C-scans are represented as 2-dimensional transverse slices at any given depth through the retina, thereby enabling visualization of the lateral extent of different structures. OCT C-scans are associated pixel-to-pixel points with simultaneously confocal images used to identify the exact position of lesions on the posterior pole. With en face OCT, localization and relation of retinal lesions can be precisely defined, especially in myopic eyes. En face OCT provides accurate imaging of retinal abnormalities in high myopia and allows width measurement and point-to-point localization of changes.Myopic posterior staphyloma, with its outward globe bulge, results in a deep concave B-scan OCT and distorted retinal structures. Because of the smaller base curvature of the horizontal section and larger base curvature of vertical section, macular structures can be defined more easily by vertical sections . Remarkably, the incidence of internal limiting membrane (ILM) detachment and macular retinoschisis was significantly higher in eyes with paravascular lamellar holes than those with other paravascular abnormalities. Paravascular lamellar holes were associated with ILM detachment in 44% and with macular retinoschisis in 20% of eyes. More than 80% of eyes with macular retinoschisis had paravascular lamellar holes.Tractional ILM detachment was reported in 2.4%–6% of highly myopic persons.In a study by Bando et alThe prevalence of myopic foveoschisis (MFS) ranges from 9% to 34% in highly myopic eyes with posterior staphylomas.23MFS tends to occur in eyes with severe myopic retinopathy.25Half of patients with MFS have been reported to develop retinal detachment or macular holes within two or more years of follow-up.26Choroidal neovascular membrane (CNV) is considered to be the most important sight-threatening complication of high myopia. Myopic CNV has been classified into active, scar and atrophic stages.In the scar stage, only the surface of the neovascular tuft shows high reflectivity and the tissue underneath is markedly attenuated. Overlying retinal layers are not disarranged or disorganized, cystic changes are also absent.In the atrophic stage, the CNV becomes flat and chorioretinal atrophy around the regressed CNV is highly reflective.29Macular holes always exist at the edge of a pigmented lesion suggestive of an old CNV. Therefore periodic OCT examinations for evaluation of macular holes or macular retinoschisis, even in patients in atrophic stages, are recommended. The retina in the area of chorioretinal atrophy around the CNV is very thin regardless of coexisting macular retinoschisis, but the retina at the edge of CNV in myopic eyes with large chorioretinal atrophy (larger than 1 disc diameter) has been reported to be significantly thinner in eyes with chorioretinal atrophy smaller than 1 disc diameter around the CNV.28Posterior retinal detachment is another consequence of ocular expansion and posterior staphyloma in high myopic eyes. Full thickness macular hole and tractional maculopathy may accompany this finding and may be its contributor .Macular hole formation in myopic eyes has multiple etiologies. Expansion of the globe, tractional components and growth of neovascular membranes through clefts in the posterior pole may create partial thickness and/ or full-thickness macular holes. In asymptomatic myopic persons, macular holes were detected in 6.26% of cases.Pathologic myopia is accompanied by thinning and stretching of the globe. With increasing age and myopia, these changes further progress.Individuals with high myopia are subject to various retinal pathologies including posterior pole and peripapillary lesions. Because of extreme retinal thinning and chorioretinal changes in these persons, simple fundus examination may miss subtle retinal pathologies. OCT is an accurate tool which can localize such anatomical changes. It seems to be the paraclinical test of choice for highly myopic eyes.


Cancer cells have altered metabolism, with an increased rate of glucose uptake, increased glycolysis, and due to the Warburg phenomenon, increased production of biomass (amino acids and nucleic acids) as metabolic byproducts. Association of these metabolic changes with breast cancer subtype and tumor gene expression has not been evaluated, nor is it well understood what biological processes and characteristics are most important in driving altered metabolic state in breast cancer.Thirty-four breast tumors and six samples of cancer-adjacent normal tissue were analyzed by liquid and gas chromatography (LC/GC) mass spectrometry (MS) to determine the expression of 379 metabolites. The same tissues were also used to perform gene expression microarrays and the resulting gene expression profiles were used to identify tumor subtype. These data combined with clinical tumor characteristics were evaluated in association with metabolic phenotype. To complement these analyses of human tissues, in vitro assays using human breast cancer cells alone and in coculture with cancer-associated fibroblasts (CAFs) were performed to study glucose uptake and utilization.Unsupervised cluster analysis on all metabolites resulted in two main clusters, with differences in the prevalence of aggressive tumor subtypes in each cluster. Characteristic of the Warburg phenomenon, four groups of metabolites showed differential levels between the two tumor clusters: amino acids, sugars, nucleic acids, and citric acid (TCA) cycle metabolites. Normal breast tissue samples were similar to less aggressive tumors with low biomass production and decreased levels of glycolysis/TCA byproducts. Parallel to these results in human tissues, glucose uptake was increased in basal-like breast cancer cells when they were cocultured with fibroblasts, but less aggressive luminal-fibroblast cultures did not show increased glucose uptake. Rather, in luminal cocultures, the fibroblasts appeared to suppress glucose uptake. Glucose utilization (oxidation and glycogen synthesis) was also higher in basal-like mono- and cocultures compared to luminal cultures. Consistent with these findings that the stroma influences metabolic phenotype, tumors that had active wound response gene expression had the more aggressive, Warburg-like metabolic profile.We observed distinct metabolic profiles associated with more aggressive breast cancer tumors. Based both on the observed patterns of glucose uptake and utilization in cocultures, and on the association between wound response activation and metabolic profile, these metabolic signatures appear to be influenced by multiple factors, including the stroma. These data suggest that integration of genomic and metabolic data provide insights for better understanding a Warburg effect in human breast cancer that may be regulated by the stroma.


Visual hallucinations (VH) represent one of the core features in discriminating dementia with Lewy bodies (DLB) from Alzheimer’s Disease (AD). Previous studies reported that in DLB patients functional alterations of the parieto-occipital regions were correlated with the presence of VH. The aim of our study was to assess whether morphological changes in specific cortical regions of DLB could be related to the presence and severity of VH. We performed a cortical thickness analysis on magnetic resonance imaging data in a cohort including 18 DLB patients, 15 AD patients and 14 healthy control subjects. Relatively to DLB group, correlation analysis between the cortical thickness and the Neuropsychiatric Inventory (NPI) hallucination item scores was also performed. Cortical thickness was reduced bilaterally in DLB compared to controls in the pericalcarine and lingual gyri, cuneus, precuneus, superior parietal gyrus. Cortical thinning was found bilaterally in AD compared to controls in temporal cortex including the superior and middle temporal gyrus, part of inferior temporal cortex, temporal pole and insula. Inferior parietal and supramarginal gyri were also affected bilaterally in AD as compared to controls. The comparison between DLB and AD evidenced cortical thinning in DLB group in the right posterior regions including superior parietal gyrus, precuneus, cuneus, pericalcarine and lingual gyri. Furthermore, the correlation analysis between cortical thickness and NPI hallucination item scores showed that the structural alteration in the dorsal visual regions including superior parietal gyrus and precuneus closely correlated with the occurrence and severity of VH. We suggest that structural changes in key regions of the dorsal visual network may play a crucial role in the physiopathology of VH in DLB patients. Dementia with Lewy bodies (DLB) is the second most common form of dementia in the elderly after Alzheimer’s Disease (AD). Despite the precise nosological relationship between DLB and AD remains uncertain, persistent visual hallucinations (VH) along with fluctuations in cognitive function and parkinsonism represent the core feature in discriminating DLB from AD and other dementias The physiopathology underlying VH in DLB is not well understood. Causative models for complex VH have suggested that the origin of this deficit may be related to a range of pathological alterations affecting both the dorsal visual stream, specialized for visuo-spatial attention and location, and the ventral visual stream, designated to object recognition In this context, structural Magnetic Resonance Imaging (MRI) could be a powerful tool to investigate the aetiology of core symptoms in DLB. To date, several studies by using voxel-based morphometry (VBM) found widespread cortical sites of grey matter atrophy in DLB patients compared to healthy controls Cortical thickness approach allows to perform cortical parcellation and to measure with great reliability the pathological thinning of the cortical grey matter. By measuring a real physical quantity that is much easier to interpret and to localize in comparison to the three-dimensional VBM, cortical thickness methods provide a better localization of structural changes because the smoothing procedure is performed on a two-dimensional manifold which reduces morphological artefacts due to ageing and pathological processes Despite the potential advantage of cortical thickness analysis, to date this approach has not been applied to DLB patients.In the present study, we performed cortical thickness analysis to verify the presence of cortical grey matter damage related to VH in DLB patients. To this aim, firstly, we compared DLB patients respect to healthy controls. Subsequently, we included a comparison against AD groups to exclude possible pathologic alterations arising as a result of a dementia process per se and not specific to DLB pathology. Finally, relatively to DLB group, we performed a correlation analysis between the cortical thickness and the Neuropsychiatric Inventory (NPI) hallucination item scores to highlight the cortical regions which were effectively related to the occurrence and severity of VH.The study was approved by the Institutional and Ethics Committee of the University “G. d’Annunzio” Chieti-Pescara (ID#157801). All procedures were conducted according to the Declaration of Helsinki and subsequent revisions All patients underwent MRI scan within six months before the inclusion in the study and dopaminergic presynaptic ligand ioflupane SPECT (DAT scan).Global tests of cognition included Clinical Dementia Rating (CDR), Mini Mental State Examination (MMSE), Dementia Rating scale-2 (DRS-2) Exclusion criteria were uncontrolled hypertension, myocardial ischemia, peripheral vascular disease and chronic kidney.1-Weighted Turbo Field-Echo sequence  was carried out. Participants with images characterized by motion artefacts were excluded from the analysis.All acquisitions were carried out with a Philips Achieva 3 T scanner  equipped with 8-channel receiver coil. After scout and reference sequences, a 3-dimensional T1-weighted image was estimated at each vertex across the brain surface using Freesurfer software package (http://surfer.nmr.mgh.harvard.edu). This method has been previously described in detail by Fischl and Dale Cortical thickness on TCortical brain regions affected by cortical thinning were classified by using the Desikan-Killiany Atlas integrated in FreeSurfer.SPSS version 14.0 was used for statistical analysis. Analysis of variance (ANOVA) among groups and Tukey’s HSD post-hoc test was performed on demographic and clinical data. Chi-squared test was carried out for gender.For each hemisphere, comparison among groups was performed by using general linear model (GLM), including cortical thickness as dependent factor and group  as independent factor. MMSE, CAF and UPDRS scores were included as covariates.https://surfer.nmr.mgh.harvard.edu/fswiki/FsgdFormat).FreeSurfer processing stream was used to assess the pairwise differences among groups  was used to individuate statistically significant regions where NPI hallucination item scores and thickness were correlated. MMSE, CAF, UPDRS scores were included as nuisance factors.In DLB group, QDEC . Next, SPSS version 14.0 was used for statistical computation and MMSE, CAF and UPDRS scores were included as nuisance factors.To perform partial correlations between NPI hallucination item scores and mean cortical thickness values in the posterior regions which were affected significantly in DLB in comparison with AD, the mean cortical thickness values were extracted from each region of interest (defined by Desikan-Killiany Atlas) by using “aparcstats2table” command line,  in cortical thickness between DLB and age-matched healthy controls. Cortical thickness was reduced in the posterior regions of DLB. Specifically, pericalcarine, lingual gyri, cuneus precuneus and superior parietal gyrus were affected bilaterally. panel B shows regional differences (corrected p<0.05) in cortical thickness between DLB and AD. Cortical thickness was reduced in DLB compared to AD in the right posterior regions including superior parietal gyrus, precuneus, cuneus, pericalcarine and lingual gyri.With the present MRI study, we have evidenced that DLB patients are characterized by marked cortical thinning in posterior regions when compared with AD and age-matched healthy controls.Posterior cortical regions play a critical role in the visual information processing, and their damage was related to recurrent VH in DLB patients Consistently with neuroimaging Although previous studies reported abnormalities related to VH in DLB or Parkinson’s Disease (PD) patients compared to control groups Additionally, despite both the dorsal and ventral attention networks were affected in DLB as compared to controls and AD, the correlation analysis between cortical thickness and NPI hallucination item scores, suggested a greater involvement of the dorsal attention regions including superior parietal gyrus and precuneus in the occurrence and severity of VH in DLB.The superior parietal region is implicated in visuo-spatial working memory Precuneus acts in concert with the parietal regions, elaborating information about motor imagery and more abstract mental imagery tasks Despite impairment of dorsal stream is well defined in DLB patients, the ventral stream resulted to be affected in both AD and DLB as compared to controls. However a different distribution of grey matter loss along the ventral pathway was found in DLB and AD. Specifically, cortical thinning in lingual and pericalcarine giri was observed in DLB, whereas a marked and diffuse cortical thinning was found in the mesial temporal cortex in AD. This latter structural alteration could be linked to greater memory impairment affecting AD patients and it is in accordance with previous morphometric studies According to previous reports from our group Different models have suggested a central role of posterior regions in manifestation of VH


Client-owned pet dogs represent exceptional translational models for advancement of Cancer Research, as they reflects the complex heterogeneity observed in human cancer. We have recently shown that a genetic vaccine targeting dog telomerase (dTERT) and based on Ad/DNA-EGT technology can induce strong cell-mediated immune responses against this tumor antigen and increase overall survival of dogs affected by B-cell lymphosarcoma (LSA) in comparison with historical controls when combined with COP chemotherapy regimen. Here, we have conducted a double arm clinical trial with an extended number of LSA patients, measured the antigen-specific immune response and evaluated potential toxic effects of the immunotherapy along with a follow up of patients survival for three years and half. The immune response was measured by ELISPOT. The expression of dTERT was quantified by quantitative PCR. Changes in hematological parameters, local/systemic toxicity or organic dysfunction and fever were monitored over time during the treatment. dTERT-specific cell mediated immune responses were induced in almost all treated animals. No adverse effects were observed in any dog patient that underwent treatment. The overall survival time of vaccine/COP treated dogs was significantly increased over the COP-only treated cohort . There was a significant association between dTERT expression levels in LSA cells and overall survival (OS) among vaccinated patients. In conclusion, Ad/DNA-EGT-based cancer vaccine against dTERT in combination with COP chemotherapy is safe and significantly prolongs the survival of LSA canine patients. These data confirm the therapeutic efficacy of dTERT vaccine and support the evaluation of this approach for other cancer types as well as the translation of this approach to human clinical trials.


Optimizing productivity and growth of recombinant Chinese hamster ovary (CHO) cells requires insight and intervention in regulatory processes. This is to some extent accomplished by several 'omics' approaches. However, many questions remain unanswered and bioprocess development is therefore still partially empirical. In this regard, the analysis of DNA methylation as one of the earliest cellular regulatory levels is increasingly gaining importance. This epigenetic process is known to influence transcriptional events when it occurs at specific genomic regions with high CpG frequencies, called CpG islands (CGIs). Being methylated, CGIs attract proteins with methyl-DNA binding domains (MBD proteins) that in turn can interact with chromatin modifying complexes, thereby leading to a transcriptionally inactive state of the associated gene . In CHO Based on the genomic and transcriptomic information available for CHO cells ,5, 21,992+, MAPK and Wnt signalling systems were comprised within the latter group and showed a large coverage by differentially methylated components. 48 hours upon butyrate addition the number of differential methylations decreased by about 90 %. COBRA analysis of the Wnt responsive myc proto-oncogene protein-like gene showed clearly detectable cleavage products  24 hours upon butyrate addition, that completely vanished another 24 hours later (Figure Butyrate treated CHO DP-12 cultures stopped proliferating and decreasing viabilities could be detected 24 hours upon addition of the SCFA Figure . Simultar Figure , confirmOur first genome-wide screening for differential DNA methylation in CHO cells shows that the epigenetic response upon butyrate treatment seems to be highly dynamic and reversible. This was confirmed by applying the bisulfite-based single-gene method COBRA to analyze a region of the myc proto-oncogene protein-like gene. Furthermore, detection of differential methylation before butyrate addition indicates that heterogeneity in DNA methylation occurs even if cells originated from the same preculture and were treated identically. This occurrence of differentially methylated genes in parallel cultivations strongly fosters the hypothesis that the culture history influences final process outcomes . It unde


Sensory cortex often undergoes population activity fluctuations due to attentional modulations and arousal changes, but activity fluctuations can also be generated intrinsically. The mechanisms that underlie these spontaneous activity fluctuations are unknown, in particular whether they arise from common inputs or more active neuronal processes. We measured the activity of tens of V1 neurons simultaneously in monkeys stimulated with oriented visual stimuli, and found that the tuning curves (TCs) of orientation selective neurons spontaneously underwent shifts and multiplicative scalings proportional to population activity while their widths remained constant Based on this observed TC activity dependence, we constructed a precise statistical model featuring Poisson-like firing, shifts and gain modulation. The model not only accounted for neuronal co-variability but also approached the performance of state-of-art decoding methods. Surprisingly, we found that decoding performance on sensory stimuli increased with population activity, despite the fact that the stimulus was identical. Therefore, spontaneous population activity fluctuations display highly non-random features, boosting TCs by shifts and multiplicative factors that gate information processing.We found that TCs are shifted and scaled by a multiplicative factor proportional to mean population activity Figure . The amoThe observed boosting properties of TCs motivated us to build a statistical model with Poisson-like neurons that includes a multiplicative scaling factor (PS). We observed that the effect of population activity on TCs is not purely multiplicative, and we introduced therefore a shift (PSS). Our models approached the performance of state-of-art decoding techniques  and provided higher accuracy than other tested decoders based on population vector and independent Poisson neurons. Experimental methods are as in .


Diverse therapeutic strategies for food allergy are under investigation including food allergen-specific and non-specific approaches. Allergen-specific approaches include immunotherapy with native food allergens, immunotherapy with mutated recombinant allergens, and diets containing extensively heated milk or egg. Native allergen immunotherapy can be administered via oral, sublingual, subcutaneous, or epicutaneous route. Oral immunotherapy trials with native food allergens such as milk, peanut, or egg reported that approximately 50-75% of the treated subjects reached and tolerated the maintenance dose. Failure of desensitization occurred in about 10-20% of the treated subjects and might be associated with the most severe and likely permanent food allergy phenotype, as opposed to the successful desensitization and tolerance that might be associated with a transient clinical phenotype and higher chances of spontaneous resolution of food allergy. Food allergens can be altered to decrease their allergenicity and lower the risk of acute adverse reactions. Immunotherapy with mutated recombinant peanut proteins, which have decreased IgE-binding activity, co-administered within heat-killed E.coli to generate maximum immune response had a strong protective effect in the mouse model of peanut anaphylaxis. This vaccine is being currently evaluated in the phase I clinical trials in adults with peanut allergy. Heating and processing changes food protein conformation and affects how food proteins are digested and transported via the gut barrier. Extensively heated (baked) milk and egg are tolerated by approximately 70% of the milk or egg allergic children. Diets containing extensively heated (baked) milk and egg represent an alternative approach to food oral immunotherapy and are already changing the paradigm of strict dietary avoidance for food-allergic patients.