Modern human activity fueled by economic development is profoundly altering our relationship with microorganisms. This altered interaction with microbes is believed to be the major driving force behind the increased rate of emerging infectious diseases from animals. The spate of recent infectious disease outbreaks, including Ebola virus disease and Middle East respiratory syndrome, emphasize the need for development of new innovative tools to manage these emerging diseases. Disseminating vaccines are one such novel approach to potentially interrupt animal to human (zoonotic) transmission of these pathogens. If at first the idea is not absurd, then there is no hope for it.Modern human activity is profoundly and irreversibly changing our natural world. In additMicrobial movement into humans from animals (zoonotic transmission) has always been responsible for the majority of EIDs, including at one time EIDs that form such present day matrix diseases as malaria falciparum, measles The task of identifying and preventing zoonoses is daunting, particularly given that the emerging pathogen may be completely unknown. The availability of limited resources and allocation of these resources over short-term funding periods further compounds the problem.,9 RecentMycobacterium bovis to cattle has had some success in the control of bovine TB in Ireland and regions of the UK . Howeveiew, see  This unaiew, see  – with liew, see In non-temperate ‘hotspot’ regions of Africa, Asia and South America, the ability to contain even known EIDs such as Ebola virus in wildlife is currently not possible. Management of diseases that involve livestock in these regions, such as Rift Valley fever and Crimean Congo hemorrhagic fever, pose similar problems,,18 in thEven with programs such as EPT, prediction of which animal pathogens will become established as globally significant EIDs within the human population still remains beyond our capability. However, pathogens emerging from an animal source are often initially poorly adapted to their new human host in terms of sustained human to human transmission. MechanisThe earliest disseminating vaccine for animals was designed to target two highly lethal rabbit-specific EIDs in the European rabbit population, myxoma virus (MV) and rabbit hemorrhagic disease virus (RHDV). The vaccIn the above studies, MV was selected as the genetic basis for the disseminating vaccine due its ability to spread through rabbit populations. High species specificity of MV for rabbits also decreased the potential for spread to ‘off-species’ targets within the environment. However, use of a normally virulent pathogen for the host species being targeted as the self-disseminating vaccine platform necessarily required use of an attenuated MV strain. This requirement had a clear impact on the disseminating capacity of the MV6918-based vaccine. More recent self-disseminating vaccine approaches have used cytomegalovirus (CMV), which is a beta-herpesvirus, as the disseminating vaccine platform. Similar to MV, CMVs are immunogenic and spread efficiently through their host species.–27 HowevAlthough not targeting an EID, over a decade of work toward the use of a murine CMV (MCMV) as a viral vectored immunocontraceptive vaccine to control mouse plagues in Australia gives some insight into the application of disseminating vaccines to target high risk pathogens for EID control. Mice directly infected with MCMV strains expressing female mouse fertility antigens develop prolonged – essentially life-long – infertility. Immunocoin vitro combined with the lower anti-PCMV antibody levels suggested a level of attenuation. However, PCMV(ΔP33:G1EGFP) was still able to induce G1-specific immunity in healthy deer mice previously infected with either PCMV(ΔP33:G1EGFP) or wild-type PCMV.[The first studies using CMV as a disseminating vaccine targeting a human EID used CMV from deer mice (Peromyscus CMV (PCMV)) to target SNV in the wild deer mouse SNV transmission species. Using a PCMV expressing the SNV envelope glycoprotein G1, the PCMV(ΔP33:G1EGFP) vaccine induced G1-specific antibodies following direct inoculation of deer mice. PCMV are also a known reservoir of Marburg virus [CTL) was shown to induce durable NP-specific immunity (>14 months).[CTL vaccinated mice showed no evidence of Ebola virus disease (EVD) following lethal Ebola virus challenge. Impressively, 5/8 mice completely controlled Ebola virus infection, with no detectable viremia; the remaining 3 mice showed a 2.8 log reduction in viremia relative to non-vaccinated controls. Protection was long-lived as mice vaccinated with a single dose of MCMV/ZEBOV-NPCTL were protected against EVD following lethal challenge 17 weeks post-vaccination – an attractive quality for a disseminating vaccine to be used in wildlife populations. Studies in the NHP Ebola virus challenge model were recently completed (manuscript under review). In this model, the key question of transmissibility of immunity in CMV seropositive animals (which cannot be assessed in the laboratory mouse model (see above)) can now be addressed in an experimental system more translatable to NHPs in the wild.A disseminating CMV-based approach is also being developed toward the control of Ebola virus in wildlife reservoir and transmission species in Africa.,39 Approrg virus ; a disserg virus ,39 In th months).,39 MCMV/Substantial evidence supports the ability of primate CMVs, including human CMV (HCMV), to superinfect the seropositive host. A 2008 study examining HCMV seropositive women showed the frequent presence of multiple glycoprotein N (gN) and/or gB variants within HCMV positive urine and blood samples suggesting that most individuals are infected with multiple HCMV strains. A subseqThe studies performed in the SIV:rhesus macaque model showed that the ability to superinfect was dependent on genes in the US2-11 region of the genome – a region which contains several genes involved in down-modulation of MHC class I antigen presentation. CD8+ T-cFurther studies are needed to ensure recombinant CMVs expressing heterologous target antigens can similarly maintain wild-type transmission and target-specific immune responses following transmission. Experience from studies exploring the use of disseminating vaccines targeting other pathogens (see above) are expected to prove invaluable for these studies, especially in regard to the importance of avoiding vaccine attenuation to maintain wild-type characteristics of CMV transmissibility. Where studied, frequencies of CMV infection from natural transmission in animal populations approach 100%. Consistent with epidemiological studies in humans, a major peak of infection occurs at an early host age, with essentially all US primate center rhesus macaques being RhCMV seropositive by the age of one year. Environmall countries has been raised to a level that enables identification and control of high-risk EIDs at source, we as a global community will be fated to reactive responses to EID outbreaks. Innovative strategies are therefore urgently required to identify and then pre-emptively control EIDs in these under-resourced ‘hotspot’ regions. Self-disseminating vaccination is one innovative approach that may potentially overcome the problems associated with use of conventional vaccines for pre-emptive pathogen control in ‘high-value’ animal populations within these challenging environments. Although still in relatively early stages, the nascent field of self-disseminating transmissible vaccines has the potential to solve many current intractable public health and conservation problems that cannot be addressed by conventional vaccines ; and UK applications No. 1508136.7, No. 1514005.6 and No. 1514034.6 (MA Jarvis and AA Murphy). The authors would like to acknowledge support by the School of Biomedical and Healthcare Sciences, Plymouth University (MA Jarvis and AA Murphy), The Marie Skłodowska-Curie Programme (MA Jarvis) and The Institute for Immunology and Infectious Diseases, Murdoch University (AJ Redwood). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.Emerging infectious diseases (EIDs) place an increasing burden on human health and infrastructure. The increasing rate of EID incidence can be directly linked to human activity.Current funding paradigms often focus on wealthy countries rather than developing countries which contain geographic EID ‘hotspots’. This funding paradigm leaves the world ill-prepared to detect and then control these diseases.Targeting zoonotic pathogens in wildlife populations by vaccination is a potentially powerful method for reducing transmission of EIDs. Direct vaccination of these animals may be hampered by a number of obstacles, including: poor infrastructure to support necessary cold chains, inaccessibility of animal species, inhospitable terrain and cost.Vaccines capable of self-dissemination may overcome many of the hurdles faced by direct administration of conventional vaccines. Self-disseminating vaccines will need to display species specificity, immunogenicity and normal transmission dynamics. Cytomegalovirus (CMV)-based viral vectors potentially fulfill many of these requirements, including being native to and endemic in target animal species.‘Proof of concept’ studies using direct inoculation of CMV-based vaccine vectors have been performed for viral pathogens such as Sin Nombre hantavirus and Ebola virus. CMV-based vaccine vectors have also been used for successful ‘proof of concept’ of viral vectored immunocontraception.Production of CMV-based vaccines with proven capacity to transmit is the next step in the development of self-disseminating vaccines.


A 15-year-old previously healthy boy was involved in a traffic accident and presented to our emergency department (ED) with complaints of chest pain and mild shortness of breath. On physical examination, he had complained for the last 2 hours of the right hemithorax pain in the fifth intercostal space radiating to the right midaxillary line. He had no past medical or surgical history. Chest x-ray demonstrated doubtful multiple thin-walled cystic lesions in the right lung (A). Computed tomography (CT) scan of thorax revealed a well- defined thin-walled, and 9.97 × 8.78 cm in diameter giant air cyst occupying the right upper lobe parenchyma with multiple air-space cysts (B). The patient was finally diagnosed as a multiple cystic lung disease and he underwent elective thoracic surgery for cyst removal. Cystic lung disease is defined as intrapulmonary air-containing multiple cysts surrounded by sharply demarcated thin walls. CT scanning of thorax is more sensitive than chest radiography in the detection and the distribution of lung cysts. Patients with cystic lung disease may be asymptomatic or present with nonspecific symptoms, such as chronic cough or shortness of breath. They are at increased risk for spontaneous pneumothorax. Surgical treatment of multiple cystic lung disease plays a crucial role in the prevention of pneumothorax.


Spinal cord ischemia is a well-known potential complication of endovascular aneurysm repair (EVAR), and it is usually manifested by paraplegia or paraparesis. We describe a case in which spinal cord ischemia after EVAR presented by isolated bladder and rectal incontinence without other neurological deficits. A 63-year-old woman presented with intermittent claudication secondary to an infrarenal abdominal aortic aneurysm (AAA), and a left common iliac artery obstruction, for which she underwent EVAR using an aorto-uniiliac (AUI) device and ilio-femoral artery bypass. On postoperative day 3, she developed urinary and fecal incontinence without signs of paraplegia or paraparesis. Magnetic resonance imaging (MRI) showed a hyper-intense signal in the spinal cord. She received hyperbaric oxygen (HBO) therapy and was discharged after 18 days when her urinary and fecal incontinence were almost resolved. This report suggests that spinal cord ischemia after EVAR for aortoiliac occlusive disease might present as bladder and rectal incontinence without other neurological manifestations. Neurologic complications due to spinal cord ischemia (SCI) after elective infrarenal abdominal aortic aneurysm (AAA) repair, while catastrophic, rarely happen. Approximately 1 in 400 cases occur after open AAA repair while 1 in 5000 occur after arterial reconstruction of occlusive aortoiliac disease . ProlongWe describe a case of a woman who developed isolated bladder and rectal incontinence without paraparesis/paraplegia after endovascular aneurysm repair, treated with hyperbaric oxygen (HBO) therapy, and with significant clinical improvement after 3 weeks.A 63-year-old woman with a history of hypertension and chronic kidney disease presented with intermittent claudication. A computed tomography (CT) revealed a 41-mm infrarenal AAA with total obstruction of the left common iliac artery. The bilateral external and right internal iliac arteries were patent. The distal left internal iliac artery was occluded but filled through collateral circulation Fig. . Given tAfter a thorough risk-benefit discussion with the patient, we performed an endovascular repair with an aorto-uniiliac (AUI) device  and ilio-femoral artery bypass using an 8-mm vascular graft (Goretex ePTFE graft) in a “hybrid” operating room under general anesthesia. After bilateral groin incision to expose the right and left common femoral artery, a bifurcated stent graft  was placed just below the renal artery branch section to the right common iliac artery through the right external iliac artery, followed by placing a converter of 24 mm in a proximal diameter ESC-24-12-80) and an iliac extender of 16 mm in a distal diameter (TFLE-16-56-ZT) for the right common iliac artery. An arteriogram confirmed adequately positioned endografts, with patent bilateral renal arteries without evidence of proximal or distal endovascular leak. Subsequently, the right external iliac artery was cut longitudinally for the inflow for the bypass. The right external iliac artery and an 8-mm vascular graft (Goretex ePTFE graft) were anastomosed by using a parachute technique with a CV6 needle. The left common femoral artery was also identically anastomosed to the graft  showed a hyper-intense signal at the lower end of the conus medullaris visible in the T2-weighted image. Fig. . CerebroSCI has been reported after endovascular aneurysm repair (EVAR) either as an immediate or a delayed finding . An analThe process leading to SCI after elective surgical management of an infrarenal AAA has not been fully understood. Factors that may contribute to it after EVAR include atheromatous embolization, interruption of the great radicular artery (artery of Adamkiewicz), or collateral circulation  , 4, 7. IAtheroembolization is a well-known complication of endovascular surgery and may lead to SCI. Rockman et al. reported two cases of paraplegia as a result of atheroembolization to the spinal cord after successful or attempted endovascular management of AAA . In the The patient’s clinical presentation is explained by the anatomy of the conus medullaris. The conus medullaris is the lower end of the spinal cord, and it usually occurs at the level of the first lumbar vertebra. The segments of spinal cord that give origin to the nerves that control the bladder and bowel sphincters are located below the third sacral segment (S3). An injury to the conus medullaris below the third sacral segment (S3) will manifest with bladder and rectal incontinence and symmetric sensory loss around the anus and sexual organs. Motor dysfunction will not develop because most of the lower extremity motor nerves originate above the S2 segment of the spinal cord . AlthougThe goal of the treatment for SCI is to augment spinal cord perfusion pressure and reduce edema. The usefulness of the COPS protocol (CSF drainage to maintain a pressure of less than 5 mmHg without limit under CSF drain status/oxygen delivery/patient status) has been reported after delayed neurologic deficit in thoracoabdominal aortic aneurysm repair . Other tHBO therapy has sporadically demonstrated a decrease in the size of infarcts . In the We describe a case of a woman who developed isolated bladder and rectal incontinence without paraparesis/paraplegia after EVAR, and HBO might be beneficial for improving the symptoms.Written informed consent was obtained from the patient for publication of this case report and any accompanying images.


The 11-year old girl presented a diagnosis of viral hepatitis A confirmed by anti-HVA IgM. Eight 8 weeks after, she developed hemorrhagic syndrome coupled with fever associated and persistent jaundice. The blood count showed aregenerative pancytopenia secondary to aplastic anemia confirmed by bone marrow biopsy. During conditioning for allogenic bone marrow grafts, the patient developed generalized seizures revealing disseminated intracerebral hemorrhage (pictures). There was a favorable outcome after blood transfusions and resuscitation. Hematological complications in hepatitis A are rare. Although intracerebral hemorrhage resulting from Hep A has never been reported in the literature; its occurrence is directly correlated with the degree of thrombocytopenia due to aplastic anemia. The prevalence of aplastic anemia in viral hepatitis is estimated at 0.1%. This complication implies systematic hematological evaluation in cases of viral hepatitis A; especially in severe forms or those associated with involving cholestasis.


Double ventricular response is a rare arrhythmia that results from simultaneous antegrade conduction over the fast and slow pathways of AV node. Double ventricular responses may lead to arrhythmia-related cardiomyopathy. As far as we know, there is not any reported reversible cardiomyopathy development that resulted from double ventricular responses to one atrial impulse after slow pathway ablation. We report a unique case of double ventricular response cardiomyopathy that has been developed 5 years after slow pathway ablation. Double ventricular response is a rare arrhythmia that results from simultaneous antegrade conduction over the fast and slow pathways of AV node. Slow-pathway conduction must be slow enough to allow his-purkinje system to recover excitability after depolarization by the first excitation over the fast pathway. Double ventricular responses may lead to tachycardia-related cardiomyopathy. We report a unique case of double ventricular response cardiomyopathy that has been developed 5 years after slow-pathway ablation.A 69-year-old patient admitted to our ambulance with severe dyspnea. Five years ago a slow-pathway cryoablation due to typical atrioventricular nodal reentry tachycardia was performed. Transthoracic echocardiography performed at admission revealed severely reduced left ventricular ejection fraction (LVEF) (30%), which was reported normal 5 years ago. In the next step, haemodynamic relevant stenoses were excluded in coronary angiography. Cardiac MRI confirmed severely reduced LVEF without the presence of myocardial fibrosis. Electrocardiography (ECG) revealed an uncommon finding of two ventricular beats after one atrial beat . An elecTo the best of our knowledge, this is the first report of reversible cardiomyopathy due to double ventricular response to one atrial impulse that developed as a late complication of slow-pathway cryoablation.Atrioventricular nodal reentrant tachycardia (AVNRT) is one of the most common sustained supraventricular arrhythmia. Catheter ablation of the slow pathway is treatment of choice for patients with recurrent drug-refractory AVNRT . The risDouble ventricular response is a rare arrhythmia that results from simultaneous antegrade conduction over the fast and slow pathways of AV node . Slow-paThe presence of his electrograms before each QRS complex and a relatively stable interval between the fast- and slow-pathway his electrograms during electrophysiological study supports the concept. However, simple junctional extrabeats as bigeminus cannot fully be excluded. The termination of the tachycardia with slow-pathway ablation is the treatment of choice.Although double ventricular responses and tachycardia-related cardiomyopathy that reverses with slow-pathway ablation has been described in the literature, our case is unique in which double ventricular responses and related cardiomyopathy occurred 5 years after slow-pathway ablation.It is known that there may be late recovery of conduction after successful ablation . In our The normalization of LV function after ablation strongly supports that cardiomyopathy was related to the dual ventricular responses.


The development of immune-based strategies for cancer for solid cancers is challenged by the scarcity of T cells with high receptor avidity for tumor-specific antigens within the patient´s lymphocyte repertoire, and by the failure of tumor cells to present antigen to T cells. Both obstacles can be bypassed by genetic modification of T cells with recombinant chimeric receptors (CARs) which redirect T cells towards a tumor surface antigen independent of antigen presentation. CAR reengineered T cells efficiently interact with tumor cells in vitro and have significant in vivo activity against tumor xenografts. Recently, first clinical trials have shown evidence for a potent antitumor activity of CD19-specific CAR T cells in leukemia. Current efforts focus on improving in vivo survival, functional persistence and potency of adoptively transferred anti-tumor T cells. The design of more effective strategies against both solid tumors and leukemias further depends on enhanced knowledge of specific mechanisms of immune escape. Moreover, rational combinations of targeted therapies with immunotherapies and optimal integration of cellular therapies into current treatment regimens may allow higher rates of durable responses.


Septo-optic dysplasia (SOD) is a heterogeneous malformation condition consisting of optic nerve hypoplasia, various types of forebrain defects and hormonal deficiencies. This study aims to expand knowledge about endocrine abnormalities in patients with SOD in ASEAN countries.Forty-eight patients  who has been diagnosed as having SOD in ASEAN countries were clinically reviewed from medical records.Clinical manifestations and endocrine abnormalities of the patients are shown in Table 1This multicenter and multinational study shows that about 20-35 % of SOD patients have endocrine abnormalities. Hypothyroidism and GH insufficiency are the most common endocrine problems associated with this condition.


Plasmodium berghei in inbred mouse strains [Babesia rossi causes cerebral disease in some cases [The favoured animal model of cerebral malaria is an artificial host-parasite combination caused by  strains . Canine me cases . This dime cases .Post mortem brains collected from 50 natural cases of canine babesiosis showing clinical signs of cerebral involvement were collected and evaluated grossly and using light and electron microscopy.Grossly visible lesions (seen in 31/50) were classified as global (16/50) or regional (34/50). Global lesions were diffuse swelling and diffuse cerebral congestion or pallor. Multifocal petechial haemorrhages and white matter malacia appeared more regional. There were 18/50 cases that had a grossly appreciable oedema. Histological lesions appeared in a spectrum of severity, and included very localized endothelial injury. Babesia-parasitised red cell sequestration was a feature in some sections. Early lesions were multifocal and strictly associated with the microvasculature. Intermediate lesions were characterized by perivascular haemorrhage and some neutrophil infiltration. Advanced lesions were locally extensive and similar in appearance to haemorrhagic infarction. Ultrastructural evidence of cytoadherence between erythrocytes and capillary endothelium was demonstrated. Endothelial cell necrosis occurred early in the development of the lesions before neuronal and glial changes.The endothelial injury, parasitized red cell packing and perivascular haemorrhage showed some similarities to the neuropathology of human CM. However the large haemorrhagic infarctions and clinical presentation with almost 100% mortality of dogs presenting with cerebral babesiosis were key differences.


Acute respiratory distress syndrome (ARDS) and ventilator induced lung injury (VILI) are characterized by neutrophil recruitment in the lung, and human neutrophil peptides (HNP) are the most abundant cationic proteins that are released into the extracellular matrix upon neutrophil activation. We and others have previously shown that high concentrations of HNP can lead to capillary-epithelial barrier damage in a mouse model of ARDS. We also demonstrated that VILI can lead to lung fibrotic development. We now extended the studies to examine the effects of HNP in lung remodeling.Examine the role of HNP on lung remodeling in a mouse model of ARDS followed by mechanical ventilation.+/+) were generated using a human neutrophil elastase promoter. The HNP+/+and the FVB wild type mice received either HCl  intratracheally, or vehicle solution as a control. Respiratory system compliance, protein concentrations and differential cell counting in bronchoalveolar lavage fluid (BALF) were determined 48 h after HCl. In a parallel experiment, the mice were randomized into 3 different groups 48 h after HCl: 1) no mechanical ventilation; 2) low pressure mechanical ventilation for 2 h; and 3) high pressure mechanical ventilation for 2 h. The mice were then observed for 14 days for evaluation of epithelial damage, Ashcroft score for fibrosis and inflammation.Mice endogenously expressing HNP (HNP+/+ mice showed significantly body weight loss (Fig +/+ mice 14 days after HCl instillation followed by mechanical ventilation at either low pressure or high pressure (Fig The HNPloss Fig , increasloss Fig  and enhaloss Fig  in BALF loss Fig  at 48 h sure Fig .Figure 1HNP can accelerate inflammation at an early phase in a mouse model of HCl-induced ARDS, which may contribute to lung remodeling at late phases, magnifying the deleterious effects of ventilator-induced lung injury. We are hoping to present a full picture regarding lung remodeling 14 days after receiving HCl/mechanical ventilation at the ESICM conference in September.


Acute Lymphoblastic Leukaemia (ALL) is the most common cancer in children. Over the past four decades, research has advanced the treatment of this cancer from a less than 60% chance of survival to over 85% today. The causal molecular mechanisms remain unclear. Here, we performed sequencing-based genomic DNA methylation profiling of eight paediatric ALL patients using archived bone marrow smear microscope slides.SOLiD™ sequencing data was collected from Methyl-Binding Domain (MBD) enriched fractions of genomic DNA. The primary tumour and remission bone marrow sample was analysed from eight patients. Four patients relapsed and the relapsed tumour was analysed. Input and MBD-enriched DNA from each sample was sequenced, aligned to the hg19 reference genome and analysed for enrichment peaks using MACS  and HOMER (Hypergeometric Optimization of Motif EnRichment). In total, 3.67 gigabases (Gb) were sequenced, 2.74 Gb were aligned to the reference genome . This dataset enables the interrogation of differential DNA methylation associated with paediatric ALL. Preliminary results reveal concordant regions of enrichment indicative of a DNA methylation signature.Our dataset represents one of the first SOLiD™MBD-Seq studies performed on paediatric ALL and is the first to utilise archival bone marrow smears. Differential DNA methylation between cancer and equivalent disease-free tissue can be identified and correlated with existing and published genomic studies. Given the rarity of paediatric haematopoietic malignancies, relative to adult counterparts, our demonstration of the utility of archived bone marrow smear samples to high-throughput methylation sequencing approaches offers tremendous potential to explore the role of DNA methylation in the aetiology of cancer. This project was approved by the Royal Children’s Hospital Human Research Ethics Committee (RCH HREC# 29140C). We have performed Methyl-Binding Domain protein 2 (MBD2) enrichment and isolated fractions of DNA from 40 individuals for sequencing on the Sequencing by Oligonucleotide Ligation and Detection (SOLiD™) sequencing platform . MBD2 has been shown to bind to double-stranded methylated DNA molecules and used to interrogate the human methylome [Genomic DNA from archived bone marrow smear microscope slides from ALL patients, cells and cell lines were extracted as previously described  and usedSingle and paired-end SOLiD™ sequencing reads were aligned using LifeScope™ Genomic Analysis Suite (Life Technologies) with default parameters against the hg19 reference genome. Alignment efficiency  ranged from 26.57% to 93.15% across all samples in this study   and HOMEThis study is unique in a number of ways. This is the first sequencing-based DNA methylation profiling study in childhood ALL using archived bone marrow samples of similar quality to formalin-fixed paraffin-embedded (FFPE) tissue samples . We haveμg and 5 μg of starting genomic DNA to determine if 1 μg of starting material was sufficient for DNA methylation enrichment. This was less than the recommended quantity but a typical amount obtainable from our primary patient samples.We performed replicate DNA methylation enrichment analysis using the JWL cell line with 1 We isolated four haematopoietic cell populations  at major stages of development corresponding to the arrested stages of development in paediatric leukaemia. This was achieved by positive selection using fluorescent-labelled antibodies and Fluorescent Activated Cell Sorting (FACS) from four individuals. This would enable us to track changes in DNA methylation between cell lineages and contrast them with leukaemic cells. After MACS enrichment peak analysis, a large proportion of peaks were common between the CD19 cells from three individuals, confirming the premise of tissue-specific DNA methylation profiles in haematopoietic cells  from the same individual, distinct enrichment peaks are seen; these are likely to correlate to disease state Figure B. The nuFor each of the samples analysed in this study, we have generated track hubs that can be uploaded and visualised on the UCSC Genome Browser. This permits the immediate visualisation of regions of differential DNA methylation with potential biological significance. Moreover, we have performed Infinium analysis on these samples, and visualisation using the Genome Browser permits direct comparison to other publicly available data such as The Cancer Genome Atlas (TCGA)  and TARGIn summary, our data represent one of the first DNA methylation enrichment analyses using SOLiD™MBD-Seq on archival bone marrow smears from children diagnosed with ALL. Such specimens are readily available in most pathology laboratories across the world and are amenable to genomic-scale analysis, as we have demonstrated here. These data should prove valuable for other DNA methylation studies in childhood ALL in haematopoeitic cell development.Supporting data is available from the GigaScience Database, GigaDB  and at NSRA Files included BioProject PRJNA272864MACS and HOMER output files of peaks and peak locationsTrack Hubs for UCSC Genome Browser


Central nervous system (CNS) injury, such as stroke, is known to increase susceptibility to infections that adversely affect clinical outcome. This impaired immune response to infection is termed CNS injury-induced immune deficiency syndrome (CIDS). Activation of the cannabinoid 2 receptor (CB2R) suppresses immune function suggesting that antagonism of this receptor may overcome CIDS. The main purpose of this study was to determine the impact of CB2R inhibition on leukocyte activation within the microcirculation following endotoxin challenge in an experimental stroke model.2). The CB2R antagonist was given 15 minutes prior to LPS administration. Intravital microscopy was carried out 2 hours after LPS administration. Brains were then extracted and stained with tetrazolium chloride to calculate the infarct volume. The primary outcome measurement in this study regarding the immune response after stroke was the quantification of leukocyte adhesion following endotoxin challenge in submucosal venules of the gut - an important organ in the development of multiorgan failure in endotoxemia and sepsis.This was a prospective, randomized animal study. Five experimental groups  were subjected to the following treatments: control; endotoxemia ; transient cerebral hypoxia-ischemia (HI) + endotoxemia; HI + endotoxemia + CB2R antagonist . HI was induced by unilateral carotid artery occlusion, followed by 50-minute exposure to a low oxygen atmosphere (8% OCompared with endotoxemic animals without CNS injury, mice subjected to HI displayed reduced leukocyte activation in intestinal submucosal venules indicative of CIDS. Administration of the CB2R antagonist in animals with CIDS challenged with endotoxin restored peripheral leukocyte recruitment without a detrimental impact on infarct size.CB2R-related modulation of leukocyte activation is involved in the impaired immune response following CNS injury. Future studies will explore the CB2R pathway in order to develop novel therapies to improve the immune response in CIDS.


In vitro studies into the role of OPG produced by breast tumor cells have demonstrated that OPG can block TNF-related apoptosis inducing ligand (TRAIL)-mediated apoptosis. Furthermore, in vivo studies show that OPG expression by breast tumors can promote tumor growth and metastasis. In addition it has been shown that OPG stimulates endothelial cell survival and tube formation thus it may indirectly promote breast tumor progression through impacting angiogenesis. This article will present a summary of the data concerning the tumor-promoting effects of OPG in breast cancer.Osteoprotegerin (OPG) is a secreted protein and member of the Tumor Necrosis Factor (TNF) Receptor superfamily. OPG has been well characterized as a regulator of bone metabolism which acts by blocking osteoclast maturation and preventing bone breakdown. Given this role, early studies on OPG in breast cancer focused on the administration of OPG in order to prevent the osteolysis observed with bone metastases. However OPG is also produced by the breast tumor cells themselves. Research focusing on OPG produced by breast tumor cells has revealed actions of OPG which promote tumor progression.  Osteoprotegerin (OPG) is a secreted protein that acts in the bone to block the maturation of bone resorbing osteoclast cells and thus prevent bone breakdown , 2. ThisOPG was initially identified through sequence homology to the tumor necrosis factor (TNF) receptor superfamily. OPG contains the four cysteine-rich domain repeats characteristic of the TNF receptor family but lacks a transmembrane region and is therefore a secreted protein , 5, 2. Iin vitro osteoclast maturation studies demonstrate that OPG acts as a negative regulator of bone metabolism and blocks osteoclast maturation [Generation of transgenic mice has allowed elucidation of the major physiological role of OPG as a regulator of bone metabolism. OPG over-expression results in a lack of mature bone-resorbing osteoclasts and a pronounced increase in bone density known as osteopetrosis . Mice inturation , 2. Substuration , 10.Given the characterization of the role of OPG as a negative regulator of bone metabolism as described above, early studies on OPG and breast cancer focused on the bone microenvironment. Bone is the most common site of breast cancer metastasis. Breast tumors tend to form osteolytic metastases where the tumor is surrounded by osteoclasts which degrade the bone tissue -13. An iThese early studies focused on actions of exogenously added, truncated OPG within the bone microenvironment. However, subsequent studies that are described below demonstrate a more extensive and complex role for OPG in breast cancer.One of the first studies to characterize OPG revealed expression in two human breast cancer cell lines – MDA-MB-436 and MCF-7 . FurtherWhile there have been no investigations focusing on OPG in normal breast epithelium, OPG knockout mice are able to maintain litters which would suggest that OPG is not required for normal mammary gland development and lactation . It appeThe studies described above demonstrate a basal level of expression of OPG in breast cancer cells and tissue. Further investigation has revealed the potential for modulation of OPG expression levels within breast tumors. Immunohistochemical staining of human breast tumor samples showed that OPG expression correlated with the presence of the Estrogen and Progesterone receptors . Howeverin vitro using human breast cancer cell lines. TRAIL-induced apoptosis in MDA-MB-436 and MDA-MB-231 human breast cancer cells could be reduced by the addition of recombinant OPG (10-1000 ng/ml). MDA-MB-436 cells which had been incubated for 72 to 120 h to allow accumulation of endogenous OPG were less susceptible to TRAIL-induced apoptosis. This effect could be reversed by the addition of excess RANKL to compete for OPG binding [TNF-related apoptosis inducing ligand (TRAIL) is secreted by normal tissues and preferentially induces apoptosis in tumor cells , 26. The binding .in vitro studies clearly demonstrate effects of the OPG-TRAIL interaction, the relevance of this interaction for breast cancer cells has yet to be demonstrated in vivo. MDA-MB-231 human breast cancer cells were transfected to over-express full length OPG. Despite showing a significant increase in OPG expression along with a reduced sensitivity to TRAIL-mediated apoptosis in vitro, there was a lack of effect in vivo. Two weeks after intra-tibial injection in mice of cells the anti-tumor effect observed with TRAIL treatment was identical between empty vector and OPG over-expressing human breast tumor cells [While or cells .in vivo due to simultaneous presence of TRAIL and RANKL. An in vitro binding study demonstrated that OPG binds TRAIL and RANKL with equal affinity [The role of OPG produced by breast cancer cells may be more complex affinity . Howeveraffinity , 24. Thiin vivo studies that indicate tumor-promoting effects of OPG on breast cancer cells, potentially via TRAIL-independent mechanisms as described below. MCF-7 human breast cancer cells were engineered to express Parathyroid Hormone allowing them to form osteolytic metastases after intra-tibial injection in nude mice. Administration of truncated recombinant Fc-OPG reduced bone destruction in agreement with previous studies. In contrast, when the MCF-7 cells were transfected to over-express full length OPG and injected into mice tibias there was an increase in osteolysis. The intra-tibial tumors from the cells over-expressing OPG had an increase in cells staining positive for the proliferation marker Ki67. These MCF7 cells over-expressing OPG produced larger tumors compared to control cells after injection into the mammary fat pad although in this context there was no difference in Ki67 staining. The MCF-7 cells over-expressing OPG did not show any alteration in sensitivity to TRAIL treatment in vitro, suggesting that there must be additional mechanisms whereby OPG can have a tumor-promoting effect [There are now a number of g effect . The cong effect . Howeverg effect .in vivo study suggests that the tumor-promoting effects of the Estrogen-Related-Receptor-Alpha (ERRα) in breast cancer may be mediated in part via OPG. ERRα expression in bone metastasizing MDA-MB-231 human breast cancer cell derivatives led to an increased expression of OPG. In agreement with this experimental observation, the analysis of 251 patient samples showed that OPG levels were significantly higher in ERRα-positive tumors compared with ERRα-negative tumors. High ERRα and high OPG expression in breast tumor tissue from patients is associated with a greater increase in risk of recurrence than high expression of ERRα only. An increase in tumor growth was observed when the ERRα expressing cells were implanted in the mouse mammary fat pad however a reduction in osteolysis was observed when these cells formed bone metastases after intravenous injection in mice [Another  in mice . TherefoThis possibility that OPG can protect the bone microenvironment from tumor growth while promoting growth in other tissues is also evident in a recent study. MDA-MB-231 human breast cancer cells were stably transfected to express OPG and injected into the tibias of nude mice. Over-expression of OPG by injected breast cancer cells protected bone from destruction and inhibited growth of the tumors within the bone. However breast cancer cells over-expressing OPG persisted to grow in the space outside of the bone. In addition, OPG over-expressing breast tumor cells injected into the bone had an increased tendency to metastasize to lungs .in vivo model of breast cancer metastasis. Knockdown of OPG expression reduced metastasis formation in chick tissues both from a primary tumor and after direct introduction of cells by intravenous injection. Our in vitro mechanistic studies show that OPG knockdown cells express lower levels of Matrix Metalloproteinase-2 and have a reduced ability to invade through a collagen matrix. This would suggest that OPG can exert an autocrine effect and promote breast cancer cell invasion and metastasis [Despite the accumulating evidence that OPG produced by breast cancer cells may promote tumor growth in sites outside of the bone microenvironment, the mechanism behind this effect is unclear. We have recently performed a study using MDA-MB-231 human breast cancer cells transfected with sh- or siRNA to knock down OPG in a chick embryo tastasis .in vivo studies suggest that while the interaction between OPG and the bone microenvironment can lead to an inhibition of bone tumor growth, OPG can promote breast tumor formation and growth at other sites.The findings of these While OPG produced by breast cancer cells may exert effects directly on breast tumor cells it appears that OPG’s interaction with other cells in the tumor microenvironment may also promote breast tumorigenesis. In particular, several studies have reported an impact of OPG on endothelial cells suggesting the potential role of OPG to regulate angiogenesis.vβ3 antibody in the co-culture medium [vβ3 signaling can promote OPG expression. This study also demonstrated that OPG (10 ng/ml) promoted tube formation by HuDMECs confirming the potential for OPG to exert pro-angiogenic effects.Human umbilical vein endothelial cells (HUVECs) and Human Dermal Microvascular Endothelial cells (HuDMECs) both express OPG. Treatment of endothelial cells with recombinant OPG (100 ng/ml) promotes cell survival and formation of cord-like structures. Analysis of human breast tumor tissues showed that OPG is expressed by endothelial cells in breast tumor tissue but not in normal surrounding tissue. There is a strong positive correlation between endothelial OPG expression and tumor grade, and a strong negative correlation between endothelial OPG expression and ER expression . Furthere medium . This woSeveral studies have investigated the mechanism whereby OPG could promote angiogenic behavior of endothelial cells. HuDMECs treated with OPG were elongated with extensive actin networks compared to untreated cells. OPG (0.5-2ug/ml) led to cytoskeletal reorganization and cell spreading; this may be mediated by Focal Adhesion Kinase (FAK) as phosphorylation of FAK Y397) was induced by treatment with OPG. OPG-mediated HuDMECs proliferation was blocked by the Erk 1/2 inhibitors U0126 and PD98059 while OPG-induced cell migration and cord-like structure formation was blocked by the Src inhibitor PP1 . The abi7 was indThe cumulative findings of these endothelial cell studies demonstrate the potential for OPG to exert tumor-promoting effects by stimulating angiogenesis in addition to the direct effects of OPG on breast cancer cells.A few studies have investigated links between OPG and breast cancer prognosis although conflicting findings have been reported. Real-time quantitative PCR was used to analyze the expression of OPG mRNA in 127 primary human breast cancer tissues. It was found patients with higher expression levels of OPG had significantly poorer overall survival as compared with patients who had lower OPG expression levels . HoweverIn addition to the expression of OPG, changes in the OPG gene have been reported in breast cancer. We analyzed the TCGA-2013 human breast invasive carcinoma data set, through the cBioPortal website, and found that OPG gene copy gain occurred in 182 out of 934 tumors in the set (19.4 %). The presence of an OPG copy number gain is a significant predictor of decreased overall survival in this cohort . The occSince its initial discovery and characterization, evidence has been accumulating that OPG plays a role in breast tumorigenesis. The reported actions of OPG in breast cancer are summarized in Fig. While OPG clearly has potential to modulate breast tumor growth and progression, further investigation is required to fully determine the mechanism of effect and overall outcomes of the different types of interactions. The data available currently would suggest that OPG can no longer be considered solely in the bone microenvironment in breast cancer and caution must be exercised in the development of systemic treatment strategies aimed at increasing OPG levels. Localized delivery of OPG to the bone may be more appropriate to prevent osteolysis associated with metastatic breast cancer. Additional studies are required to determine whether strategies to block OPG signaling would be effective in blocking the growth and progression of primary breast tumors.


Clinical examination revealed a loud continuous murmur with diastolic accentuation in the left precordium, noncollapsing pulse, and no differential pulses in the upper and lower limbs. The ECG showed normal sinus rhythm with PR interval of 220 milliseconds.A 40 year-old man with no significant past history except for asymptomatic 1Transthoracic echocardiogram Figure  showed ast degree atrioventricular (AV) block. Coronary arteriovenous fistula (CAVF) arises as a persistence of sinusoidal connections between the lumens of the primitive tubular heart in the early embryologic period. It is reported in 0.1%–0.2% of all cases of selective coronary angiography[st degree AV block in our case may reflect a flow limitation to atrioventricular (AV) node directly caused by the CAVF.This case illustrated a rare saccular aneurysm formation in a right coronary artery to right atrium  fistula associated with 1giography or penetCT angiogram has now emerged as an excellent imaging technique to delineate the lesion anatomy-6 CAVF g


Neurons of the cerebellar nuclei convey the final output of the cerebellum to their targets in various parts of the brain. Within the cerebellum their direct upstream connections originate from inhibitory Purkinje neurons. Purkinje neurons have a complex firing pattern of regular spikes interrupted by intermittent pauses of variable length. How can the cerebellar nucleus process this complex input pattern? In this modeling study, we investigate different forms of Purkinje neuron simple spike pause synchrony and its influence on candidate coding strategies in the cerebellar nuclei. That is, we investigate how different alignments of synchronous pauses in synthetic Purkinje neuron spike trains affect either time-locking or rate-changes in the downstream nuclei. We find that Purkinje neuron synchrony is mainly represented by changes in the firing rate of cerebellar nuclei neurons. Pause beginning synchronization produced a unique effect on nuclei neuron firing, while the effect of pause ending and pause overlapping synchronization could not be distinguished from each other. Pause beginning synchronization produced better time-locking of nuclear neurons for short length pauses. We also characterize the effect of pause length and spike jitter on the nuclear neuron firing. Additionally, we find that the rate of rebound responses in nuclear neurons after a synchronous pause is controlled by the firing rate of Purkinje neurons preceding it. Neurons can transmit information by two different coding strategies: Rate coding, where the firing rate of the neuron is vital, and time coding where timing of individual spikes carries relevant information. In this study we analyze the importance of brief cessations in firing of the presynaptic neuron (pauses) on the spiking of the postsynaptic neuron. We perform this analysis on the inhibitory synaptic connection between Purkinje neurons (presynaptic) and nuclear neurons (postsynaptic) of the cerebellum. We employ a computational model of nuclear neurons and “synthetic” Purkinje neuron spike trains to study the effect of synchronous pauses on the spiking responses of nuclear neurons. We find that synchronous pauses can cause both well-timed spikes and increased firing rate in the nuclear neuron. In addition, we characterize the effect of pause length, amount and type of pause synchrony, and spike jitter. As such, we conclude that nuclear cells use both rate and time coding to relay upstream spiking information. Cerebellar nucleus (CN) neurons are crucial to the olivo-cerebellar circuit as they provide the sole output of the entire cerebellum ,2. CN nePNs exhibit an elaborate firing pattern characterized by simple spikes and complex spikes . Simple Several coding strategies have been proposed for the CN neurons. Broadly, these strategies can be categorized as rate coding or time coding. For a long time it was assumed that CN neurons receive information from PNs by means of a rate code. In cats, CN neurons (recorded from the anterior interpositus nucleus) exhibited rate modulations during locomotion . SimultaSystematic analysis of simple spikes from neighboring PNs in anaesthetized rats revealed that spikes associated with pauses in firing are far more synchronized (±2 ms precision) than regular firing simple spikes , which iBased on the experimental study of pause synchronization , we defiin vitro [In vivo, the firing of these neurons becomes irregular [in vivo data, we tuned both the gain of inhibitory and excitatory synapses impinging on the CN neurons so that the neurons fire spontaneously around 37–38 Hz, which is the observed mean firing rate in vivo of normal rats [The firing of deep CN neurons is highly regular in vitro . In vivorregular  mediatedmal rats . The anamal rats . The excin vivo [Synaptic input had maximum impact on the regularity firing of the neuron at the high gain condition . A CN nein vivo , the higIn order to analyze the influence of PN synchrony on the firing response of the CN neuron, we characterized the neuron’s response to synchronous PN pauses see . SimulatWe quantified the increase in firing rate of the CN neuron during synchronous pauses with spikes synchronized at the beginning or ending, or overlapping pauses (where no spikes are synchronized), for different amount of synchronization. Synchronous pauses were generated by randomly selecting a pause greater than threshold and temporally aligning it with pauses from other Purkinje neuron spike trains according to the desired amount of synchronization. Pause ending and pause overlapping synchronization consistently mediated greater firing rate increases compared to pause beginning synchronization. This could be seen for both pause values of 20 and 40 ms . For allWe also simulated the effect of a mixed-type synchronization where PN pauses were synchronized with all three types of synchronization  in equal proportion. This resulted in increased firing rate modulation of CN neurons, which was greater than that of the pause beginning synchronization, but less than the pause overlapping condition.In order to elucidate the membrane mechanisms underlying increased firing of CN neurons during the synchronous PN pauses, we blocked the rebound conductances  in the CN neuron model. Next we investigated the effect of the amount of synchrony on the rate modulation of CN neurons. Increase in PN input synchronization resulted in an increased rate modulation for all pause synchrony conditions . We starWe observed similar results for the pause overlapping synchronization condition: a 40 ms pause represents the amount of presynaptic PN pause synchrony better . One notvivo and in vitro [According to Person and Raman, PN synchronization causes brief periods of relief from inhibition and this time-locks the CN neuron’s response to the input synchronization . This noin vitro . By varySynchronous pause beginning spikes effectively time-locked the CN neuron’s response to the synchronized event . For almIn the previous section we demonstrated how synchronous PN pauses elicit time-locked CN neuron spikes thereby transmitting the timing information in a reliable manner to downstream targets of CN neuron. We compared the degree of time-locking caused by pause beginning or pause overlapping synchronization by looking at their spike latency distributions in detail. This analysis revealed that time-locking by pause beginning spikes is more accurate than that of pause overlapping condition for a 20 ms  synchronIn order to elucidate membrane mechanisms causing the time-locking of CN neuron spiking to PN pause synchronization, we blocked the rebound conductances in CN neuron model and analyzed its effect on time-locking. Rebound conductances had an insignificant effect on the time-locking of the CN neuron . There wUp to this point, we used precise synchrony of spikes without any jitter. Because this is a rather idealized situation, we also investigated whether jittered pause spikes can still evoke time-locking of CN neuron’s responses. Therefore, we quantified the time-locking behavior of the neuron when presented with synchronous pause differing in the amount of synchronization and pause beginning spikes jittered with various values of jitter. We generated synchronous PN pauses in the same way as described in previous sections but with jittered pause beginning spikes and a pause duration of 40 ms.When the CN neuron's input gain is high, the strong inhibition prevents the neuron to spike during the jitter period. This can be seen in For the low gain condition, the CN neuron often fired during the jitter period because the inhibition was too weak . We compSo far our analysis was limited to the effect of synchronous pauses on the firing modulation or time-locking within the pause. Our analysis has revealed that, on average, rebound conductances have no or little effect on this firing modulation or time-locking. CN neurons exhibit rebound firing when the membrane is hyperpolarized for a considerable amount of time and subsequently released from it . De SchuAverage PN firing rate  during a period of Δt before the synchronous pause was forced to a particular frequency 'fPN' see . We sampIn this study we investigated the effect of PN synchrony in the context of pauses on the spiking response of CN neurons. We stress the importance of pause synchrony in our simulations because the cerebellum is known to control the motor output pattern by transient disinhibition of CN neurons through corresponding pausing of PN firing ,24. A wiin vitro study [in vivo optogenetic study highlighted the importance of input synchrony through transient ‘pausing’ of PNs by photostimulation of molecular layer interneurons expressing channelrhodopsin [We find that pause overlapping and pause ending spike synchronies mediate greater firing rate modulation during the synchronous pause than their pause beginning counterparts. It is worth noting that the synchrony types are not uniquely encoded in the firing rate of these neurons as their standard deviations overlap . Similarro study , we alsohodopsin . They shCN neurons exhibit both rate and time coding, but it remains unclear how these neurons can switch between these two different coding schemes. We reasoned that synchronous pause length could play a vital role in controlling the coding strategy of CN neuron. Therefore we compared the effect of pause length, by measuring the effect of synchronous pauses of 20 or 40 ms. For pause beginning synchronization, a 40 ms pause sequence evoked greater firing rate increases than a similar 20 ms pause sequence . ConversWith respect to time-locking of CN neuron spikes to PN pause synchrony, we found that pause beginning synchronization cause more accurate time-locking than pause overlapping or pause ending type synchronies. A similar phenomenon was reported earlier by Person and Raman  where thThe amount of PN synchrony reaching a CN neuron is unknown . This isWe conclude that the multiplexed coding scheme  of the PThe CN neuron model used in our study is based on a previously published model . Three dCN neurons receive inhibitory inputs from PNs and excitatory inputs from mossy fibers. Both synaptic pathways are included in our model and described below. Climbing fiber inputs were not simulated.τrise and τdecay are rise and decay time constant respectively. gmax is peak synaptic conductance, and N is a normalization factor that makes the maximum of Gsyn(t) equal to gmax.PNs inhibit CN neurons. The number of PN synapses on a single CN neuron is highly debated and values in the literature range from 40  to 860 ,6,31–33.max = 11.7 nS and synaptic reversal potential of -75 mV [10 values for synaptic current kinetics (Q10 = 2)[Rss) and time constant of depression (τ) was fitted to experimental data [PN to CN neuron IPSCs time constants are based on published experimental data . IPSCs rf -75 mV . The syn(Q10 = 2),36. The (Q10 = 2),37. Syna(Q10 = 2) and steatal data . The equNMDA) that shows weak voltage dependence and much slower component (SNMDA) that shows strong voltage dependence. NMDA voltage dependence is modeled [NMDA, SNMDA time constants and peak synaptic conductance values, parameters s1 and s2 of voltage dependence were obtained from [10 values for synaptic current kinetics (Q10 = 2) [CN neurons are excited by mossy fibers . The mos modeled  accordinned from  (S1 Tablned from . AMPA anQ10 = 2) ,36.et al [PN simple spike trains contain highly regular inter-spike intervals (ISIs) and hence cannot be described accurately by a Poisson process . Shin etet al  showed tSynthetic PN simple spikes were generated based on the method described in  using spvivo [Pause synchrony is defined by either having the pause beginning or pause ending spike synchronized or no spikes (overlapping) synchronized across the population of PN inputs. We introduce synchrony by randomly selecting a pause that was longer than the threshold (either 20 or 40 ms) and temporally aligning it with pauses from other spike trains. The pause thresholds were selected so that they are greater than the spontaneous PN simple spike ISIs in vivo . We usedFor simulations where we tested the role of PN firing rate before the pause on CN neuron’s firing rate post pause onset, average PN firing rate during each trial was forced to a particular frequency, ‘fPN'. We used the same number of trials for each fPN. Average PN firing rate for a trial was forced to fPN in the following way: For each PN participating in the synchronous pause, a pause exceeding a threshold of 20 ms and whose preceding spiking for a time period of 100 ms has a firing frequency of fPN, was randomly selected. We then temporally aligned both the pause and preceding spiking with their corresponding counterparts from other PN spike trains. For certain PNs where no desired fPN could be found before a pause, a pause exceeding the threshold was randomly selected.Neuronal spike times were recorded from the simulated CN neurons. The firing rate during synchronous pauses was computed and compared to a control condition  to investigate changes in the firing rate caused by the release of inhibition. We measured the spike latency with respect to the onset of synchronous pause, the standard deviation/median absolute deviation of which was then used to further assess the degree of time-locking.To avoid the stochastic influences inherent to the synthetic spike trains, we performed 100 or more simulations for each input regime and computed the average change in firing rate, as well as the standard deviation/median absolute deviation of spike latency. We used standard deviation of latency to first spike within the synchronous pause if the distribution of spike latencies of both distributions followed the normal distribution. If either one of the distribution was not normal, the median absolute deviation was used. The normality of the distribution was established by Lilliefors test  using 99Vector strength ,42 quantcov is the covariance and σ the standard deviation.For simulations where we tested the role of PN firing rate before the synchronous pause on the CN neuron’s firing rate following the immediate pause, the CN neuron’s firing rate was computed for a period of 1s from synchronous pause onset and compared to a control condition (model run without synchronized spike and pause sequence). Linear regression was used to fit a straight line through the data points. Correlation was measured with Pearson’s correlation asS1 FigAnalysis for 20 ms (A-D) and 40 ms (E-H) synchronous pauses. (A&E) Population spike timing histogram of all PNs projecting onto the CN neuron. (B&F) Increase in firing rate of CN neuron quantified for both pause ending and pause overlapping conditions and for low input gain. (C&G) Same for medium input gain. (D&H) Same for high input gain.(TIF)Click here for additional data file.S2 Fig(A&E) Population spike timing histogram of all Purkinje cells projecting onto CN neuron. Pause beginning type synchronization is illustrated in A and pause overlapping type is explained in E. (B&F) Increase in firing rate of the CN neuron during synchronous pause of length 20 ms and 40 ms and for low gain condition. (C&G) Same for medium gain condition. (D&H) Same for high gain condition. In all panels red color stands for 20 ms global pause period, blue for 40 ms global pause period. Error bars are represented by shaded region. (*) represents pairwise comparisons between results of different amount of input synchronization  that are significant (p<0.05) and (^) represents insignificant comparisons (p> = 0.05).(TIF)Click here for additional data file.S3 Fig(A&E) Population spike timing histogram of all PNs projecting onto the CN neuron. Note the presence of synchronous pause of length 20 ms (A) and 40 ms (E) for both types of synchronization. (B&F) Variability of latency calculated from 100 trials for pause ending and pause overlapping type synchronization and low gain condition. (C&G) Same for medium gain condition. (D&H) Same for high gain condition. In all panels red color stands for pause ending condition, blue for pause overlapping condition. (*) represents comparisons of variability in latency between pause ending and pause overlapping type synchronization that are significant (p<0.05) and (^) represents insignificant comparisons (p> = 0.05).(TIF)Click here for additional data file.S4 Fig(A&E) Population spike timing histogram of all Purkinje cells projecting onto the nuclear neuron. Note the presence of synchronous pause beginning spikes and pause length of 20 ms (or 40 ms). Plots B,C and D represents quantification of variability in latency of CN neuron’s spiking for low (B), medium (C) and high input gain (D) condition respectively. Plots F,G and H represents quantification of increase in firing rate of the nuclear neuron for low gain (F), medium gain (G) and high gain (H) condition respectively. (*) represents appropriate comparisons that are significant (p<0.05) and (^) represents insignificant comparisons (p> = 0.05).(TIF)Click here for additional data file.S5 FigSynthetic PN spikes were generated using a three-step process: First we segregated the experimental ISIs into regular firing patterns  and pause ISIs (red) (step 1). For each regular firing pattern a corresponding synthetic regular firing pattern was generated based on gamma distribution statistics from  (step 2)(TIF)Click here for additional data file.S6 FigA-I: Distribution of experimental and synthetic PN ISIs for nine randomly selected PNs. The similarity between experimental and synthetic ISIs was determined by Kolmogorov-Smirnov test. None of the generated synthetic ISIs was significantly different from experimental ones .(TIF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.


Oxidative stress can cause DNA breaks which induce activation of the DNA nick sensor enzyme poly(ADP-ribose) polymerase-1 (PARP-1), part of the 17 member PARP enzyme family. PARP-1 modifies target proteins by attaching to them several NAD-derived ADP-ribose units forming poly(ADP-ribose) (PAR) polymers. PARylation controls many cellular processes while intense PARylation may also lead to cell death by various mechanisms. Here we summarize the modes of activation, inhibitors and modulators of PARP-1 and review the cellular functions regulated by the enzyme. Poly(ADP-ribosyl)ation (PARylation) is a protein modification catalyzed by poly(ADP-ribose) polymerase (PARP) enzymes. The enzyme family was created based on database screening using the highly conserved PARP signature motif. Whereas the PARP enzyme family consists of 17 members Here we review the mechanisms by which activity of PARP-1 can be stimulated, inhibited or modulated. We also aim to summarize the cellular functions that are regulated by PARP-1.PARP-1 has originally been described as a DNA nick sensor enzyme activated by DNA single and double strand breaks The findings that stimuli other than broken DNA can also activate PARP-1  led to ai values remains to be determined.Several classes of compounds with PARP inhibitory properties have been developed and characterized. Their discussion would go way beyond the scope of this review so we only refer to excellent reviews on PARP inhibitors and their therapeutic potentials A much more interesting issue is the interference between certain signaling events and PARP activation . Above wUntil recently calcium was the only ion considered to serve a second messenger role. However, similar roles have later been assigned to magnesium and zinc ions The role of several metabolites connected to PARP-1 including its substrate (NAD), a key metabolite required for NAD synthesis (ATP) and a product (nicotinamide) should not be disregarded as they have also been shown to affect PARP activity The effects of PARP-1 and PARylation are translated to cellular responses in various different ways . On the The consequences of PARylation, PAR signaling, interaction of PARP-1 with partner proteins and substrate competition affect many cellular events ranging from genome organization + and ATP consumption has recently been challenged. Dawson's group has elegantly demonstrated [From these intertwining networks of molecular events and cellular functions here we would like to highlight one aspect bearing the highest relevance to oxidative stress: regulation of oxidative stress-induced cell death. The generally held view that in severe oxidative stress situations unrepairable DNA damage induces excessive PAR production and consequent NADnstrated  that PARAuthors declare no conflict of interest.


The objectives of the research were to enhance our understanding of the phenotype (water holding capacity) and search for biomarkers of this economically significant pork quality attribute. Major changes in protein abundance across nine phenotype-by-time conditions were observed. Proteomic patterns were dominated by post mortem ageing timepoint. Using a machine learning algorithm (l1-regularized logistic regression), a model was derived with the ability to discriminate between high drip and low drip phenotypes using a subset of 25 proteins with an accuracy of 63%. Models discriminating between divergent phenotypes with accuracy of 72% and 73% were also derived comparing respectively, high drip plus intermediate phenotype (considered as one phenotype) versus low drip and comparing low drip plus intermediate phenotype (considered as one phenotype) versus high drip. In all comparisons, the general classes of discriminatory proteins identified include metabolic enzymes, stress response, transport and structural proteins. In this research we have enhanced our understanding of the protein related processes underpinning this phenotype and provided strong data to work toward development of protein biomarkers for water holding capacity.Two dimensional Difference Gel Electrophoresis (2-D DIGE) and mass spectrometry were applied to investigate the changes in metabolic proteins that occur over a seven day   Numerous proteomic studies have shown that very many of the observed post mortem proteome changes relate to these protein degradation processes  .puma.princeton.edu) . The oupost mortem respectively were identified. Details of the significant spots at day 3 and 7 post mortem  are presented in et al., .analysis . cCompar(DOC)Click here for additional data file.


Heme peroxidases are found in all kingdoms of life and typically catalyze the one- and two-electron oxidation of a myriad of organic and inorganic substrates. In addition to this peroxidatic activity distinct (sub)families show pronounced catalase, cyclooxygenase, chlorite dismutase or peroxygenase activities. Here we describe the phylogeny of these four superfamilies and present the most important sequence signatures and active site architectures. The classification of families is described as well as important turning points in evolution. We show that at least three heme peroxidase superfamilies have ancient prokaryotic roots with several alternative ways of divergent evolution. In later evolutionary steps, they almost always produced highly evolved and specialized clades of peroxidases in eukaryotic kingdoms with a significant portion of such genes involved in coding various fusion proteins with novel physiological functions.Four heme peroxidase superfamilies  arose independently during evolution, which differ in overall fold, active site architecture and enzymatic activities. The redox cofactor is heme  During turnover H2O2 is reduced to water and one-electron donors (AH2) are oxidized to the respective radicals (•AH) (−) are oxidized to the corresponding hypohalous acids (HOX)   whereas ds (HOX) . Besidesreaction  and use reaction .(ReactioIn the last decade – due to the application of powerful sequencing techniques – an ever increasing amount of protein sequences (including numerous heme peroxidases) were automatically assigned to related protein families due to typical conserved motifs This review demonstrates the independent evolution of four heme peroxidase superfamilies, namely of the (1) peroxidase–catalase superfamily, (2) peroxidase–cyclooxygenase superfamily, (3) peroxidase–chlorite dismutase superfamily and the (4) peroxidase–peroxygenase superfamily. We present an update of their reconstructed phylogeny and introduce their representative sequence signatures and essential amino acids in the heme cavity. From http://pfam.xfam.org) this universally distributed superfamily is annotated as PF00141.The peroxidase–catalase superfamily previously known as the “superfamily of bacterial, fungal and plant heme peroxidases” families in analogy with all other heme peroxidase superfamilies (see below). Novel clades representing hybrid enzymes between the previously defined classes are well distinguishable in this reconstruction. R-XX-W/F-H-X- and the proximal triad His-Trp/Phe-Asp (these amino acids are not neighbored in the primary sequence). The typical overall globular fold of representatives of this superfamily consists of twelve α-helices and was already acquired from the beginning. It was only slightly modified in the later steps of ongoing divergent evolution. Generally, the level of structural conservation is higher than the conservation of amino acid sequences Here we have selected 500 representative full-length sequences that were used for an updated phylogenetic reconstruction based on the maximum likelihood (ML) method. c peroxidases catalatic activity according to c peroxidases are typical monofunctional peroxidases . The main function of Family I peroxidases seems to be scavenging of excess H2O2.Family I was previously defined to contain intracellular bacterial catalase–peroxidases c peroxidases from Choanoflagellida predecessors of fungi and animals Family I is the most divergent one among the three structural classes. It contains eukaryotic intracellular A peculiar minor descendant clade containing yet putative secretable ascorbate peroxidases from Unikonts/Metazoan lineages represents another evolutionary turning point . A majorThe highest number of members are found in Family III that is comprised of numerous plant secretory peroxidases like horseradish peroxidase(s) (HRP) e.g.,  but alsoThe peroxidase–cyclooxygenase superfamily has Pfam accession PF03098 and its members are widely distributed among all domains of life In contrast to the peroxidase–catalase superfamily, in many cases members of the peroxidase–cyclooxygenase superfamily are multidomain proteins with one heme peroxidase domain of predominantly α-helical fold with a central heme-containing core of five α-helices. Moreover, this superfamily is unique in having the prosthetic heme group posttranslationally modified Q-X-X-D-H-D-X- which includes the distal catalytic histidine that is neighbored by two aspartates, the first being involved in ester bond formation and the second in Ca2+-binding. The highly conserved glutamine seems to be involved in halide binding R-X-X-E-X-  the early evolved heme peroxidases to cope with reactive oxygen species produced by ancient oxygenic photosynthesis The ancestral clade of short bacterial peroxicins  is supposed to contain the oldest representatives of this superfamily , namely One evolutionary trend led to the emergence of the well known animal peroxidases like myeloperoxidase (MPO), eosinophil peroxidase (EPO), lactoperoxidase (LPO) and thyroid peroxidase (TPO) and started with bacterial peroxidockerins (Family 6). The cyanobacterial LspPOX is a typical representative of early stage in this direction and it was already investigated on protein level The second evolutionary direction leads towards bacterial and animal cyclooxygenases (Family 4) . AlthougThe alternative evolutionary trend leading from long peroxicins towards Families 3–1 occurred stepwise. First steps led over  peroxidockerins (Family 6). It is important to note that evolutionary descendants of variable length emerged not only in eubacteria but can be also found in genomes of eukaryotic Amoebozoa, Heterolobosea and Chromalveolata . This fa2O2 but the function of the modified peroxidase domain in this large multidomain protein remains obscure Dual oxidases (Family 7) were segregated from peroxidockerins at the level of primitive eukaryotes as stated before . These mIn a further step peroxinectins (Family 3) evolved. They were shown to exhibit cell adhesion function(s) and to be involved in invertebrate immune response by production of hypohalous acids according to Next evolutionary event led to division in peroxidasins (Family 2) and a sister family of Chordata peroxidases (Family 1). They diverged in several Chordata subfamilies including the separated subfamily of thyroid peroxidases, minor subfamilies of fish peroxidases and three closely connected subfamilies of mammalian lactoperoxidases (LPO), myeloperoxidases (MPO) and eosinophil peroxidases (EPO)  that areIn contrast, we can observe a broad and rather abundant distribution of peroxidasin isozymes in various Eumetazoan phyla . They weThis superfamily was first constituted by Goblirsch et al. I-P-V/I-K/R-K-X- that includes proximal residues  and PF06778 (for Cld), they are classified in a common protein clan CL0032 (SCOP 54909) that comprises up to 18 member families, but most of them without the ability to bind the prosthetic heme group. It is important to note that in contrast to the two previously described heme peroxidase superfamilies, the peroxidase–chlorite dismutase family appears to cover only a small part of a very large dimeric α/β barrel structural superfamily . The modification of such a fold to acquire the ability to bind the heme group and exploit it evolved probably only in one evolutionary event. The whole dimeric α/β barrel structural superfamily is outside of the scope of this issue.2, whereas Cld-like enzymes neither exhibit a peroxidase nor a chlorite degrading activity. Moreover, Cld-like proteins show a diminished affinity for the prosthetic group.DyP heme peroxidases are abundant not only in many bacterial phyla Mucor circinelloides  did not yield any output for DyP peroxidase genes. This supports the theory of HGT from autotrophic prokaryotes to heterotrophic eukaryotes. Anyway, upcoming new genomes of early diverging fungi must be also screened with blast in this respect. Their codon usage has to be analyzed and compared.The ancestral clade of the whole superfamily is probably located in DyP-subfamily A, namely among sequences from thermophilic facultatively anaerobic bacteria of the division Firmicutes . This woDictyostelium discoideum or Naegleria gruberi DyP-peroxidases occur. Most of here presented protein sequences from this subfamily stem from Proteobacteria therefore also in subfamily B HGT events appeared probably several times during the evolution. Even more surprising is the occurrence of DyP-type B genes in the Trematode parasite Schistosoma mansoni. Also in this multicellular flatworm DyP genes were acquired by HGT from lower eukaryotes but their functionality must be proven experimentally. Thus it appears that subfamily B is the most divergent one of all four described DyP subfamilies leading to unique evolution of DyP peroxidases within Unicont genomes. Turning points in this superfamily’s evolution lie unequivocally in this subfamily clades that besides several HGT events enabled also further evolution towards chlorite-dismutase like proteins.Fungal DyP sequences occur not only in subfamily D but also in subfamily B, where also several other sequences from lower eukaryotes like b prosthetic group are segregated in two distinct clades (6). Clade 1 is almost completely formed by proteobacterial penta- or hexameric representatives whereas Clade 2 contains dimeric proteobacterial as well as numerous cyanobacterial enzymes. The N-terminus of Clade 2 proteins is truncated  efficiently incorporate peroxide-derived oxygen into substrate molecules. Additionally their proximal heme ligand is cysteine instead of histidine  fumago has been in the focus of research only. In the last decade other members of this superfamily (Pfam accession number PF01328) were found and started to be analyzed.This heme peroxidase superfamily is unique since its members can  – included in the genomes of ancestral, primitive fungi already from the beginning of the evolution of the fungal kingdom. The monophyletic clade of Stramenopiles enzymes diverged apparently very early in parallel with the evolution of “classical” but minor fungal enzymes represented by CfuHalPrx from the ascomycete Caldariomyces fumago with known 3D structure Auricularia delicata or Coprinellus radians are results of later evolutionary events from Mucor peroxidases via repeated gene duplication and speciation events (Auricularia heme-thiolate peroxidase (AaeAPO) reveal slight differences in the heme pocket if compared to CfumagHTP Mucor clade no sequences from phyla other than dikaryotic fungal subkingdom are known in the four above mentioned abundant clades yet. As most here presented sequences are only putative, the future research shall uncover their peculiar enzymatic properties not only for their peroxidatic activity but for oxyfunctionalization of organic molecules in general The ancestral clade appears to contain n events . The dev


The correlation between headache and cocaine is controversial and still understimated.As many as 14-21 million people worldwide  use cocaine , seems eWritten informed consent to publish was obtained from the patient(s).None.


Simultaneous PET and MR acquisitions have now become possible with the new hybrid Biograph Molecular MR (mMR) scanner from Siemens. The purpose of this work is to create a platform for mMR 3D and 4D PET image reconstruction which would be freely accessible to the community as well as fully adjustable in order to obtain optimal images for a given research task in PET imaging. The proposed platform is envisaged to prove useful in developing novel and robust image bio-markers which could then be adapted for use on the mMR scanner.STIR (Software for Tomographic Image Reconstruction ), an opeConversion of the Siemens Dicom files to the STIR interfile format.Histogramming the emission data.Component-based normalisation.µ-maps [Estimation and correction for attenuation and scatter using atlas based µ-maps .Correction for randoms.Iterative image reconstruction –5)..5]).The reconstruction pipeline involves the following:µ-map including the bed component (bottom).Two datasets were used: uniform cylinder phantom and FDG-PET human brain scan. Figure The reconstruction of the FDG human brain dataset is presented in Figure It was demonstrated that STIR image reconstruction of PET mMR data is possible paving the way to more advanced models included in the pipeline of 4D PET reconstruction.


Postoperative new-onset atrial fibrillation (PNAF) is the most common complication after cardiac surgery. Inflammation as an underlying mechanism has been studied by various inflammatory markers, and white blood cell count (WBC) is the only present consequent inflammatory marker predicting PNAF . This stpost-hoc cohort study. The WBC was prospectively measured preoperatively and once during each of the first four postoperative days. Development of PNAF was evaluated with continuous 12-lead ECG monitoring the first 5 days postoperatively.Patients >18 years undergoing elective cardiac surgery with a sinus rhythm preoperatively were recruited from the Dexamethasone for Cardiac Surgery-PNAF trial for this P < 0.001), CABG plus valve surgery , single valve surgery  and other surgery  were correlated with the occurrence of PNAF.A total of 657 patients were included in this trial, 277 developed PNAF. The WBC was significantly higher in the PNAF group on day 2 and day 4 Figure . HoweverPreoperative and postoperative WBC were not associated with development of PNAF.


Patients previously prescribed one epinephrine auto-injector device may be switched to an alternative device by their pharmacist or physician - sometimes without training on the new device. It is unclear whether “device switches” without retraining compromise the ability to deliver epinephrine.TM. ISRCTN29175528We evaluated mothers of food-allergic children participating in a UK study of epinephrine auto-injectors (EAI), 1 year after they were first trained to use an EAI, either Anapen or Epipen (old design). Participants' ability to deliver epinephrine using their device was assessed using a simulated anaphylaxis scenario. Participants then underwent repeat assessment using a different EAI device, randomly allocated, without training on the new device. The UK-approved EAIs Epipen (new/old designs), JEXT or Anapen were used, or Intelliject, an EAI with audio/visual prompts approved in North America as AuviQ(tm) and AllerjectWe evaluated ability to deliver epinephrine in 108 participants. Overall success rates were similar using their original EAI 68/108 (63%) to the new device 65/108 . However the outcome differed significantly for different types of device switch. Success rates were lower when switching between Anapen and either old Epipen, new Epipen or JEXT  compared with switching from old Epipen to either new Epipen or JEXT . Success rates were highest when switching from Anapen or old Epipen to Intelliject  compared with switching to other EAIs .The safety of EAI device switches varies according to the specific device. Switches to Intelliject appear to be safer than other forms of device switch.


Rheumatology key messageUS-guided injection improves patients’ experience with this intervention, which may contribute to improved response rates.ir, US-guided intra-articular and soft tissue steroid injections are common procedures in rheumatology and are taking the place of fluoroscopic and CT-guided injections. Little is known about patients’ views about US-guided steroid injections despite several efficacy studies comparing blind to US-guided injections [Sjections . In the jections . We condRheumatology Online).Fifty questionnaires that included balanced Likert scale questions were distributed to rheumatology patients who underwent a US-guided procedure between January 2011 and January 2012. Survey receipt was concluded in April 2012 relating to a post-injection period ranging from 4 to 16 months. Of the 50 questionnaires distributed, 30 (60%) were returned and 26 (50%) were completed and included for data analysis  performed all the procedures. During the procedure the sonographer explained to the patient the anatomical features of the diseased target site, power Doppler activity and dynamic needle progression through the superficial tissues into the target site. A total of 34 joints/tendon regions were injected in 26 patients . All procedures were part of the patients’ normal care pathways.All patients felt that seeing the US images was very helpful or helpful in understanding the procedure A. EightyThis pilot study has some limitations. No validated psychometric questionnaire was available for this specialized purpose, therefore we used a non-validated questionnaire to obtain retrospective views of patients. Furthermore, such a retrospective survey is vulnerable to response bias, potentially enhancing the number of overtly positive or negative responses.Observing US images during the procedure improved the overall experience of this intervention. Observing the US images in real-time improved patients’ understanding and tolerability of the procedure and reduced patients’ anxiety. This is consistent with a randomised controlled study that suggested US guidance improved pain scores (p < 0.001) and overall response rate (p < 0.01) compared with traditional palpation-guided injection . AnxietyA larger study is required to confirm our preliminary findings that US-guided injection improves the tolerability of the procedure and reduces patients’ anxiety during the procedure. Further issues that require investigation include whether visualising real-time images during US scanning improves patients’ understanding of disease pathology, which could lead to indirect benefits such as improving therapy adherence and improved pain management strategy.Funding: Arthritis Research UK (Grant 17767) provided funding for the US Unit.Disclosure statement: The authors have declared no conflicts of interest.Supplementary data are available at Rheumatology Online.


Rituximab (RTX) is a monoclonal antibody; recently it has been use as a new treatment strategy in patients with high-degree steroid-dependent nephrotic syndrome. It provokes a lymphocyte B depletion in some cases that interacts with Immunoglobulins creating decreased plasma IgG levels and hypogammaglobulinemia.2. Baseline analysis before RTX treatment evidenced low IgG levels, although these values are not conclusive because they were taken during nephrotic state. Twenty days later, the patient presented severe hypogammaglobulinemia with IgG levels of 149 mg/dL, without nephrotic proteinuria. We decided to start intravenous gammaglobulin (IVIG) every 4-6 weeks. Analysis showed low levels of lymphocytes (predominantly B type) four days after initiated the treatment. Follow up demonstrated complete recovery of lymphocyte count two years later. After 3 years of RTX administration the patient continues to have hypogammaglobulimenia, requiring IVIG until now.We report the case of a 2.5 year old male diagnosed of idiopathic nephrotic syndrome. He presented resistance to systemic steroids as well as second line treatments (cyclophosphamide and mycophenolate). Patient started with intravenous infussion of rituximab at 375mg/mThe cause of immunodeficiency in this patient has not yet been established. Is it because of the nephrotic syndrome itself? An adverse effect of the drug? Maybe a pre-existing primary immunodeficiency?Use of RTX has reported cases of hypogammaglobulinemia values usually normalizing after 11 months; persistent hypogammaglobulinemia is not a common side effect. We only found one series describing the association of persistent hypogammaglobulinemia in nephrotic patients treated with RTX, suggesting a certain predisposition in patients with low baseline IgG levels previous to the treatment with this monoclonal antibody.Starting a full immune panel previous to treatment with RTX could be a good way to predict possible persistent hypogammaglobulinemia and to rule out immunologic pathology before making any therapeutic decision.


Extensive DNA damage and inefficient DNA repair might be responsible for some of the pathogenic features in patients suffering from asthma. To determine whether DNA adducts can be used as a “dosimeter” for asthma disease severity, we measured the DNA adducts level in lung of mouse with house dust mite (HDM)-induced allergic airway inflammation, as the disease progresses. Apoptosis of airway epithelial cells is one of the most critical pathophysiological factors in the development of chronic asthma. As repairing of DNA lesions is important in preventing apoptosis, we propose that DNA repair plays an important pathophysiologic role in regulating lung epithelial cell DNA damage response.We immunofluorescence-stained mice asthmatic lung tissue sections and observed an increase in DNA double strand break (DBS) markers, γH2AX and 53BP1 as compared to control. Level of DNA repair proteins that involved in homologous recombination and non-homologous end joining, were up-regulated substantially as early as 1 day-post last challenge. TUNEL assay revealed high level of DNA strand breaks in bronchial epithelium. DNA damage signaling pathway PCR array showed a reproducible increase in expression of multiple genes involved in DNA damage and repair. Treatment with glucocorticoid significantly reduced cell infiltration into airway as well as DNA damage and repair markers. To elucidate the role of DNA repair in regulating disease outcome, we treated mice with NU7441, a DNA-dependent protein kinases inhibitor, and had observed an increased DNA damage makers expression in lung and increased apoptotic level in bronchial epithelium as compare to no-drug treatment control. When DNA repair was obstructed, apoptosis of airway epithelium cells was enhanced. This indicates the important role of DNA repair in airway inflammation.


Significant abnormal hemispheric cerebral physiology resulting in neurological injury can occur without knowing because the current monitoring system could not detect the abnormal hemispheric abnormality. Cerebral oximetry can provide a rapid, non-invasive detection of each hemisphere's cerebral physiologic changes during ictal/post-ictal phases. The aim was to study left and right rSO2 values in patients in the pre and post seizure periods and in nonseizing controls.During ictal/post-ictal events, altered cerebral physiology occurs: increased neuronal activity causes significant increase in cerebral metabolism with changes in ipsilateral cerebral blood flow. Standard PED seizure monitoring is by O2 readings compared with nonseizure patients.An observational study of seizing and nonseizing patient's left and right rSO2 readings across ages. See Figure No difference for ictal left and right rSO2 values were significantly decreased. In patients with focal seizures, the ipsilateral rSO2 values were significantly different from the contralateral rSO2 readings and correlated to the hemisphere experiencing the focal seizure. In certain patients, during the ictal phase their rSO2 readings rose and stayed or rose then dropped. Overall, cerebral oximetry has shown great monitoring potential for actively seizing patients in the emergency department.We have demonstrated abnormal hemispheric cerebral physiology during focal or generalized ictal activity. In patients with generalized seizures, the left and right rSO


We hypothesized that the region of latest mechanical contraction would correspond to the region of latest electrical activation but the time between mechanical and electrical activation would vary between patients.Numerous studies have shown that placing the left ventricular (LV) pacing lead in the latest contracting region  or the latest activated region  of the LV improves patient response to Cardiac Resynchronization Therapy (CRT). However, no studies have examined electrical and mechanical dyssynchrony Mechanical dyssynchrony: High temporal resolution steady-state free precession (SSFP) cine images were acquired with 60 frames per cardiac cycle in ten patients enrolled for CRT. The endocardial boundary was delineated in a stack of short-axis images. Radial contraction curves (RDCs) were generated for 100 locations on the endocardial boundary for each slice. The delay between each RDC and a patient-specific reference was determined using cross-correlation analysis. Mechanical delay times throughout the LV were mapped to the AHA 17-segment model. Electrical dyssynchrony: Regional electrical delay times were determined from local electrograms (EGMs) acquired during CRT device implantation. EGMs were acquired simultaneously from the RV lead (placed in the apex of the RV) and LV lead . 35 EGM recordings were obtained at several potential pacing sites in the 10 patients. The electrical delay was defined as the peak-to-peak difference between the EGMs from the RV and LV leads. Lead localization: Dual-plane fluoroscopic projection images were acquired at each LV lead recording site and was mapped onto the AHA 17-segment model. Mechanical delay times within a region the size of an AHA segment centered at the LV pacing location were averaged to obtain a corresponding mechanical delay time. Analysis: Mechanical delays (from MRI bullseye) and electrical delays (from EGM) were correlated on a patient-by-patient basis.2=0.85 ± 0.21). The linear best fit in each patient yielded a positive slope (1.7 ± 1.3) and the slope varied greatly between patients (Range: 0.06-4.11). The site that exhibited the largest electrical delay always corresponded to the site of latest mechanical delay. The location of this latest activated/contracting site varied significantly between patients .There was a strong linear relationship between electrical and mechanical delay times over all patients (RThe latest electrically activated site always corresponded to the latest mechanically contracting site; therefore either mechanical or electrically dyssynchronous regions can be targeted in an attempt to improve patient response to CRT.This study was funded in part by the National Science Foundation Graduate Research Fellowship Program.


A 36-year old man presents to the emergency room five days after undergoing extracorporeal shock wave lithotripsy (ESWL) for a symptomatic 11mm left renal pelvis stone. The patient has persistent symptoms of severe left flank pain at presentation.steinstrassen)  .1 This pFor asymptomatic cases of steinstrasse, conservative management is preferred with observation and serial imaging with plain radiography to follow progression of the stones and ultrasonography to evaluate for hydronephrosis. Sixty-five percent of steinstrassen will pass spontaneously over days to weeks.This particular patient underwent urgent ureteroscopy with stent placement the following day and had an uneventful recovery thereafter.


The adoptive transfer of tumor infiltrating lymphocytes (TIL) can mediate the regression of metastatic melanoma. In addition, the adoptive transfer of lymphocytes genetically modified to express tumor reactive T cell receptors (TCRs) can mediate tumor regression. Many T cells from TIL recognize mutated antigens expressed only on the autologous patient's tumors. Therefore, we attempted to isolate TCRs reactive with unique mutated antigens so that we might eventually be able to treat patients with autologous T cells genetically modified to express those TCRs.in vitro transcribed and electroporated into autologous dendritic cells (DCs). Recognition of TMGs by TIL was evaluated on the basis of IFN-γ secretion and CD137 expression after overnight coculture with the electroporated DCs. Subsequently, mutation reactive T cells were enriched from TIL by sorting for CD137+ T cells after overnight coculture with the electroporated DCs and were expanded in vitro with anti-CD3 and IL-2. Dominant TCR α and β chain sequences were identified in the enriched mutation reactive populations, and retroviruses encoding those TCRs were used to transduce human PBL to determine if they mediated recognition of the mutated antigen.Mutations in tumors were identified using whole exome sequencing and/or RNA sequencing. Tandem minigene (TMG) constructs containing 12-24 minigenes were synthesized, each encoding the mutated amino acid flanked by 12 amino acids on both sides. RNAs encoding the TMGs were Thus far, using these techniques we have isolated mutation reactive TCRs from 6 different patients with metastatic melanoma as described in the attached table. We are currently extending these techniques to identify mutation reactive TCRs for patients with other cancers including those of the gastrointestinal tract, breast, and ovaries. We are also developing clinical reagents to treat patients with TCRs that recognize unique mutations on autologous tumor cells.