We further showthat cells which have switched to [+PSI] haveimproved viability during chronological ageing which requires active autophagy.[+PSI] stains show increased autophagic fluxwhich correlates with increased viability and decreased levels of cellularprotein aggregation. Taken together, our data indicate that the frequency of[+PSI] prion formation increases duringyeast chronological ageing, and switching to the[+PSI] form can exert beneficial effects via thepromotion of autophagic flux.Ageing involves a time-dependent decline in a variety of intracellular mechanismsand is associated with cellular senescence. This can be exacerbated by priondiseases which can occur in a sporadic manner, predominantly during the laterstages of life. Prions are infectious, self-templating proteins responsible forseveral neurodegenerative diseases in mammals and several prion-forming proteinshave been found in yeast. We show here that the frequency of formation of theyeast [ Biological ageing involves a progressive decline in the ability of an organism tosurvive stress and disease. It is a complex process which is influenced by bothgenetic and environmental factors C) into a prion form (PrPsc) that is amyloidogenic.The amyloid form of the protein can subsequently convert other soluble moleculesinto the prion form thus resulting in the accumulation of the aberrant proteins inneuronal cells 6PSI+] and [PIN+], whichare formed from the Sup35 and Rnq1 proteins, respectively 9PSI+] is the altered conformation of the Sup35protein, which normally functions as a translation termination factor during proteinsynthesis. The de novo formation of[PSI+] is enhanced by the presence of the[PIN+] prion, which is the altered form of the Rnq1protein whose native protein function is unknown Prions are aberrant, infectious protein conformations which can self-replicate de novo [+PSI] prion formation14316We have previously shown that autophagy protects against Sup35 aggregation andPSI+] prion formation using the yeast CLS model andfound that the frequency of prion formation is increased during ageing. Furthermore,this frequency is elevated in an autophagy mutant suggesting that autophagy normallyacts to suppress age-dependent prion formation. We show that the prion-status ofcells influences CLS in an autophagy-dependent manner suggesting that prions can bebeneficial in aged populations of yeast cells.Yeast cells can survive for prolonged periods of time in culture and have been usedas a model of the chronological life span (CLS) of mammals, particularly for tissuescomposed of non-dividing populations 19+PSI] appearance during ageing. Cultureswere grown to stationary phase in liquid SCD media and prion formation measuredover time. [PSI+] prion formation was quantifiedusing the ade1-14 mutant allele which confers adenineauxotrophy and prions differentiated from nuclear gene mutations by theirirreversible elimination in guanidine hydrochloride (GdnHCl). The frequency ofde novo [PSI+] prion formationin a control[PIN+][-psi] straingrown to stationary phase was estimated to be 1.1 x 10-5 comparableto previously reported frequencies 12PSI+] prionformation in an atg1 autophagy mutant. Atg1 is aserine/threonine kinase which is responsible for the initiation of autophagy13PSI+] prionformation was further elevated in the atg1 mutant compared witha wild-type strain, suggesting that autophagy normally acts to suppress[PSI+] prion formation during ageing .We examined yeast CLS to determine whether there is an increased frequency of[PSI+] prion status of cells influenceslongevity. For this analysis, flow cytometry was used to monitor propidiumiodide uptake to assess yeast cell death PIN+][PSI+] strainshowed a modest increase in maximal lifespan compared with a[PIN+][psi-] strain. Additionally, cell death was lower in the[PIN+][PSI+] strainover the entire lifespan compared with the[PIN+][psi-] strainsuggesting that the [PSI+] prion improves viabilityduring ageing. To verify this difference in ageing between[PIN+][PSI+] and[PIN+][psi-]strains, viability was monitored using colony formation assays. Whilst lifespanmeasured using the colony forming assay was shorter compared with the propidiumiodide uptake assay, it confirmed that the [PSI+]strain maintained viability longer and had an increased lifespan compared withthe [psi-] strain .We next examined cell survival to determine whether the[PSI+] and all known prions. Curing the[PIN+][PSI+] strainwith GdnHCl dramatically decreased maximal lifespan to 10 days suggesting thatprions are beneficial during CLS . It should be noted however, thatGdnHCl treatment can also potentially affect Hsp10-related processes which areunrelated to prions. Autophagy is known to be required for chronologicallongevity and for example loss of ATG1 reduces CLS ATG1 reduced CLS in the 74D-694 yeast strain used for ourstudies . Interestingly, longevity was comparable in the[PIN+][PSI+],[PIN+][psi-] andcured strains indicating that active autophagy is required for prion-dependenteffects on longevity.Treating cells with GdnHCl blocks the propagation of most yeast prions byinhibiting the key ATPase activity of Hsp104, a molecular chaperone that isabsolutely required for yeast prion propagation 25+PSI]-prion status of cells affectsCLS in an autophagy-dependent manner, we examined whether autophagy is alteredin [+PSI] cells. We utilized a GFP-Atg8 fusionconstruct to follow the autophagy-dependent proteolytic liberation of GFP fromGFP-Atg8, which is indicative of autophagic flux PIN+][PSI+] straincompared with the[PIN+][psi-] andcured [pin-][psi-]strains indicative of increased autophagic flux . By day three, therewas also an increase in autophagic flux in the[PIN+][psi-] andcured [pin-][psi-]strains detected as an increase in free GFP. As expected, no autophagic activitywas observed for the atg1 knockout mutants. This suggests thatcells carrying the [+PSI] prion have increasedautophagic activity which may be beneficial during CLS.Given that the [PIN+][PSI+] strainaffects amorphous protein aggregation, which seemed likely given the previousstudies which have suggested that autophagy plays a role in the clearance ofmisfolded and aggregated proteins 16PIN+][PSI+] straincompared with the[PIN+][psi-] andcured [pin-][psi-]strains . This reduction in protein aggregation required autophagysince the levels of protein aggregation were comparable in theatg1-mutant version strains.We next examined whether the increased autophagic activity in the[de novo [+PSI] formation duringyeast chronological ageing, with the frequency of spontaneous formation increasedapproximately 40-fold in aged cells. Yeast has emerged as a powerful model toinvestigate the stochasticity of the ageing process and its contributing factors.The yeast CLS model more closely resembles the ageing of non-dividing cells such asneurons de novo formation of [+PSI] wasexacerbated in an atg1 mutant suggesting that autophagy normallyacts to suppress spontaneous prion formation during chronological ageing. Similarly,loss of autophagy has been found to cause neurodegenerative diseases in mice 16The majority of prion disease cases in humans occur in a sporadic manner,predominantly manifesting during later stages of life PSI+] prionconfers a beneficial advantage during yeast chronological ageing, which correlateswith enhanced autophagic flux. Given that the[PSI+]-status of cells improves viability during ageing,this may result in selection for [PSI+] in aged cells.Our data indicate that the presence of the [PSI+] prionacts to simulate autophagy which results in improved viability during ageing. Thereis previous evidence to suggest that the increased formation and accumulation ofprotein aggregates may exert a stimulatory effect on the autophagy pathway. Forexample, there is a correlation between the accumulation of PrPSC and theenhanced activity of quality control pathways including endoplasmic reticulumchaperones, the unfolded protein response and autophagy PSI+] strain, suggesting thatautophagy provides a beneficial effect during chronological ageing by removingpotentially harmful protein aggregates, including both amorphous and amyloid forms.Increasing autophagic flux would also presumably act to prevent further amyloidaggregation, potentially protecting against any negative impact of[PSI+] aggregation altering translation terminationefficiency.We found that that the presence of the [PSI+] prion causes a loss of function phenotypewhere translation termination activity is reduced due to the aggregation of thenormally soluble Sup35 protein PSI+] prion is thought to allow cells toreprogram gene expression such that new genetic traits become uncovered which mayaid survival during altered conditions 3132PSI+] prion can improve viability during ageing viamodulation of autophagic flux. It is unclear what triggers the increased frequencyof [PSI+] prion formation during ageing. One possibilityis oxidative stress, since ROS-induced protein aggregation and mitochondrialdysfunction is a common feature in age-related diseases 34PSI+] formation observed during chronologicalageing. Further research will be required to elucidate the exact signaling pathwaysand the range of quality control mechanisms that may be modulated through the directand indirect action of the [PSI+] prion during yeastageing.The [+PIN][PSI+],[+PIN][psi−] and[-pin][psi−]derivatives of the wild-type yeast strain 74D-694  were used for all experiments. Thestrain deleted for ATG1 (atg1::HIS3) has beendescribed previously  prion formation was scored by growth in theabsence of adenine as described previously PSI+] formation was calculatedbased on the mean of at least three independent biological repeatexperiments.[CLS experiments were performed according to Protein extracts were electrophoresed under reducing conditions on SDS-PAGEminigels and electroblotted onto PVDF membrane (Amersham Pharmacia Biotech).Bound antibody was visualised using WesternSure® Chemiluminescent Reagents(LI-COR) and a C-DiGit® Blot Scanner (LI-COR). Insoluble protein aggregates wereisolated as previously described 38


Five datasets were constructed from ligand and bioassay result data from the literature. These datasets include bioassay results from the Ames mutagenicity assay, Greenscreen GADD-45a-GFP assay, Syrian Hamster Embryo (SHE) assay, and 2 year rat carcinogenicity assay results. These datasets provide information about chemical mutagenicity, genotoxicity and carcinogenicity. Specifications TableValue of the data•This article contains the largest public collection of ligands and results for the GreenScreen GADDα-45 and Syrian Hamster Embryonic Cell Transformation assays collated from previous literature to date.•A benchmark dataset of pharmaceutically relevant ligands for use in rat carcinogenicity QSAR models is presented and compared with ligands from regulatory domains.•Physiochemical descriptors were calculated from the SMILES structures and selected for QSAR model performance.1in vivo carcinogenicity datasets in this article are solely rat carcinogenicity outcomes due to previous literature finding the rat carcinogenicity bioassay produces better endpoint reliability in comparison to the mouse carcinogenicity bioassay. This article presents two rat carcinogenicity datasets from the regulatory toxicology and pharmaceutical safety chemical domains.The creation of a QSAR model for the 2-year rodent carcinogenicity bioassay is highly desirable since it is the gold standard for assessing potential chemical carcinogenicity. However, previous modelling efforts have been hampered due to data availability and reliability issues stemming from bioassay limitations such as low throughput, high cost, and modest reproducibility between laboratories and rodent species. The in vitro bioassays with larger libraries of screened molecules in comparison to in vivo rodent carcinogenicity bioassay data. QSAR models capable of utilizing this data in combination with rodent carcinogenicity data may address the limited applicability domain of the in vivo data. The data was exhaustively collated from the in vitro GreenScreen GADDin vivo rodent carcinogenicity outcomes. The Ames Bacterial Mutagenicity Benchmark Dataset has also been included for comparison.Genotoxicity occurs from chemicals acting with genomic mechanisms of toxicity and this has been associated with potential carcinogenicity. This endpoint type features many 22.1in vivo rat carcinogenicity from ISSCAN: 854 chemical database of in vivo rodent carcinogenicity results on pharmaceutical chemicals from PHARM: GreenScreen: 1415 GADD-45a-GFP assay results from Syrian Hamster Embryonic: Data on 1415 chemicals extracted from Ames: 6512 Ames results from 2.2SMILES structures were generated using ChemAxon JChem for Office from CAS Numbers or chemical names.These structures were curated using ChemAxon Standardizer to remove salts and solvents and aromatized.2.3The CfsSubsetEval algorithm 2.4knn-based distance The applicability domain of each dataset compared to the PHARM dataset was quantified using leverage, Euclidean distance from centroid, and a variable 2.5Attribute Importance Scores were calculated from each RandomForest QSAR model , Table 7


Mycobacteriumtuberculosis.We read this report with interest,The pulmonary nodules identified by CT chest in the index patient (case 1) may have cavitated as part of an immune reconstitution syndrome. It would be useful to know if any respiratory symptoms developed during the admission or subsequently; the timing of the original CT in relation to the immune reconstitution syndrome; if any additional chest imaging occurred; and if respiratory specimens were obtained for mycobacterial culture at any stage.,Patients with HIV infection may be sputum culture positive despite a normal chest x-ray.M. tuberculosis transmission during aerosol generating procedures, including the irrigation of abscesses.There have been previous reports of M. tuberculosis is probably from individuals with pulmonary disease not yet on effective treatment (largely undiagnosed).A better mechanistic understanding of which procedures are potentially aerosol generating might help guide practice. Ruling-out pulmonary disease is non-trivial, particularly in the context of HIV co-infection. Most transmission of DAB is funded through a Wellcome Trust PHD studentship grant ref 105165/Z/14/A.Conflict of interest: None declared.


Objectives: Tourette Syndrome (TS) and Obsessive-Compulsive Disorders (OCD) share many clinical similarities and show a strong comorbidity. Current theories view a frontal-striatal dysfunction as the underlying cause of many clinical aspects of both disorders.This study sought to investigate mechanisms of conceptual integration and attention in both disorders. We hypothesized that the processing of stimuli with interfering aspects would be altered in a similar way while attentional mechanisms could differ.Methods: Event-related brain potentials (ERPs) were recorded in a modified STROOP-paradigm in groups of TS and OCD patients and in a control group. The paradigm involved the presentation of color words in a range of different colors. The subjects had to respond to words of matching word content and color and to ignore mismatching stimuli.Results: Incongruent stimuli elicited a frontal negative component (“N450”) which was enhanced in amplitude and prolonged in latency in both patient groups. Matching stimuli evoked enhanced N2 and P3b components representing target evaluation mechanisms. The OCD group alone displayed a larger P3b amplitude in comparison to both other groups.Conclusions: The data are interpreted to indicate that frontal inhibitory mechanisms are altered alike in TS and OCD. In contrast, only the OCD group showed evidence for aberrant target evaluation.


Blood lactate concentration (BLC) is widely used to guide resuscitation in critically ill patients . MusculaAn adult patient presented with a total body surface area burn of 70% after a suicide attempt by self-immolation. A triple-lumen central venous catheter (CVC) with an integrated microdialysis function was placed in the left internal jugular vein. A second CVC was placed in the right femoral vein. Figure To summarize, this case illustrates that continuous monitoring could allow sensitive detection of rapid and short-lasting changes in BLC after a 500-ml RL infusion. This could be used to assess lactate clearance. Further studies are needed to validate this concept.


E. coli) followed by repeated rounds of nickel column affinity chromatography, failed to generate recombinant amelogenin at single band purity. This was because following an initial round of nickel column affinity chromatography, subsequent cleavage of the His-tag was not 100% efficient. A second round of nickel column affinity chromatography, used in attempts to separate the cleaved His-tag free recombinant from uncleaved His-tagged contaminants, was still unsatisfactory as cleaved recombinant amelogenin exhibited significant affinity for the nickel column. To solve this problem, we used preparative SDS PAGE to successfully purify cleaved recombinant amelogenins to single band purity on silver stained SDS PAGE. The resolving power of preparative SDS PAGE was such that His-tag based purification of recombinant amelogenin becomes redundant. We suggest that acetic acid extraction of recombinant amelogenin and subsequent purification using preparative SDS PAGE provides a simple route to highly purified His-tag free amelogenin for use in structure-function experiments and beyond.Recombinant protein technology provides an invaluable source of proteins for use in structure-function studies, as immunogens, and in the development of therapeutics. Recombinant proteins are typically engineered with “tags” that allow the protein to be purified from crude host cell extracts using affinity based chromatography techniques. Amelogenin is the principal component of the developing enamel matrix and a frequent focus for biomineralization researchers. Several groups have reported the successful production of recombinant amelogenins but the production of recombinant amelogenin free of any tags, and at single band purity on silver stained SDS PAGE is technically challenging. This is important, as rigorous structure-function research frequently demands a high degree of protein purity and fidelity of protein sequence. Our aim was to generate His-tagged recombinant amelogenin at single band purity on silver stained SDS PAGE for use in functionality studies after His-tag cleavage. An acetic acid extraction technique (previously reported to produce recombinant amelogenin at 95% purity directly from  Amelogenesis involves the incremental secretion of an extracellular protein matrix by ameloblasts. Amelogenin, present as a range of related molecules generated through extracellular proteolysis of several alternatively spliced gene products, comprises ~90% of the total extracellular protein matrix and is essential for normal enamel biomineralisation. Amelogenin mutations can lead to amelogenesis imperfecta (AI) in humans  mutation promotes intracellular amelogenin retention, stalls the secretory pathway, and triggers the UPR to switch into pro-apoptotic mode resulting in ameloblast death, thus classifying this case of AI as a conformational disease  on amelogenin aggregation/assembly and ameloblast secretory trafficking. Wild-type (WT) amelogenin begins to self-assemble during transit through the ameloblast secretory pathway  is via recombinant technology.E. coli based expression systems are the most widely used methodology for producing recombinant amelogenin even though post translational phosphorylation of serine 16 will be absent. Baculo virus . Once the cells reached an optical density of 0.6–1.2 recombinant protein expression was induced by adding isopropyl β-D-1-thiogalactopyranoside (IPTG). Induction of expression was confirmed by analytical SDS PAGE. After shaking overnight incubation at 37°C, the cells were harvested by centrifugation and amelogenin-enriched fractions obtained by extraction in acetic acid as previously reported  modified with a HRV 3C protease site in Rosetta DE3 Lyophilized crude extracts of recombinant amelogenin were dissolved in a minimum volume of nickel column binding buffer , loaded on to a nickel affinity chromatography column  up to a maximum protein load of 40 mg and eluted at a flow rate of 4 mL/min. Unbound proteins (column flow through) were collected. Bound recombinant amelogenin was eluted by increasing the imidazole concentration to 200 mM. The recombinant amelogenin was then buffer exchanged into 125 mM formic acid using size exclusion chromatography  and the fractions containing recombinant amelogenin collected and lyophilized. Fractions were analyzed by analytical SDS PAGE and anti-amelogenin Western blotting using rabbit IgGs raised against a peptide corresponding to the amelogenin hydrophilic C-terminal . The His-tag was removed enzymatically by adding recombinant restriction grade protease  at a concentration of 3 μL enzyme solution per mg recombinant protein. Cleavage was carried out over 20 h at 4°C. The resulting cleavage reaction mixture comprising cleaved recombinant, any uncleaved recombinant, free His-tag and other contaminants was finally buffer-exchanged into nickel column binding buffer by size exclusion chromatography ready for a second round of nickel affinity column chromatography.The lyophilized recombinant amelogenin, prepared as described above, was dissolved at 2 mg/mL in either 50 mM Tris-HCl, pH = 8 or 0.1 M NaA second round of His-tag cleavage was employed to remove cleaved His-tag and other contaminants present in the cleavage reaction mixture including any uncleaved recombinant. Cleavage products in binding buffer were loaded on to the nickel affinity column as described in the Section entitled Affinity Nickel Column Purification of His-Tagged Recombinant Amelogenins Extracted Using Acetic Acid. Unbound proteins (expected to contain the cleaved recombinant amelogenin) were collected in the column flow through. Bound proteins  were eluted using a stepped imidazole gradient  in order to optimize the separation. Fractions were analyzed by analytical SDS PAGE as described in the Section entitled Analytical SDS PAGE and Western Blotting.Analytical SDS PAGE was carried out based upon the method of Laemmli  except 2Following electrophoresis, proteins were detected using Coomassie Blue  or silver staining  or were transferred to nitrocellulose membranes  for Western blot analysis. After blocking and washing, the membrane was incubated with primary antibody  at a concentration of 5–7mg/1.5 mL. Preparative electrophoresis was carried out using a Model 491 Prep Cell . A 12% resolving gel was cast in the 28 mm internal diameter gel tube to a height of 9.5 cm with a 2 cm 4% stacking gel. The gel was run at a constant 12 W power at room temperature using the circulating cooling pump as recommended by the manufacturer. Once the tracker dye reached the bottom of the gel, 2.5 mL fractions were collected at a flow rate of 0.8 mL/min. Fractions of interest were identified by subjecting every third fraction to analytical SDS PAGE. All fractions of interest were then subjected to analytical SDS PAGE to accurately determine any fractions containing cleaved recombinant amelogenin at single band purity on silver staining.The methodologies described above are summarized in Figure E. coli both immediately after the addition of IPTG and 1 h post induction to determine the presence of any induced recombinant protein expression using analytical SDS PAGE and Coomassie Blue staining , free His-tag and the cleavage enzyme itself. Following analytical SDS PAGE, it was clear that that cleavage was only around 50% efficient at either pH 8 or 9 Figure . In an aGiven that a second round of nickel column purification failed to provide cleaved recombinant amelogenin at single band purity, we pursued an alternative strategy using preparative SDS PAGE. Fractions collected from preparative SDS PAGE were characterized using analytical SDS PAGE and silver staining Figure . Using tThe production of recombinant proteins is essential to the study of protein function and particularly in the case of amelogenins, which are difficult to purify from native tissue due to their aggregative behavior, multiplicity of isoforms and their degradation products and scarcity of material in the most commonly used animal models. In addition, without recourse to recombinant expression, it would be almost impossible to study the effects of specific mutations on amelogenin behavior.Despite much effort to obtain highly purified recombinant amelogenin using His-tag affinity chromatography we were unable to produce recombinant amelogenin of single band purity when analyzed by silver stained SDS PAGE. Apparent single band purity on SDS PAGE can be achieved by reducing sample load or by employing less sensitive gel staining techniques such as Coomassie Blue but this merely places any contaminants below the limit of stain detection. SDS PAGE is a standard technique for assessing degrees of protein heterogeneity in sample mixtures but additional analyses such as isoelectric focusing or mass spectroscopy would be needed to confirm absolute purity. With that caveat in mind, our goal was to produce recombinant amelogenin exhibiting single band purity on silver stained SDS PAGE gels. The challenges were twofold: (i) removing contaminating bacterial proteins and (ii) separating contaminating His-tagged recombinant amelogenin from His-tag-free recombinant amelogenin following His-tag cleavage, which is <100% efficient.E. coli at >95% purity in a single purification step  that are commonly co-purified when using immobilized metal affinity chromatography due to their metal chelating properties or clusters of exposed histidine residues  to a second round of nickel column affinity chromatography. In theory, the cleaved recombinant amelogenin should have little affinity for the column after removal of the His-tag ligand and would be easily collected in the flow through or at low imidazole concentration while the uncleaved recombinant amelogenin, free His-tag and HRV3C cleavage enzyme (which is itself His-tagged) would remain bound to the nickel column. However, cleaved recombinant amelogenin still exhibited significant affinity for the nickel column. This may be due to the high histidine content of amelogenin which includes a tri-histidine domain (Snead et al., To solve this problem we decided to employ preparative SDS PAGE rather than using nickel column affinity chromatography. This method was able to separate cleaved recombinant amelogenin from all contaminants to single band purity on analytical silver stained SDS PAGE. Substituting nickel column affinity purification with preparative SDS PAGE reduces the number of steps and leads to a much purer end product. Indeed, the excellent resolving power of preparative SDS PAGE, coupled with the acetic acid extraction technique makes the need for nickel column purification of His-tagged recombinant amelogenin redundant. We suggest that recombinant amelogenin without any His-tag could be purified to single band purity using preparative SDS PAGE Figure . In compThe preparative electrophoretic equipment employed here is not restricted to purifying proteins using SDS PAGE which separates proteins based on their molecular size. Separations can also be achieved using native PAGE buffer systems where proteins, in the absence of SDS, are separated based on their net charge which in turns depends on pH. The manufacturer's instructions provide a range of buffer formulations covering the pH range 3.8–10.2 which can be further adapted by the addition of chaotropic agents and uncharged detergents to aid protein solubility. Although, this additional flexibility is not required in the application reported here, it may prove useful if using preparative electrophoresis for other application such as purifying native enamel proteins extracted from developing enamel.SB, JK and CG made substantial contributions to the conception and analysis of data; drafted the paper and/or revised it critically for intellectual content; provided final approval of the version to be published; agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved. CG acquired the data.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Angiomyolipoma is a well described but relatively uncommon benign renal neoplasm composed of varying admixtures of mature adipose tissue, smooth muscle, and thick-walled blood vessels.[1] The incidence of angiomyolipoma is about 0.3% overall.[2] It frequently occurs in patients with tuberous sclerosis. Even more uncommon is the simultaneous occurrence of angiomyolipoma and renal cell cancer in the same kidney in a patient without tuberous sclerosis.


The recent elucidation of the mechanistic basis for transpersulfuration in the CsdA-CsdE model system has paved the way for a better understanding of its role under oxidative stress. Herein we present the crystal structure of the oxidized, inactivated CsdE dimer at 2.4 Å resolution. The structure sheds light into the activation of the Cys61 nucleophile on its way from a solvent-secluded position in free CsdE to a fully extended conformation in the persulfurated CsdA-CsdE complex. Molecular dynamics simulations of available CsdE structures allow to delineate the sequence of conformational changes underwent by CsdE and to pinpoint the key role played by the deprotonation of the Cys61 thiol. The low-energy subunit orientation in the disulfide-bridged CsdE dimer demonstrates the likely physiologic relevance of this oxidative dead-end form of CsdE, suggesting that CsdE could act as a redox sensor in vivo.Sulfur trafficking in living organisms relies on transpersulfuration reactions consisting in the enzyme-catalyzed transfer of S atoms  Escherichia coli comprises two large gene systems termed ISC (Iron-Sulfur Clusters) and SUF (Sulfur Mobilization) . Cys61 alpha carbon atom is shown as a yellow sphere.(MP4)Click here for additional data file.S8 Fig(PDF)Click here for additional data file.S9 Fig(PDF)Click here for additional data file.


An 84-year-old female status post-Mohs micrographic surgery (MMS) presented to the emergency department (ED) for evaluation after a syncopal episode. Surgical excision of a scalp basal cell carcinoma occurred immediately prior to arrival . HemostaPoint-of-care cardiac ultrasound suggested acute air embolism with right heart strain as the cause of the patient’s syncope. An air embolism is a rare but serious complication of any procedure that may involve venous or arterial vasculature. Air emboli in the setting of MMS has previously been cited in a dermatology case report.6To our knowledge, this is the first case within the ED setting to capture acute air emboli causing hemodynamic compromise on point-of-care ultrasound (POCUS). We placed the patient in reverse Trendelenburg while providing supplemental oxygen which lead to real-time clinical improvement with echocardiographic evidence . Compute6CPC-EM CapsuleWhat do we already know about this clinical entity?Acute air emboli may result in coronary or cerebral infarct, cardiovascular collapse and death without appropriate diagnosis and treatment.What is the major impact of image(s)?To our knowledge, these are the first ED ultrasound images to capture air emboli causing hemodynamic compromise as well as real-time clinical improvement with echocardiographic evidence.How might this improve emergency medicine practice?EM physicians must consider air embolus when approaching a hemodynamically unstable patient, particularly after procedures involving venous or arterial vasculature.


A 23-year-old female was involved in a motor vehicle collision with multiple injuries, including a right acetabular fracture, but no pelvic fracture. Urology consultation was obtained due to difficulty placing a urethral catheter. Examination revealed a longitudinal urethral tear with vaginal laceration extending 2 cm from the urethral meatus proximally toward the bladder neck. The longitudinal urethral tear was repaired primarily. Traumatic female urethral injury in the absence of a pelvic fracture is an exceedingly rare occurrence.


However, more evidence is necessary to in order verify the role of the NMDA receptor subunit NR1 and ROS in anesthetic-induced neurodegeneration.Prolonged exposure of developing mammals to general anesthetics affects the N-methyl-D-aspartate (NMDA)–type glutamate or γ-aminobutyric acid (GABA) receptor systems and enhances neuronal toxicity. Stimulation of immature neurons by NMDA antagonists or GABA agonists is thought to increase overall nervous system excitability and may contribute to abnormal neuronal cell death during development. Although the precise mechanisms by which NMDA antagonists or GABA agonists cause neuronal cell death are still not completely understood, up-regulation of the NMDA receptor subunit NR1 may be an initiative factor in neuronal cell death. It is increasingly apparent that mitochondria lie at the center of the cell death regulation process. Evidence for the role of oxidative stress in anesthetic-induced neurotoxicity has been generated in studies that apply oxidative stress blockers. Prevention of neuronal death by catalase and superoxide dismutase


The tractography showed absence of major crossing pathways within the pons and midbrain, and normal interhemispheric connections in the corpus callosum (A 13-year-old patient was referred for surgical treatment of scoliosis. Her neurological examination revealed the complete loss of horizontal eye movements with normal vertical gaze, suggesting diagnosis of horizontal gaze palsy and progressive scoliosis. The magnetic resonance imaging revealed a hypoplastic pons and medulla with a prominent midline cleft, therefore resulting in characteristic “butterfly” configuration.callosum . Surgicacallosum .Horizontal gaze palsy with progressive scoliosis is a rare autosomal recessive disorder characterized by congenital absence of normal horizontal eye movements and progressive scoliosis during childhood and adolescence. Heterozygous mutations in ROBO3 are reported as a cause of this disorder.


Polycomb-based repressive chromatin.The first genome-wide examination of the chromatin landscape at the periphery of the plant cell nucleus reveals substantial enrichment of heterochromatin and  Arabidopsis NUCLEOPORIN1 (NUP1) protein, an inner subunit of the nuclear pore complex (NPC) [The cell nucleus is a highly structured subcellular organelle that functionally houses the genome. The double-membrane nuclear envelope provides a physical interface that contributes to the compartmentalization of chromosomal domains that have distinct chromatin states and activities within the nuclear space. Our knowledge of chromatin organization and function in plant systems is rapidly expanding, and the roles of different nuclear peripheral components in structuring chromatin are also beginning to emerge . In metaex (NPC)  responsiArabidopsis cells, most heterochromatin typically aggregates around the centromeric and peri-centromeric regions, forming conspicuous subnuclear foci referred to as chromocenters which are thought to tether transposable elements (TEs) from distant genomic domains [Arabidopsis chromocenters are usually located near the nuclear periphery [Arabidopsis NPCs enhanced its expression [CAB (CHLOROPHYLL A/B-BINDING PROTEINS) genes are repositioned from the interior to the periphery of Arabidopsis cotyledon nuclei when induced by light signaling [In  domains   are frequently involved in long-range interactions between distant loci in plants [Arabidopsis chromosomes also form nucleolus-associated domains (NADs) that are enriched in transcribed ribosomal DNA genes and telomeric regions. Interestingly, NADs also contain heterochromatic TEs and silent protein-coding genes that might be sequestered in the nucleolus away from the RNA Pol II machinery (reviewed in [The integration of genome-wide ChIP-seq and Chromosome Conformation Capture 3C) approaches has recently extended linear epigenomic information on successive chromatin states towards a three-dimensional model of intra-chromosomal associations in n plants  and mammn plants . ArabidoC approacThe study by Bi and colleagues  offers aArabidopsis CAB genes [Polycomb-repressed genes (Fig. Bi and colleagues also report that NUP1 RE-CHIP pulled down chromatin fragments that were enriched in acetylated histone H3 and other chromatin modifications characteristic of transcribed genes. The proximity between transcribed genes and NPCs might facilitate mRNA export via the so-called 'gene-gating process' proposed both for animal cells  and for nes Fig. . DetermiArabidopsis. On the one hand, the association of transcriptionally active chromatin with NPCs can promote efficient mRNA surveillance and export [Polycomb-repressed silent regions at the nuclear periphery may provide an efficient way to organize or stabilize higher-order chromatin architecture. A potential contribution of H3K27me3-marked domains in tethering long-range euchromatin interactions around the nuclear periphery offers a novel and exciting working hypothesis for further investigations.The study by Bi and colleagues  adds to d export  while, oArabidopsis nuclei. PRC2 subnuclear foci are not exclusively enriched at the nuclear periphery, neither is RNA Polymerase II excluded from this area [A further question that is raised is whether there is a gradient of transcriptionally active chromatin domains from the interior to the periphery in his area , possiblhis area . This mohis area . AdditioArabidopsis to be extended, and uncover the molecular drivers of plant nuclear architecture diversity.Cell cycle and cellular responses to endogenous and environmental signals trigger dramatic changes in nuclear architecture whose functional impact on genome expression programs remains poorly understood. Future investigations of peripheral chromatin dynamics during cellular transitions thus offer exciting perspectives that can be used to unravel the functional plasticity of plant cell nuclear organization. Exploration of how plant lamina-like components and chromatin-bound factors mediate the anchoring of distinct chromosomal domains in plant species with diverse genomic and chromosomal arrangements should additionally allow the work of Bi and colleagues in


Mycobacterium avium subsp. paratuberculosis (Map) is a potential human pathogen known to be present in raw milk from infected dairy herds. Current pasteurisation regimes do not totally inactivate Map resulting in the possibility of viable cells being present in pasteurised milk used for Cheddar cheese production. A laboratory-based method, ensuring strict safety precautions, was developed to manufacture 800-g Cheddar blocks, experimentally contaminated (postpasteurisation) with two different strains of Map. The composition of the model Cheddar produced was consistent with commercial product. Syneresis of the cheese curd caused a 1 log10 concentration of Map numbers from milk to cheese for a strain isolated from pasteurised milk. The type strain NCTC 8578 did not show a similar concentration effect, but did however survive the Cheddar manufacturing process. A small percentage (<5%) of the Map load for each strain was recovered in the whey fraction during the process.


This study aims to understand the relative roles of external forcing versus internal climate variability in causing the observed Barents Sea winter sea ice extent (SIE) decline since 1979. We identify major discrepancies in the spatial patterns of winter Northern Hemisphere sea ice concentration trends over the satellite period between observations and CMIP5 multi-model mean externally forced response. The CMIP5 externally forced decline in Barents Sea winter SIE is much weaker than that observed. Across CMIP5 ensemble members, March Barents Sea SIE trends have little correlation with global mean surface air temperature trends, but are strongly anti-correlated with trends in Atlantic heat transport across the Barents Sea Opening (BSO). Further comparison with control simulations from coupled climate models suggests that enhanced Atlantic heat transport across the BSO associated with regional internal variability may have played a leading role in the observed decline in winter Barents Sea SIE since 1979. The relative roles of external forcing versus internal variability in the Barents Sea winter sea ice decline since 1979 remain unconstrained. Here, the authors show that enhanced Atlantic heat transport into the Barents Sea associated with regional internal variability may have played a leading role. The decline of Arctic sea ice over the satellite period has often been attributed in large part to anthropogenic climate warming. Were the observed decline to continue at its recent rate, the Arctic would likely become nearly ice-free during summer within the next decade12Meanwhile, substantial multi-decadal/centennial variability of Arctic sea ice cover has been inferred from high-resolution multi-century algal proxy681014161318The Barents Sea is located on one of the main corridors of Atlantic inflow entering the Arctic. Strong coupling between winter Barents Sea SIE and the Atlantic heat transport into the Arctic through the Barents Sea Opening (BSO) are found in both observations and modelling studies1319202123In this study, we would like to address the following questions: What are the relative roles of changes in external radiative forcing versus internal variability in causing the observed winter Barents Sea SIE decline over the satellite period? Are there any substantial discrepancies between the observed and CMIP5-simulated externally forced response in winter NH sea ice decline, particularly given that the internal variability of Atlantic heat transport into the Arctic is largely removed in the CMIP5 multi-model ensemble mean forced response? The work presented here aims to provide insights on these questions by comparing observations with the CMIP5 multi-model ensemble simulations forced by changes in external radiative forcing as well as with long pre-industrial control simulations under constant radiative forcing. We show that the CMIP5 externally forced winter NH sea ice concentration (SIC) trend in individual regions  differs substantially from that observed. Our analysis suggests that enhanced Atlantic heat transport into the Barents Sea associated with regional internal variability may have played a leading role in the observed winter Barents Sea SIE decline since 1979.2 per 37 years) is weaker than that observed (−1.51 million km2 per 37 years), but still accounts for a substantial part (68%) of the observed declining trend. On the other hand, the observed March Barents Sea SIE declining trend is not well captured by the CMIP5 multi-model ensemble mean. Both the observed March Barents Sea SIE declining trend and anomalies often stay outside of the ±1 s.d. of the CMIP5 multi-model ensemble mean (2 per 37 years) is much weaker than that observed (−0.36 million km2 per 37 years), accounting for only a modest part (28%) of the observed declining trend.In this study, we included 79 ensemble members from 32 CMIP5 models (‘Methods' section), and the multi-model ensemble mean represents the response to changes in external radiative forcings in the models, as the internal variability in individual ensemble members tends to cancel out in the multi-model ensemble mean. The CMIP5 multi-model ensemble mean March NH SIE shows a gradual decline over the 37-year period from 1979 to 2015 due to changes in external radiative forcing . The obsble mean . The CMIDespite the weaker than observed declining trends in March NH and Barents Sea SIEs in the CMIP5 multi-model ensemble mean, the annual global mean SAT simulated by the CMIP5 multi-model ensemble mean displays a warming trend (0.86 K per 37 years) stronger than that observed (0.51 K per 37 years) . Both thWhy does the CMIP5 externally forced response account for a substantial part of the observed March NH SIE trend, but only a modest part of the observed March Barents Sea SIE trend ? We inter=−0.2) between annual global mean SAT trends and March Barents Sea SIE trends over the 37-year period across the CMIP5 ensemble members (BSO)  between the trends in annual mean net upward surface heat flux integrated over the Barents Sea region (FSFC) and trends in HTBSO, and strong anti-correlation (r=−0.64) between trends in FSFC and trends in March Barents Sea SIE, over the 37-year period across the CMIP5 ensemble members. Hence, an enhanced HTBSO leads to a reduced winter sea ice cover in the Barents Sea, and more net upward surface heat flux is released into the atmosphere through larger winter open water area in the Barents Sea2026SFC provide a negative feedback to changes in winter Barents Sea SIE. The positive correlation between HTBSO and FSFC trends and the negative correlation between FSFC and March Barents Sea SIE trends suggests that at multi-decadal time scales, changes in HTBSO are forcing changes in Barents Sea SIE and FSFC in the models, not the other way around. If the net upward surface heat flux (FSFC) were driving the winter Barents Sea SIE changes, one would expect a positive correlation between them , which is opposite to the behaviour simulated in the models.There is no significant anti-correlation  is thus too small to cause the observed decline in winter Barents Sea SIE  due to regional internal variability is necessary to explain the dramatic decrease of winter Barents Sea SIE seen in recent decades, in addition to the externally forced response. Here internal HTBSO changes could be affected both by local variability in surface winds through changes in volume transport across the BSO, as well as by remote variability in the large scale ocean circulation, such as AMOC, through changes in both volume transport across the BSO and ocean temperature of the Atlantic inflow.The CMIP5 multi-model ensemble mean forced response in HTter plot . This is Sea SIE . Based o Sea SIE , we estiBSO), we seek insights from long-term pre-industrial control simulations that are available from three coupled climate models . As discussed in the previous section, we estimate that an increase in HTBSO on the order of 30 TW per 37 years due to regional internal variability is necessary to explain the dramatic decrease of winter Barents Sea SIE seen in recent decades. The simulated March Barents Sea SIC declines in response to a positive trend of HTBSO  in all three models due to internal variability , respectively. Even for GFDL CM3, only 38 out of all 3,564 available 37-year segments have such large HTBSO trends (⩾30 TW per 37 years), suggesting a rare occurrence. The implications are that either current climate models have unrealistically small amplitudes of low-frequency internal variability in HTBSO, or the recent observed decline of March Barents Sea SIE is a remarkably rare event. We speculate that it is more likely that models have unrealistically small amplitudes of low-frequency internal variability in HTBSO due to common systematic model deficiencies and mean state biases, rather than that the recent observed decline is a rare event. The underestimation of mean state HTBSO is implied from the simulated excessive mean state winter Barents Sea SIE  in CMIP5 models . It also exhibits a weak declining trend over a broader region in the NH, and thus opposite to the observed increasing trend in central Arctic. Hence, the CMIP5 externally forced response in March Barents Sea SIE exhibits a large discrepancy  compared to that observed, but the discrepancy is less severe in March NH SIE because the discrepancies in the spatial pattern are partially masked by summing over the entire NH. The large discrepancies in the spatial pattern of March NH SIC trends and in March Barents Sea SIE trend suggest that the observed winter NH sea ice trend over the satellite period cannot be well explained by the externally forced response alone, and that the observed rapid declining trend in winter Barents Sea SIE is not predominantly due to external radiative forcing.BSO) has played a key role in March Barents Sea SIE decline, while the warming in global mean SAT has been relatively unimportant for March Barents Sea SIE decline in individual CMIP5 ensemble members. We identify a significant high anti-correlation between winter Barents Sea SIE trends and HTBSO trends throughout individual CMIP5 ensemble members. This suggests that at multi-decadal time scales, changes in HTBSO are forcing changes in Barents Sea SIE and in net upward surface heat flux over the Barents Sea in these models. The CMIP5 externally forced HTBSO increase is too small to cause the observed decline in winter Barents Sea SIE. Based on CMIP5 results and observations, we estimate that a total 35 TW per 37 years trend in HTBSO is needed to explain the observed winter Barents Sea SIE decline over the satellite period. Among this, an increase in HTBSO on the order of 30 TW per 37 years due to regional internal variability is necessary to explain the majority of the observed winter Barents Sea SIE decline seen in recent decades and its discrepancies with the CMIP5 multi-model mean forced response. Results from long pre-industrial control simulations of three coupled models show that the simulated March Barents Sea SIC declines in response to a positive trend of HTBSO  are much larger than the CMIP5 multi-model ensemble mean forced response and are on the same order as that observed.Our analysis further suggests that an enhanced Atlantic heat transport across the BSO  from the National Snow and Ice Data Center (NSIDC)The observed global mean SAT is derived from the averaged SAT of three reanalysis data sets: NCEP/NCARBSO) using monthly mean ocean velocities and potential temperature (reference temperature being 0 °C) from the selected CMIP5 ensemble members.To compare with the observations, we combined CMIP5 twentieth century all-forcing  simulations with CMIP5 representative concentration pathway (RCP) 4.5 future emission scenario simulations to cover the period from 1979 to 2015. The RCP4.5 pathway is chosen from among the four available future emission scenarios to represent the period after 2005, noting that these scenarios deviate little from each other during the short period 2006–2015. A total of 79 ensemble members from 32 CMIP5 models are selected, based on the criteria that all variables needed in our analyses are available in a selected ensemble member, and that the ensemble member has both historical and RCP4.5 simulations so that the they can be combined. Due to the large number of selected ensemble members, the multi-model ensemble means represent the response to changes in external radiative forcings in the models, as the internal variability in individual ensemble members tends to cancel out in the multi-model ensemble means. In this study, both observed and CMIP5-simulated anomalies are referenced to climatological means for 1979–2015. The correlation between March and November–April averaged Barents Sea SIE trends over the 37-year period across CMIP5 ensemble members is 0.94, and the correlation between March and February–April averaged Barents Sea SIE trends over the 37-year period across CMIP5 ensemble members is 0.99. These confirm that trends in March SIE are representative for trends in the winter SIE in this study. To study the oceanic impact on the Barents Sea winter SIE, we calculated the annual mean advective Atlantic heat transport across the BSO   Coupled Model version 2.1 (CM2.1)36BSO, 500 random samples of 37-year segments (same length as 1979–2015) are drawn from the control simulations, and the 500 pairs of 37-year trends in SIC and HTBSO are obtained for each model, respectively. Then SIC trends are regressed onto HTBSO trends for each grid point to obtain the spatial pattern of SIC changes associated with HTBSO changes due to simulated internal variability in each model.Additionally, long-term control simulations available from three coupled climate models under constant pre-industrial radiative forcing are employed to assess the impact of internal variability in HThttps://pcmdi.llnl.gov/search/cmip5. NSIDC sea ice satellite sea ice can be downloaded from http://nsidc.org/data/NSIDC-0051. NCEP/NCAR reanalysis data can be downloaded from http://www.esrl.noaa.gov/psd/data/gridded/data.ncep.reanalysis.html. Other data and code are available from the authors on request.All CMIP5 model output are available from How to cite this article: Li, D. et al. On the discrepancy between observed and CMIP5 multi-model simulated Barents Sea winter sea ice decline. Nat. Commun.8, 14991 doi: 10.1038/ncomms14991 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


Aspergillusflavus to displace aflatoxin-producers during crop development. The displacement results in lower aflatoxin concentrations in harvested grain. The current study evaluated in field performances of two atoxigenic strains of A. flavus endemic to Italy in artificially inoculated maize ears and in naturally contaminated maize. Co-inoculation of atoxigenic strains with aflatoxin producers resulted in highly significant reductions in aflatoxin concentrations (>90%) in both years only with atoxigenic strain A2085. The average percent reduction in aflatoxin B1 concentration in naturally contaminated maize fields was 92.3%, without significant differences in fumonisins between treated and control maize. The vegetative compatibility group of A2085 was the most frequently recovered A. flavus in both treated and control plots . A2085 was therefore selected as an active ingredient for biocontrol products and deposited under provisions of the Budapest Treaty in the Belgian Co-Ordinated Collections of Micro-Organisms (BCCM/MUCL) collection (accession MUCL54911). Further work on development of A2085 as a tool for preventing aflatoxin contamination in maize produced in Italy is ongoing with the commercial product named AF-X1™.Since 2003, non-compliant aflatoxin concentrations have been detected in maize produced in Italy. The most successful worldwide experiments in aflatoxin prevention resulted from distribution of atoxigenic strains of  Data on aflatoxin B1 and percent reduction were ln and arcsin transformed, respectively, prior to analyses. Fields with aflatoxin B1 concentrations below 1 μg/kg in both treatments were not considered for the analyses. Aspergillus flavus populations in the maize flour are reported as log (10) transformed colony forming units (CFU)/g dry maize flour.Percent reduction in aflatoxin B


Heart failure (HF) is a condition characterized by increased filling pressures in the left ventricle due to altered cardiopulmonary blood flow. However, there are no clinically available non-invasive quantitative methods to measure filling pressures in HF. The pulmonary blood volume variation (PBVV) indexed to stroke volume (SV) by cardiac magnetic resonance (CMR) could be a novel non-invasive measure of HF. However, the relationship between PBVV/SV and increased filling pressures has not been validated. We have previously shown that the effect of increased filling pressures and redistribution of pulmonary blood flow can objectively be quantified using perfusion gradients derived from ventilation/perfusion single-photon emission computed tomography (V/P SPECT). V/P SPECT perfusion gradients have a high positive predictive value to diagnose decompensated HF.The aim of this study was therefore to investigate the relationship between the PBVV/SV and perfusion gradients in patients with heart failure.Fifteen patients with HF  were included. All patients underwent CMR at 1.5T and V/P SPECT. Four patients were under consideration for heart transplantation and 11 patients were candidates for cardiac resynchronization therapy. By measuring the blood flow in the pulmonary trunk and in one pulmonary vein, the PBVV was calculated, defined as the maximum cumulative difference in pulmonary blood volume over one heart beat. The SV by CMR was measured as the integral of the flow measurement over one heart beat in the pulmonary trunk. Dorso-ventral perfusion gradients (%-counts/cm) were automatically derived from V/P SPECT images using a user independent algorithm previously developed and validated in our department.There was a positive relationship between PBVV/SV and perfusion gradients Figure  suggestiPBVV/SV measured by CMR, as well as perfusion gradients by V/P SPECT have the potential to serve as non-invasive measures of HF. Further validation work with comparison to invasive pressure measurements are needed to assess the accuracy of the methods.


Clinical scales to detect large vessel occlusion (LVO) may help to determine the optimal transport destination for patients with suspected acute ischemic stroke (AIS). The clinical benefit associated with improved diagnostic accuracy of these scales has not been quantified.We used a previously reported conditional model to estimate the probability of good outcome  for patients with AIS and unknown vessel status occurring in regions with greater proximity to a primary than to a comprehensive stroke center. Optimal rapid arterial occlusion evaluation (RACE) scale cutoff scores were calculated based on time-dependent effect-size estimates from recent randomized controlled trials. Probabilities of good outcome were compared between a triage strategy based on these cutoffs and a strategy based on a hypothetical perfect LVO detection tool with 100% diagnostic accuracy.In our model, the additional benefit of a perfect LVO detection tool as compared to optimal transport-time dependent RACE cutoff scores ranges from 0 to 5%. It is largest for patients with medium stroke symptom severity (RACE score 5) and in geographic environments with longer transfer time between the primary and comprehensive stroke center.Based on a probabilistic conditional model, the results of our simulation indicate that more accurate prehospital clinical LVO detections scales may be associated with only modest improvements in the expected probability of good outcome for patients with suspected acute ischemic stroke and unknown vessel status.The online version of this article (10.1186/s12883-018-1021-8) contains supplementary material, which is available to authorized users. Patients with acute ischemic stroke (AIS) due to large vessel occlusion (LVO) have better outcomes if they receive endovascular therapy (EVT) in addition to thrombolysis.  It is cuOur analysis is based on a probabilistic model built to estimate the probability of good outcome  for patients with AIS and unknown vessel status as a function of stroke severity, transport times to the nearest PSC and CSC, and transfer times between PSC and CSC. Details of the model have been published previously.  Briefly,Absolute probabilities of good outcome using a prehospital triage strategy based on optimal RACE cutoff scores are displayed in Fig. p of LVO: ~ 50%; corresponding binary Shannon entropy -p x log2 p – (1-p) x log2 (1-p): ~ 1.00; Additional file We quantify the clinical benefit potentially achievable with improved prehospital LVO detection scales. Depending on stroke symptom severity, the geographic environment, and the location of stroke incidence relative to the surrounding stroke centers, the probability of good outcome increases by 0–5%. The largest difference is seen for medium severity strokes  the relatively large number of patients with mild stroke symptoms in an unselected sample who are less likely to have LVO and have a higher absolute probability of good outcome and b) the fact that the sensitivity of existing LVO detection scales is already high (85% for a RACE cutoff score ≥ 5). Availability of clinical scales with better discriminatory power would be nonetheless highly desirable. First, each falsely negative triage decision to a PSC may be associated with delayed access to EVT and permanent disability for an individual patient and should therefore be avoided. Second, from a societal perspective, a hypothetical scale with perfect accuracy would eliminate negative consequences of prehospital triage caused by low specificity and negative predictive value of currently available LVO detection scales, such as overburdening of CSCs with non-LVO infarcts, intracerebral hemorrhages and stroke mimics and deskilling and deprioritizing of PSCs due to lower patient volume. , 13The limitations of our model have been described previously.  Briefly,In conclusion, our results indicate that more accurate prehospital clinical LVO detections scales may be associated with only modest improvements in the expected probability of good outcome for patients with suspected acute ischemic stroke and unknown vessel status. Development of LVO detection scales with higher specificity could help to minimize the negative effects on patient distribution and resource use associated with currently available triage scales.Additional file 1:Panel A: Displayed are distributions of point-wise differences between the estimated probabilities of good outcome  associated with a prehospital triage strategy based on a hypothetical perfect large vessel occlusion (LVO) detection tool and the estimated probabilities of good outcome associated with a prehospital triage strategy based on optimal rapid arterial occlusion evaluation (RACE) scale cutoff scores, for different stroke severities and different transfer time settings . Panel B: Displayed are distributions of point-wise differences between the estimated probabilities of good outcome  associated with a prehospital triage strategy based on a hypothetical perfect LVO detection tool and the estimated probabilities of good outcome associated with a prehospital triage strategy based on a RACE cutoff score ≥ 5, for different stroke severities and different transfer time settings . Colors represent different treatment scenarios, with the possibility for treatment options defined by standard treatment time windows: symptom onset-tothrombolysis – 270 min; symptom onset-to-groin puncture – 360 min): I, both thrombolysis at the comprehensive stroke center (CSC) under the mothership approach and secondary transfer for endovascular therapy (EVT) under the drip and ship approach possible. II, thrombolysis at the CSC under the mothership approach not possible, secondary transfer for EVT under the drip and ship approach possible. III, thrombolysis at the CSC under the mothership approach possible, secondary transfer for EVT under the drip and ship approach not possible. IV, neither thrombolysis at the CSC under the mothership approach nor secondary transfer for EVT under the drip and ship approach possible. Horizontal lines indicate means. (TIFF 2376 kb)Incremental benefit of a hypothetical perfect large vessel occlusion detection scale according to treatment scenario I – IV. Additional file 2:-p x log2 p - (1-p) x log2 (1-p). Here, p denotes the probability of the presence of LVO among patients with ischemic stroke in a given RACE score category. Incremental benefits of a hypothetical perfect LVO tool correspond to average values in a geographic environment with a transfer time between PSC and CSC of 120 minutes. Labels indicate RACE score categories. (TIFF 1292 kb)Relationship between the uncertainty with regards to the presence of large vessel occlusion and the incremental benefit of a hypothetical perfect large vessel occlusion detection tool over optimal rapid arterial occlusion evaluation scale cutoff scores. Uncertainty with regards to the presence of large vessel occlusion (LVO) among patients with ischemic stroke in each rapid arterial occlusion evaluation (RACE) scale score category is quantified using the Shannon entropy


Treatment-resistant schizophrenia (TRS) is a major cause of disability and functional impairment worldwide. Approximately 30% of patients with schizophrenia will develop TRS at some point during their illness course. Despite the staggering financial and emotional costs associated with TRS, this severe disorder is poorly understood. The pathophysiological basis of TRS is posited in part to have neurodevelopmental roots. If early brain development (<2 years of age) influences TRS, then cortical gyrification, which is often complete by 2 years of life, could be abnormal in TRS when compared to non-TRS subjects. Subtle but diffuse pathological changes that occur during early development are postulated to disrupt the maturational relationship (covariance) among brain regions, even if no localised morphological changes are seen in adult life. The disrupted structural covariance resulting from diffuse developmental dyscoordination in early life can be quantified using gyrification-based connectomes obtained using graph theory. We applied this method to baseline MRI data collected during first contact with mental health services for psychosis to predict the emergence of TRS in the next 5 years.70 patients with first episode schizophrenia spectrum disorder who presented to mental health services between 2005 and 2010 were followed up for 5 years using electronic case notes. Psychopathology was assessed at baseline with the Positive and Negative Syndrome Scale (PANSS) and symptom dimensions were derived using Wallwork’s model. TRS was defined according to Health and Clinical Excellence guidelines. Structural MRI images were obtained at baseline, with minimal exposure to antipsychotics (<3 months). Local gyrification indices were computed using Schaer’s method for 68 contiguous cortical regions (34 in each hemisphere) using Freesurfer’s Desikan atlas. After adjusting for age, gender and intracranial volume, group-based structural covariance was estimated (68x68 correlation indices) and each subject’s contribution to the covariance was quantified using a jack-knife procedure, providing one distance matrix for each subject. These matrices were used to construct distance-based gyrification connectomes using Graph Analysis Toolbox. We used a functional data analysis approach across a range of cost-thresholds to reduce multiple testing when comparing TRS and non-TRS groups.17 (24.3%) of patients with first episode schizophrenia spectrum disorder met criteria for TRS at the end of the 5 years of follow up; 53 (75.7%) were non-TRS. TRS subjects had a significant reduction in small-worldness compared to non-TRS group  and reduced clustering coefficient  with increased path length .The positive symptoms were positively correlated  with higher small-worldness  suggesting that a predominantly hyperdopaminergic status that induces positive symptoms may relate to preserved small-worldness seen in non-TRS individuals, while subtle developmental changes resulting in reduced small-worldness may underlie TRS.These changes suggest that in the presence of TRS, the cortex-wide covariance in folding patterns become less organized, with reduced regional segregation as well as reduced overall integration of the morphological connectome. Such an effect may result from weakening of the tensions that arise from inter-regional connectivity in the neonatal brain. The emergence of TRS may be characterised by a neurodevelopmentally driven abnormality in structural organisation of the human cortex in those who develop schizophrenia.


The TRT data consists of measured inlet and outlet temperatures, fluid flow and injected heat rate recorded every 10 min. The field dataset is made available to enable model verification studies.This data article comprises records from five Thermal Response Tests (TRT) of quadratic cross section pile heat exchangers. Pile heat exchangers, typically referred to as energy piles, consist of traditional foundation piles with embedded heat exchanger pipes. The data presented in this article are related to the research article entitled  Specifications TableValue of the data•Each TRT is presented in an individual Excel sheet.•These data can be used to validate thermal models of pile heat exchangers.•There are not publicly available full-scale TRT datasets for pile heat exchangers.•Sharing data will support the development of this type of ground heat exchangers.•These data can supplement other data sets to assist the development of thermal dimensioning guidelines for pile heat exchanger foundations.1Dimensioning of Ground Source Heat Pump installations typically relies on thermal response testing (TRT) of one or more ground heat exchangers. The dataset of this article provides raw TRT data of several precast pile heat exchangers, described in 2During the TRT, the heat carrier fluid (water) is circulated in the ground heat exchanger while being continuously heated at a specified rate. Heat dissipates to the ground heat exchanger and subsequently to the ground. The test records fluid inlet- and outlet temperatures, the fluid flow rate and energy consumption and logs them in 10-min intervals for at least 48 h.The shared data consists of five TRTs of square cross section precast pile heat exchangers. The five energy piles have different lengths and pipe configurations. The tests have been carried out at two different locations in Denmark.0 [°C], the average soil thermal conductivity λs [W/m/K] over the length of the heat exchanger and the thermal resistance the ground heat exchanger Rb [K m/W] Model interpretation of the measured TRT temperatures yield estimates of the undisturbed soil temperature T3], inlet temperature T1 [°C], outlet temperature T2 [°C], flow [l/h] and injection heat rate or effect [kW]. Notice the data is given from the closest in time to the most distant.The TRT sets are compiled in a single Excel file, separated in sheets named by the pile IDs (refer to Table 2 in The TRT equipment is produced by UBeG, Ref.


This exhaustion, accompanied by increased cell surface expression of check point inhibitors was originally attributed to chronic TA-driven stimulation of CD8+ tumor infiltrating lymphocytes (TILs)[+ TILs, directed to antigens not present within the TME, contradicts this notion but rather points towards a different mechanism.At least in experimental animals the efficacy of active immunotherapy of cancer can be increased by metabolic reprogramming of tumor antigen (TA) specific CD8ycolysis . In a nues (TILs). Exhaust+ T cells that survive the harsh TME and are still present in advanced tumors have managed to switch from glucose to FA catabolism [T cells during activation in the periphery switch to glycolysis to meet their demand for energy and biomass production. Upon entering solid tumors they encounter an environment that is metabolically challenging and provides opposing cues. Lack of oxygen due to insufficient vessel formation in rapidly growing tumors increases signaling through the hypoxia-induced factor (HIF)-1a pathway, which promotes glycolysis. Glycolysis obviously requires consumption of glucose, which is commonly scarce in solid tumors, as it is also the cancer cells' preferred fuel. As a result, T cells conditioned towards glycolysis may starve within the TME. The interstitial fluid of solid tumors is commonly rich in fatty acids (FAs), which activate signaling through the peroxisome proliferator-activated receptor (PPAR)-α, a transcription factor that augments lipid metabolism. Indeed CD8tabolism .+ T cells within the TME during tumor progression suggests that many T cells are unable to make this crucial metabolic switch. Like in any war it helps to train the troops before sending them into battle. CD8+ T cells taught to gain energy through FAs rather than glucose before they enter the TME fare better that those that are untutored; their functions remain preserved and they are more effective at delaying tumor progression [The loss of activated CD8gression .There are many ways to change a cell's metabolism. Knockdown of HIF-1α or other key factors of the glycolysis pathway reduces a cell's reliance on glucose. The same effect could be achieved by reinforcing PD-1 signaling, which through blockade of the Akt/mTor pathways decreases glycolysis and increases lipid catabolism. FAO can be enhanced directly through strengthening PPAR-α signaling or genetically increasing the activity of rate-limiting steps of FAO, such as Carnitine Palmitoyltransferase (CPT 1), an enzyme that is essential to transfer long-chained FAs into mitochondria. Production of ketone bodies, which may provide crucial nutrients in hypoglycemic and hypoxic solid tumors, could be intensified by knock-in of 3-Hydroxy-3-Methylglutaryl-CoA Synthase (HMGCS) 2, the rate-limiting enzyme of ketogenesis.+ T cells stimulated in mice treated with fenofibrate are more efficacious at reducing tumor burden than those activated without the drug. Fenofibrate upon entering mitochondria reversibly inhibits complex I of the electron transfer chain[+ T cells and should thus be explored further (unpublished)*.Our efforts thus far have focused in fenofibrate , a PPAR-fer chain. In seru+ T cells to better prepare them for the hostile conditions of a TME. Treatment would need to be targeted to the unique metabolic conditions with different types of cancer.Recent years have shown tremendous progress in active immunotherapy of cancers through TA-specific vaccines, adoptiv


A 66-year-old woman presented with 4 years of progressive speech difficulty. She had nonfluent speech with phonemic errors but intact single-word comprehension and object knowledge. Her grammar was impaired in both speech and writing, and she exhibited orofacial apraxia. A clinico-radiologic see  diagnosi1 It is most frequently due to tau pathology, and clinicians should be alert to the potential development of progressive supranuclear palsy or corticobasal syndrome.2Nonfluent variant primary progressive aphasia is a neurodegenerative disease within the spectrum of frontotemporal dementia, characterized by the typical language and brain atrophy patterns seen here.


Deviation in level of intrinsic neural activity  is observed in psychosis. Neurophysiological models have proposed this physiological indicator as a genetically mediated core deviation in psychosis. Translational models of intrinsic activity deviations promise to identifying multiple distinct physiological mechanisms for psychosis manifestation. Intrinsic activity deviations may masquerade as higher levels of neural response in sensory cortices, but ultimately may lead to poor signal-to-noise ratios, particularly when psychosis cases are required to identify stimulus salience.Why do we not hear more about intrinsic activity as a core biomarker for any psychosis variation? An explanation is provided by the current project. The Bipolar-Schizophrenia Network for Intermediate Phenotypes (B-SNIP) published a means for categorizing psychoses by neurobiological homology via use of multiple biomarkers (psychosis Biotypes) rather than by clinical features. B-SNIP demonstrated the superiority of Biotypes versus DSM diagnoses for capturing neurobiological similarity through multiple external validating measures . Independent analyses since the initial publication have provided additional support for the usefulness of psychosis Biotypes.For this project, we analyzed ongoing neural activity from 64 EEG sensors during 150 intervals of 10 sec duration from over 1450 B-SNIP subjects. These data (never before published) were from the inter-trial interval (ITI) of an auditory paired-stimuli task used in Biotypes construction (these ITI data themselves were not used). Although the subjects were engaged in a task (counting the number of stimulus pairs), the data used here were not part of the task itself. Data were evaluated for single trial power  as a function of frequency of neural oscillations (from 2–50 Hz) over the whole head. Data were then averaged over single trials to yield an estimate of the overall strength of nonspecific (unrelated to sensory processing) neural activity.When evaluated by DSM diagnoses , the 95% confidence intervals for all groups overlapped the healthy group means across all frequencies. When considered by psychosis Biotypes, differences were obvious and statistically significant. In comparison to healthy persons, Biotype-2 probands  were notably high on nonspecific neural activity, and Biotype-1 probands  were notably low. Group separations on this metric were better than those obtained with original intrinsic EEG measure used in psychosis Biotypes construction, indicating this more pure intrinsic activity measure is capturing a meaningful component of Biotype neurophysiology. This was true across a range of oscillatory frequencies for the probands. The first-degree relatives of the Biotype probands showed similar patterns, although higher frequency oscillations (above 20 Hz) better differentiated relatives from healthy persons.Intrinsic activity deviation is a promising biomarker for translational research programs aimed at differential treatment development, but using DSM psychosis diagnoses would obscure its importance for understanding psychosis.


Candida albicans is a virulent human opportunistic pathogen. It evades innate immune surveillance by masking an immunogenic cell wall polysaccharide, β-glucan, from recognition by the immunoreceptor Dectin-1. Glucan unmasking by the antifungal drug caspofungin leads to changes in the nanostructure of glucan exposure accessible to Dectin-1. The physical mechanism that regulates glucan exposure is poorly understood, but it controls the nanobiology of fungal pathogen recognition. We created computational models to simulate hypothetical physical processes of unmasking glucan in a biologically realistic distribution of cell wall glucan fibrils. We tested the predicted glucan exposure nanostructural features arising from these models against experimentally measured values. A completely spatially random unmasking process, reflective of random environmental damage to the cell wall, cannot account for experimental observations of glucan unmasking. However, the introduction of partially edge biased unmasking processes, consistent with an unmasking contribution from active, local remodeling at glucan exposure sites, produces markedly more accurate predictions of experimentally observed glucan nanoexposures in untreated and caspofungin-treated yeast. These findings suggest a model of glucan unmasking wherein cell wall remodeling processes in the local nanoscale neighborhood of glucan exposure sites are an important contributor to the physical process of drug-induced glucan unmasking in C. albicans. Candida albicans is a normal commensal fungus living on various human mucosal surfaces. It is also an opportunistic pathogen that can cause superficial or systemic infections. Healthcare costs associated with treatment of candidiasis exceed $1 billion annually in the US alone . Given that, we chose to simply use the random unmasking model  in subsequent studies as it seemed to better mimic the experimental processes we expected might be present. The above values were calculated from n = 100 runs for each condition.(TIF)Click here for additional data file.S2 FigCellwall represents the 200 x 200 pixelated simulation space . Boundary collects the  boundary pixels of existing glucan exposure sites within the masked stripes. Masked keeps track of which pixels are available to be chosen, however, only those in non-fixed regions cover masked glucan and so can be exposed. The actual MATLAB algorithm was written in such a way that it can be run on multiple processors simultaneously.(DOCX)Click here for additional data file.


Here, we presented new original data on the effects of iron depletion on the circulating lipid profile in B6HCV mice, a murine model of HCV-related dyslipidemia. Male adult B6HCV mice were subjected to non-invasive iron depletion by low iron diet. Serum iron concentration was assessed for evaluating the effects of the dietary iron depletion. Concentrations of circulating triglycerides, total cholesterol, Low Density Lipoproteins (LDLs), High Density Lipoproteins (HDLs) were analyzed and reported by using stacked line charts. The present data indicated that low serum iron concentration is associated to i) lower serum triglycerides concentrations and ii) increased circulating LDLs. The presented original data have not been published elsewhere. Specifications TableValue of the data•The presented data indicated that low-iron diet i) reduced serum triglycerides concentrations and ii) increased levels of circulating LDLs in HCV transgenic mouse model. No differences have been found on cholesterol and HDLs concentrations in low iron feed mice respect to controls.•These data are the further evidence on the interplay between the iron and lipid metabolism.•The data in this article could be useful to optimize further clinical trials to evaluate the potential therapeutic effect of dietary iron depletion on dyslipidaemia in patients with Hepatitis C infection and hepatic iron overload.1Mice were fed a low-iron diet for thirty days and reduction in blood iron concentration was assessed by biochemical analysis . During 2To date, many evidences showed that iron overload plays a causal role in the hepatic fat accumulation. In particular, studies on HCV (hepatitis-C-virus) transgenic mice fed an excess-iron diet demonstrated that hepatic iron overload exacerbates the imbalance of lipid metabolism by both lipogenesis activation and reduced oxidation activity of fatty acids 32 asphyxiation and blood collected by post-mortem cardiac puncture. No animals became severely ill or died prior to the experimental endpoint. Euthanasia in mice was performed by carbon dioxide asphyxiation.B6HCV transgenic mice were from Markus Heim laboratory at Department of Biomedicine University Hospital of Basel t-test. The statistical significance of correlation was determined by the use of a simple regression analysis. A P value of 0.05 was considered to be significant.Blood biochemical analysis were performed on an automated hematology analyzer using mouse-specific algorithms and parameters (ProCyte DX IDEXX and Abaxis VetScan5). Total cholesterol was measured by the CHOD-PAP method adapted to automated analyzer (Chem 400), non-complexed HDL was measured using the CHOD-PAP method. For triglyceride determinations GPO-PAP method was used. Quantitative values are expressed as mean±SD . Data beMIUR Ministero dell’Università e Ricerca Scientifica .This work was supported by grants from


Atypical porcine pestivirus (APPV) was recently reported to be associated with neurologic disorders in newborn piglets. Investigations of 1,460 serum samples of apparently healthy pigs from different parts of Europe and Asia demonstrate a geographically wide distribution of genetically highly variable APPV and high APPV genome and antibody detection rates. Flaviviridae. The recently discovered atypical porcine pestivirus (APPV) is capable of inducing neurologic disorder in its host, like other pathogens of this family . Several recently published reports demonstrate that APPV is a prominent cause of virus-induced congenital tremor in pigs  3 encoding region and confirmed specificity of amplification by gel electrophoresis . Genome We used individual samples with high genome loads to generate amplicons in a seminested PCR and subsequently performed Sanger sequencing . We generated 20 different APPV NS3 sequences from apparently healthy pigs of all countries (sequences deposited into GenBank under accession nos. MF279213–32). Genetic differences reflect geographic origin to a low degree ; genetic,In addition to the NS3 fragments, we determined APPV complete coding sequences (CDS) from 1 sample from a healthy pig from China (deposited into GenBank under accession no. MF167292) and 2 samples (accession nos. MF167290 and MF167291) obtained from pigs during outbreaks of congenital tremors in Germany ,.rns antibody ELISA, as described (>60% of the animals showed intermediate to high reactivity in the antibody ELISA (We applied an indirect APPV Edy ELISA , which idy ELISA . We founOur findings indicate that the recently discovered APPV is abundant on several continents. APPV must be regarded as a pig pathogen of likely worldwide relevance.Additional information from genetic analysis of atypical porcine pestivirus in animals from Europe and Asia


Intracranial arteriovenous malformations can be further complicated by the development of aneurysms, which themselves carry the risk of rupture and hemorrhage. New endovascular techniques allow for more treatment options for these lesions in the setting of arteriovenous malformations. Here we present the case of a patient who developed an aneurysm in the setting of an arteriovenous malformation and subsequently underwent successful endovascular treatment via percutaneous access of the vertebral artery along with reviewing the literature on further treatment options and developments. Intracranial arteriovenous malformations (AVMs) are rare vascular lesions associated with a risk of hemorrhage of up to 4% per year . AVMs caA 63-year-old female with no significant past medical history presented to our facility with headache and neck stiffness. The headache progressed to being the most severe pain she had ever experienced. She was also mildly lethargic but did not exhibit any focal deficits. She denied history of trauma or use of aspirin or blood thinners. Initial brain computerized tomography (CT) demonstrated evidence of scattered subarachnoid hemorrhage and diffuse cerebral edema which was most prevalent within the posterior fossa. Subsequent brain magnetic resonance imaging (MRI) with and without contrast demonstrated persistent right superior cerebellar hemisphere subarachnoid hemorrhage with associated stable parenchymal hematoma and evidence of a distal flow-related right superior cerebellar artery aneurysm secondary to an underlying posterior fossa AVM. Cerebral angiography showed two ruptured flow-related aneurysms on an enlarged branch of the right superior cerebellar artery associated with a small tectal AVM Figure .However, initial attempts at endovascular treatment were unsuccessful due to profound tortuosity of the left vertebral artery, which was the only potential transfemoral or transradial access to the flow-related aneurysm Figure .Given the marked tortuosity of the vessels, the patient underwent treatment of the flow-related aneurysms beginning with percutaneous access of the vertebral artery at the C1 level, obtained using roadmap guidance from selective catheterization of the ostia of the left vertebral artery Figure .Using a standard microcatheter system over a microcatheter guidewire, a super-selective catheterization of the two flow-related aneurysms was performed Figure .Multiple detachable platinum coils were then deposited into the aneurysmal branch resulting in occlusion of the vessel. Final control angiography revealed marked stasis in the two flow-related aneurysms well into the venous phase suggesting that they will rapidly thrombose Figure .The patient subsequently underwent surgical resection of the AVM and made complete neurological recovery. Repeat angiographic evaluation at six months showed no evidence of vascular abnormality Figure .The incidence of AVM-associated aneurysms has been reported in the literature ranging from 2.7 to 58% with a prevalence of 10-20% being cited in the largest case series -5. It isAVM-associated aneurysms are a rare but dangerous complication of AVMs. The emergence of new endovascular techniques has allowed for broader management of AVM-associated aneurysms. Prompt treatment is necessary to prevent aneurysmal rupture, hemorrhage, and fatality. The patient we presented underwent successful endovascular treatment for an AVM-associated aneurysm using a novel approach. It is important for physicians to recognize and treat AVM-associated aneurysms promptly while taking into account the unique neurovascular anatomy for each patient in order to avoid potentially dangerous complications.


To assess accuracy and precision of the positron emission tomography-computed tomography (PET-CT) carotid standardized uptake values (SUV) of 18F–fluorodeoxyglucose (18FDG) as an inflammatory biomarker for determining cerebrovascular diseases such as stroke, methodology and statistical issues should be taken into account. Otherwise, misleading messages will be the main outcome of such research. Briefly, confusing accuracy and precision will mainly produce misleading messages.18F–fluorodeoxyglucose (18FDG) have been proposed as an inflammatory biomarker for determining cerebrovascular diseases such as stroke. Consideration of varying methodological approaches and software packages is critical to the calculation of accurate SUVs in cross-sectional and longitudinal patient studies. They aimed to investigate whether or not carotid atherosclerotic plaque SUVs are consistent and reproducible between software packages [18FDG-PET SUVs of carotids were taken in 101 patients using two different software packages [n = 23) of the ROI locations, the differences between the SUV measurements from each software package were found to be statistically significant. They highlighted the importance of standardizing all aspects of methodological approaches to ensure accuracy and reproducibility.I was interested to read the paper by Giannotti N and colleagues published in the Dec 2017 issue of EJNMMI Res [However, reproducibility  and accuracy  are two completely different methodological issues –8. The m


Carcinosarcomas are rare tumours containing both malignant mesenchymal and epithelial elements. This reports presents a 70-year-old man with carcinosarcoma of the urinary bladder, which is proven histologically.


A 61-year-old alcoholic man presented to the emergency department because of vomiting and dyspnea after discharged from hospital due to appendectomy two days ago. He smoked heavily and his past medical history revealed gastritis. On the examination, he was hypotension (50/00 mmHg) and lethargy. Cardiopulmonary resuscitation was initiated aggressively. His chest x-ray showed right-sided tension pneumothorax (A). Chest drainage tube was inserted immediately and his clinical status improved spectacularly. On the fifth hospital day, the presence of food particles  (red arrows) in the chest tube was found accidentally (B). Suspecting for esophageal perforation, the oral methylene blue test showed that the fluid in the chest tube turned bluish (C). Boerhaave's syndrome was established. An operation undertaken revealed a perforation of lower esophagus which was marked with nasogastric tube (yellow arrow) (D). However, his condition became worsen with nosocomial infection and died after that. The contents in the chest tube which can be an important diagnostic clue should be checked closely in the clinical practice.


With significant improvements in percutaneous coronary intervention (PCI) technology, complex high risk PCI is increasingly offered to patients with limited revascularization options. Percutaneous mechanical circulatory support devices are often utilized for hemodynamic support during these complex procedures. Veno-arterial extracorporeal membrane oxygenation (ECMO) allows full hemodynamic support and provides systemic oxygenation. We describe a case of left main bifurcation stenting performed at our center with ECMO support in an awake patient without general anesthesia.  Significant improvements in percutaneous coronary intervention (PCI) technology has led to complex high-risk PCI being offered to patients with limited revascularization options. Percutaneous mechanical circulatory support (MCS) devices, including veno-arterial extracorporeal membrane oxygenation (VA-ECMO), are often utilized for hemodynamic support during these complex procedures. VA-ECMO provides cardiopulmonary support and is comprised of a venous cannula to drain the deoxygenated blood, centrifugal nonpulsatile pump, membrane for gas exchange and an arterial cannula to return the oxygenated blood to the body . This alThe definition of high-risk PCI is currently evolving. Patient, lesion and clinical presentation risk factors need to be considered when making the decision about need for percutaneous hemodynamic support . CommonlA 54-year old male with a history of ischemic heart disease and hypertension presented with an acute coronary syndrome (ACS). Coronary angiography demonstrated a left dominant system with a 90% stenosis in the distal left main (LM) extending into the left circumflex (Cx) and left anterior descending (LAD) arteries, in addition to a previously documented 100% mid-LAD occlusion Figure , 1B. LefThe patient was lightly sedated with midazolam and fentanyl. PCI was performed via the right femoral artery using a 7F sheath. ECMO cannulation Figure  was via  With significant improvements in PCI technology, complex high-risk PCI is increasingly offered to patients with limited revascularization options. Percutaneous MCS devices are often utilized for hemodynamic support during these complex procedures.IABP is the most common type of MCS used during PCI, mainly due to its ease of use. A single double lumen catheter with a balloon at the end is inserted and attached to a pump. Helium inflates the balloon at the beginning of diastole. IABP increases diastolic blood pressure and coronary artery perfusion decreases afterload and myocardial oxygen consumption and may increase cardiac output slightly . AdvantaAnother type of MCS is the Impella , which is a device placed across the aortic valve. When optimally positioned, it propels blood from the LV to the aorta. The Impella decreases end-diastolic wall stress while increasing diastolic compliance as well as aortic and intracoronary pressures and coronary flow velocity reserve .  AdvantWhile traditionally, IABP and Impella are used, the former might not provide adequate hemodynamic support  and the Percutaneous mechanical circulatory support devices, including awake VA-ECMO, can be safely performed in appropriate cases of high-risk PCI.


A feared complication of ventricular access for drainage or shunting is ventriculitis. Early diagnosis and treatment is vital to prevent morbidity and mortality. Efficacy of directed antibiotic therapy in ventriculitis is limited by increasing multidrug resistant microorganisms and insufficient systemic antibiotic absorption into the cerebrospinal fluid. Treatment may involve intravenous and/or intrathecal antibiotics as well as external ventricular drainage.We present the first case report suggesting a potential role of a novel technique – direct ventricular catheter-mediated continuous saline irrigation and serial prone patient positioning – to treat a fulminant bacterial ventriculitis. This novel technique promotes egress of purulence from the ventricles and may result in more rapid control of intraventricular infectious burden. Ventriculitis is a feared complication associated with cerebrospinal fluid (CSF) diversion and invasive intracranial pressure monitoring . A largeFirst-line empiric treatment for ventriculitis includes a third generation cephalosporin in addition to vancomycin . UnfortuHerein, we report the case of 48-year-old man with hydrocephalus and a history of numerous ventriculoperitoneal shunt (VPS) placements and revisions. He developed ventriculitis after undergoing VPS placement and required removal of the VPS and placement of temporary CSF diversion. We present a novel technique of catheter-mediated direct continuous saline irrigation into the ventricle paired with intrathecal antibiotic administration and serial prone positioning to promote ventricular drainage.The patient is a 48-year-old man who presented with recurrent, worsening symptoms of hydrocephalus following failed endoscopic third ventriculostomy. He subsequently underwent placement of a right frontal VPS. On post-operative day 4, the patient developed a high fever and altered mental status and underwent a shunt tap revealing grossly purulent fluid Figure . UltimatAt this time, we provided a novel treatment strategy for the patient that involved the following:1. Placement of bilateral  EVDs.2. Preservative-free normal saline was instilled in the right frontal EVD at 20 cc/hr with the left frontal EVD left open at 2 cm H2O.3. Intraventricular gentamicin 8 mg daily was administered with both EVDs clamped for one hour after administration of the gentamicin.4. Gentamicin to be administered intrathecally daily until three to five days after culture clearance.5. Serial prone positioning for 30 minutes every three hours was carried out.6. Care was taken to pad all pressure points well and a standard face cushion was applied which was identical to the cushion utilized in prone positioned cases in the operating room.At the outset of therapy, a computed tomography (CT) scan of the brain was obtained  so the bilateral EVD were removed and a new shunt was implanted on Day 14. IV ceftriaxone was ultimately continued for a total of 21 days. The patient had no untoward effects from his bilateral EVD placement, intrathecal antibiotic administration, or serial prone positioning.Bacterial ventriculitis is linked to significant morbidity and mortality following shunt surgery. Rapid drainage of purulent material may be beneficial in curtailing inflammation. Indeed, insufficient drainage may raise intracranial pressure and exacerbate intracranial infection . This isThere have been previous reports of intraventricular irrigation utilized to treat ventriculitis. A case report by Wada, et al. in 2000 reported a successful use of multiple intraventricular lavages for the treatment of post-traumatic ventriculitis in a 44-year-old male following traumatic brain injury . In addiThis is the first report of the use of intraventricular catheter-mediated continuous saline irrigation paired with intraventricular antibiotic administration and serial prone patient positioning in the treatment of ventriculitis. Our case report demonstrates the safety and efficacy of this technique.