Structure of a C1/C4-oxidizing AA9 lytic polysaccharide monooxygenase from the thermophilic fungus Malbranchea cinnamomea

The thermophilic fungus Malbranchea cinnamomea contains a host of enzymes that enable its ability as an efficient degrader of plant biomass and that could be mined for industrial applications. This thermophilic fungus has been studied and found to encode eight lytic polysaccharide monooxygenases (LPMOs) from auxiliary activity family 9 (AA9), which collectively possess different substrate specificities for a range of plant cell-wall-related polysaccharides and oligosaccharides. To gain greater insight into the molecular determinants defining the different specificities, structural studies were pursued and the structure of McAA9F was determined. The enzyme contains the immunoglobulin-like fold typical of previously solved AA9 LPMO structures, but contains prominent differences in the loop regions found on the surface of the substrate-binding site. Most significantly, McAA9F has a broad substrate specificity, with activity on both crystalline and soluble polysaccharides. Moreover, it contains a small loop in a region where a large loop has been proposed to govern specificity towards oligosaccharides. The presence of the small loop leads to a considerably flatter and more open surface that is likely to enable the broad specificity of the enzyme. The enzyme contains a succinimide residue substitution, arising from intramolecular cyclization of Asp10, at a position where several homologous members contain an equivalent residue but cyclization has not previously been observed. This first structure of an AA9 LPMO from M. cinnamomea aids both the understanding of this family of enzymes and the exploration of the repertoire of industrially relevant lignocellulolytic enzymes from this fungus

RF-Energized Intracoronary Guidewire to Enhance Bipolar Ablation of the Interventricular Septum: In-silico Feasibility Study

"This is an Accepted Manuscript of an article published by Taylor & Francis in International Journal of Hyperthermiaon [date of publication], available online: https://doi.org/10.1080/02656736.2018.1425487"[EN] Purpose: Although bipolar radiofrequency (RF) ablation (RFA) is broadly used to eliminate ventricular tachycardias in the interventricular septum wall, it can fail to create transmural lesions in thick ventricular walls. To solve this problem, we explored whether an RF-energised guidewire inserted into the ventricular wall would enhance bipolar RFA in the creation of transmural lesions through the ventricular wall.Methods: We built three-dimensional computational models including two irrigated electrodes placed on opposing sides of the interventricular septum and a metal guidewire inserted into the septum. Computer simulations were conducted to compare the temperature distributions obtained with two ablation modes: bipolar mode (RF power delivered between both irrigated electrode) and time-division multiplexing (TDM) technique, which consists of activating the bipolar mode for 90% of the time and applying RF power between the guidewire and both irrigated electrodes during the remaining time.Results: The TDM technique was the most suitable in terms of creating wider lesions through the entire ventricular wall, avoiding the hour-glass shape of thermal lesions associated with the bipolar mode. This was especially apparent in the case of thick walls (15mm). Furthermore, the TDM technique was able to create transmural lesions even when the guidewire was displaced from the midplane of the wall.Conclusions: An RF-energised guidewire could enhance bipolar RFA by allowing transmural lesions to be made through thick ventricular walls. However, the safety of this new approach must be assessed in future pre-clinical studies, especially in terms of the risk of stenosis and its clinical impact.This work was supported by the Spanish Ministerio de Economia, Industria y Competitividad under "Plan Estatal de Investigacion, Desarrollo e Innovacion Orientada a los Retos de la Sociedad" Grant "TEC2014-52383-C3 (TEC2014-52383-C3-1-R)". A. Gonzalez-Suarez has a "Juan de la Cierva-formacion" Postdoctoral Grant (FJCI-2015-27202) supported by the Spanish Ministerio de Economia, Industria y Competitividad, Secretaria de Estado de Investigacion, Desarrollo e Innovacion.Pérez, JJ.; González Suárez, A.; D Avila, A.; Berjano, E. (2018). RF-Energized Intracoronary Guidewire to Enhance Bipolar Ablation of the Interventricular Septum: In-silico Feasibility Study. International Journal of Hyperthermia. 34(8):1202-1212. https://doi.org/10.1080/02656736.2018.1425487S12021212348Baszko, A., Telec, W., Kałmucki, P., Iwachów, P., Kochman, K., Szymański, R., … Siminiak, T. (2016). Bipolar irrigated radiofrequency ablation of resistant ventricular tachycardia with a septal intramural origin: the initial experience and a description of the method. Clinical Case Reports, 4(10), 957-961. doi:10.1002/ccr3.648Gizurarson, S., Spears, D., Sivagangabalan, G., Farid, T., Ha, A. C. T., Masse, S., … Nanthakumar, K. (2014). Bipolar ablation for deep intra-myocardial circuits: human ex vivo development and in vivo experience. Europace, 16(11), 1684-1688. doi:10.1093/europace/euu001Koruth, J. S., Dukkipati, S., Miller, M. A., Neuzil, P., d’ Avila, A., & Reddy, V. Y. (2012). Bipolar irrigated radiofrequency ablation: A therapeutic option for refractory intramural atrial and ventricular tachycardia circuits. Heart Rhythm, 9(12), 1932-1941. doi:10.1016/j.hrthm.2012.08.001Baldinger, S. H., Kumar, S., Barbhaiya, C. R., Mahida, S., Epstein, L. M., Michaud, G. F., … Stevenson, W. G. (2015). Epicardial Radiofrequency Ablation Failure During Ablation Procedures for Ventricular Arrhythmias. Circulation: Arrhythmia and Electrophysiology, 8(6), 1422-1432. doi:10.1161/circep.115.003202Santangeli, P., Shaw, G. C., & Marchlinski, F. E. (2017). Radiofrequency Wire Facilitated Interventricular Septal Access for Catheter Ablation of Ventricular Tachycardia in a Patient With Aortic and Mitral Mechanical Valves. Circulation: Arrhythmia and Electrophysiology, 10(1). doi:10.1161/circep.116.004771Berjano, E. J., Hornero, F., Atienza, F., & Montero, A. (2003). Long electrodes for radio frequency ablation: comparative study of surface versus intramural application. Medical Engineering & Physics, 25(10), 869-877. doi:10.1016/s1350-4533(03)00125-5McLELLAN, A. J. A., ELLIMS, A. 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Calculation of electric fields in conductive media. Medical Physics, 9(4), 566-573. doi:10.1118/1.595107PÉREZ, J. J., D’AVILA, A., ARYANA, A., & BERJANO, E. (2015). Electrical and Thermal Effects of Esophageal Temperature Probes on Radiofrequency Catheter Ablation of Atrial Fibrillation: Results from a Computational Modeling Study. Journal of Cardiovascular Electrophysiology, 26(5), 556-564. doi:10.1111/jce.12630Jo, B., & Aksan, A. (2010). Prediction of the extent of thermal damage in the cornea during conductive keratoplasty. Journal of Thermal Biology, 35(4), 167-174. doi:10.1016/j.jtherbio.2010.02.004Gonzalez-Suarez, A., & Berjano, E. (2016). Comparative Analysis of Different Methods of Modeling the Thermal Effect of Circulating Blood Flow During RF Cardiac Ablation. IEEE Transactions on Biomedical Engineering, 63(2), 250-259. doi:10.1109/tbme.2015.2451178WINTERFIELD, J. R., JENSEN, J., GILBERT, T., MARCHLINSKI, F., NATALE, A., PACKER, D., … WILBER, D. J. (2015). Lesion Size and Safety Comparison Between the Novel Flex Tip on the FlexAbility Ablation Catheter and the Solid Tips on the ThermoCool and ThermoCool SF Ablation Catheters. Journal of Cardiovascular Electrophysiology, 27(1), 102-109. doi:10.1111/jce.12835PÉREZ, J. J., D’AVILA, A., ARYANA, A., TRUJILLO, M., & BERJANO, E. (2016). Can Fat Deposition After Myocardial Infarction Alter the Performance of RF Catheter Ablation of Scar-Related Ventricular Tachycardia?: Results from a Computer Modeling Study. Journal of Cardiovascular Electrophysiology, 27(8), 947-952. doi:10.1111/jce.13006Haines, D. E. (2011). Letter by Haines Regarding Article, «Direct Measurement of the Lethal Isotherm for Radiofrequency Ablation of Myocardial Tissue». Circulation: Arrhythmia and Electrophysiology, 4(5). doi:10.1161/circep.111.965459González-Suárez, A., Trujillo, M., Koruth, J., d’ Avila, A., & Berjano, E. (2014). Radiofrequency cardiac ablation with catheters placed on opposing sides of the ventricular wall: Computer modelling comparing bipolar and unipolar modes. International Journal of Hyperthermia, 30(6), 372-384. doi:10.3109/02656736.2014.949878Gianni, C., Mohanty, S., Trivedi, C., Di Biase, L., Al-Ahmad, A., Natale, A., & David Burkhardt, J. (2017). Alternative Approaches for Ablation of Resistant Ventricular Tachycardia. Cardiac Electrophysiology Clinics, 9(1), 93-98. doi:10.1016/j.ccep.2016.10.006Boll, D. T., Lewin, J. S., Duerk, J. L., & Merkle, E. M. (2003). Do Surgical Clips Interfere with Radiofrequency Thermal Ablation? American Journal of Roentgenology, 180(6), 1557-1560. doi:10.2214/ajr.180.6.1801557Eung Je Woo, Tungjitkusolmun, S., Hong Cao, Jang-Zem Tsai, Webster, J. G., Vorperian, V. R., & Will, J. A. (2000). A new catheter design using needle electrode for subendocardial RF ablation of ventricular muscles: finite element analysis and in vitro experiments. 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Retrograde Coronary Venous Ethanol Infusion for Ablation of Refractory Ventricular Tachycardia. Circulation: Arrhythmia and Electrophysiology, 9(7). doi:10.1161/circep.116.00435

Effectiveness and reach of a directed-population approach to improving dental health and reducing inequalities: a cross sectional study

Background Childsmile School adopts a directed-population approach to target fluoride varnish applications to 20% of the primary one (P1) population in priority schools selected on the basis of the proportion of enrolled children considered to be at increased-risk of developing dental caries. The study sought to compare the effectiveness of four different methods for identifying individuals most in need when a directed-population approach is taken. <p></p> Methods The 2008 Basic National Dental Inspection Programme (BNDIP) cross-sectional P1 Scottish epidemiological survey dataset was used to model four methods and test three definitions of increased-risk. Effectiveness was determined by the positive predictive value (PPV) and explored in relation to 1-sensitivity and 1-specificity. <p></p> Results Complete data was available on 43470 children (87% of the survey). At the Scotland level, at least half (50%) of the children targeted were at increased-risk irrespective of the method used to target or the definition of increased-risk. There was no one method across all definitions of <i>increased-risk</i> that maximised PPV. Instead, PPV was highest when the targeting method complimented the definition of <i>increased-risk</i>. There was a higher percentage of children at <i>increased-risk</i> who were not targeted (1-sensitivity) when caries experience (rather than deprivation) was used to define <i>increased-risk</i>, irrespective of the method used for targeting. Over all three definitions of <i>increased-risk</i>, there was no one method that minimised (1-sensitivity) although this was lowest when the method and definition of <i>increased-risk</i> were complimentary. The false positive rate (1-specificity) for all methods and all definitions of <i>increased-risk</i> was consistently low (<20%), again being lowest when the method and definition of <i>increased-risk</i> were complimentary. <p></p> Conclusion Developing a method to reach all (or even the vast majority) of individuals at <i>increased-risk</i> defined by either caries experience or deprivation is difficult using a directed-population approach at a group level. There is a need for a wider debate between politicians and public health experts to decide how best to reach those most at need of intervention to improve health and reduce inequalities. <p></p&gt

Mirroring everyday clinical practice in clinical trial design: a new concept to improve the external validity of randomized double-blind placebo-controlled trials in the pharmacological treatment of major depression

Background: Randomized, double-blind, placebo-controlled trials constitute the gold standard in clinical research when testing the efficacy of new psychopharmacological interventions in the treatment of major depression. However, the blinded use of placebo has been found to influence clinical trial outcomes and may bias patient selection. Discussion: To improve clinical trial design in major depression so as to reflect clinical practice more closely we propose to present patients with a balanced view of the benefits of study participation irrespective of their assignment to placebo or active treatment. In addition every participant should be given the option to finally receive the active medication. A research agenda is outlined to evaluate the impact of the proposed changes on the efficacy of the drug to be evaluated and on the demographic and clinical characteristics of the enrollment fraction with regard to its representativeness of the eligible population. Summary: We propose a list of measures to be taken to improve the external validity of double-blind, placebocontrolled trials in major depression. The recommended changes to clinical trial design may also be relevant for other psychiatric as well as medical disorders in which expectations regarding treatment outcome may affect the outcome itself

Hedging in the ethanol and sugar production: integrating financial and production decisions

Abstract Agricultural producers face financial risk at the moment of final products selling. This imposes the use of instruments to reduce risks in order to assure prices and production process economic feasibility. This paper examines the problem of creating hedging strategies with production constraints and proposes a deterministic multi-period optimization model to solve it. Uncertainty was introduced in the model through scenario trees and risk was analyzed according to the traditional mean variance approach. The model was analyzed for the sugar and ethanol market in order to aid in the financial management of a sugar cane refinery

Standardisation of rates using logistic regression: a comparison with the direct method

<p>Abstract</p> <p>Background</p> <p>Standardisation of rates in health services research is generally undertaken using the direct and indirect arithmetic methods. These methods can produce unreliable estimates when the calculations are based on small numbers. Regression based methods are available but are rarely applied in practice. This study demonstrates the advantages of using logistic regression to obtain smoothed standardised estimates of the prevalence of rare disease in the presence of covariates.</p> <p>Methods</p> <p>Step by step worked examples of the logistic and direct methods are presented utilising data from BETS, an observational study designed to estimate the prevalence of subclinical thyroid disease in the elderly. Rates calculated by the direct method were standardised by sex and age categories, whereas rates by the logistic method were standardised by sex and age as a continuous variable.</p> <p>Results</p> <p>The two methods produce estimates of similar magnitude when standardising by age and sex. The standard errors produced by the logistic method were lower than the conventional direct method.</p> <p>Conclusion</p> <p>Regression based standardisation is a practical alternative to the direct method. It produces more reliable estimates than the direct or indirect method when the calculations are based on small numbers. It has greater flexibility in factor selection and allows standardisation by both continuous and categorical variables. It therefore allows standardisation to be performed in situations where the direct method would give unreliable results.</p

Electrocardiographic changes in a rare case of flecainide poisoning: a case report

Flecainide is a class Ic anti - arrhythmic drug with sodium channel blocking activities. We report a case of a 57 year - old woman who attempted a suicide by ingesting approximately 1,8 gr of flecainide. On the surface electrocardiogram this results in a large QRS complex and in prolongation of the QTc interval. Overdose with a class Ic drug is very uncommon, its management is difficult and the mortality high

Identification of Giardia lamblia DHHC Proteins and the Role of Protein S-palmitoylation in the Encystation Process

Protein S-palmitoylation, a hydrophobic post-translational modification, is performed by protein acyltransferases that have a common DHHC Cys-rich domain (DHHC proteins), and provides a regulatory switch for protein membrane association. In this work, we analyzed the presence of DHHC proteins in the protozoa parasite Giardia lamblia and the function of the reversible S-palmitoylation of proteins during parasite differentiation into cyst. Two specific events were observed: encysting cells displayed a larger amount of palmitoylated proteins, and parasites treated with palmitoylation inhibitors produced a reduced number of mature cysts. With bioinformatics tools, we found nine DHHC proteins, potential protein acyltransferases, in the Giardia proteome. These proteins displayed a conserved structure when compared to different organisms and are distributed in different monophyletic clades. Although all Giardia DHHC proteins were found to be present in trophozoites and encysting cells, these proteins showed a different intracellular localization in trophozoites and seemed to be differently involved in the encystation process when they were overexpressed. dhhc transgenic parasites showed a different pattern of cyst wall protein expression and yielded different amounts of mature cysts when they were induced to encyst. Our findings disclosed some important issues regarding the role of DHHC proteins and palmitoylation during Giardia encystation.Fil: Merino, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Zamponi, Nahuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Vranych, Cecilia Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Touz, Maria Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Ropolo, Andrea Silvana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentin

Familial Longevity Is Marked by Lower Diurnal Salivary Cortisol Levels: The Leiden Longevity Study

BACKGROUND: Reported findings are inconsistent whether hypothalamic-pituitary-adrenal (HPA) signaling becomes hyperactive with increasing age, resulting in increasing levels of cortisol. Our previous research strongly suggests that offspring from long-lived families are biologically younger. In this study we assessed whether these offspring have a lower HPA axis activity, as measured by lower levels of cortisol and higher cortisol feedback sensitivity. METHODS: Salivary cortisol levels were measured at four time points within the first hour upon awakening and at two time points in the evening in a cohort comprising 149 offspring and 154 partners from the Leiden Longevity Study. A dexamethasone suppression test was performed as a measure of cortisol feedback sensitivity. Age, gender and body mass index, smoking and disease history (type 2 diabetes and hypertension) were considered as possible confounding factors. RESULTS: Salivary cortisol secretion was lower in offspring compared to partners in the morning (Area Under the Curve = 15.6 versus 17.1 nmol/L, respectively; p = 0.048) and in the evening (Area Under the Curve = 3.32 versus 3.82 nmol/L, respectively; p = 0.024). Salivary cortisol levels were not different after dexamethasone (0.5 mg) suppression between offspring and partners (4.82 versus 5.26 nmol/L, respectively; p = 0.28). CONCLUSION: Offspring of nonagenarian siblings are marked by a lower HPA axis activity (reflected by lower diurnal salivary cortisol levels), but not by a difference in cortisol feedback sensitivity. Further in-depth studies aimed at characterizing the HPA axis in offspring and partners are needed