Coordination of metronidazole to Cu(II): Structural characterization of a mononuclear square-planar compound Joshua H.

The reaction between metronidazole [1-(2-hydroxyethyl)-2-methyl-5-nitroimidazole, MET] and CuCl2•2H2O in methanol solution has allowed isolation of blue crystals of composition Cu(MET)2Cl2•MeOH. These crystals have been shown by X-ray diffraction to consist of mononuclear square-planar trans-Cu(MET)2Cl2 molecules in which the metronidazole ligands are trans to each other, as are the Cl ligands. The structure of this compound is very different from other compounds that have been obtained from the reaction between CuCl2•2H2O and metronidazole, namely [Cu(MET)2(μ-Cl)Cl]2 and [Cu(MET)2(μ-Cl)(OH2)]2[Cl]2, which are dimers featuring bridging chloride ligands

Crystal structure of hexa-μ-chlorido-μ4-oxido-tetrakis{[1-(2-hydroxyethyl)-2- methyl-5-nitro-1H-imidazole-κN3]copper(II)} containing short NO2· · ·NO2 contacts

The title tetranuclear copper complex, [Cu4Cl6O(C6H9N3O3)4] or [Cu4Cl6O- (MET)4][MET is 1-(2-hydroxyethyl)-2-methyl-5-nitro-1Η-imidazole or metronidazole], contains a tetrahedral arrangement of copper(II) ions. Each copper atom is also linked to the other three copper atoms in the tetrahedron via bridging chloride ions. A fifth coordination position on each metal atom is occupied by a nitrogen atom of the monodentate MET ligand. The result is a distorted CuCl3NO trigonal–bipyramidal coordination polyhedron with the axial positions occupied by oxygen and nitrogen atoms. The extended structure displays O− H ⋅ ⋅ ⋅O hydrogen bonding, as well as unusual short O⋅ ⋅ ⋅ N interactions [2.775 (4) A ˚ ] between the nitro groups of adjacent clusters that are oriented perpendicular to each other. The scattering contribution of disordered water and methanol solvent molecules was removed using the SQUEEZE procedure [Spek (2015). Acta Cryst. C71, 9–16] in PLATON [Spek (2009). Acta Cryst. D65, 148–155]

Eicosapentaenoic Acid Modulates Trichomonas 1 vaginalis Activity

Trichomonas vaginalis is a sexually transmitted parasite and, while it is often asymptomatic in 50 males, the parasite is associated with disease in both sexes. Metronidazole is an effective 51 treatment for trichomoniasis, but resistant strains have evolved and, thus, it has become 52 necessary to investigate other possible therapies. In this study, we examined the effects of native 53 and oxidized forms of the sodium salts of eicosapentaenoic, docosahexaenoic and arachidonic 54 acids on T. vaginalis activity. Eicosapentaenoic acid was the most toxic with 190 μM and 380 55 μM causing approximately 90% cell death in Casu2 and ATCC 50142 strains, respectively. In 56 contrast, oxidized eicosapentaenoic acid was the least toxic, requiring \u3e3 mM to inhibit activity, 57 while low levels (10μM) were associated with increased parasite density. Mass spectrometric 58 analysis of oxidized eicosapentaenoic acid revealed C20 products containing one to six 59 additional oxygen atoms and various degrees of bond saturation. These results indicate that 60 eicosapentaenoic acid has different effects on T. vaginalis survival, depending on whether it is 61 present in the native or oxidized form. A better understanding of lipid metabolism in T. vaginalis 62 may facilitate the design of synthetic fatty acids that are effective for the treatment of 63 metronidazole-resistant T. vaginalis

Bioremediation of petroleum-contaminated soils: mathematical modelling as a tool for the simulation of alternative strategies

POCI-01-0145-FEDER-016575; ERC Grant n.º 323009; UID/BIO/04469/2013; POCI -01-0145-FEDER-006684; NORTE-01-0145-FEDER-000004; FCOMP-01-0124- FEDER-027462; SFRH/BPD/80528/2011info:eu-repo/semantics/publishedVersio

Outcomes of Patients on Extracorporeal Membrane Oxygenation (ECMO) for Periods of Time without Anticoagulation at LVHN in the Past 3 Years

Outcomes of Patients on Extracorporeal Membrane Oxygenation (ECMO) for Periods of Time Without Anticoagulation at LVHN in the Past 3 Years Bryan Auvil, Rosalie Mattiola, Rita Pechulis, MD, James Wu, MD Department of Surgery, Division of Cardiothoracic Surgery Lehigh Valley Health Network, Allentown, PA Abstract Extracorporeal membrane oxygenation (ECMO) is an aggressive life support technique that utilizes extracorporeal circuits and oxygenators to support severe lung injury such as acute respiratory distress syndrome (ARDS), as well as severe cardiac disorders like cardiogenic shock and cardiac arrest. Veno-Venous (VV) ECMO is primarily used in respiratory distress. Clots can form within the oxygenator or circuit, which can develop embolic complications or flow problems. To prevent this, patients are anticoagulated, usually with heparin. However patients can develop bleeding complications, and therefore must have their AC discontinued. A retrospective chart review was done on 57 VV ECMO patients, out of which 19 had their heparin held for at least one period of 24 hours or longer. Temporarily non-heparinized patients required far more oxygenator and circuit replacements than control (58% compared to 8% of patients), had more DVTs (37% compared to 21%), and had a higher mortality rate (47% compared to 29%), however the rates of DVT per day on ECMO were very similar. Patients in the non-heparinized cohort died when the families withdrew care because of prolonged ECMO without improvement, worsening sepsis, and/or severe bleeding complications. Based on these findings there is no direct evidence that temporarily discontinuing AC resulted in increased patient mortality. This was more likely due to increased severity of illness and bleeding complications, evidenced by the increased time on ECMO. Keywords Extracorporeal membrane oxygenation, ECMO, Veno-venous ECMO, VV ECMO, heparin, anticoagulation, DVT, thrombosis, thrombotic complication Introduction Extracorporeal membrane oxygenation (ECMO), also known as extracorporeal life support (ECLS) is a very aggressive technique that utilizes extracorporeal circuits and oxygenators to support severe lung injury such as acute respiratory distress syndrome (ARDS), as well as cardiac support for cardiogenic shock and cardiac arrest. There are two major categories of ECMO: Veno-Venous ECMO is primarily used in respiratory distress, using a right internal jugular vein access; and Veno-Arterial ECMO is primarily used for cardiac or cardiopulmonary support, using femoral vein and artery access1. Extra Corporeal Life Support (ECLS) is sometimes used synonymously. Blood that comes into contact with foreign surfaces tends to coagulate. Clots that form within the oxygenator and extracorporeal tubing can be pushed into the patient and ultimately cause a thrombus, or in severe cases, a stroke. Therefore, as part of the protocol for extracorporeal circuits, patients undergo anticoagulation (AC) therapy and are uniformly heparinized with a partial thromboplastin time (PTT) in the range of 50-70 seconds, and an activated clotting time (ACT) of 180-200 sec2. Some of these severely ill patients can develop bleeding complications involving the oral pharyngeal cavity, abdominal cavity, thoracic cavity, and cannulation sites. In these cases, because of ongoing bleeding and exploratory surgery, AC needs to be stopped, sometimes for prolonged periods of time. Other patients are coagulopathic due to their illness, and therefore don’t receive therapeutic AC. Major bleeding is defined as clinically overt bleeding associated with a hemoglobin (Hgb) fall of at least 2 g/dl in a 24 hour period, greater than 20 ml/kg over a 24 hour period, or a transfusion requirement of one or more 10 ml/kg packed red blood cell (PRBC) transfusions over that same time period. In addition, bleeding that is retroperitoneal, pulmonary or involves the central nervous system, or bleeding that requires surgical intervention would also be considered major bleeding. Minor bleeding would be considered less than 20 ml/kg/day and require transfusion of one 10 ml/kg PRBC transfusion, or less. This is significant, because hemorrhagic complications and the requirement for greater red blood cell transfusion volumes are associated with increased mortality in both cardiac and non-cardiac ECLS2. The ultimate goal of this project was to review the safety and outcome of patients on ECMO in whom therapeutic anticoagulation is held secondary to bleeding complications, as there is no significant body of literature regarding this topic. Methods This retrospective study involved 57 VV ECMO patients (47% male, average age: 48) selected from the hospital database. Patients were sorted based off of the following inclusion criteria: (1) Patient had to have been on VV ECMO at LVHN since 2013, and (2) Patient had to be at least 18 years old. Of the 57 patients, 19 had their heparin held for at least one period of 24 hours or longer. All patients’ electronic medical records (EHMR) were examined to determine incidence of upper extremity (UE) and lower extremity (LE) deep vein thrombosis (DVT), patient mortality, and possible link between thrombotic complications and patient outcomes. Thrombotic complications include stroke, ischemic limb, kidney and liver injury, and surgical procedures for bleeding from body cavities and insertion sites. Records were also examined to determine if/how many oxygenator or circuit changes were necessary. For patients whose heparin was held, the source of bleeding was determined. Incidence of DVT, as well as need for oxygenator/circuit changes were calculated both as a percentage of the population, and as a rate of total events per total days on ECMO for that group. This was done in an attempt to normalize the data to account for the large difference between groups in average time spent on ECMO. Results The 19 patients whose heparin was held for a period of at least 24 hours (non-heparinized patients) spent an average of 25 days on ECMO, with an average AC hold time of 175 hours, while the 38 control patients spent an average of 10 days on ECMO (Table 1). The non-heparinized group was 47% male, with an average age of 46, and the control group was 42% male with an average age of 49 (Table 1). 58% of non-heparinized patients required oxygenator or circuit changes, at a rate of 0.034 changes per total day on ECMO (16 changes/475 days), while only 7.8% of control patients required changes, at a rate of 0.0079 (3 changes/379 days) (Table 1). 26% of non-heparinized patients and 13% of control patients experienced a LE DVT, but the rates were more similar at 0.017 (8 DVTs/475 days) and 0.013 (5 DVTs/379 days), respectively (Table 1). This trend continued with UE DVTs; although 26% of non-heparinized patients experienced an UE DVT compared to 11% of control patients, non-heparinized patients’ rate of 0.017 (8 DVTs/475 days) was actually lower than control patients’ rate of 0.026 (10 DVTs/379 days) (Table 1). The mortality rate for non-heparinized patients was 47% (9/19), with 6 out of 9 patients’ families deciding to withdraw care. The mortality rate for control patients was 29% (11/38), with 7 out of 11 patients’ families deciding to withdraw care (Table 1). 11% (2/19) of non-heparinized patients were discharged home and 42% (8/19) were discharged to rehab facilities, while 18% (7/38) of control patients were discharged home and 55% (21/38) were discharged to rehab facilities (Table 1). Reasons for discontinuing heparin or other AC included hemoptysis (7/19) and oropharyngeal (5/19), tracheotomy site (6/19), and cannula site (6/19) bleeding. VV ECMO patient etiology included H1N1 (28%), pneumonia (16%), aspiration (14%), Legionella (9%), post-operative complications (7%), MI and cardiac arrest (5%), and other illnesses (9%). Etiology was unknown for 12% of patients (Figure 3). Non-heparinized patients died due to severe bleeding, sepsis, hypoxia-related brain death, and family withdrawing care due to prolonged ECMO treatment without improvement (Table 3). Conclusions Patients whose heparin was held for period(s) of at least 24 hours required oxygenator and circuit changes much more frequently. They also experienced more DVTs and had a higher mortality rate; however, their overall rate of DVTs per day on ECMO is similar that of the control group, indicating that the increased incidence of DVT is likely due to the significantly longer average amount of time those patients spent on ECMO relative to control patients. Patients in the non-heparinized cohort mostly died when the families withdrew care because of prolonged ECMO without improvement, worsening sepsis, and/or severe bleeding complications. There were no obvious systemic thrombotic complications. Based on these findings there is no direct evidence that temporarily discontinuing AC resulted in increased patient mortality. This was more likely due to increased severity of illness and bleeding complications, requiring more time on ECMO. The study has quite a few limitations. It cannot necessarily be broadly applied due to the small sample size of only 57 total patients over three years, with an experimental group of only 19 patients. Furthermore, only statistics from a single hospital system were examined. However, the findings indicate that at least in this patient cohort, there was no obvious link between withholding anticoagulation therapy while on ECMO and increased patient mortality. This suggests that in contrast to standard ECMO protocol, it might be reasonable to withhold heparin from ECMO patients - at least temporarily - in the case of moderate to severe bleeding2. Further study is indicated to determine whether the results of this study are generalizable to a wider patient population, and whether discontinuing AC therapy in ECMO patients does in fact affect patient mortality. Acknowledgements James Wu, MD, Rita Pechulis, MD, Hope Kincaid, MPH, CPH, Jane Scott, Hubert Huang, Ph. D., Jordan Williams, Amanda Broderick, Alex Maryshina, Joseph Napolitano, Ph.D. References D Brodie, M Bacchetta. Extracorporeal Membrane Oxygenation for ARDS in Adults. N Engl J Med 2011;365:1905-14. ELSO Anticoagulation Guideline 2014 Table 1: LVHN VV ECMO data 2013-2015 Table 2: Reasons for discontinuing heparin Figure 1: Etiology of LVHN VV ECMO patients 2013-2015 Table 3: Causes of death for heparin-held patients Figure 2: Patient complications and outcomes as percentage of total group Figure 3: Rate of patient complications (total # of events per total days on EC

MethylC-analyzer: A comprehensive downstream pipeline for the analysis of genome-wide DNA methylation

DNA methylation is a crucial epigenetic modification involved in multiple biological processes and diseases. Current approaches for measuring genome-wide DNA methylation via bisulfite sequencing (BS-seq) include whole-genome bisulfite sequencing (WGBS), reduced representation bisulfite sequencing (RRBS), and enzymatic methyl-seq (EM-seq). The computational analysis tools available for BS-seq data include customized aligners for mapping bisulfite-converted reads and computational pipelines for downstream data analysis. Current post-alignment methylation tools are specialized for the interpretation of CG methylation, which is known to dominate mammalian genomes, however, non-CG methylation (CHG and CHH, where H refers to A, C, or T) is commonly observed in plants and fungi and is closely associated with gene regulation, transposon silencing, and plant development. Thus, we have developed a MethylC-analyzer to analyze and visualize post-alignment WGBS, RRBS, and EM-seq data focusing on CG. The tool is able to also analyze non-CG sites to enhance deciphering genomes of plants and fungi. By processing aligned data and gene location files, MethylC-analyzer generates a genome-wide view of methylation levels and methylation in user-specified genomic regions. The meta-plot, for example, allows the investigation of DNA methylation within specific genomic elements. Moreover, our tool identifies differentially methylated regions (DMRs) and investigates the enrichment of genomic features associated with variable methylation. MethylC-analyzer functionality is not limited to specific genomes, and we demonstrated its performance on both plant and human BS-seq data. MethylC-analyzer is a Python- and R-based program designed to perform comprehensive downstream analyses of methylation data, providing an intuitive analysis platform for scientists unfamiliar with DNA methylation analysis. It is available as either a standalone version for command-line uses or a graphical user interface (GUI) and is publicly accessible at https://github.com/RitataLU/MethylC-analyzer