sj-docx-1-cpc-10.1177_10556656221151096 - Supplemental material for Design and Validation of a 3D Printed Cranio-Facial Simulator: A Novel Tool for Surgical Education

Supplemental material, sj-docx-1-cpc-10.1177_10556656221151096 for Design and Validation of a 3D Printed Cranio-Facial Simulator: A Novel Tool for Surgical Education by Joshua M. Wright, Jonathan M. Ford, Fatima Qamar, Matthew Lee, Jordan N. Halsey, Matthew D. Smyth, Summer J. Decker and S. Alex Rottgers in The Cleft Palate Craniofacial Journal</p

Relative Contributions of <i>Dehalobacter</i> and Zerovalent Iron in the Degradation of Chlorinated Methanes

The role of bacteria and zerovalent iron (Fe⁰) in the degradation of chlorinated solvents in subsurface environments is of interest to researchers and remediation practitioners alike. Fe⁰ used in reactive iron barriers for groundwater remediation positively interacted with enrichment cultures containing <i>Dehalobacter</i> strains in the transformation of halogenated methanes. Chloroform transformation and dichloromethane formation was up to 8-fold faster and 14 times higher, respectively, when a <i>Dehalobacter</i>-containing enrichment culture was combined with Fe⁰ compared with Fe⁰ alone. The dichloromethane-fermenting culture transformed dichloromethane up to three times faster with Fe⁰ compared to without. Compound-specific isotope analysis was employed to compare abiotic and biotic chloroform and dichloromethane degradation. The isotope enrichment factor for the abiotic chloroform/Fe⁰ reaction was large at −29.4 ± 2.1‰, while that for chloroform respiration by <i>Dehalobacter</i> was minimal at −4.3 ± 0.45‰. The combined abiotic/biotic dechlorination was −8.3 ± 0.7‰, confirming the predominance of biotic dechlorination. The enrichment factor for dichloromethane fermentation was −15.5 ± 1.5‰; however, in the presence of Fe⁰ the factor increased to −23.5 ± 2.1‰, suggesting multiple mechanisms were contributing to dichloromethane degradation. Together the results show that chlorinated methane-metabolizing organisms introduced into reactive iron barriers can have a significant impact on trichloromethane and dichloromethane degradation and that compound-specific isotope analysis can be employed to distinguish between the biotic and abiotic reactions involved

Heterologous Production and Purification of a Functional Chloroform Reductive Dehalogenase

Reductive dehalogenases (RDases) are key enzymes involved in the respiratory process of anaerobic organohalide respiring bacteria (ORB). Heterologous expression of respiratory RDases is desirable for structural and functional studies; however, there are few reports of successful expression of these enzymes. <i>Dehalobacter</i> sp. strain UNSWDHB is an ORB, whose preferred electron acceptor is chloroform. This study describes efforts to express recombinant reductive dehalogenase (TmrA), derived from UNSW DHB, using the heterologous hosts <i>Escherichia coli</i> and <i>Bacillus megaterium</i>. Here, we report the recombinant expression of soluble and functional TmrA, using <i>B. megaterium</i> as an expression host under a xylose-inducible promoter. Successful incorporation of iron–sulfur clusters and a corrinoid cofactor was demonstrated using UV–vis spectroscopic analyses. <i>In vitro</i> dehalogenation of chloroform using purified recombinant TmrA was demonstrated. This is the first known report of heterologous expression and purification of a respiratory reductive dehalogenase from an obligate organohalide respiring bacterium

Structure-Based Design of Novel Class II c-Met Inhibitors: 1. Identification of Pyrazolone-Based Derivatives

Deregulation of c-Met receptor tyrosine kinase activity leads to tumorigenesis and metastasis in animal models. More importantly, the identification of activating mutations in c-Met, as well as <i>MET</i> gene amplification in human cancers, points to c-Met as an important target for cancer therapy. We have previously described two classes of c-Met kinase inhibitors (class I and class II) that differ in their binding modes and selectivity profiles. The class II inhibitors tend to have activities on multiple kinases. Knowledge of the binding mode of these molecules in the c-Met protein led to the design and evaluation of several new class II c-Met inhibitors that utilize various 5-membered cyclic carboxamides to conformationally restrain key pharmacophoric groups within the molecule. These investigations resulted in the identification of a potent and novel class of pyrazolone c-Met inhibitors with good in vivo activity

Structure-Based Design of Novel Class II c-Met Inhibitors: 2. SAR and Kinase Selectivity Profiles of the Pyrazolone Series

As part of our effort toward developing an effective therapeutic agent for c-Met-dependent tumors, a pyrazolone-based class II c-Met inhibitor, <i>N</i>-(4-((6,7-dimethoxyquinolin-4-yl)­oxy)-3-fluorophenyl)-1,5-dimethyl-3-oxo-2-phenyl-2,3-dihydro-1<i>H</i>-pyrazole-4-carboxamide (<b>1</b>), was identified. Knowledge of the binding mode of this molecule in both c-Met and VEGFR-2 proteins led to a novel strategy for designing more selective analogues of <b>1</b>. Along with detailed SAR information, we demonstrate that the low kinase selectivity associated with class II c-Met inhibitors can be improved significantly. This work resulted in the discovery of potent c-Met inhibitors with improved selectivity profiles over VEGFR-2 and IGF-1R that could serve as useful tools to probe the relationship between kinase selectivity and in vivo efficacy in tumor xenograft models. Compound <b>59e</b> (AMG 458) was ultimately advanced into preclinical safety studies