Comparative Functional Dynamics Studies on the Enzyme Nano-bio Interface

Citation: Thomas, S. E., Comer, J., Kim, M. J., Marroquin, S., Murthy, V., Ramani, M., … DeLong, R. K. (2018, August 8). Comparative functional dynamics studies on the enzyme nano-bio interface. https://doi.org/10.2147/IJN.S152222Comparative functional dynamics studies on the enzyme nano-bio interface Spencer E Thomas,1,2,* Jeffrey Comer,1,* Min Jung Kim,1 Shanna Marroquin,1 Vaibhav Murthy,1 Meghana Ramani,1 Tabetha Gaile Hopke,2 Jayden McCall,1 Seong-O Choi,1 Robert K DeLong1 1Nanotechnology Innovation Center of Kansas State, Department of Anatomy and Physiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS, USA; 2Department of Biomedical Science, Missouri State University, Springfield, MO, USA *These authors contributed equally to this work Introduction: Biomedical applications of nanoparticles (NPs) as enzyme inhibitors have recently come to light. Oxides of metals native to the physiological environment (eg, Fe, Zn, Mg, etc.) are of particular interest—especially the functional consequences of their enzyme interaction. Materials and methods: Here, Fe2O3, zinc oxide (ZnO), magnesium oxide (MgO) and nickel oxide (NiO) NPs are compared to copper (Cu) and boron carbide (B4C) NPs. The functional impact of NP interaction to the model enzyme luciferase is determined by 2-dimensional fluorescence difference spectroscopy (2-D FDS) and 2-dimensional photoluminescence difference spectroscopy (2-D PLDS). By 2-D FDS analysis, the change in maximal intensity and in 2-D FDS area under the curve (AUC) is in the order Cu~B4C>ZnO>NiO>>Fe2O3>MgO. The induced changes in protein conformation are confirmed by tryptic digests and gel electrophoresis. Results: Analysis of possible trypsin cleavage sites suggest that cleavage mostly occurs in the range of residues 112–155 and 372–439, giving a major 45 kDa band. By 2-D PLDS, it is found that B4C NPs completely ablate bioluminescence, while Cu and Fe2O3 NPs yield a unique bimodal negative decay rate, -7.67×103 and -3.50×101 relative light units respectively. Cu NPs, in particular, give a remarkable 271% change in enzyme activity. Molecular dynamics simulations in water predicted that the surfaces of metal oxide NPs become capped with metal hydroxide groups under physiological conditions, while the surface of B4C becomes populated with boronic acid or borinic acid groups. These predictions are supported by the experimentally determined zeta potential. Thin layer chromatography patterns further support this conception of the NP surfaces, where stabilizing interactions were in the order ionic>polar>non-polar for the series tested. Conclusion: Overall the results suggest that B4C and Cu NP functional dynamics on enzyme biochemistry are unique and should be examined further for potential ramifications on other model, physiological or disease-relevant enzymes. Keywords: 2-dimensional fluorescence difference, 2-D FDS, AUC, corrected light intensity emitted, emission wavelength, excitation wavelength, wavelength of max intensity, RL

Comparison of abdominal computed tomography to ultrasound in the diagnosis of canine biliary disease manifesting as acute abdominal signs

Biliary diseases are uncommon, potentially fatal causes of acute abdomen in dogs. Little information is present comparing the performance of computed tomography (CT) to ultrasound in identifying canine biliary pathology. Thirty-five client-owned dogs presenting for acute abdomen signs received an abdominal ultrasound and contrast-enhanced abdominal CT. Two authors reviewed the randomized, anonymized CT and ultrasound studies. Twenty-eight dogs had biliary pathology and seven dogs serving as controls had no evidence of biliary disease. The final diagnoses of patients with biliary pathology included cholelithiasis, gallbladder mucoceles, cholangiohepatitis/cholangitis, extrahepatic biliary obstruction, gallbladder wall edema, gallbladder wall mass, and cystic mucosal hyperplasia. Computed tomography was more accurate in identifying cholelithiasis than ultrasound. No statistical difference in the odds to identify other biliary pathology was identified between ultrasound and CT. Findings from this study suggest CT may be used in place of ultrasound in canine patients presenting for acute abdominal signs of biliary origin