Antimicrobial and Heavy Metal Sequestration Capacities of Graphene Polymer Nanofilms
Membrane bioreactors (MBR) are important components in the production of effluent in wastewater treatment systems. However, MBR are susceptible to biofouling, a process by which bacteria colonize the surface of the membrane in contact with water. Graphene could be a solution to biofilm formation. In this study, the graphene polymer nanocomposite’s antimicrobial and heavy metal removal properties and the mechanisms behind the properties were investigated. Five different films of nanocomposites with a form of graphene and a polymer were synthesized: Graphene, Graphene Oxide, PVK-GO, PVK-G, PVK. A Büchner funnel and a vacuum pump were used to coat membrane filters with solutions of each nanomaterial. Using the Büchner funnel, E. coli and B. subtilis bacteria were filtered through the filter and both the filtrate and the filter were examined for bacterial content. Similarly, a Pb2+ solution was filtered through the coated filters and percentage removal of the ion was calculated using Atomic Absorbtion Spectrometry. Further analysis from SEM data, ATR-IR, and an Oxidative Stress test revealed that the PVK-GO nanocomposite inactivates bacteria by causing oxidative stress and the carboxyl group binds to lead ions. PVK-GO was most effective at removing the highest percentage of heavy metal and inactivated the most bacteria and displayed the most antimicrobial properties. PVK-GO coatings provide an efficient and economical alternative to the current wastewater industry standard and can save millions of dollars and reduce environmental waste. Also, the coatings have applications in indwelling medical devices and can reduce the risks associated with biofilm formational and bacterial infections.
An investigation of the inhibitory potential of Dronedarone on CYP2J2 mediated astemizole metabolism
Dronedarone is an anti-arrhythmic drug approved in 2009 for paroxysmal and persistent atrial fibrillation. It is less toxic than its predecessor Amiodarone as it does not cause systemic toxicity but has the same pharmacological activity. However the administration of dronedarone to permanent AF and heart failure patients leads to increased risk of stroke and cardiac death. The exact mechanism of the toxicity is currently unknown. Extrahepatic Cytochrome P450 enzymes play a dominant role in organ-specific drug metabolism and toxicity. Cytochrome P450 2J2 (CYP2J2) enzyme, a predominant enzyme found in human cardiac myocytes, metabolizes endogenous arachidonic acid (AA) into epoxyeicosatrienoic acids (EETs) which play an important role in maintaining normal cardiac physiology. Inhibition of CYP2J2 and perturbation of AA metabolic pathway could result in exacerbation of cardiac failure. This research aims to find out whether dronedarone inhibits CYP2J2 in a suitable cell model (H9C2) using astemizole as a probe substrate. Our in-house studies using recombinant CYP2J2 enzyme have shown that dronedarone potently inhibits CYP2J2. Rat myoblast cells (H9C2) will be seeded in 12-well plate and differentiated for 4 days. The cells will be then treated with different concentrations of astemizole and incubated for 24 h. The cells will then be harvested, lysed, and the cell lysate will be analyzed using liquid chromatography-mass spectrometry (LCMS). Using multi-reaction monitoring (MRM) on the LCMS, astemizole concentration as well as its CYP2J2-specific metabolite O-desmethylastemizole concentrations will be measured. The presence of O-desmethylastemizole confirms the metabolism of astemizole by CYP2J2 in H9C2 cells. By plotting a Michaelis-Menten kinetics curve, we will be able to determine the Michaelis constant (KM) and maximum rate of reaction (Vmax). H9C2 cells will be then treated with fixed concentration of astemizole while varying the dronedarone concentration. A decrease in metabolite O-desmethylastemizole conce ntration, indicates inhibition of CYP2J2 metabolism by dronedarone. Using this data, Lineweaver-Burke graph will be plotted, to determine the mode and potency of the inhibition. Our preliminary studies showed that the KM value was 2.7μM. This study will be useful in understanding if dronedarone inhibits CYP2J2 which may lead to clinically significant drug-drug interactions, one of the dangers of polypharmacy. Finally this study will shed a new light on the mechanisms for dronedarone mediated cardiac failure exacerbation.
Findings of new oscillations in BR reaction
The Briggs Rauscher reaction, i. e., BR reaction, which is one of the oscillation reactions, produces iodide ion and iodine repeatedly. Continual color changes of the solution from colorless to deep blue, and vice versa, are observed during the reaction due to the so-called “iodine test” reaction. In this work, we studied the effects of the presence of the redox active indicators on the oscillation behavior of the BR reaction. To the reaction mixture of KIO3, H2SO4, H2O2, C3H4O4, MnSO4, and starch, which are used for the general BR reaction as added a redox active reagent (indicator). Then, the changes in color and voltage of the reaction solution were recorded by a photosensor of the LEGO MINDSTORMS and a voltmeter using Pt electrodes. Under general reaction conditions, the oscillation reaction continued for ca. 5 minutes, including 18 times of oscillations. When an indicator, such as BTB, was added instead of starch to the reaction solution, splits of the voltage wave were observed, which should be a kind of new oscillation. Moreover, we found that the addition of K3[Fe(CN)6], which exhibits high redox activity, in the reaction solution instead of starch made the life-time and the numbers of the oscillation in the reaction greater by 3 times (14 min.) in time and more than 4 times (81 times) in the frequency. It’s also a kind of new oscillation. These results suggested that the oxidation-reduction reactions by the addition of ferricyanate ion effectively promotes the redox process of iodine and iodide ion. The experiments we wrote above were conducted without starch. Thus, as a reference, we conducted the same experiments under the presence of starch and got interesting results. We also studied the effects of K4[Fe(CN)6], suggeting that not only redox reaction between ferricyanide and ferrocyanide ion, but also the redox reaction with BR solution should occur in these reactions.