The Role of Rigid Residues in Modulating TEM-1 β-Lactamase Function and Thermostability

The relationship between protein motions (i.e., dynamics) and enzymatic function has begun to be explored in β-lactamases as a way to advance our understanding of these proteins. In a recent study, we analyzed the dynamic profiles of TEM-1 (a ubiquitous class A β-lactamase) and several ancestrally reconstructed homologues. A chief finding of this work was that rigid residues that were allosterically coupled to the active site appeared to have profound effects on enzyme function, even when separated from the active site by many angstroms. In the present work, our aim was to further explore the implications of protein dynamics on β-lactamase function by altering the dynamic profile of TEM-1 using computational protein design methods. The Rosetta software suite was used to mutate amino acids surrounding either rigid residues that are highly coupled to the active site or to flexible residues with no apparent communication with the active site. Experimental characterization of ten designed proteins indicated that alteration of residues surrounding rigid, highly coupled residues, substantially affected both enzymatic activity and stability; in contrast, native-like activities and stabilities were maintained when flexible, uncoupled residues, were targeted. Our results provide additional insight into the structure-function relationship present in the TEM family of β-lactamases. Furthermore, the integration of computational protein design methods with analyses of protein dynamics represents a general approach that could be used to extend our understanding of the relationship between dynamics and function in other enzyme classes.     

Electrokinetic and antibacterial properties of needle like‐TiO2/polyrhodanine core/shell hybrid nanostructures

The aim of this study was to fabricate needle like‐TiO₂/polyrhodanine nanostructures by polymerizing rhodanine monomer on the TiO₂ nanoparticles' surfaces and investigate their antibacterial activities. The structural, thermal, morphological, surface and electrical properties of non‐covalently functionalized nanoparticles were characterized by using FTIR, XPS, elemental analysis, TGA, XRD, SEM‐EDX, TEM, contact angle, and conductivity measurements. Characterization results confirmed the formation of needle like‐TiO₂/polyrhodanine (PRh) core/shell hybrid nanostructures. Alterations on the surface and electrokinetic properties of the materials were characterized by zeta (ζ)‐potential measurements with the presence of various salts and surfactants. The ζ‐potential of needle like‐TiO₂ was observed to increase from ?7.6 mV to +28.4 mV after forming a core/shell needle like‐TiO₂/PRh nanocomposite structure and with the presence of cetyltrimethyl ammonium bromide (CTAB) surfactant. Thereby colloidally more stable dispersions were formed. Antibacterial properties of needle like‐TiO₂/PRh were also tested against Staphylococcus aureus, Klebsiella pneumoniae, and Escherichia coli by various methods and they showed good antibacterial activity. The highest killing efficiency was determined for needle like‐TiO₂/PRh against E. coli by colony‐counting method as 0.95. TEM experiments also showed the immobilizations of the nanoparticles on E. coli and revealed the interactions between E. coli and the nanoparticles. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41554.     

Flocculation of coal suspension with mono/dual polymer systems and contribution of Ca(II)/Mg(II) ions

In the experimental study carried out on mono-flocculation, anionic and especially non-ionic polymers strongly flocculated the coal suspension than cationic polymer. The dual-flocculation of the coal suspension with cationic and anionic polymer combination was also studied and the high performances of flocculation were also reached with dual-polymer approach. Furthermore, dual-polymer system required lower concentrations for the successful flocculation of coal particles. On the other hand, the pre-destabilization of the coal suspension with calcium and magnesium ions at particular pH and concentrations highly improved the flocculation of fine coal particles with these polymers.     

Effective electrodeposition of poly(3,4-ethylenedioxythiophene)-based organic coating on metallic food packaging for active corrosion protection

Journal of Applied Electrochemistry - Modern technologies continuously need special materials with specific properties to adopt the desired application. Recently, numerous researches have been...     

Investigation of bio-ethanol steam reforming over cobalt-based catalysts

The catalytic performance of cobalt catalysts supported on γ-Al₂O₃, TiO₂, ZrO₂ were studied for bio-ethanol steam reforming (BESR) reaction. The supported catalysts (10 wt%Co) were prepared by impregnation and characterized through Thermogravimetric analysis (TGA), H₂ chemisorption, laser Raman Spectroscopy, Diffuse Reflectance Infrared Fourier Transform Spectroscopy (DRIFTS), and temperature-programmed reaction (TPRxn). The metallic cobalt sites were found to correlate with the BESR reaction activity. The reaction and H₂ chemisorption showed that ZrO₂ supported catalyst showed the best dispersion and best catalytic activity. Over the 10% Co/ZrO₂ catalyst, using a H₂O:EtOH:inert molar ratio of 10:1:75 and a GHSV = 5000 h⁻¹, 100% ethanol conversion and a yield of 5.5 mol H₂/mol EtOH were obtained at 550 °C and atmospheric pressure.     

Modulus of elasticity of substandard and normal concretes

Modulus of elasticity is an important parameter used for the structural assessment and retrofitting of concrete structures. In this study, an experimental investigation on the modulus of elasticity of concrete was carried out, the results of which is presented. 60 mixtures were prepared, in which the effects of water/cement ratio, maximum size of the aggregate, aggregate type, and fly ash content were investigated. In order to obtain very low quality concrete, the water/cement ratios were increased to as high as 2.2. Modulus of elasticity of the concretes was obtained according to ASTM C469. Compressive strength and ultrasound pulse velocities of the concretes were also recorded. The experimentally obtained modulus of elasticity values were compared with those calculated by using prediction models given in a selection of codes of practice. Dynamic moduli of elasticity of the concretes were also obtained.     

Misalignment tolerance analysis of free-space optical interconnects via statistical methods

System-level packaging is one of the critical issues that need to be addressed for free space optical interconnections (FSOI) to become useful in desktop systems. The performance of FSOI, e.g., in terms of system bit-error rate, is greatly affected by misalignments in the optical system. Therefore tolerancing, i.e., the ability to analyze and predict the effects of misalignments in the system, is of prime importance to system designers. We introduce an approach in which we study the effects of optical misalignments and other tolerance factors using statistical methods. We use Monte Carlo simulations, design of the experiments, and regression techniques to fit a polynomial equation that expresses the relationship between the system performance and the tolerance factors. This prediction model can be used for design, cost optimization, and quality control purposes. In addition, we perform a sensitivity analysis to determine those tolerance variables that have the greatest effect on system performance.     

Studies of floating dosage forms of furosemide: in vitro and in vivo evaluations of bilayer tablet formulations

For the purpose of enhancement the bioavailability of furosemide (FR), a floating dosage form with controlled release of FR was designed in this study. Because of the lower solubility of active material in the gastric medium, it was first enhanced by preparing an inclusion complex of FR with beta-cyclodextrin (beta-CD) in a 1:1 proportion using the kneading method. Following the design of dosage form, bilayer floating tablets were prepared. After dissolution rate studies were performed using the continuous flow-through cell method, the formulation that provided delivery of active material near the target profile was given to six healthy male volunteer subjects, and in vivo tests were performed. It was determined by radiographs that floating tablets prepared by adding BaSO4 stayed in the stomach for 6 hr. Further, values of the area under the plasma concentration-time curve (AUC) obtained with the floating dosage form were about 1.8 times those of the conventional FR tablet in blood analyses; maximum and minimum plasma concentrations were also found to be between the desired limits. In urine analyses, the peak diuretic effect seen in classical preparations was decreased and prolonged in floating dosage forms. Also, a considerably significant correlation was detected between in vivo results and in vitro data of the dissolution rate, and it was concluded that the modified continuous flow-through cell method is usable for in vitro dissolution rate tests of floating dosage forms.