Musa Kaleem Baloch

Effect of Temperature, Polymer, and Salts on the Interfacial and Micellization Behavior of 3-Dodecyl-1-Methyl-1H-Imidazol-3-Ium-Bromide: A Dispersion of a Long-Chain Ionic Liquid

Owing to low melting point (<100°C), negligible vapor pressure, and broad spectrum of applications, ionic liquids have gained much attention in the recent years. In this paper, the effects of temperature, polyvinyl pyrrolidone (PVP) polymer, and salts on the interfacial and bulk properties of aqueous solution of long-chain ionic liquid [i.e., 3-dodecyl-1-methyl-1H-imidazol-3-ium-bromide (C12mimBr)] were investigated using tensiometry. This long-chain ionic liquid was found to behave like a conventional surfactant, and thus it can be called surface active ionic liquid (SAIL). An inflection point in surface tension versus concentration curve at specific concentration was observed, which is termed the critical micelle concentration (CMC) of C12mimBr. The pre-micellar region was used to calculate various surface active parameters whereas the CMC was used to investigate its micellar behavior. Both surface activity and CMC of SAIL increased with the increase in temperature. No prominent effect was observed with the addition of PVP, whereas NaCl and CuNO3 addition distinctly changed the CMC. Similarly, surface effectiveness (πcmc) of C12mimBr decreased with the addition of NaCl and increased with the addition of CuNO3, whereas addition of PVP showed no effect on it. It shows that PVP has negligible interaction with C12mimBr. Further, the process of micellization was found to be entropy driven. GRAPHICAL ABSTRACT

Analysis of biophysical studies of metal binding to zinc α2 glycoprotein (ZAG) using fluorescence

A critical element in a phage display sorting experiment is immobilization of the target protein in functional conformation. There are inherent issues associated with the current methods of immobilization, especially for proteins of limited stability. To circumvent these problems, we have developed a mutant form (N5A) of calmodulin binding peptide (CBP) as an immobilization tag for phage display sorting. The immobilization relies on the ultra high affinity of calmodulin to the mutant CBP in the presence of calcium, which can then be reversed by adding calcium chelators like EDTA allowing “capture and release” of the specific binders in a controlled manner. To evaluate the capabilities of this system, we chose a set of challenging cases that had failed in selection using traditional immobilization. In virtually all cases, we were able to generate synthetic antibodies (sABs) for these targets using CBP fused constructs in selection campaigns. The sABs are of high affinity and have been successfully used to selectively recognize antigens in cell-based experiments. Some of these targets were problematic even without any tag, so the fact that all led to successful selection endpoints means that borderline cases can be worked on with a high probability of a positive outcome. Taken together, we feel the evidence indicates that the CBP tag embodies all the attributes of covalent immobilization tags, but do not suffer from some of their well-documented drawbacks.E kinetic analysis of drug interactions has remained the same for nearly a century and has essentially been supplanted by the use IC50 and EC50 in drug analysis. The fall from favor of enzyme kinetic analysis can be primarily attributed to the difficulties associated with kinetic modeling and the absence of relevance, inhibitory classifications, have on therapeutic development. However, the problems with enzyme kinetic analysis can also be attributed to a lack of clear distinction between binding constants and terms defining the effect produced by the compounds under investigation. The most basic inhibitory equations; competitive, noncompetitive and mixed non-competitive inhibition, define inhibitory effect using the disassociation constant (Ki) and this necessitates the use of separate equations to model the inhibitory effects attributed to each equation. By designating the Ki as simply, a binding term like the Kd in receptor interactions the effects compounds have on enzyme activity can be defined separately producing a simple empirical equation for activators and inhibitors. This treatment unifies and simplifies kinetic analysis providing an intuitive way of conceptualizing the modulation of complex catalytic regulatory processes. For example, the modulation of substrate activation and substrate inhibition associated with amyloid precursor protein processing by gamma-secretase can be simple, concise manner. While IC50 values may be sufficient for characterizing the majority of drug interactions, the complex interactions at the center of diseases that have stubbornly resisted therapeutic progress may benefit from this empirical modeling approach.A total of 407 samples from western region of Saudi Arabia were collected. These samples were collected from both soil samples and dead larvae of Spodoptera littoralis (Lepidoptera) and they were examined for the presence of Bacillus thuringiensis. The bacterium was isolated by acetate-selective enrichment medium and plating. Identification of isolates performed by microscopic examination and analysis of 16S rRNA genes by DNA sequencing for PCR products. The confirmed Bacillus thuringiensis isolates are 22 in total were recovered from 4.6% of soil samples and from 6.6% of dead larvae. Although Bacillus thuringiensis was not found to be abundant in soil habitats in Makkah Province, the results suggest that the bacterium is part of the indigenous microflora of the area we have explored. The 88 kDa parasporin protein was secreted by Bacillus thuringiensis during the stationary phase of growth. Isolated strains were screened for the presence of parasporin genes by Polymerase Chain Reaction (PCR) amplification with only four strains producing the desired bands of parasporin. The amplified fragments were cloned in pGEM-vector, sequenced and analyzed. The nucleotide sequences of parasporin were given Gene-bank accession numbers: KJ576792 and showed 99% identity with the previously isolated genes in neucleotide level while it was 98% identity in amino acid level. The full length gene was sub-cloned into pET-30a expression vector and overexpressed in E. coli under the control of the inducible T7 promoter. The heterologously produced of parasporin protein (#30% of total protein) was found in both soluble and insoluble forms. Expressed protein was been purified.