A whole-cell assay for the high throughput screening of calmodulin antagonists

Cell-based screening systems for pharmaceuticals are desired over molecular biosensing systems because of the information they provide on toxicity and bioavailability. However, the majority of sensing systems developed are molecular biosensing type screening systems and cannot be easily adapted to cell-based screening. In this study, we demonstrate that protein-based molecular sensing systems that employ a fluorescent protein as a signal transducer are amenable to cell-based sensing by expressing the protein molecular sensing system in the cell and employing these cells for screening of desired molecules. To achieve this, we expressed a molecular sensing system based on the fusion protein of calmodulin (CaM) and enhanced green fluorescent protein (EGFP) in bacterial cells, and utilized these cells for the screening of CaM antagonists. In the presence of Ca²⁺, CaM undergoes a conformational change exposing a hydrophobic pocket that interacts with CaM-binding proteins, peptides, and drugs. This conformational change induced in CaM leads to a change in the microenvironment of EGFP, resulting in a change in its fluorescence intensity. The observed change in fluorescence intensity of EGFP can be correlated to the concentration of the analyte present in the sample. Dose-response curves for various tricyclic antidepressants were generated using cells containing CaM-EGFP fusion protein. Additionally, we demonstrate the versatility of our system for studying protein-protein interactions by using cells to study the binding of a peptide to CaM. The study showed that the CaM-EGFP fusion protein within the intact cells responds similarly to that of the isolated fusion protein, hence eliminating the need for any isolation and purification steps. We have demonstrated that this system can be used for the rapid screening of various CaM antagonists that are potential antipsychotic drugs.     

On minimum metric dimension of honeycomb networks

A minimum metric basis is a minimum set W of vertices of a graph G(V,E) such that for every pair of vertices u and v of G, there exists a vertex wW with the condition that the length of a shortest path from u to w is different from the length of a shortest path from v to w. The honeycomb and hexagonal networks are popular mesh-derived parallel architectures. Using the duality of these networks we determine minimum metric bases for hexagonal and honeycomb networks.     

Production of Oils from Acetic Acid by the Oleaginous Yeast Cryptococcus curvatus

The feasibility of the conversion of acetic acid, a metabolite commonly obtained during anaerobic fermentation processes, into oils using the yeast Cryptococcus curvatus was reported. This microorganism exhibited very slow growth rates on acetate as carbon source, which led to design a two-stage cultivation process. The first consisted of cell growth on glucose as carbon source until its complete exhaustion. The second step involved the use of acetate as carbon source under nitrogen limitation in order to induce lipid accumulation. A typical experiment performed in a bioreactor involved a preliminary yeast growth with a glucose initial concentration of 15?g/L glucose. Further additions of acetate and nitrogen source allowed a final lipid accumulation up to 50% (w/w). These promising results demonstrated the suitability of the technique proposed.     

Synthetic study on the relationship between structure and sweet taste properties of steviol glycosides

The structure activity relationship between the C??-C?? methylene double bond on the aglycone of steviol glycosides and the corresponding impact on their sweet taste has been reported here for the first time. It has been observed that converting stevioside and rebaudioside A to their corresponding ketones by switching the doubly bonded methylene on C-17 for a ketone group actually removes the sweet taste properties of these molecules completely. Regenerating the original molecules tends to restore the sweet taste of both the steviol glycosides. Thus this C??-C?? methylene double bond in rebaudioside A and stevioside can be regarded as a pharmacophore essential for the sweetness property of these molecules.     

Effect of iron oxide addition on structural properties of calcium silico phosphate glass/glass-ceramics

Glasses with nominal composition 34SiO₂–(45 ? x) CaO–16 P₂O₅–4.5 MgO–0.5 CaF₂–x Fe₂O₃ (where x = 5, 10, 15, 20 wt.%) have been synthesized by melt quench technique. These have been investigated for structural features by using Fourier transform infra-red (FTIR) spectroscopy and X-ray photoelectron spectroscopy (XPS). Results have shown an increase in fraction of non‐bridging oxygen in glasses with an increase in iron oxide content up to 15 wt.% and subsequently decreases with further increase in iron oxide content to 20 wt.%. These effects are originated by the incorporation of Fe₂O₃ into the silica network. Iron oxide behaves as a network modifier at low concentration and stabilizes the glass network at higher content. The glass-ceramics exhibit an increase in the formation of magnetite phase with an increase in iron oxide. The glass phase in the glass-ceramics matrix, controls the surface dissolution, which in turn decides the response of the material in-vitro. The glass-ceramics with 15 wt.% iron oxide has shown optimum response in simulated body fluid.     

Atomistic properties of γ uranium

The properties of the body-centered cubic γ phase of uranium (U) are calculated using atomistic simulations. First, a modified embedded-atom method interatomic potential is developed for the high temperature body-centered cubic (γ) phase of U. This phase is stable only at high temperatures and is thus relatively inaccessible to first principles calculations and room temperature experiments. Using this potential, equilibrium volume and elastic constants are calculated at 0 K and found to be in close agreement with previous first principles calculations. Further, the melting point, heat capacity, enthalpy of fusion, thermal expansion and volume change upon melting are calculated and found to be in reasonable agreement with experiment. The low temperature mechanical instability of γ U is correctly predicted and investigated as a function of pressure. The mechanical instability is suppressed at pressures greater than 17.2 GPa. The vacancy formation energy is analyzed as a function of pressure and shows a linear trend, allowing for the calculation of the extrapolated zero pressure vacancy formation energy. Finally, the self-defect formation energy is analyzed as a function of temperature. This is the first atomistic calculation of γ U properties above 0 K with interatomic potentials.     

4D Biofabrication of Mechanically Stable Tubular Constructs Using Shape Morphing Porous Bilayers for Vascularization Application

This study reports the fabrication of highly porous electrospun self-folding bilayers, which fold into tubular structures with excellent mechanical stability, allowing them to be easily manipulated and handled. Two kinds of bilayers based on biocompatible and biodegradable soft (PCL, polycaprolactone) and hard (PHB, poly-hydroxybutyrate) thermoplastic polymers have been fabricated and compared. Multi-scroll structures with tunable diameter are obtained after the shape transformation of the bilayer in aqueous media, where PCL-based bilayer rolled longitudinally and PHB-based one rolled transversely with respect to the fiber direction. A combination of higher elastic modulus and transverse orientation of fibers with respect to rolling direction allowed precise temporal control of shape transformation of PHB-bilayer – stress produced by swollen methacrylated hyaluronic acid (HA-MA) do not relax with time and folding is not affected by the fact that bilayer is fixed in unfolded state in cell culture medium for more than 1 h. This property of PHB-bilayer allowed cell culturing without a negative effect on its shape transformation ability. Moreover, PHB-based tubular structure demonstrated superior mechanical stability compared to PCL-based ones and do not collapse during manipulations that happened to PCL-based one. Additionally, PHB/HA-MA bilayers showed superior biocompatibility, degradability, and long-term stability compared to PCL/HA-MA.     

Catalytic reduction of 4-nitrophenol using synthesized and characterized CoS@MorphcdtH/CoS@4-MPipzcdtH nanoparticles

CoS@MorphcdtH NPs and CoS@4-MPipzcdtH NPs were synthesized by precipitation method involving three mechanisms: inclusion, occlusion, and adsorption. The synthesized NPs were characterized with the help of UV-Vis spectroscopy, FESEM-EDAX, powder x-ray diffraction, TEM, ESIMS, TG/DSC analysis. The morphology of the CoS@MorphcdtH NPs and CoS@4-MPipzcdtH NPs were hexagonal and rectangular, and the particles were in the range 7–12 nm. UV–visible spectral measurements showed surface plasmon resonance at 320 nm–340 nm with band gap of 3.65 eV–3.86 eV. The catalytically active CoSNPs called were investigated for the reduction of 4-nitrophenol (4-NP) via hydrogenation using sodium borohydride (NaBH₄) as a reducing agent. Both the CoS NPs successfully reduced 4-NP to 4- aminophenol (4-AP) in a short time, catalytic performances are almost unchanged for the first five cycles. Herein, we report the preparation and characterizations of efficient active CoS NPs consisting carbodithioic acid framework as a support/capping material, along with catalytic property.     

The Playground Shade Index: A New Design Metric for Measuring Shade and Seasonal Ultraviolet Protection Characteristics of Parks and Playgrounds

Current shading strategies used to protect outdoor playgrounds from harmful solar radiation include the placement of artificial cloth weaves or permanent roofing over a playground site, planting trees in proximity to playground equipment, and using vegetation or surface texture variations to cool playground surfaces. How and where an artificial shade structure is placed or a tree is planted to maximize the shade protection over specific playground areas, requires careful assessment of local seasonal sun exposure patterns. The Playground Shade Index (PSI) is introduced here as a design metric to enable shade and solar ultraviolet exposure patterns to be derived in an outdoor space using conventional aerial views of suburban park maps. The implementation of the PSI is demonstrated by incorporating a machine learning design tool to classify the position of trees from an aerial image, thus enabling the mapping of seasonal shade and ultraviolet exposure patterns within an existing 7180 m² parkland. This is achieved by modeling the relative position of the sun with respect to nearby buildings, shade structures, and the identified evergreen and deciduous tree species surrounding an outdoor playground.