Iron(III) and ruthenium(II) porphyrin complex-catalyzed selective olefination of aldehydes with ethyl diazoacetate

The commercially available Fe(III) and Ru(II) porphyrin complexes Fe(TPP)Cl and Ru(TPP)(CO) are efficient catalysts for selective olefination of a variety of aldehydes with ethyl diazoacetate in the presence of triphenylphosphine. The reactions were carried out under mild conditions in a one-pot fashion with the use of a stoichiometrical amount of EDA, which proceeded with excellent yields and high (E)-selectivity. Air atmosphere, low catalyst loadings, and functional group tolerance were also demonstrated.     

Imaging the stomatal physiology of somatic embryo-derived peanut leaves by scanning electrochemical microscopy

The stomatal physiology, chlorophyll distribution and photosynthetic activity of somatic embryo (SE)- and seedling-derived peanut plants grown in vitro (test tube-grown) and extra vitrum (soil-grown) are investigated using scanning electrochemical microscopy (SECM). This SECM imaging is performed in two different feedback modes, corresponding to oxygen evolution and chlorophyll distribution. More specifically, the oxygen evolution profiles of the in vitro leaves indicate important differences in leaf anatomy between the SE- and seedling-derived leaves. On the other hand, the chlorophyll distribution images show individual stomata of size ca. 27 ± 5μm. Further studies on senescing (aged) leaves reveal interesting voltammograms that vary widely over the stomatal complexes and the surrounding tissues, probably due to the release of electroactive metabolites during chlorophyll breakdown when the leaves turn yellow. Thus, the present investigation could open up new opportunities for characterizing botanical systems using electroanalytical techniques. In addition, it could provide further insights into various areas of current relevance, including signal transduction, cell fate/differentiation and developmental biology. Schematic representation of SECM imaging used in this investigation. The SECM probe is a Pt UME disk (25 μm diameter) embedded in an insulating glass sheath so that the ratio of the diameter of the death to that of the electrode surface (RG) is 7. RE denotes the reference electrode Ag/AgCl, sat. KCl and CE refers to the counter electrode, a Pt wire. Oxygen evolving from the leaf surface during photosynthesis diffuses into the electrolyte (0.1 M KCl) and gets reduced at the Pt UME, biased to a potential of −0.5 V, at a diffusion-limited rate to produce a change in the tip-current     

Dynamic Expectation Maximization Algorithm for Estimation of Linear Systems with Colored Noise

The free energy principle from neuroscience has recently gained traction as one of the most prominent brain theories that can emulate the brain’s perception and action in a bio-inspired manner. This renders the theory with the potential to hold the key for general artificial intelligence. Leveraging this potential, this paper aims to bridge the gap between neuroscience and robotics by reformulating an FEP-based inference scheme—Dynamic Expectation Maximization—into an algorithm that can perform simultaneous state, input, parameter, and noise hyperparameter estimation of any stable linear state space system subjected to colored noises. The resulting estimator was proved to be of the form of an augmented coupled linear estimator. Using this mathematical formulation, we proved that the estimation steps have theoretical guarantees of convergence. The algorithm was rigorously tested in simulation on a wide variety of linear systems with colored noises. The paper concludes by demonstrating the superior performance of DEM for parameter estimation under colored noise in simulation, when compared to the state-of-the-art estimators like Sub Space method, Prediction Error Minimization (PEM), and Expectation Maximization (EM) algorithm. These results contribute to the applicability of DEM as a robust learning algorithm for safe robotic applications.     

Purification of chloroperoxidase from Musa paradisiaca stem juice

Chloroperoxidase from Musa paradisiaca stem juice has been purified to homogeneity using a concentration obtained by ultrafiltration and anion exchange chromatography on diethylaminoethyl (DEAE) cellulose. The purified enzyme gave a single protein band in SDS‐PAGE analysis corresponding to molecular mass of 43 kDa. The native PAGE analysis result has also given a single protein band, confirming the purity of the enzyme. The purified enzyme was chlorinated and brominated with monochlorodimedone, the substrate used for measuring the halogenating activity of chloroperoxidases. The K_m and k_cat values using monochlorodimedone as the substrate were 20 μM and 1.64 s^?1, respectively, giving a k_cat/K_m value of 8.2 × 10⁴ M^?1 s^?1. The pH and temperature optima of the chlorinating activity were 3.0 and 25°C, respectively. The K_m values for the peroxidase activity using pyragallol and H₂O₂ as the variable substrates were 89 and 120 μM, respectively. The pH and temperature optima of the peroxidase activity using pyrogalllol as the substrate were the same as the pH and temperature optima of the halogenating activity. The peroxidase activity of the enzyme is competitively inhibited by sodium azide, indicating that it is a hemeperoxidase different from nonheme peroxidases. © 2012 Wiley Periodicals, Inc. Int J Chem Kinet 45: 92–100, 2013     

Capture and Recycling of Sortase A through Site‐Specific Labeling with Lithocholic Acid

Enzyme‐mediated protein modification often requires large amounts of biocatalyst, adding significant costs to the process and limiting industrial applications. Herein, we demonstrate a scalable and straightforward strategy for the efficient capture and recycling of enzymes using a small‐molecule affinity tag. A proline variant of an evolved sortase A (SrtA 7M) was N‐terminally labeled with lithocholic acid (LA)—an inexpensive bile acid that exhibits strong binding to β‐cyclodextrin (βCD). Capture and recycling of the LA‐Pro‐SrtA 7M conjugate was achieved using βCD‐modified sepharose resin. The LA‐Pro‐SrtA 7M conjugate retained full enzymatic activity, even after multiple rounds of recycling.     

Polymer Embedded Nanocrystalline Titania: A New Generation Sorbent for the Separation of 77As from Ge for Biomedical Applications

The present work demonstrates the utility of polymer embedded nanocrystalline titania (TiP) as a new, accessible and viable solid sorbent for the chromatographic separation of high specific activity ⁷⁷As from neutron irradiated natural GeO₂. Experimental parameters such as distribution ratios (K _d), equilibrium sorption capacity, breakthrough capacity and separation yields were determined. A two-step ion-exchange procedure was developed to avail ⁷⁷As, from irradiated Ge in alkaline medium. The first step involved removal of the bulk Ge from ⁷⁷As by selective sorption of Ge ions on a TiP column at pH 13. Subsequently, the effluent solution containing ⁷⁷As was further purified and concentrated by sorption on a small TiP column at pH 10. ⁷⁷As could be eluted from the second column in 2?C3?mL of 0.1?M NaOH solution with >80% radiochemical yield. The ⁷⁷As obtained by this method was found to have insignificantly small level of radiocontaminants.     

An Alumina Based 99Mo–99mTc Generator to Produce 99mTc in Organic Medium Suitable for Industrial Radiotracer Applications

An industrial chromatographic ⁹⁹Mo–^99mTc generator has been developed with the aid of chromatographic alumina to obtain ^99mTc in a non-aqueous medium. This generator system takes advantage of tributyl phosphate to extract ^99mTc selectively with appreciable yield and in high radiochemical and radionuclidic purity. This facile, versatile and efficient approach provides ^99mTc at industrial sites in a medium soluble in hydrocarbon solvents, for radiotracer applications.     

Radiosynthesis and in vitro evaluation of 99mTc(CO)3-labeled folic acid derivative

The over-expression of folate receptors in variety of neoplastic tissues makes radiolabeled folate conjugates potential agents for imaging and therapy of such cancers. With the aim of preparing an imaging agent for targeting folate receptors, folic acid has been conjugated with homocysteine for complexation with [^99mTc(CO)₃(H₂O)₃]⁺ core. The radiolabeled complex of the homocysteine-folate could be obtained in >95% radiochemical yield as observed by HPLC. Stability of complex in saline was studied and challenge studies with histidine and cysteine revealed kinetic stability of the complex. Lipophilicity of the radiolabeled complex (log P) was found to be 0.45. In vitro uptake of ^99mTc(CO)₃-labeled folic acid derivative was studied in KB cells and inhibition studies were carried out using ³H-folic acid and cold homocysteine–folate conjugate. The in vitro studies indicated loss of binding affinity of the derivative towards folate receptors.     

Separation of Clinical Grade 188Re from 188W Using Polymer Embedded Nanocrystalline Titania

Currently, ¹⁸⁸Re is obtained from ¹⁸⁸W/¹⁸⁸Re chromatographic generator containing alumina which has a limited capacity (~80 mg Wg⁻¹) for ¹⁸⁸W. This results in high bolus volumes of ¹⁸⁸Re, which often needs to be concentrated before radiolabeling. We have demonstrated the feasibility of using polymer embedded nano crystalline titania (TiP), a novel high capacity sorbent material (~300 mg Wg⁻¹), for developing a ¹⁸⁸W/¹⁸⁸Re chromatographic generator. A TiP based chromatographic ¹⁸⁸W/¹⁸⁸Re generator was developed in which ¹⁸⁸Re could be eluted with 0.9% saline solution. About 90% of the ¹⁸⁸Re could be recovered in the first 4–5 mL of total activity with more than 80% yield. The purity of ¹⁸⁸Re is adequate for clinical applications.