High throughput screening for neurodegeneration and complex disease phenotypes

High throughput screening (HTS) for complex diseases is challenging. This stems from the fact that complex phenotypes are difficult to adapt to rapid, high throughput assays. We describe the recent development of high throughput and high-content screens (HCS) for neurodegenerative diseases, with a focus on inherited neurodegenerative disorders, such as Huntington's disease. We describe, among others, HTS assays based on protein aggregation, neuronal death, caspase activation and mutant protein clearance. Furthermore, we describe high-content screens that are being used to prioritize hits identified in such HTS assays. These assays and screening approaches should accelerate drug discovery for neurodegenerative disorders and guide the development of screening approaches for other complex disease phenotypes.     

Indium Trichloride (InCl3) Catalyzed Synthesis of Fused 7‐Azaindole Derivatives Using Domino Knoevenagel‐Michael Reaction

An efficient and high yielding methodology developed for the synthesis of fused 7‐azaindole derivatives via one pot multicomponent assembly process of cyclic 1,3‐dicarbonyls with substituted aldehydes and 5‐amino‐1‐tert‐butyl‐1H‐pyrrole‐3‐carbonitrile. The transformation occurs via domino Knoevenagel‐ Michael reaction followed by intramolecular cyclization in the presence of catalytic amount of InCl₃ (10 mol %). Mild reaction conditions, easy isolation of products, and good to excellent yields in a shorter period of time are the silent features of present methodology. Structures of all the newly prepared compounds have been corroborated by various spectroscopic methods.     

A catalyst-and solvent-free multicomponent synthesis of 7-azagramine analogues via a Mannich type reaction

A catalyst-and solvent-free protocol for the synthesis of 7-azagramine analogues is described via a three-component Mannich type reaction between 7-azaindole, aromatic aldehydes and heterocyclic amines in acceptable to excellent yields. Structures of the compounds were confirmed satisfactorily by ¹H NMR, IR, mass, TOCSY, HSQC and HMBC spectral analyses.     

Ni(II) and Cu(II) complexes of phenoxy-ketimine ligands: Synthesis,structures and their utility in bulk ring-opening polymerization (ROP) of l-lactide

Synthetic, structural and catalysis studies of Ni(II) and Cu(II) complexes of a series of phenoxy-ketimine ligands with controlled variations of sterics, namely 2-[1-(2,6-diethylphenylimino)ethyl]phenol (1a), 2-[1-(2,6-dimethylphenylimino)ethyl]phenol (1b) and 2-[1-(2-methylphenylimino)ethyl]phenol (1c), are reported. Specifically, the ligands 1a, 1b and 1c were synthesized by the TiCl₄ mediated condensation reactions of the respective anilines with o-hydroxyacetophenone in 21–23% yield. The nickel complexes, {2-[1-(2,6-diethylphenylimino)ethyl]phenoxy}₂Ni(II) (2a) and {2-[1-(2,6-dimethylphenylimino)ethyl]phenoxy}₂Ni(II) (2b), were synthesized by the reaction of the respective ligands 1a and 1b with Ni(OAc)₂ · 4H₂O in the presence of NEt₃ as a base in 71–75% yield. The copper complexes, {2-[1-(2,6-diethylphenylimino)ethyl]phenoxy}₂Cu(II) (3a), {2-[1-(2,6-dimethylphenylimino)ethyl]phenoxy}₂Cu(II) (3b) and {2-[1-(2-methylphenylimino)ethyl]phenoxy}₂Cu(II) (3c) were synthesized analogously by the reactions of the ligands 1a, 1b and 1c with Cu(OAc)₂ · H₂O in 70–87% yield. The molecular structures of the nickel and copper complexes 2a, 2b, 3a, 3b and 3c have been determined by X-ray diffraction studies. Structural comparisons revealed that the nickel centers in 2a and 2b are in square planar geometries while the geometry around the copper varied from being square planar in 3a and 3c to distorted square planar in 3b. The catalysis studies revealed that while the copper complexes 3a, 3b and 3c efficiently catalyze ring-opening polymerization (ROP) of l-lactide at elevated temperatures under solvent-free melt conditions, producing polylactide polymers of moderate molecular weights with narrow molecular weight distributions, the nickel counterparts 2a and 2b failed to yield the polylactide polymer.     

A roe-average algorithm for a granular-gas model with non-conservative terms

A Roe-average algorithm has been derived for a granular-gas model, proposed by Goldshtein and Shapiro [Goldshtein, Shapiro, Mechanics of collisional motion of granular materials: Part 1. General hydrodynamic equations, J. Fluid Mech. 282 (1995) 75–114], which contains non-conservative terms in the Euler-like hyperbolic governing equations apart from sink terms, which arise from inelastic collision of granules and are present only in the energy equation. The non-conservative terms introduce non-isentropic effects in acoustic-wave propagation within granular media and they also contribute to the Rankine–Hugoniot relations across a discontinuity. A Roe-average algorithm, based on the same granular-gas model, was derived in the literature [V. Kamenetsky, A. Goldshtein, M. Shapiro, D. Degani, Evolution of a shock wave in a granular gas, Phys. Fluids, 12 (2000) 3036–3049] which then required the implementation of a shock-fitting technique at a discontinuity. In the present work, Roe-averaged variables have been obtained from the Rankine–Hugoniot jump relations and the non-conservative terms have been incorporated in the numerical flux formula consistent with upwind principles associated with the granular speed of sound. Results for unsteady one-dimensional granular flows, colliding with a wall, demonstrate the capability of the proposed algorithm to capture strong shocks in addition to flow features not found in molecular gases, such as a fluidized region downstream of the shock and a compacted solid-block region adjacent to the wall.     

Genetic individualization of <Emphasis Type="Italic">Cannabis sativa</Emphasis> by a short tandem repeat multiplex system

Cannabis sativa is the most frequently used of all illicit drugs in the USA. Cannabis has been used throughout history for its stems in the production of hemp fiber, seed for oil and food, and buds and leaves as a psychoactive drug. Short tandem repeats (STRs) were chosen as molecular markers owing to their distinct advantages over other genetic methods. STRs are codominant, can be standardized such that reproducibility between laboratories can be easily achieved, have a high discrimination power, and can be multiplexed. In this study, six STR markers previously described for C. sativa were multiplexed into one reaction. The multiplex reaction was able to individualize 98 cannabis samples (14 hemp and 84 marijuana, authenticated as originating from 33 of the 50 states of the USA) and detect 29 alleles averaging 4.8 alleles per loci. The data did not relate the samples from the same state to each other. This is the first study to report a single-reaction sixplex and apply it to the analysis of almost 100 cannabis samples of known geographic origin. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Metal-catalyzed reduction of HCONR'2, R' = Me (DMF), Et (DEF), by silanes to produce R'2NMe and disiloxanes: a mechanism unraveled

We demonstrate that using Mo(CO)(6), Mo(CO)(5)NMe(3), and (η(5)-C(5)H(5))Mn(CO)(3) as catalysts for the silane, R(3)SiH, reduction of N,N-dimethylformamide (DMF), and N,N-diethylformamide (DEF), we can observe, intercept, and isolate, the important siloxymethylamine intermediates, R(3)SiOCH(2)NR'(2), R' = Me, Et, for the first time. In the presence of excess DMF such intermediates thermally react with a variety of silanes to form the corresponding disiloxanes in the absence of a metal catalyst. We also show that the germanium hydrides, Et(3)GeH and Bu(3)GeH, also reduce DMF to form trimethylamine and the corresponding digermoxane but observe no intermediates R(3)GeOCH(2)NMe(2). Bu(3)SnH reduces DMF, but along with the low yields of Bu(3)SnOSnBu(3) (but no Bu(3)SnOCH(2)NMe(2)) significant side products are obtained including (Bu(3)Sn)(2) and Bu(4)Sn. In the absence of DMF the siloxymethylamines can undergo metal-catalyzed reactions with silanes, germanes and stannanes to form disiloxanes, and R(3)SiOER(3) E = Ge, Sn, respectively. To date, the most efficient catalyst for this latter process is (η(5)-C(5)H(5))Mo(CO)(3)CH(3) via a photochemical reaction.     

Range‐independent background subtraction algorithm for recovery of Raman spectra of biological tissue

An important requirement for the use of Raman spectroscopy for tissue diagnostic applications is an appropriate algorithm that can faithfully retrieve weak tissue Raman signals from the measured raw Raman spectra. Although iterative modified polynomial‐fitting‐based automated algorithms are widely used, these are sensitive to the choice of the fitting range, thereby leading to significantly different Raman spectra for different start and stop wavenumber selection. We report here an algorithm for automated recovery of the weak Raman signal, which is range independent. Given a raw Raman spectrum and the choice of the start and the stop wavenumbers, the algorithm first truncates the spectrum to include the raw data within this wavenumber range, linearly extrapolates the truncated raw spectrum beyond the points of truncation on the two sides by using coefficients of linear least‐square fit, adds two Gaussian peaks of appropriate height and width on the extrapolated linear wings on either side and then iteratively smoothens the data with all these add‐ons such that the smaller of the ordinate values of the smoothed and the starting raw data serve as the input to each successive round of iterative smoothing until the added Gaussian peaks are fully recovered. The algorithm was compared with the modified polynomial‐based algorithms using mathematically simulated Raman spectrum as well as experimentally measured Raman spectra from various biological samples and was found to yield consistently range‐independent and artifact‐free Raman signal with zero baseline. Copyright © 2012 John Wiley & Sons, Ltd.     

Integrated Perspective for Effective Bioremediation

Identification of factors which can influence the natural attenuation process with available microbial genetic capacities can support the bioremediation which has been viewed as the safest procedure to combat with anthropogenic compounds in ecosystems. With the advent of molecular techniques, assimilatory capacity of an ecosystem can be defined with changing community dynamics, and if required, the essential genetic potential can be met through bioaugmentation. At the same time, intensification of microbial processes with nutrient balancing, expressing and enhancing the degradative capacities, could reduce the time frame of restoration of the ecosystem. The new concept of ecosystems biology has added greatly to conceptualize the networking of the evolving microbiota of the niche that helps in effective application of bioremediation tools to manage pollutants as additional carbon source.