Pathway-Selective Sensitization of Mycobacterium tuberculosis for Target-Based Whole-Cell Screening

Whole-cell screening of Mycobacterium tuberculosis (Mtb) remains a mainstay of drug discovery, but subsequent target elucidation often proves difficult. Conditional mutants that underexpress essential genes have been used to identify compounds with known mechanism of action by target-based whole-cell screening (TB-WCS). Here, the feasibility of TB-WCS in Mtb was assessed by generating mutants that conditionally express pantothenate synthetase (panC), diaminopimelate decarboxylase (lysA), and isocitrate lyase (icl1). The essentiality of panC and lysA, and conditional essentiality of icl1 for growth on fatty acids, was confirmed. Depletion of PanC and Icl1 rendered mutants hypersensitive to target-specific inhibitors. Stable reporter strains were generated for use in high-throughput screening, and their utility was demonstrated by identifying compounds that display greater potency against a PanC-depleted strain. These findings illustrate the power of TB-WCS as a tool for tuberculosis drug discovery.     

Thermal [1,5] hydrogen sigmatropic shifts in cis,cis-1,3-cyclononadienes probed by gas-phase kinetic studies and density functional theory calculations

The kinetics of gas-phase thermal [1,5] hydrogen shifts interconverting the five isomeric mono-deuterium-labeled cis,cis-1,3-cyclononadienes have been followed at four temperatures from 240 to 287 degrees C. The activation parameters found were Ea = 37.1 +/- 0.8 kcal/mol, log A = 11.6 +/- 0.3, DeltaH++ = 36.0 +/- 0.8 kcal/mol, and DeltaS++ = -9.0 +/- 0.3 eu. Density functional theory based calculations have provided geometries and energies for the ground-state cyclononadiene conformational isomers, for the transition states linking one to another, and for the transition states for [1,5] hydrogen shifts responsible for isomerizations among the five labeled dienes. A generalized formulation of the Winstein-Holness equation is presented and applied to the complex system, one that involves 11 ground-state conformers, 10 transition states separating them, and five transition states for [1,5] hydrogen shifts. The value for the empirical Ea derived from calculated mole fractions of ground-state conformers and calculated energies for specific ground-state conformers and [1,5] hydrogen shift transition structures was 37.5 kcal/mol, in excellent agreement with the experimentally obtained activation energy. The significance of conformational options in various ground states and transition structures for the [1,5] hydrogen shifts is considerable, an inference that may well have general applicability.     

Complex amphiphilic networks derived from diamine‐terminated poly(ethylene glycol) and benzylic chloride‐functionalized hyperbranched fluoropolymers

Amphiphilic copolymer networks were prepared from hyperbranched fluoropolymer (HBFP*, M_n = 38 kDa, by atom transfer radical‐self condensing vinyl copolymerization) and linear diamine‐terminated poly(ethylene glycol) (DA‐PEG, M_n = 1,630 Da). Model studies found that the crosslinking mechanism occurred at ambient temperature as a result of reaction between DA‐PEG and the benzylic chlorides of HBFP*. These networks underwent covalent attachment to glass microscope slides derivatized with 3‐aminopropyltriethoxysilane, whereupon gel percent studies at various weight percentages of DA‐PEG to HBFP* found that curing could be achieved at lower temperatures and shortened time periods relative to the previously reported parent HBFP–PEG system. Thermogravimetric analysis revealed that the crosslinked materials gave no evident mass loss up to 250 °C. Differential scanning calorimetry of the complex amphiphilic networks showed a suppressed glass transition temperature, relative to that observed for neat HBFP*, and multiple melting DA‐PEG endotherm(s) near 30 °C. The films possessed a topographically‐complex surface with features that increased in tandem with an increase in the ratio of DA‐PEG to HBFP*, as detected by atomic force microscopy and quantified by increased rms roughness values. Internal reflection infrared imaging revealed a heterogeneous surface composition and confirmed that the domain sizes increased as the weight percent of DA‐PEG increased. © 2006 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 44: 4782–4794, 2006     

A rapid field detection method for arsenic in drinking water.

In this study, a rapid colorimetric method for arsenic detection was developed. Different reagents containing magnesium turnings in combination with a series of acids were tested for arsine generation. The arsine was then allowed to react with auric chloride on Whatman filter paper No. 3, which in turn changed color. The detection time and detection limit were measured for each acid. Oxalic acid was found to be the most appropriate acid among all the acids used for detection in this study. It took 10 min to detect 10 ppb arsenic concentration and only 1 min to detect concentrations higher than 50 ppb. This method thus reduced the detection time for arsenic and has the potential to develop better field kit.     

Chemical composition and antibacterial activity of essential oil from Salvia mukerjeei

The composition of steam volatile oil from aerial parts of Salvia mukerjeei Bennet & Raizada (Lamiaceae) was analyzed by capillary GC and GC-MS. The oil was rich in sesquiterpene hydrocarbons (67.3%). Among 71 identified constituents representing 91.7% of the oil, beta-caryophyllene (28.7%), gamma-muurolene (15.5%) and dehydro-aromadendrane (9.5%), were the principal constituents. The oil was tested against ten bacterial strains and was active against Enterococcus faecalis, Erwinia chrysanthemi and Agrobacterium tumefaciens.     

Tailoring nanocarriers for intracellular protein delivery

Proteins play a crucial role in life, taking part in all vital processes in the body. In the past decade, there was increasing interest in delivering active forms of proteins to specific cells and organs. Intracellular protein delivery holds enormous promise for biological and medical applications, including cancer therapy, vaccination, regenerative medicine, treatment for loss-of-function genetic diseases and imaging. This tutorial review surveys recent developments in intracellular protein delivery using various nanocarriers. Methods such as lipid-mediated colloidal systems, polymeric nanocarriers, inorganic systems and protein-mediated carriers are reviewed. Advantages and limitations of current strategies, as well as future opportunities and challenges are also discussed.     

Synthesis and thermoluminescence characteristics of γ-irradiated K3Ca2(SO4)3F:Eu or Ce fluoride

New halophosphor K₃Ca₂(SO₄)₃F activated by Eu and Ce has been synthesized by a co-precipitation method and characterized according to its thermoluminescence. The formation of traps in rare earth doped K₃Ca₂(SO₄)₃F and the effects of γ-radiation dose on the glow curve are discussed. The glow curve of K₃Ca₂(SO₄)₃F:Ce shows a prominent single peak at 150°C, whereas K₃Ca₂(SO₄)₃F:Eu and K₃Ca₂(SO₄)₃F:Ce,Eu at 142°C and 192°C, respectively. A single glow peak indicates that there is only one set of trap being activated within the particular temperature range. The presented phosphors are also studied because of its fading, reusability and trapping parameters. There was just 2% fading during a period of 10 days, indicating no serious fading problem. Trapping parameters such as order of kinetics (b), activation energy (E) and frequency factor (S) were calculated by using Chen's half-width method. The observations presented in this paper are good for lamp phosphors as well as solid-state dosimeter.