Lead selection and characterization of antitubercular compounds using the Nested Chemical Library

Discovering new drugs to treat tuberculosis more efficiently and to overcome multidrug resistance is a world health priority. To find novel antitubercular agents several approaches have been used in various institutions worldwide, including target-based approaches against several validated mycobacterial enzymes and phenotypic screens. We screened more than 17,000 compounds from Vichem's Nested Chemical Library(TM) using an integrated strategy involving whole cell-based assays with Corynebacterium glutamicum and Mycobacterium tuberculosis, and target-based assays with protein kinases PknA, PknB and PknG as well as other targets such as PimA and bacterial topoisomerases simultaneously. With the help of the target-based approach we have found very potent hits inhibiting the selected target enzymes, but good minimal inhibitory concentrations (MIC) against M. tuberculosis were not achieved. Focussing on the whole cell-based approach several potent hits were found which displayed minimal inhibitory concentrations (MIC) against M. tuberculosis below 10 mu M and were non-mutagenic, non-cytotoxic and the targets of some of the hits were also identified. The most active hits represented various scaffolds. Medicinal chemistry-based lead optimization was performed applying various strategies and, as a consequence, a series of novel potent compounds were synthesized. These efforts resulted in some effective potential antitubercular lead compounds which were confirmed in phenotypic assays. (C) 2015 Elsevier Ltd. All rights reserved.

Evolution of high-pressure mafic granulites and pelitic gneisses from NE Madagascar: Tectonic implications

The occurrence of high-pressure mafic-ultramafic bodies within major shear zones is one of the indicators of paleo-subduction. In mafic granulites of the Andriamena complex (north-eastern Madagascar) we document unusual textures including garnet-clinopyroxene-quartz coronas that formed after the breakdown of orthopyroxene-plagioclase-ilmenite. Textural evidence and isochemical phase diagram calculations in the Na2O-CaO-K2O-FeO-MgO-Al2O3-SiO2-H2O-TiO2 system indicate a pressure-temperature (P-T) evolution from an isothermal (780 degrees C) pressure up to c. 24 kbar to decompression and cooling. Such a P-T trajectory is typically attained in a subduction zone setting where a gabbroic/ultramafic complex is subducted and later exhumed to the present crustal level during oceanic closure and final continental collision. The present results suggest that the presence of such deeply subducted rocks of the Andriamena complex is related to formation of the Betsimisaraka suture. LA-ICPMS U-Pb zircon dating of pelitic gneisses from the Betsimisaraka suture yields low Th/U ratios and protolith ages ranging from 2535 to 2625 Ma. A granitic gneiss from the Alaotra complex yields a zircon crystallization age of ca. 818 Ma and Th/U ratios vary from 1.08 to 2.09. K-Ar dating of muscovite and biotite from biotite-kyanite-sillimanite gneiss and garnet-biotite gneiss yields age of 486 +/- 9 Ma and 459 +/- 9 Ma respectively. We have estimated regional crustal thicknesses in NE Madagascar using a flexural inversion technique, which indicates the presence of an anomalously thick crust (c. 43 km) beneath the Antananarivo block. This result is consistent with the present concept that subduction beneath the Antananarivo block resulted in a more competent and thicker crust. The textural data, thermodynamic model, and geophysical evidence together provide a new insight to the subduction history, crustal thickening and evolution of the high-pressure Andriamena complex and its link to the terminal formation of the Betsimisaraka suture in north-eastern Madagascar. (C) 2015 Elsevier B.V. All rights reserved.

Organic-inorganic hybrid PtCo nanoparticle with high electrocatalytic activity and durability for oxygen reduction

In Pt-transition metal (TM) alloy catalysts, the electron transfer from the TM to Pt is retarded owing to the inevitable oxidation of the TM surface by oxygen. In addition, acidic electrolytes such as those employed in fuel cells accelerate the dissolution of the surface TM oxide, which leads to catalyst degradation. Herein, we propose a novel synthesis strategy that selectively modifies the electronic structure of surface Co atoms with N-containing polymers, resulting in highly active and durable PtCo nanoparticle catalysts useful for the oxygen reduction reaction (ORR). The polymer, which is functionalized on carbon black, selectively interacts with the Co precursor, resulting in Co-N bond formation on the PtCo nanoparticle surface. Electron transfer from Co to Pt in the PtCo nanoparticles modified by the polymer is enhanced by the increase in the difference in electronegativity between Pt and Co compared with that in bare PtCo nanoparticles with the TM surface oxides. In addition, the dissolution of Co and Pt is prevented by the selective passivation of surface Co atoms and the decrease in the O-binding energy of surface Pt atoms. As a result, the catalytic activity and durability of PtCo nanoparticles for the ORR are significantly improved by the electronic ensemble effects. The proposed organic/inorganic hybrid concept will provide new insights into the tuning of nanomaterials consisting of heterogeneous metallic elements for various electrochemical and chemical applications.

Regulatory quality and the regulatory system in the UK

Busca identificar como o sistema regulatório britânico se desenvolveu nos últimos anos contribuindo para a melhoria da regulação no país. Aborda o tema da qualidade legislativa no Reino Unido.

Estimating age of spotted and spinner dolphins (Stenella attenuata and Stenella longirostris) from teeth

This paper is an account of preparation and examination techniques and criteria used to estimate age in decalcified and stained tooth thin sections from spinner and spotted dolphins. A dentinal growth layer group (GLG), composed of two thin light and two thicker dark-stained layers, is deposited annually. The GLG component layers are variably visible, but the "ideal" pattern and successive thinning of dentinal GLGs are used as a guide to determine GLG limits. Age-specific thicknesses of dentinal GLGs found in Hawaiian spinner dolphin teeth seem to be applicable to teeth of spotted dolphins and can be used as an aid in locating GLG boundaries. Cementa1 GLGs are composed of a dark-stained and alightly stained layer and usually are deposited at a rate of one per year, but may be deposited every other year or two or three times per year. Two slightly different methods of counting dentinal GLGs are presented, along with guidelines for determining whether dentinal or cementa1 GLG counts provide the best estimate of age for a specimen. (PDF contains 23 pages.)

Effect of coal-over-coal reburn on furnace temperature and heat flux distributions in 1 MW tangentially fired furnace

The furnace temperature and heat flux distributions of 1 MW tangentially fired furnace were studied during coal-over-coal reburn, and the influences of the position of reburn nozzle and reburn fuel fraction on furnace temperature and heat flux distributions were investigated. Compared with the baseline, the flue gas temperature is 70–90 C lower in primary combustion and 130–150 C higher at furnace exit, and the variations of the flue gas temperature distributions along furnace height are slower. The temperature distribution along the width of furnace wall decreases with the increase of the relative furnace height. In the primary combustion zone and the reburn zone, the temperature and heat flux distributions of furnace wall are much non-uniform and asymmetric along the width of furnace wall, those of furnace wall in the burnout zone are relatively uniform, and the temperature non-uniformity coefficients of the primary combustion zone, the reburn zone and the burnout zone are 0.290, 0.100 and 0.031, respectively.

Application of microwave diagnostics to copper chloride and carbon dioxide lasers

Part A

A problem restricting the development of the CuCl laser has been the decrease in output power with increases of tube temperature above 400°C. At that temperature the CuCl vapor pressure is about .1 torr. This is a small fraction of the buffer gas pressure (He at 10 torr).

The aim of the project was to measure the peak radiation temperature (assumed related to the mean energy of electrons) in the laser discharge as a function of the tube temperature. A 24 gHz gated microwave radiometer was used.

It was found that at the tube temperatures at which the output power began to deteriorate, the electron radiation temperature showed a sharp increase (compared with radiation temperature in pure buffer).

Using the above result, we have postulated that this sudden increase is a result of Penning ionization of the Cu atoms. As a consequence of this process the number of Cu atoms available for lasing decrease.

PART B

The aim of the project was to study the dissociation of CO₂ in the glow discharge of flowing CO₂ lasers.

A TM₀₁₁ microwave (3 gHz) cavity was used to measure the radially averaged electron density n_e and the electron-neutral collision frequency in the laser discharge. An estimate of the electric field is made from these two measurements. A gas chromatograph was used to measure the chemical composition of the gases after going through the discharge. This instrument was checked against a mass spectrometer for accuracy and sensitivity.

Several typical laser mixtures were .used: CO₂-N₂-He (1,3,16), (1,3,0), (1,0,16), (1,2,10), (1,2,0), (1,0,10), (2,3,15), (2,3,0), (2,0,15), (1,3,16)+ H₂O and pure CO₂. Results show that for the conditions studied the dissociation as a function of the electron density is uniquely determined by the STP partial flow rate of CO₂, regardless of the amount of N₂ and/or He present. The presence of water vapor in the discharge decreased the degree of dissociation.

A simple theoretical model was developed using thermodynamic equilibrium. The electrons were replaced in the calculations by a distributed heat source.

The results are analyzed with a simple kinetic model.

Computational Predictions of G Protein-Coupled Receptor Structures and Binding Sites

G protein-coupled receptors (GPCRs) are the largest family of proteins within the human genome. They consist of seven transmembrane (TM) helices, with a N-terminal region of varying length and structure on the extracellular side, and a C-terminus on the intracellular side. GPCRs are involved in transmitting extracellular signals to cells, and as such are crucial drug targets. Designing pharmaceuticals to target GPCRs is greatly aided by full-atom structural information of the proteins. In particular, the TM region of GPCRs is where small molecule ligands (much more bioavailable than peptide ligands) typically bind to the receptors. In recent years nearly thirty distinct GPCR TM regions have been crystallized. However, there are more than 1,000 GPCRs, leaving the vast majority of GPCRs with limited structural information. Additionally, GPCRs are known to exist in a myriad of conformational states in the body, rendering the static x-ray crystal structures an incomplete reflection of GPCR structures. In order to obtain an ensemble of GPCR structures, we have developed the GEnSeMBLE procedure to rapidly sample a large number of variations of GPCR helix rotations and tilts. The lowest energy GEnSeMBLE structures are then docked to small molecule ligands and optimized. The GPCR family consists of five subfamilies with little to no sequence homology between them: class A, B1, B2, C, and Frizzled/Taste2. Almost all of the GPCR crystal structures have been of class A GPCRs, and much is known about their conserved interactions and binding sites. In this work we particularly focus on class B1 GPCRs, and aim to understand that family’s interactions and binding sites both to small molecules and their native peptide ligands. Specifically, we predict the full atom structure and peptide binding site of the glucagon-like peptide receptor and the TM region and small molecule binding sites for eight other class B1 GPCRs: CALRL, CRFR1, GIPR, GLR, PACR, PTH1R, VIPR1, and VIPR2. Our class B1 work reveals multiple conserved interactions across the B1 subfamily as well as a consistent small molecule binding site centrally located in the TM bundle. Both the interactions and the binding sites are distinct from those seen in the more well-characterized class A GPCRs, and as such our work provides a strong starting point for drug design targeting class B1 proteins. We also predict the full structure of CXCR4 bound to a small molecule, a class A GPCR that was not closely related to any of the class A GPCRs at the time of the work.

Mössbauer effect investigations of the hyperfine interactions and relaxation phenomena in salts of thulium

Two new phenomena have been observed in Mössbauer spectra: a temperature-dependent shift of the center of gravity of the spectrum, and an asymmetric broadening of the spectrum peaks. Both phenomena were observed in thulium salts. In the temperature range 1˚K ≤ T ≤ 5˚K the observed shift has an approximate inverse temperature dependence. We explain this on the basis of a Van Vleck type of interaction between the magnetic moment of two nearly degenerate electronic levels and the magnetic moment of the nucleus. From the size of the shift we are able to deduce an “effective magnetic field” H = (6.0 ± 0.1) x 10⁶ Gauss, which is proportional to ‹r^-3›_M‹G|J|E› where ‹r^-3›_M is an effective magnetic radial integral for the 4f electrons and |G› and |E› are the lowest 4f electronic states in Tm Cl₃·6H₂O. From the temperature dependence of the shift we have derived a preliminary value of 1 cm^-1 for the splitting of these two states. The observed asymmetric line broadening is independent of temperature in the range 1˚K ≤ T ≤ 5˚K, but is dependent on the concentration of thulium ions in the crystal. We explain this broadening on the basis of spin-spin interactions between thulium ions. From size and concentration dependence of the broadening we are able to deduce a spin-spin relaxation time for Tm Cl₃·6H₂O of the order of 10^-11 sec.

The resolution of the thermodynamic paradox and the theory of guided wave propagation in anisotropic media

The resolution of the so-called thermodynamic paradox is presented in this paper. It is shown, in direct contradiction to the results of several previously published papers, that the cutoff modes (evanescent modes having complex propagation constants) can carry power in a waveguide containing ferrite. The errors in all previous “proofs” which purport to show that the cutoff modes cannot carry power are uncovered. The boundary value problem underlying the paradox is studied in detail; it is shown that, although the solution is somewhat complicated, there is nothing paradoxical about it.

The general problem of electromagnetic wave propagation through rectangular guides filled inhomogeneously in cross-section with transversely magnetized ferrite is also studied. Application of the standard waveguide techniques reduces the TM part to the well-known self-adjoint Sturm Liouville eigenvalue equation. The TE part, however, leads in general to a non-self-adjoint eigenvalue equation. This equation and the associated expansion problem are studied in detail. Expansion coefficients and actual fields are determined for a particular problem.

Part I. Regulation of development in the cellular slime mold Dictyostelium discoideum. Part II. Polysomes and RNA synthesis during early development of the surf clam Spisula solidissima

Part I. The cellular slime mold Dictyostelium discoideum is a simple eukaryote which undergoes a multi-cellular developmental process. Single cell myxamoebae divide vegetatively in the presence of a food source. When the food is depleted or removed, the cells aggregate, forming a migrating pseudoplasmodium which differentiates into a fruiting body containing stalk and spore cells. I have shown that during the developmental cycle glycogen phosphorylase, aminopeptidase, and alanine transaminase are developmentally regulated, that is their specific activities increased at a specific time in the developmental cycle. Phosphorylase activity is undetectable in developing cells until mid-aggregation whereupon it increases and reaches a maximum at mid-culmination. Thereafter the enzyme disappears. Actinomycin D and cycloheximide studies as well as studies with morphologically aberrant and temporally deranged mutants indicate that prior RNA and concomitant protein synthesis are necessary for the rise and decrease in activity and support the view that the appearance of the enzyme is regulated at the transcriptional level. Aminopeptidase and alanine transaminase increase 3 fold starting at starvation and reach maximum activity at 18 and 5 hours respectively.

The cellular DNA s of D. discoideum were characterized by CsC1 buoyant density gradient centrifugation and by renaturation kinetics. Whole cell DNA exhibits three bands in CsCl: ρ = 1.676 g/cc (nuclear main band), 1.687 (nuclear satellite), and 1.682 (mitochondrial). Reassociation kinetics at a criterion of Tm -23°C indicates that the nuclear reiterated sequences make up 30% of the genome (Cot_1/2 (pure) 0.28) and the single-copy DNA 70% (Cot_1/2(pure) 70). The complexity of the nuclear genome is 30 x 10⁹ daltons and that of the mitochondrial DNA is 35-40 x 10⁶ daltons (Cot_1/2 0.15). rRNA cistrons constitute 2.2% of nuclear DNA and have a ρ = 1.682.

RNA extracted from 4 stages during developmental cycle of Dictyostelium was hybridized with purified single-copy nuclear DNA. The hybrids had properties indicative of single-copy DNA-RNA hybrids. These studies indicate that there are, during development, qualitative and quantitative changes in the portion of the single-copy of the genome transcribed. Overall, 56% of the genome is represented by transcripts between the amoeba and mid-culmination stages. Some 19% are sequences which are represented at all stages while 37% of the genome consists of stage specific sequences.

Part II. RNA and protein synthesis and polysome formation were studied during early development of the surf clam Spisula solidissima embryos. The oocyte has a small number of polysomes and a low but measurable rate of protein synthesis (leucine-³H incorporation). After fertilization, there is a continual increase in the percentage of ribosomes sedimenting in the polysome region. Newly synthesized RNA (uridine-5-³H incorporation) was found in polysomes as early as the 2-cell stage. During cleavage, the newly formed RNA is associated mainly with the light polysomes.

RNA extracted from polysomes labeled at the 4-cell stage is polydisperse, nonribosomal, and non-4 S. Actinomycin D causes a reduction of about 30% of the polysomes formed between fertilization and the 16-cell stage.

In the early cleavage stages the light polysomes are mostly affected by actinomycin.

Electromagnetic fields and transmission properties in tapered hollow metallic waveguides

We analyze the electromagnetic spatital distributions and address an important issue of the transmission properties of spherical transverse-electric (TE) and transverse-magnetic (TM) eigenmodes within a tapered hollow metallic waveguide in detail. Explicit analytical expressions for the spatital distributions of electromagnetic field components, attenuation constant, phase constant and wave impedance are derived. Accurate eigenvalues obtained numerically are used to study the dependences of the transmission properties on the taper angle, the mode as well as the length of the waveguide. It is shown that all modes run continuously from a propagating through a transition to an evanescent region and the value of the attenuation increases as the distance from the cone vertex and the cone angle decrease. A strict distinction between pure propagating and pure evanescent modes cannot be achieved. One mode after the other reaches cutoff in the tapered hollow metallic waveguide as the distance from the cone vertex desreases. (C) 2008 Optical Society of America

Structure and properties of amorphous thin film for optical data storage

In this paper the magnetic and magneto-optical properties of amorphous rare earth-transition metal (RE-TM) alloys as well as the magnetic coupling in the multi-layer thin films for high density optical data storage are presented. Using magnetic effect in scanning tunneling microscopy the clusters structure of amorphous RE-TM thin films has been observed and the perpendicular magnetic anisotropy in amorphous RE-TM thin films has been interpreted. Experimental results of quick phase transformation under short pulse laser irradiation of amorphous semiconductor and metallic alloy thin films for phase change optical recording are reported. A step-by-step phase transformation process through metastable states has been observed. The waveform of crystallization propagation in micro-size spot during laser recording in amorphous semiconductor thin films is characterized and quick recording and erasing mechanism for optical data storage with high performance are discussed. The nonlinear optical effects in amorphous alloy thin films have been studied. By photo-thermal effect or third order optical nonlinearity, the optical self-focusing is observed in amorphous mask thin films. The application of amorphous thin films with super-resolution near field structure for high-density optical data storage is performed. (c) 2007 Elsevier B.V. All rights reserved.