Role of a PAS sensor domain in the Mycobacterium tuberculosis transcription regulator Rv1364c

The Mycobacterium tuberculosis transcriptional regulator Rv1364c regulates the activity of the stress response sigma factor sigma(F). This multi-domain protein has several components: a signaling PAS domain and an effector segment comprising of a phosphatase, a kinase and an anti-anti-sigma factor domain. Based on Small Angle X-ray Scattering (SAXS) data, Rv1364c was recently shown to be a homo-dimer and adopt an elongated conformation in solution. The PAS domain could not be modeled into the structural envelope due to poor sequence similarity with known PAS proteins. The crystal structure of the PAS domain described here provides a structural basis for the dimerization of Rv1364c. It thus appears likely that the PAS domain regulates the anti-sigma activity of Rv1364c by oligomerization. A structural comparison with other characterized PAS domains reveal several sequence and conformational features that could facilitate ligand binding - a feature which suggests that the function of Rv1364c could potentially be governed by specific cellular signals or metabolic cues. (C) 2010 Elsevier Inc. All rights reserved.

Structure-Reactivity Correlations of Benzoin Alkyl Ethers. Structures of 2-Methoxy-1,2-diphenylethanone (I) and 2-Isopropoxy-1,2-diphenylethanone (II)

(I): C15H1402, Mr---226.27, triclinic, Pi,a=8.441 (2), b= 10.276 (1), c= 15.342 (2)A, a=91.02 (2), ~ t= 79.26 (2), y= 105.88 (2) °, V=1256.8 (4)A 3, Z=4, D,,= 1.209 (flotation in KI),D x - 1.195 g cm -3, #(Mo, 2 = 0.7107/~) = 0.44 cm -~,F(000) = 480, T= 293 K, R -- 0.060 for 1793 significant reflections. (II): C~THlsO2, Mr= 254.83, orthorhombic, Pca21, a=8.476 (1); b= 16.098 (3), c=10.802(3)A, V=1473.9 (5) A s, Z=4, Dm=1.161 (flotation in KI), Dx= 1.148gem -3, /~(Mo, 2=0.7107 A) =0.41 cm -~, F(000) = 544, T= 293 K, R = 0.071 for 867 significant reflections. Both (I) and (II) crystallize in a cisoid conformation for the carbonyl group and alkoxy groups. Compounds (I) and (II) are photostable on irradiation in the solid state in spite of the favourable conformation of the functional groups for intramolecular H abstraction. Absence of photoreaction of (I)and (II) in the solid state is rationalized in the light of unfavourable intramolecular geometry.

High Starting Torque Drive For Step Motors

A bi-level voltage drive circuit for step motors that can provide the required high starting torque is described. In this circuit, microprocessor 8085 and parallel port interface 8255 are used for generating the code sequence. The inverter buffer 74LS06 provides enough drive to a darlington pair transistor. The comparator LM339 is used to compare the required voltage for step motor with the set value. This circuit can be effectively used for step motors having maximum rated current of less than 15 A with proper heat sink.

Directional solidification of aluminum-nickel eutectic alloys using electroslag remelting

Attempts were made to produce directionally solidified, specifically grain aligned Al-6 wt pct Ni eutectic alloy using a laboratory scale ESR unit. For this purpose sand cast alloy electrodes were electroslag remelted under different mold conditions. The grain structure of the ingots obtained from these meltings showed that insulated silica molds gave the best vertical alignment of grains along the length of the ingot. The NiAl3 fibers within the grains tended to fan out and there was only a preferred alignment of fibers along the growth direction under the conditions of our experiments. The ESR parameters most suitable for vertical alignment of eutectic grains have been identified. In some electroslag remelting trials ingots were grown on a seed ingot. This resulted in a fewer vertical grains compared to the case when no seed ingot was used. The sand cast specimen of the eutectic exhibited a maximum tensile strength of around 88.2 MN/m2 (9.0 kg/mm2) whereas conventional ESR using water cooled mold gave strength value of 98.0 MN/m2 (10 kg/mm2). The directionally solidified ESR material showed longitudinal tensile strength as high as 213.7 MN/m2 (21.8 kg/mm2) which could be further increased to 220.6 MN/m2 (22.5 kg/mm2) by using the seed ingot. The average growth rate was varied between 5 to 25 mm/min during electroslag remelting in this study. The flow stresses, tangent modulus and ultimate tensile strength of directionally solidified eutectic increased with increasing growth rates.

Structure of pregna-4,16-diene-7[alpha],14[alpha]-diol-3,20-dione

7a,14a-Dihydroxypregna-4,16-diene-3,20- dione, C21H2804, M r = 344.45, orthorhombic, P212121, a = 7.136 (1), b = 12.342 (1), c = 20.049 (3)/k, V= 1765.7 (3)/k 3, Z = 4, Dx = 1.295 g cm -3, A(Cu Kte) = 1.5418/k, /z = 6.7 cm- a, F(000) = 744, T = 293 K, R = 0.048 for 1345 observations. The A ring may be described as in a l a,2flhalf- chair conformation or a l a-sofa conformation. The B and C rings adopt normal chair conformations and the D ring has a 14a-envelope conforma tion. The molecules are held together by a hydrogen bond [0(3)...0(7)= 2.767 A].

Polyelectrolyte microcapsules for sustained delivery of water-soluble drugs

Polyelectrolyte capsules composed of weak polyelectrolytes are introduced as a simple and efficient system for spontaneous encapsulation of low molecular weight water-soluble drugs. Polyelectrolyte capsules were prepared by layer-by-layer (LbL) assembling of weak polyelectrolytes, poly(allylamine hydrochloride) (PAH) and poly (methacrylic acid) (PMA) on polystyrene sulfonate (PSS) doped CaCO3 particles followed by core removal with ethylene-diaminetetraacetic add (EDTA). The loading process was observed by confocal laser scanning microscopy (CLSM) using tetramethylrhodamineisothiocyanate labeled dextran (TRITC-dextran) as a fluorescent probe. The intensity of fluorescent probe inside the capsule decreased with increase in cross-linking time. Ciprofloxacin hydrochloride (a model water-soluble drug) was spontaneously deposited into PAH/PMA capsules and their morphological changes were investigated by scanning electron microscopy (SEM) and atomic force microscopy (AFM). The quantitative study of drug loading was also elucidated which showed that drug loading increased with initial drug concentration, but decreased with increase in pH. The loaded drug was released in a sustained manner for 6 h, which could be further extended by cross-linking the capsule wall. The released drug showed significant antibacterial activity against E. coli. These findings indicate that such capsules can be potential carriers for water-soluble drugs in sustained/controlled drug delivery applications. (C) 2010 Elsevier B.V. All rights reserved.

Social selection and the evolution of cooperative groups: The example of the cellular slime moulds

In social selection the phenotype of an individual depends on its own genotype as well as on the phenotypes, and so genotypes, of other individuals. This makes it impossible to associate an invariant phenotype with a genotype: the social context is crucial. Descriptions of metazoan development, which often is viewed as the acme of cooperative social behaviour, ignore or downplay this fact. The implicit justification for doing so is based on a group-selectionist point of view. Namely, embryos are clones, therefore all cells have the same evolutionary interest, and the visible differences between cells result from a common strategy. The reasoning is flawed, because phenotypic heterogeneity within groups can result from contingent choices made by cells from a flexible repertoire as in multicellular development. What makes that possible is phenotypic plasticity, namely the ability of a genotype to exhibit different phenotypes. However, co-operative social behaviour with division of labour requires that different phenotypes interact appropriately, not that they belong to the same genotype, or have overlapping genetic interests. We sketch a possible route to the evolution of social groups that involves many steps: (a) individuals that happen to be in spatial proximity benefit simply by virtue of their number; (b) traits that are already present act as preadaptations and improve the efficiency of the group; and (c) new adaptations evolve under selection in the social context-that is, via interactions between individuals-and further strengthen group behaviour. The Dictyostelid or cellular slime mould amoebae (CSMs) become multicellular in an unusual way, by the aggregation of free-living cells. In nature the resulting group can be genetically homogeneous (clonal) or heterogeneous (polyclonal); in either case its development, which displays strong cooperation between cells (to the extent of so-called altruism) is not affected. This makes the CSMs exemplars for the study of social behaviour.

Impact of metal binding on the antitumor activity and cellular imaging of a metal chelator cationic imidazopyridine derivative

A new water soluble cationic imidazopyridine species, viz. (1E)-1-((pyridin-2-yl)methyleneamino)-3-(3(pyridin-2-yl) imidazo1,5-a]pyridin-2(3H)-yl)propan-2-ol (1), as a metal chelator is prepared as its PF6 salt and characterized. Compound 1 shows fluorescence at 438 nm on excitation at 342 nm in Tris-HCl buffer giving a fluorescence quantum yield (phi) of 0.105 and a life-time of 5.4 ns. Compound 1, as an avid DNA minor groove binder, shows pUC19 DNA cleavage activity in UV-A light of 365 nm forming singlet oxygen species in a type-II pathway. The photonuclease potential of 1 gets enhanced in the presence of Fe2+, Cu2+ or Zn2+. Compound 1 itself displays anticancer activity in HeLa, HepG2 and Jurkat cells with an enhancement on addition of the metal ions. Photodynamic effect of 1 at 365 nm also gets enhanced in the presence of Fe2+ and Zn2+. Fluorescence-based cell cycle analysis shows a significant dead cell population in the sub-G1 phase of the cell cycle suggesting apoptosis via ROS generation. A significant change in the nuclear morphology is observed from Hoechst 33258 and an acridine orange/ethidium bromide (AO/EB) dual nuclear staining suggesting apoptosis in cells when treated with 1 alone or in the presence of the metal ions. Apoptosis is found to be caspase-dependent. Fluorescence imaging to monitor the distribution of 1 in cells shows that 1 in the presence of metal ions accumulates predominantly in the cytoplasm. Enhanced uptake of 1 into the cells within 12 h is observed in the presence of Fe2+ and Zn2+.

Spatial gradients of calcium in the slug of Dictyostelium discoideum

Starved amoebae of D. discoideum aggregate and give rise to a long and thin multicellular structure called the slug. The cells within the slug eventually differentiate according to a simple anterior/posterior dichotomy. This motivates a search for gradients of putative morphogens along its axis. Calcium may be one such morphogen. On the basis of observations made by using the calcium-sensitive fluorescent dyes fura-2 and chlortetracyline, we report that there are spatial gradients in cytoplasmic and sequestered calcium in the slug. Anteriorly located and genetically defined prestalk cells (ecmA/pstA, ecmB/pstAB) contain significantly higher levels of calcium than the prespore cells in the posterior. However, the proportion of 'calcium-rich' cells in the slug is greater than that of the subset of prestalk cells defined by the expression of the ecmA or ecmB genes.

Mutations in the Res subunit of the EcoPI restriction enzyme that affect ATP-dependent reactions

The Res subunits of the type III restriction-modification enzymes share a statistically significant amino acid sequence similarity with several RNA and DNA helicases of the so-called DEAD family. It was postulated that in type III restriction enzymes a DNA helicase activity may be required for local unwinding at the cleavage site. The members of this family share seven conserved motifs, all of which are found in the Res subunit of the type III restriction enzymes. To determine the contribution, if any, of these motifs in DNA cleavage by EcoPI, a type III restriction enzyme, we have made changes in motifs I and II. While mutations in motif I (GTGKT) clearly affected ATP hydrolysis and resulted in loss of DNA cleavage activity, mutation in motif II (DEPH) significantly decreased ATP hydrolysis but had no effect on DNA cleavage. The double mutant R.EcoPIK90R-H229K showed no significant ATPase or DNA restriction activity though ATP binding was not affected. These results imply that there are at least two ATPase reaction centres in EcoPI restriction enzyme. Motif I appears to be involved in coupling DNA restriction to ATP hydrolysis. Our results indicate that EcoPI restriction enzyme does not have a strand separation activity. We suggest that these motifs play a role in the ATP-dependent translocation that has been proposed to occur in the type III restriction enzymes. (C) 1997 Academic Press Limited.

Permeability and consolidation behavior of fly ashes

A knowledge of permeability and consolidation is essential in a number of engineering problems such as settlement, seepage, and stability of the structures. Since fly ash is used very widely for several geotechnical applications, there is a need to understand its permeability and consolidation behavior. This paper presents a detailed study conducted on two Indian fly ashes. It brings out the role of chemical composition (free lime) on the permeability and consolidation behavior of fly ashes. It is found that the permeability values computed based on grain-size distribution agree well with those obtained based on test data.

Role of oxygen transients in the facile scission of C-O bonds of alcohols on Zn surfaces

The alkoxy species produced by the interaction of alcohols with Zn surfaces undergoes C-O bond scission at 150 K giving hydrocarbon species, but this transformation occurs even at 80 K when alcohol-oxygen mixtures are coadsorbed, due to the oxygen transients.

A ring closing metathesis-based approach to (+/-)-herbertene, (+/-)-alpha-herbertenol, (+/-)-beta-herbertenol and (+/-)-herbertenediol

An efficient methodology for the synthesis of the aromatic sesquiterpenes (+/-)-herbertene, (+/-)-alpha-herbertenol, (+/-)-beta-herbertenol (+/-)-herbertenediol and (+/-)-alpha-cuparenone, employing a combination of Claisen rearrangement and ring closing metathesis reactions, is described.