Molecular mechanism for the specific inhibition of reverse transcriptase of rous sarcoma virus by the copper complexes of isonicotinic acid hydrazide

Isonicotinic acid hydrazide (isoniazid), one of the most potent antitubercular drugs, was recently shown, in our laboratory, to form two different complexes with copper, depending upon the oxidation state of the metal ion. Both the complexes have been shown to possess antiviral activity against Rous sarcoma virus, an RNA tumor virus. The antiviral activity of the complexes has been attributed to their ability to inhibit the endogenous reverse transcriptase activity of RSV. More recent studies in our laboratory indicate that both these complexes inhibit both endogenous and exogenous reactions. As low a final concentration as 50 μM of the cupric and the cuprous complexes inhibits the endogenous reaction to the extent of 93 and 75 per cent respectively. Inhibition of the exogenous reaction varies with the templates. The inhibition can be reversed by either β-mercaptoethanol or ethylene-diamine-tetra-acetic acid. The specificity of this inhibition has been ascertained by using a synthetic primer-template, −(dG)not, vert, similar15−(rCm)n, which is highly specific for reverse transcriptases. The inhibition is found to be template specific. The studies carried out, using various synthetic primer-templates, show the inhibition of both the steps of reverse transcription by the copper complexes of isoniazid.

A Jacobian-Based Algorithm for Planning Attitude Maneuvers Using Forward and Reverse Rotations

Algorithms for planning quasistatic attitude maneuvers based on the Jacobian of the forward kinematic mapping of fully-reversed (FR) sequences of rotations are proposed in this paper. An FR sequence of rotations is a series of finite rotations that consists of initial rotations about the axes of a body-fixed coordinate frame and subsequent rotations that undo these initial rotations. Unlike the Jacobian of conventional systems such as a robot manipulator, the Jacobian of the system manipulated through FR rotations is a null matrix at the identity, which leads to a total breakdown of the traditional Jacobian formulation. Therefore, the Jacobian algorithm is reformulated and implemented so as to synthesize an FR sequence for a desired rotational displacement. The Jacobian-based algorithm presented in this paper identifies particular six-rotation FR sequences that synthesize desired orientations. We developed the single-step and the multiple-step Jacobian methods to accomplish a given task using six-rotation FR sequences. The single-step Jacobian method identifies a specific FR sequence for a given desired orientation and the multiple-step Jacobian algorithm synthesizes physically feasible FR rotations on an optimal path. A comparison with existing algorithms verifies the fast convergence ability of the Jacobian-based algorithm. Unlike closed-form solutions to the inverse kinematics problem, the Jacobian-based algorithm determines the most efficient FR sequence that yields a desired rotational displacement through a simple and inexpensive numerical calculation. The procedure presented here is useful for those motion planning problems wherein the Jacobian is singular or null.

Inhibition of avian myeloblastosis virus reverse transcriptase and virus inactivation by metal complexes of isonicotinic acid hydrazide

The cupric and ferric complexes of isonicotinic acid hydrazide (INH) inhibit the DNA synthesis catalysed by avian myeloblastosis virus (AMV) reverse transcriptase. The inhibition was to the extent of 95% by 50 μM of cupric-INH complex and 55% by 100 μM of ferric-INH complex. These complexes have been found to bind preferentially to the enzyme than to the template-primer. Kinetic analysis showed that the cupric-INH complex is a non-competitive inhibitor with respect to dTTP. The time course of inhibition has revealed that the complexes are inhibitory even after the initiation of polynucleotide synthesis. In vivo toxicity studies in 1-day-old chicks have shown that the complexes are not toxic up to a concentration of 500 μg per chick. Infection of the 1-day-old chicks with AMV pretreated with 150 μg of either of the complexes prevented symptoms of leukemia due to virus inactivation.

Effect of human telomerase reverse transcriptase transfection on differentiation in BeWo choriocarcinoma cells

Arrest of proliferation is one of the prerequisites for differentiation of cytotrophoblasts into syncytiotrophoblasts, and thus during differentiation telomerase activity, as well as human telomerase reverse transcriptase (hTERT) expression, is down-regulated. Considering this, it is of interest to investigate whether syncytium formation can be delayed by prolonging the expression of telomerase in cytotrophoblasts. BeWo cells were transfected with pLPC-hTERT retroviral vector and the reverse transcription-polymerase chain reaction analysis for hTERT mRNA concentrations in the transfected cells revealed a several-fold increase in hTERT mRNA compared with the cells transfected with empty vector, and this confirmed that the transfection was successful. An increase in the proliferation, as assessed by bromodeoxyuridine incorporation assay, as well as an increase in mRNA and protein concentration of various cyclins and proliferating cell nuclear antigen, was noticed. The effect of hTERT transfection was also assessed after the addition of forskolin to induce differentiation and it was observed that cell–cell fusion was delayed and differentiation did not occur in hTERT-transfected cells. However, the effects seen were only transient as stable transfection was not possible and the cells were undergoing apoptosis after 72 h, which suggested that apart from hTERT other factors might be important for immortalization of BeWo cells.

Reverse Channel Training For Reciprocal Mimo Systems With Spatial Multiplexing

This paper investigates the problem of designing reverse channel training sequences for a TDD-MIMO spatial-multiplexing system. Assuming perfect channel state information at the receiver and spatial multiplexing at the transmitter with equal power allocation to them dominant modes of the estimated channel, the pilot is designed to ensure an stimate of the channel which improves the forward link capacity. Using perturbation techniques, a lower bound on the forward link capacity is derived with respect to which the training sequence is optimized. Thus, the reverse channel training sequence makes use of the channel knowledge at the receiver. The performance of orthogonal training sequence with MMSE estimation at the transmitter and the proposed training sequence are compared. Simulation results show a significant improvement in performance.

Effect of cupric-isonicotinic acid hydrazide complex on avian myeloblastosis virus reverse transcriptase and its associated enzyme activities

Cupric complex of isonicotinic acid hydrazide inhibits DNA synthesis by avian myloblastosis virus reverse transcriptase. This inhibition occurs in the presence of either ribonucleotide or deoxyribonucleotide templates. The inhibition of reverse transcriptase by cupric-INH complex is considerably reduced when stored or proteolytically cleaved enzyme was used in the reaction. The complex also inhibits the reverse transciptase-associated RNase H activity. The cupric-isonicotinic acid hydrazide complex cleaves pBR 322 from I DNA into smaller molecules in the presence or absence of reverse transcriptase-associated endonuclease. However, in the presence of the enzyme the DNA is cleaved to a greater extent.

Highly regio-, chemo- and diastereoselective synthesis of oxa-bridged spirocycles: A novel observation of reverse selectivity

The reverse regio- and diastereoselectivities are observed between the reactions involving 5- and 6-membered-ring cyclic carbonyl ylide dipoles with alpha-methylene ketones. A mild catalytic route to synthesize spirocyclic systems with high regio-, chemo- and diastereoselectivities is described.

Reverse bias capacitance measurements on copper-doped germanium p-n junctions in the breakdown region

Results of measurements at a high frequency on reverse bias capacitance of copper-doped germanium junctions are reported. Phenomenal increase in capacitance is found in the breakdown region, particularly at low temperatures.

Large-scale structures in turbulent and reverse-transitional sink flow boundary layers

Aspects of large-scale organized structures in sink flow turbulent and reverse-transitional boundary layers are studied experimentally using hot-wire anemometry. Each of the present sink flow boundary layers is in a state of 'perfect equilibrium' or 'exact self-preservation' in the sense of Townsend (The Structure of Turbulent Shear Flow, 1st and 2nd edns, 1956, 1976, Cambridge University Press) and Rotta (Progr. Aeronaut. Sci., vol. 2, 1962, pp. 1-220) and conforms to the notion of 'pure wall-flow' (Coles, J. Aerosp. Sci., vol. 24, 1957, pp. 495-506), at least for the turbulent cases. It is found that the characteristic inclination angle of the structure undergoes a systematic decrease with the increase in strength of the streamwise favourable pressure gradient. Detectable wall-normal extent of the structure is found to be typically half of the boundary layer thickness. Streamwise extent of the structure shows marked increase as the favourable pressure gradient is made progressively severe. Proposals for the typical eddy forms in sink flow turbulent and reverse-transitional flows are presented, and the possibility of structural self-organization (i.e. individual hairpin vortices forming streamwise coherent hairpin packets) in these flows is also discussed. It is further indicated that these structural ideas may be used to explain, from a structural viewpoint, the phenomenon of soft relaminarization or reverse transition of turbulent boundary layers when subjected to strong streamwise favourable pressure gradients. Taylor's 'frozen turbulence' hypothesis is experimentally shown to be valid for flows in the present study even though large streamwise accelerations are involved, the flow being even reverse transitional in some cases. Possible conditions, which are required to be satisfied for the safe use of Taylor's hypothesis in pressure-gradient-driven flows, are also outlined. Measured convection velocities are found to be fairly close to the local mean velocities (typically 90% or more) suggesting that the structure gets convected downstream almost along with the mean flow.

Simulation of precipitation reactions in reverse micelles

Precipitation involving mixing of two sets of reverse micellar solutions-containing a reactant and precipitant respectively-has been analyzed. Particle formation in such systems has been simulated by a Monte Carlo (MC) scheme (Li, Y.; Park, C. W. Langmuir 1999, 15, 952), which however is very restrictive in its approach. We have simulated particle formation by developing a general Monte Carlo scheme, using the interval of quiescence technique (IQ). It uses Poisson distribution with realistic, low micellar occupancies of reactants, Brownian collision of micelles with coalescence efficiency, fission of dimers with binomial redispersion of solutes, finite nucleation rate of particles with critical number of molecules, and instantaneous particle growth. With the incorporation of these features, the previous work becomes a special case of our simulation. The present scheme was then used to predict experimental data on two systems. The first is the experimental results of Lianos and Thomas (Chem. Phys. Lett. 1986, 125, 299, J. Colloid Interface Sci. 1987, 117, 505) on formation of CdS nanoparticles. They reported the number of molecules in a particle as a function of micellar size and reactant concentrations, which have been predicted very well. The second is on the formation of Fe(OH)(3) nanoparticles, reported by Li and Park. Our simulation in this case provides a better prediction of the experimental particle size range than the prediction of the authors. The present simulation scheme is general and can be applied to explain nanoparticle formation in other systems.

Applications of reverse saturable absorbers in laser science

A variety of applications exist for reverse saturable absorbers (RSAs) in the area of optical pulse processing and computing. An RSA can be used as power limiter/pulse smoother and energy limiter/pulse shortner of laser pulses. A combination of RSA and saturable absorber (SA) can be used for mode locking and pulse shaping between high power laser amplifiers in oscillator amplifier chain. Also, an RSA can be used for the construction of a molecular spatial light modulator (SLM) which acts as an input/output device in optical computers. A detailed review of the theoretical studies of these processes is presented. Current efforts to find RSAs at desired wavelength for testing these theoretical predictions are also discussed.

Modeling of precipitation in reverse micellar systems

A model of the precipitation process in reverse micelles has been developed to calculate the size of fine particles obtained therein. While the method shares several features of particle nucleation and growth common to precipitation in large systems, complexities arise in describing the processes of nucleation, due to the extremely small size of a micelle and of particle growth caused by fusion among the micelles. Occupancy of micelles by solubilized molecules is governed by Poisson statistics, implying most of them are empty and cannot nucleate of its own. The model therefore specifies the minimum number of solubilized molecules required to form a nucleus which is used to calculate the homogeneous nucleation rate. Simultaneously, interaction between micelles is assumed to occur by Brownian collision and instantaneous fusion. Analysis of time scales of various events shows growth of particles to be very fast compared to other phenomena occurring. This implies that nonempty micelles either are supersaturated or contain a single precipitated particle and allows application of deterministic population balance equations to describe the evolution of the system with time. The model successfully predicts the experimental measurements of Kandori ct al.(3) on the size of precipitated CaCO3 particles, obtained by carbonation of reverse micelles containing aqueous Ca(OH)(2) solution.