Characterization of Mycobacterium leprae RecA intein, a LAGLIDADG homing endonuclease, reveals a unique mode of DNA-binding, helical distortion and cleavage, compared to a canonical LAGLIDADG homing endonuclease

Mycobacterium leprae, which has undergone reductive evolution leaving behind a minimal set of essential genes, has retained intervening sequences in four of its genes implicating a vital role for them in the survival of the leprosy bacillus. A single in-frame intervening sequence has been found embedded within its recA gene. Comparison of M. leprae recA intervening sequence with the known intervening sequences indicated that it has the consensus amino acid sequence necessary for being a LAGLIDADG-type homing endonuclease. In light of massive gene decay and function loss in the leprosy bacillus, we sought to investigate whether its recA intervening sequence encodes a catalytically active homing endonuclease. Here we show that the purified M. leprae RecA intein (PI-MleI) binds to cognate DNA and displays endonuclease activity in the presence of alternative divalent cations, Mg2+ or Mn2+. A combination of approaches including four complementary footprinting assays such as DNase I, Cu/phenanthroline, methylation protection and KMnO4, enhancement of 2-aminopurine fluorescence and mapping of the cleavage site revealed that PI-MleI binds to cognate DNA flanking its insertion site, induces helical distortion at the cleavage site and generates two staggered double-strand breaks. Taken together, these results implicate that PI-MleI possess a modular structure with separate domains for DNA target recognition and cleavage, each with distinct sequence preferences. From a biological standpoint, it is tempting to speculate that our findings have implications for understanding the evolution of LAGLIDADG family of homing endonucleases

Dislocation after total hip replacement: there is no such thing as a safe zone for socket placement with the posterior approach

Introduction Malorientation of the socket contributes to instability after hip arthroplasty but the optimal orientation of the cup in relation to the pelvis has not been unequivocally described. Large radiological studies are few and problems occur with film standardisation, measurement methodology used and alternative definitions of describing acetabular orientation. Methods A cohort of 1,578 patients from a single institution is studied where all patient data was collected prospectively. Risk factors for patients undergoing surgery are analysed. Radiological data was compared between a series of non-dislocating hips and dislocating cases matched 2:1 by operation type, age and diagnosis. Results The overall dislocation rate for all 1,578 cases was 3.23% but the rate varied according to the type of surgery performed. The rate in uncomplicated primary cases was 2.4% which increased to 9.3% for second stage implantation for a two stage procedure for infection. There was no significant difference in the variability of the dislocating and non-dislocating groups for either inclination (p = 0.393) or anteversion (p = 0.661). Conclusions A “safe zone” for socket orientation to avoid dislocation could not be defined. The cause of dislocation is multifactorial, re-establishing the anatomic centre of rotation, balancing soft tissues and avoidance of impingement around the hip are important considerations.

Tabulation of total marriages, births, deaths, bar mitzvahs

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Harmonic response of a shock mount employing dual-phase damping

In this note, the application of dual-phase damping to a simple shock mount experiencing a harmonic input is described. The damping ratio is a function of the relative displacement between the foundation and the mounted mass. The purpose of employing such a damping is to reduce the absolute transmissibility over the whole frequency range.

Impulse Voltage Breakdown Characteristics of Point-Plane Gaps at Low Pressures

Practical applications of vacuum as an insulator necessitated determining the low-pressure breakdown characteristics of long gap lengths of a point-plane electrode system. The breakdown voltage has been found to vary as the square root of the gap length. Further, with the point electrode as the anode, the values of the breakdown voltages obtained have been found to be larger than those obtained with a plane-parallel electrode system at a corresponding gap length. By applying the theory of the anode heating mechanism as the cause for breakdown, the results have been justified, and by utilizing a field efficiency factor which is the ratio of the average to maximum field, an empirical criterion has been developed. This criterion helps in calculating the breakdown voltage of a nonuniform gap system by the knowledge of the breakdown voltage of a plane-parallel electrode system.

Rapid synthetic routes to prepare LiNi1/3Mn1/3Co1/3O2 as a high voltage, high-capacity Li-ion battery cathode material

LiNi1/3Mn1/3Co1/3O2, a high voltage and high-capacity cathode material for Li-ion batteries, has been synthesized by three different rapid synthetic methods. viz. nitrate-melt decomposition, combustion and sol-gel methods. The first two methods are ultra rapid and a time period as small as 15 min is sufficient to prepare nano-crystalline LiNi1/3Mn1/3Co1/3O2. The processing parameters in obtaining the best performing materials are optimized for each process and their electrochemical performance is evaluated in Li-ion cells. The combustion-derived LiNi1/3Mn1/3Co1/3O2 sample exhibits large extent of cation mixing (10%) while the other two methods yield LiNi1/3Mn1/3Co1/3O2 with cation mixing <5%. LiNi1/3Mn1/3Co1/3O2 prepared by nitrate-melt decomposition method exhibits superior performance as Li-ion battery cathode material.

Essays on Economic Integration and Labour Demand

This dissertation consists of an introductory section and three essays investigating the effects of economic integration on labour demand by using theoretical models and by empirical analysis. The essays adopt an intra-industry trade approach to specify a theoretical framework of estimation for determining the effects of economic integration on employment. In all the essays the empirical aim is to explore the labour demand consequences of European integration. The first essay analyzes how labour-demand elasticities with own price have changed during the process of economic integration. As a theoretical result, intensified trade competition increases labour-demand elasticity, whereas better advantage of economies of scale decreases labour-demand elasticity by decreasing the elasticity of substitution between differentiated products. Furthermore, if integration gives rise to an increase in input-substitutability and/or outsourcing activities, labour demand will become more elastic. Using data from the manufacturing sector from 1975 to 2002, the empirical results provide support for the hypothesis that European integration has contributed to increased elasticities of total labour demand in Finland. The second essay analyzes how economic integration affects the impact of welfare poli-cies on employment. The essay considers the viability of financing the public sector, i.e. public consumption and social security expenses, by general labour taxation in an economy which has become more integrated into international product markets. The theoretical results of the second essay indicate that, as increased trade competition crowds out better economies of scale, it becomes more costly to maintain welfare systems financed by labour taxation. Using data from European countries for the years 1975 to 2004, the empirical results provide inconsistent evidence for the hypothesis that economic integration has contributed to the distortion effects of welfare policies on employment. The third essay analyzes the impact of profit sharing on employment as a way to introduce wage flexibility into the process of economic integration. The results of the essay suggest that, in theory, the effects of economic integration on the impact of profit sharing on employment clearly depend on a trade-off between intensified competition and better advantage of economies of scale. If product market competition increases, the ability of profit sharing to improve employment through economic integration increases with moderated wages. While, the economic integration associating with market power in turn decrease the possibilities of profit sharing with higher wages to improve employment. Using data from the manufacturing sector for the years 1996 to 2004, the empirical results show that profit-sharing has a positive impact on employment during the process of European integration, but can have ambiguous effects on the stability of employment in Finland.

Tools for compositional data with a total

Compositional data analysis usually deals with relative information between parts where the total (abundances, mass, amount, etc.) is unknown or uninformative. This article addresses the question of what to do when the total is known and is of interest. Tools used in this case are reviewed and analysed, in particular the relationship between the positive orthant of D-dimensional real space, the product space of the real line times the D-part simplex, and their Euclidean space structures. The first alternative corresponds to data analysis taking logarithms on each component, and the second one to treat a log-transformed total jointly with a composition describing the distribution of component amounts. Real data about total abundances of phytoplankton in an Australian river motivated the present study and are used for illustration.

Rapid distortion of axisymmetric turbulence

A generalization of the isotropic theory of Batchelor & Proudman (1954) is developed to estimate the effect of sudden but arbitrary three-dimensional distortion on homogeneous, initially axisymmetric turbulence. The energy changes due to distortion are expressed in terms of the Fourier coefficients of an expansion in zonal harmonics of the two independent scalar functions that describe the axisymmetric spectral tensor. However, for two special but non-trivial forms of this tensor, which represent possibly the simplest kinds of non-isotropic turbulence and specify the angular distribution but not the wavenumber dependence, the energy ratios have been determined in closed form. The deviation of the ratio from its isotropic value is the product of a factor containing R, the initial value of the ratio of the longitudinal to the transverse energy component, and another factor depending only on the geometry of the distortion. It is found that, in axisymmetric and large two-dimensional contractions, the isotropic theory gives nearly the correct longitudinal energy, but (when R > 1) over-estimates the increase in the transverse energy; the product of the two intensities varies little unless the distortion is very large, thus accounting for the stress-freezing observed in rapidly accelerated shear flows.Comparisons with available experimental data for the spectra and for the energy ratios show reasonable agreement. The different ansatzes predict results in broad qualitative agreement with a simple strategem suggested by Reynolds & Tucker (1975), but the quantitative differences are not always negligible.

Time-Domain Measurement of Voltage Induction by Transient EM Fields

An efficient measurement technique for studying the effect of transient electromagnetic fields under controlled conditions has been described. Broad-band TEM fields with a rise-time of a few nanoseconds were generated using a stripline method. Theoretical results are obtained and experimental measurements which confirm these results are described. The work will form the basis for a study of the susceptibility of digital integrated circuits and their interconnections to transient electromagnetic fields.

Significance of Branching Angles in Streamer Discharges

The phenomenon of branching at specific angles in streamer breakdown studies is found to be more universal than was ever thought. The angles measured in the breakdown of gases show that the coefficient of field distortion, K, lies in the range from 1 to less than 0.1. The values of K, so obtained agree well with those envisaged in the criterion of the streamer mechanism. It is hoped that branching angles observed in various types of breakdown may, possibly, be explained similarly.

Improvement in the voltage grading (axial and radial) of the generator column of a Van de Graaf generator by the use of a resistor chain

It is well known that the use of a series of resistors, connected between the equipotential rings of a Van de Graaff generator, improves the axial voltage grading of the generator. The work reported in this paper shows how the resistor chain also improves the radial voltage gradient. The electrolytic field mapping technique was adopted in the present work.

Dissipative dynamics of a harmonic oscillator: A nonperturbative approach

Starting from a microscopic theory, we derive a master equation for a harmonic oscillator coupled to a bath of noninteracting oscillators. We follow a nonperturbative approach, proposed earlier by us for the free Brownian particle. The diffusion constants are calculated analytically and the positivity of the master equation is shown to hold above a critical temperature. We compare the long time behavior of the average kinetic and potential energies with known thermodynamic results. In the limit of vanishing oscillator frequency of the system, we recover the results of the free Brownian particle.

Specific polyadenylation and purification of total messenger RNA from Escherichia coli

Obtaining pure mRNA preparations from prokaryotes has been difficult, if not impossible, for want of a poly(A) tail on these messages, We have used poly(A) polymerase from yeast to effect specific polyadenylation of Escherichia coli polysomal mRNA in the presence of magnesium and manganese, The polyadenylated total mRNA, which could be subsequently purified by binding to and elution from oligo(dT) beads, had a size range of 0.4-4.0 kb. We have used hybridization to a specific plasmid-encoded gene to further confirm that the polyadenylated species represented mRNA, Withdrawal of Mg2+ from the polyadenylation reaction rRNA despite the presence of Mn2+, indicating the vital role of Mg2+ in maintaining the native structure of polysomes, Complete dissociation of polysomes into ribosomal subunits resulted in quantitative polyadenylation of both 16S and 23S rRNA species, Chromosomal lacZ gene-derived messages were quantitatively recovered in the oligo(dT)-bound fraction, as demonstrated by RT-PCR analysis, Potential advantages that accrue from the availability of pure total mRNA from prokaryotes is discussed.

Light- and cytokinin-induced changes in the levels of leucine and tyrosine isoaccepting tRNA species and modified nucleotide contents of total tRNA in cucumber seedlings

We have determined relative levels of chloroplast leucine and tyrosine isoaccepting tRNAs and modified nucleotide contents from total tRNAs isolated from dark-grown, light-grown, N6-isopentenyladenine (i6A)-treated dark-grown and i6A-treated light-grown cucumber seedlings. Significant increases in the relative amounts of tRNA(Leu)2 and tRNA(Leu)3 were observed in the i6A-treated dark-grown seedlings compared to dark-grown, light-grown and i6A-treated light-grown seedlings. On the other hand, i6A-treated light-grown seedlings tRNA(Tyr)1 increased to 85% of total tRNAs(Tyr) from about 9% in light-grown seedlings and tRNA(Tyr)2 decreased to 15% compared with 91% in light-grown seedlings. Analysis of modified nucleotide of total tRNAs indicated that pT, pI, pm1A, pm5C, pGm, pm1G, pm2G and pm7G contents were significantly higher in the total tRNA of i6A-treated dark-grown seedlings than those from untreated dark-grown seedlings. Illumination of 8-day-old dark-grown seedlings for 12 h increased the contents of pT, pI, pGm and pm1G when compared to 8-day-old dark-grown seedlings with extended growth for 12 h in dark. On the contrary, i6A had no stimulatory effect in the contents of modified nucleotide in the light-grown seedlings.