Transformation in heterokaryons of Neurospora crassa is nuclear rather than cellular phenomenon

In Neurospora crassa, multinucleate macroconidia are used for genetic transformation. The barrier for such a transformation can be either at the cell membrane level or at the nuclear membrane level. For assessment of these possibilities, a forced heterokaryon (containing two genetically marked nuclei and auxotrophic for histidine) of Neurospora crassa was transformed with a plasmid containing his-3(+) gene. The transformants, which could grow without histidine supplementation, were then resolved into component homokaryons to determine into which nucleus or nuclei the plasmid had entered. Our results suggest that the barrier for transformation in Neurospora crassa is at the nuclear level, not at the cell membrane level. In a heterokaryon containing two genetically distinct nuclei, plasmid DNA integrated into only one of the nuclear types at any instance, but never into both nuclear types. Thus, in Neurospora crassa, the competent nucleus is essential for the transformation event to take place, and at a given time only one type of nucleus is competent to take up the exogenous DNA. Genomic Southern analysis showed that the transformants harbor both ectopic and homologous integrations of the plasmid DNA. The type and number of integrations were reflected at the post-translational level, since the specific activity of histidinol dehydrogenase (the translation product of his-3+ gene) was variable among several transformants and always less than the level of the wild type.

Localisation of discrete change in a transformer winding: a network-function-loci approach

An entirely different approach for localisation of winding deformation based on terminal measurements is presented. Within the context of this study, winding deformation means, a discrete and specific change externally imposed at a particular position on the winding. The proposed method is based on pre-computing and plotting the complex network-function loci e.g. driving-point impedance (DPI)] at a selected frequency, for a meaningful range of values for each element (increasing and decreasing) of the ladder network which represents the winding. This loci diagram is called the nomogram. After introducing a discrete change, amplitude and phase of DPI are measured. By plotting this single measurement on the nomogram, it is possible to estimate the location and identify the extent of change. In contrast to the existing approach, the proposed method is fast, non-iterative and yields reasonably good localisation. Experimental results for actual transformer windings (interleaved and continuous disc) are presented.

Discrimination of Nuclear Grade Steel Samples Subjected to Different Heat Treatment Procedures Based on Acoustic Emission (AE) Data Profiling at Crack Initiation

The present work is an attempt to study crack initiation in nuclear grade, 9Cr-1Mo ferritic steel using AE as an online NDE tool. Laboratory experiments were conducted on 5 heat treated Compact Tension (CT) specimens made out of nuclear grade 9Cr-1Mo ferritic steel by subjecting them to cyclic tensile load. The CT Specimens were of 12.5 mm thickness. The Acoustic emission test system was setup to acquire the data continuously during the test by mounting AE sensor on one of the surfaces of the specimen. This was done to characterize AE data pertaining to crack initiation and then discriminate the samples in terms of their heat treatment processes based on AE data. The AE signatures at crack initiation could conclusively bring to fore the heat treatment distinction on a sample to sample basis in a qualitative sense.Thus, the results obtained through these investigations establish a step forward in utilizing AE technique as an on-line measurement tool for accurate detection and understanding of crack initiation and its profile in 9Cr-1Mo nuclear grade steel subjected to different processes of heat treatment.

Subdiffusion as a model of transport through the nuclear pore complex

Cargo transport through the nuclear pore complex continues to be a subject of considerable interest to experimentalists and theorists alike. Several recent studies have revealed details of the process that have still to be fully understood, among them the apparent nonlinearity between cargo size and the pore crossing time, the skewed, asymmetric nature of the distribution of such crossing times, and the non-exponentiality in the decay profile of the dynamic autocorrelation function of cargo positions. In this paper, we show that a model of pore transport based on subdiffusive particle motion is in qualitative agreement with many of these observations. The model corresponds to a process of stochastic binding and release of the particle as it moves through the channel. It suggests that the phenylalanine-glycine repeat units that form an entangled polymer mesh across the channel may be involved in translocation, since these units have the potential to intermittently bind to hydrophobic receptor sites on the transporter protein. (C) 2011 American Institute of Physics. [doi:10.1063/1.3651100]

Investigation on the High Vacuum Tribological Characteristics of Surface Treated Nuclear Grade Stainless Steel Type AISI 316 LN at 25 to 500 degrees C

Although some researchers have published friction and wear data of Plasma Nitride (PN) coatings, the tribological behavior of PN/PN Pairs in high vacuum environment has not been published so far In order to bridge this knowledge gap, tribological tests under dry conditions have been conducted on PN/PN Pairs for varying temperatures of 25, 200, 400 and 500 degrees C in high vacuum (1.6 x 10(-4) bar) environment. The PN coatings showed good wear resistance layer on the ring surface. The PN coatings were removed only from the pin surface for all the tests since it contacts at a point. The friction and wear were low at lower temperatures and it eliminated adhesion between the contact surfaces until the coating was completely removed from the pin surface. (C) 2011 Journal of Mechanical Engineering. All rights reserved.

Experimental implementation of quantumUlam’s problem in a nuclear magnetic resonance quantum information processor

The Ulam’s problem is a two person game in which one of the player tries to search, in minimum queries, a number thought by the other player. Classically the problem scales polynomially with the size of the number. The quantum version of the Ulam’s problem has a query complexity that is independent of the dimension of the search space. The experimental implementation of the quantum Ulam’s problem in a Nuclear Magnetic Resonance Information Processor with 3 quantum bits is reported here.

Functional regulation of PVBV Nuclear Inclusion protein-a protease activity upon interaction with Viral Protein genome-linked and phosphorylation

Regulation of NIa-Pro is crucial for polyprotein processing and hence, for successful infection of potyviruses. We have examined two novel mechanisms that could regulate NIa-Pro activity. Firstly, the influence of VPg domain on the proteolytic activity of NIa-Pro was investigated. It was shown that the turnover number of the protease increases when these two domains interact (as: two-fold; trans: seven-fold) with each other. Secondly, the protease activity of NIa-Pro could also be modulated by phosphorylation at Ser129. A mutation of this residue either to aspartate (phosphorylation-mimic) or alanine (phosphorylation-deficient) drastically reduces the protease activity. Based on these observations and molecular modeling studies, we propose that interaction with VPg as well as phosphorylation of Ser129 could relay a signal through Trp143 present at the protein surface to the active site pocket by subtle conformational changes, thus modulating protease activity of NIa-Pro. (C) 2011 Elsevier Inc. All rights reserved.

Novel ATPase activity of the polyprotein intermediate, Viral Protein genome-linked-Nuclear Inclusion-a protease, of Pepper vein banding potyvirus

Potyviruses temporally regulate their protein function by polyprotein processing. Previous studies have shown that VPg (Viral Protein genome-linked) of Pepper vein banding virus interacts with the NIa-Pro (Nuclear Inclusion-a protease) domain, and modulates the kinetics of the protease. In the present study, we report for the first time that VPg harbors the Walker motifs A and B, and the presence of NIa-Pro, especially in cis (cleavage site (E191A) VPg-Pro mutant), is essential for manifestation of the ATPase activity. Mutation of Lys47 (Walker motif A) and Asp88:Glu89 (Walker motif B) to alanine in E191A VPg-Pro lead to reduced ATPase activity, confirming that this activity was inherent to VPg. We propose that potyviral VPg, established as an intrinsically disordered domain, undergoes plausible structural alterations upon interaction with globular NIa-Pro which induces the ATPase activity. (C) 2012 Elsevier Inc. All rights reserved.

Nuclear targeting terpyridine iron(II) complexes for cellular imaging and remarkable photocytotoxicity

Iron(II) complexes Fe(L)(2)](2+) as perchlorate (1-3) and chloride (1a-3a) salts, where L is 4'-phenyl-2,2':6',2 `'-terpyridine (phtpy in 1, 1a), 4'-(9-anthracenyl)-2,2':6',2 `'-terpyridine (antpy in 2, 2a) and 4'-(1-pyrenyl)-2,2':6',2 `'-terpyridine (pytpy in 3, 3a), were prepared and their photocytotoxicity studied. The diamagnetic complexes 1-3 having an FeN6 core showed an Fe(III)-Fe(II) redox couple near 1.0 V vs. saturated calomel electrode in MeCN-0.1 M tetrabutylammonium perchlorate. Complexes 2 and 3, in addition, displayed a quasi-reversible ligand-based redox process near 0.0 V. The redox and spectral properties are rationalized from the theoretical studies. The complexes bind to DNA in a partial intercalative mode. The pytpy complex efficiently photo-cleaves DNA in green light via superoxide and hydroxyl radical formation. The antpy and pytpy complexes exhibited a remarkable photocytotoxic effect in HeLa cancer cells (IC50, similar to 9 mu M) in visible light (400-700 nm), while remaining essentially nontoxic in dark (IC50, similar to 90 mu M). Formation of reactive oxygen species (ROS) inside the HeLa cells was evidenced from the fluorescence enhancement of dichlorofluorescein upon treatment with the pytpy complex followed by photo-exposure. The antpy and pytpy complexes were used for cellular imaging. Confocal imaging and dual staining study using propidium iodide (PI) showed nuclear localization of the complexes. (c) 2012 Elsevier Inc. All rights reserved.

Observation of the nuclear magnetic octupole moment of Yb-173 from precise measurements of the hyperfine structure in the P-3(2) state

We measure hyperfine structure in the metastable P-3(2) state of Yb-173 and extract the nuclear magnetic octupole moment. We populate the state using dipole-allowed transitions through the P-3(1) and S-3(1) states. We measure frequencies of hyperfine transitions of the P-3(2) -> S-3(1) line at 770 nm using a Rb-stabilized ring cavity resonator with a precision of 200 kHz. Second-order corrections due to perturbations from the nearby P-3(1) and P-1(1) states are below 30 kHz. We obtain the hyperfine coefficients as A = -742.11(2) MHz and B = 1339.2(2) MHz, which represent a two orders-of-magnitude improvement in precision, and C = 0.54(2) MHz. From atomic structure calculations, we obtain the nuclear moments quadrupole Q = 2.46(12) b and octupole Omega = -34.4(21) b x mu(N). DOI: 10.1103/PhysRevA.87.012512