FRAP analysis of nucleocytoplasmic dynamics of the vitamin D receptor splice variant VDRB1: preferential targeting to nuclear speckles

Although the key components of the cellular nuclear transport machinery have largely been characterized through extensive efforts in recent years, in vivo measurements of the kinetics of nuclear protein import/export are patently few. The present study applies the approach of FRAP (fluorescence recovery after photobleaching) to examine the nucleocytoplasmic flux of a novel human VDRB1 (vitamin D receptor B I) isoform in living cells. Through an N-terminal extension containing a consensus nuclear targeting sequence, VDRB1 is capable of localizing in nuclear speckles adjacent to SC-35 (35 kDa splicing component)containing speckles as well as in the nucleoplasm, dependent on ligand. Investigation of VDRB1 nucleocytoplasmic transport using FRAP indicates for the first time that the VDRB1 has a serum-modulated, active nuclear-import mechanism. There is no evidence of an efficient, active export mechanism for VDRB1, probably as a result of nuclear retention. VDRB1 nuclear import in the absence of serum occurred more rapidly and to a greater extent to nuclear speckles compared with import to other nuclear sites. This preferential transport from the cytoplasm to and accumulation within nuclear speckles is consistent with the idea that the latter represent dynamic centres of VDRB1 interaction with other nuclear proteins. The results are consistent with the existence of specialized pathways to target proteins to nuclear subdomains.

Homologues of the maize rust resistance gene Rp1-D are genetically associated with a major rust resistance QTL in sorghum

As part of a comparative mapping study between sugarcane and sorghum, a sugarcane cDNA clone with homology to the maize Rp1-D rust resistance gene was mapped in sorghum. The cDNA probe hybridised to multiple loci, including one on sorghum linkage group (LG) E in a region where a major rust resistance QTL had been previously mapped. Partial sorghum Rp1-D homologues were isolated from genomic DNA of rust-resistant and -susceptible progeny selected from a sorghum mapping population. Sequencing of the Rp1-D homologues revealed five discrete sequence classes: three from resistant progeny and two from susceptible progeny. PCR primers specific to each sequence class were used to amplify products from the progeny and confirmed that the five sequence classes mapped to the same locus on LG E. Cluster analysis of these sorghum sequences and available sugarcane, maize and sorghum Rp1-D homologue sequences showed that the maize Rp1-D sequence and the partial sugarcane Rp1-D homologue were clustered with one of the sorghum resistant progeny sequence classes, while previously published sorghum Rp1-D homologue sequences clustered with the susceptible progeny sequence classes. Full-length sequence information was obtained for one member of a resistant progeny sequence class (Rp1-SO) and compared with the maize Rp1-D sequence and a previously identified sorghum Rp1 homologue (Rph1-2). There was considerable similarity between the two sorghum sequences and less similarity between the sorghum and maize sequences. These results suggest a conservation of function and gene sequence homology at the Rp1 loci of maize and sorghum and provide a basis for convenient PCR-based screening tools for putative rust resistance alleles in sorghum.

Comments on alternative calculations of the broadening of spectral lines of neutral sodium by H-atom collisions

With the exception of the sodium D-lines, recent calculations of line broadening cross sections for several multiplets of sodium by Leininger et al (Leininger T, Gadea F X and Dickinson A 2000 J. Phys. B: At. Mol. Opt. Phys. 33 1805) are in substantial disagreement with cross sections interpolated from the tables of Anstee and O'Mara (Anstee and O'Mara 1995 Mon. Not. R. Astron. Soc. 276 859) and Barklem and O'Mara (Barklem P S and O'Mara B J 1997 Mon. Not. R. Astron. Soc. 290 102). The discrepancy is as large as a factor of 3 for the 3p-4d multiplet. The two theories are tested by using the results of each to synthesize lines in the solar spectrum. It is found that generally the data from the theory of Anstee, Barklem and O'Mara produce the best match to the observed solar spectrum. It is found, using a simple model for reflection of the optical electron by the potential barrier between the two atoms, that the reflection coefficient is too large for avoided crossings with the upper states of subordinate lines to contribute to line broadening, supporting the neglect of avoided ionic crossings by Anstee, Barklem and O'Mara for these lines. The large discrepancies between the two sets of calculations is a result of an approximate treatment of avoided ionic crossings for these lines by Leininger et al (Leininger T, Gadea F X and Dickinson A 2000 J. Phys. B: At. Mol. Opt. Phys. 33 1805).

Microstructural evolution of the Serido Belt, NE-Brazil: the effect of two tectonic events on development of c-axis preferred orientation in quartz

The polyphase evolution of the Serido Belt (NE-Brazil) includes D, crust formation at 2.3-2.1 Ga, D-2 thrust tectonics at 1.9 Ga and crustal reworking by D-3 strike-slip shear zones at 600 Ma. Microstructural investigations within mylonites associated with D-2 and D-3 events were used to constrain the tectono-thermal evolution of the belt. D-2 shear zones commenced at deeper crustal levels and high amphibolite facies conditions (600-650 degreesC) through grain boundary migration, subgrain rotation and operation of quartz Q-prism slip. Continued shearing and exhumation of the terrain forced the re-equilibration of high-T fabrics and the switching of slip systems from (c)-prism to positive and negative (a)-rhombs. During D-3, enhancement of ductility by dissipation of heat that came from syn-D-3 granites developed wide belts of amphibolite facies mylonites. Continued shearing, uplift and cooling of the region induced D-3 shear zones to act in ductile-brittle regimes, marked by fracturing and development of thinner belts of greenschist facies mylonites. During this event, switching from (a)-prism to a basal slip indicates a thermal path from 600 to 350 degreesC. Therefore, microstructures and quartz c-axis fabrics in polydeformed rocks from the Serido Belt preserve the record of two major events, which includes contrasting deformation mechanisms and thermal paths. (C) 2003 Elsevier Ltd. All rights reserved.

Implications of non-response of older women to a short form of the Center for Epidemiologic Studies Depression Scale

The Center for Epidemiologic Studies Depression Scale (CES-D) is frequently used in epidemiological surveys to screen for depression, especially among older adults. This article addresses the problem of non-completion of a short form of the CES-D (CESD-10) in a mailed survey of 73- to 78-year-old women enrolled in the Australian Longitudinal Study on Women's Health. Completers of the CESD-10 had more education, found it easier to manage on available income and reported better physical and mental health. The Medical Outcomes Study Short Form Health Survey (SF-36) scores for non-completers were intermediate between those for women classified as depressed and not depressed using the CESD-10. Indicators of depression had an inverted U-shaped relationship with the number of missing CESD- 10 items and were most frequent for women with two to seven items missing. Future research should pay particular attention to the level of missing data in depression scales and report its potential impact on estimates of depression.

Experimental and theoretical investigation of the structural, chemical, electronic, and high frequency dielectric properties of barium cadmium tantalate-based ceramics

Single-phase Ba(Cd1/3Ta2/3)O-3 powder was produced using conventional solid state reaction methods. Ba(Cd1/3Ta2/3)O-3 ceramics with 2 wt % ZnO as sintering additive sintered at 1550 degreesC exhibited a dielectric constant of similar to32 and loss tangent of 5x10(-5) at 2 GHz. X-ray diffraction and thermogravimetric measurements were used to characterize the structural and thermodynamic properties of the material. Ab initio electronic structure calculations were used to give insight into the unusual properties of Ba(Cd1/3Ta2/3)O-3, as well as a similar and more widely used material Ba(Zn1/3Ta2/3)O-3. While both compounds have a hexagonal Bravais lattice, the P321 space group of Ba(Cd1/3Ta2/3)O-3 is reduced from P (3) under bar m1 of Ba(Zn1/3Ta2/3)O-3 as a result of a distortion of oxygen away from the symmetric position between the Ta and Cd ions. Both of the compounds have a conduction band minimum and valence band maximum composed of mostly weakly itinerant Ta 5d and Zn 3d/Cd 4d levels, respectively. The covalent nature of the directional d-electron bonding in these high-Z oxides plays an important role in producing a more rigid lattice with higher melting points and enhanced phonon energies, and is suggested to play an important role in producing materials with a high dielectric constant and low microwave loss. (C) 2005 American Institute of Physics.

Wolbachia infections and the expression of cytoplasmic incompatibility in Drosophila sechellia and D. mauritiana

Various stocks of Drosophila mauritiana and D. sechellia were found to be infected with Wolbachia, a Rickettsia-like bacterium that is known to cause cytoplasmic incompatibility and other reproductive abnormalities in arthropods. Testing for the expression of cytoplasmic incompatibility in these two species showed partial incompatibility in D. sechellia but no expression of incompatibility in D. mauritiana. To determine whether absence of cytoplasmic incompatibility in D. mauritiana was due to either the bacterial or host genome, we transferred bacteria from D. mauritiana into an uninfected strain of D. simulans, a host species known to express high levels of incompatibility with endogenous Wolbachia. We also performed the reciprocal transfer of the natural D. simulans Riverside infection into a tetracycline-treated stock of D. mauritiana. In each case, the ability to express incompatibility was unaltered by the different host genetic background. These experiments indicate that in D. simulans and D. mauritiana expression of the cytoplasmic incompatibility phenotype is determined by the bacterial strain and that D. mauritiana harbors a neutral strain of Wolbachia.

Cytotoxicity of pilosulin 1, a peptide from the venom of the jumper ant Myrmecia pilosula

The synthetic peptide pilosulin 1, corresponding to the largest defined allergenic polypeptide found in the venom of the jumper ant Myrmecia pilosula, inhibited the incorporation of [methyl-H-3]thymidine into proliferating Epstein-Barr transformed (EBV) B-cells. The LD50 was four-fold lower in concentration than melittin, a cytotoxic peptide found in honey bee venom. Loss of cell viability was assessed by flow cytometry by measuring the proportion of cells that fluoresced in the presence of the fluorescent dye 7-aminoactinomycin D. Examination of proliferating EBV B-cells indicated that the cells lost viability within a few minutes exposure to pilosulin 1. Partial peptides of pilosulin 1 were less efficient in causing loss of cell viability and the results suggest that the 22 N-terminal residues are critical to the cytotoxic activity of pilosulin 1. Normal blood white cells were also labile to pilosulin I. T- and B-lymphocytes, monocytes and natural killer cells, however, were more labile than granulocytes. Analysis of pilosulin I using circular dichroism indicated that, in common with melittin and other Hymenoptera venom toxins, it had the potential to adopt an cc-helical secondary structure. (C) 1998 Elsevier Science B.V, All rights reserved.

Amphilactams A-D: Novel nematocides from southern Australian marine sponges of the genus Amphimedon

Bioassay-directed fractionation of the ethanol extracts of two Amphimedon spp. collected during trawling operations in the Great Australian Eight yielded four new macrocyclic lactone/lactams, amphilactams A-D (1-4). The amphilactams possess potent in vitro nematocidal properties, and their structures were assigned on the basis of detailed spectroscopic analysis and comparison with synthetic model compounds. The amphilactams feature both carbon skeletons and an enamino lactone/lactam moiety unprecedented in the natural products literature.

D-amino acid residue in the C-type natriuretic peptide from the venom of the mammal, Ornithorhynchus anatinus, the Australian platypus

The C-type natriuretic peptide from the platypus venom (OvCNP) exists in two forms, OvCNPa and OvCNPb, whose amino acid sequences are identical. Through the use of nuclear magnetic resonance, mass spectrometry, and peptidase digestion studies, we discovered that OvCNPb incorporates a D-amino acid at position 2 in the primary structure. Peptides containing a D-amino acid have been found in lower forms of organism, but this report is the first for a D-amino acid in a biologically active peptide from a mammal. The result implies the existence of a specific isomerase in the platypus that converts an L-amino acid residue in the protein to the D-configuration. (C) 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.