Interferon Gamma Regulation of De Novo Protein Synthesis in Human Dermal Fibroblasts in Culture Is Anatomic Site Dependent

The propensity of the skin of the lower anterior leg to be involved in Graves' dermopathy prompted an examination of the specific protein synthesis and response to interferon gamma in cultured fibroblasts from this area. Confluent cultures from normal skin of the lower leg and from the abdomen of the same three donors were pulse labeled with [35S]methionine for 3 h and subjected to two-dimensional protein gel electrophoresis and fluorography. Protein spots were mapped using a computer-driven program and the relative densities of the resolvable spots analyzed. Fibroblasts from the two anatomic sites display distinct patterns of de novo protein synthesis. Of the 157 abundant spots arbitrarily chosen for analysis, 31% varied substantially in levels of expression between the sites. A number of proteins appear to be expressed only in cultures derived from one of the two anatomic sites. Interferon gamma (100 U/ml) present in the culture medium for 48 h influenced the abundance of a number of proteins in a site-specific manner. Among them, plasminogen activator inhibitor type-I was induced three to five times in the leg cultures, whereas this same polypeptide was down-regulated in abdominal fibroblasts. A 54-kD protein was induced in interferon-treated cultures from both sites at least 50 times. It appears that fibroblasts from different regions of the integument are intrinsically distinct in terms of both their protein synthetic programs and their responses to cytokines

Fast, scalable master equation solution algorithms. III. Direct time propagation accelerated by a diffusion approximation preconditioned iterative solver

In this paper we propose a novel fast and linearly scalable method for solving master equations arising in the context of gas-phase reactive systems, based on an existent stiff ordinary differential equation integrator. The required solution of a linear system involving the Jacobian matrix is achieved using the GMRES iteration preconditioned using the diffusion approximation to the master equation. In this way we avoid the cubic scaling of traditional master equationsolution methods and maintain the low temperature robustness of numerical integration. The method is tested using a master equation modelling the formation of propargyl from the reaction of singlet methylene with acetylene, proceeding through long lived isomerizing intermediates

Attitude-behaviour consistency: the role of group norms, attitude accessibility, and mode of behavioural decision-making

This is the author's post-print version of an article published in the European Journal of Social Psychology, Vol 33, No 5: pp.591-608. The definitive version is available at www3.interscience.wiley.comThe interplay between two perspectives that have recently been applied in the attitude area—the social identity approach to attitude-behaviour relations (Terry & Hogg, 1996) and the MODE model (Fazio, 1990a)—was examined in the present research. Two experimental studies were conducted to examine the role of group norms, group identification, attitude accessibility, and mode of behavioural decision-making in the attitude-behaviour relationship. In Study 1 (Nu=u211), the effects of norms and identification on attitude-behaviour consistency as a function of attitude accessibility and mood were investigated. Study 2 (Nu=u354) replicated and extended the first experiment by using time pressure to manipulate mode of behavioural decision-making. As expected, the effects of norm congruency varied as a function of identification and mode of behavioural decision-making. Under conditions assumed to promote deliberative processing (neutral mood/low time pressure), high identifiers behaved in a manner consistent with the norm. No effects emerged under positive mood and high time pressure conditions. In Study 2, there was evidence that exposure to an attitude-incongruent norm resulted in attitude change only under low accessibility conditions. The results of these studies highlight the powerful role of group norms in directing individual behaviour and suggest limited support for the MODE model in this context

Rapid detection of Listeria monocytogenes in food using culture enrichment combined with real-time PCR.

A rapid method for the detection of Listeria monocytogenes in foods combining culture enrichment and real-time PCR was compared to the ISO 11290-1 standard method. The culture enrichment component of the rapid method is based on the ISO standard and includes 24h incubation in half-Fraser broth, 4h incubation in Fraser broth followed by DNA extraction and real-time PCR detection of the ssrA gene of L. monocytogenes. An internal amplification control, which is co-amplified with the same primers as the L. monocytogenes DNA, was also included in the assay. The method has a limit of detection of 1-5CFU/25g food sample and can be performed in 2 working days compared to up to 7days for the ISO standard. A variety of food samples from retail outlets and food processing plants (n=175) and controls (n=31) were tested using rapid and conventional methods. The rapid method was 99.44% specific, 96.15% sensitive and 99.03% accurate when compared to the standard method. This method has the potential to be used as an alternative to the standard method for food quality assurance providing rapid detection of L. monocytogenes in food

A model for tetrapyrrole synthesis as the primary mechanism for plastid-to-nucleus signaling during chloroplast biogenesis.

Chloroplast biogenesis involves the co-ordinated expression of the chloroplast and nuclear genomes, requiring information to be sent from the developing chloroplasts to the nucleus. This is achieved through retrograde signaling pathways and can be demonstrated experimentally using the photobleaching herbicide, norflurazon, which in seedlings results in chloroplast damage and the reduced expression of many photosynthesis-related, nuclear genes. Genetic analysis of this pathway points to a major role for tetrapyrrole synthesis in retrograde signaling, as well as a strong interaction with light signaling pathways. Currently, the best model to explain the genetic data is that a specific heme pool generated by flux through ferrochelatase-1 functions as a positive signal to promote the expression of genes required for chloroplast development. We propose that this heme-related signal is the primary positive signal during chloroplast biogenesis, and that treatments and mutations affecting chloroplast transcription, RNA editing, translation, or protein import all impact on the synthesis and/or processing of this signal. A positive signal is consistent with the need to provide information on chloroplast status at all times. We further propose that GUN1 normally serves to restrict the production of the heme signal. In addition to a positive signal re-enforcing chloroplast development under normal conditions, aberrant chloroplast development may produce a negative signal due to accumulation of unbound chlorophyll biosynthesis intermediates, such as Mg-porphyrins. Under these conditions a rapid shut-down of tetrapyrrole synthesis is required. We propose that accumulation of these intermediates results in a rapid light-dependent inhibition of nuclear gene expression that is most likely mediated via singlet oxygen generated by photo-excitation of Mg-porphyrins. Thus, the tetrapyrrole pathway may provide both positive and inhibitory signals to control expression of nuclear genes

tmRNA - a novel high-copy-number RNA diagnostic target - its application for Staphylococcus aureus detection using real-time NASBA

A real-time nucleic acid sequence-based amplification assay, targeting tmRNA, was designed for the rapid identification of Staphylococcus aureus. The selectivity of the assay was confirmed against a panel of 76 Staphylococcus strains and species and 22 other bacterial species. A detection limit of 1 cell equivalent was determined for the assay. A chimeric in vitro transcribed internal amplification control was developed and included in the assay. Application of the assay in natural and artificially contaminated unpasteurized (raw) milk enabled detection of 1-10 CFUS. aureus mL(-1) in 3-4 h, without the need for culture enrichment. Staphylococcus aureus was detected in all artificially contaminated milk samples (n=20) and none of the natural milk samples (n=20). Microbiological analysis of the natural milk samples was performed in parallel according to ISO 6888-3 and confirmed the absence of S. aureus. The method developed in this study has the potential to enable the specific detection of S. aureus in raw milk in a significantly shorter time frame than current standard methods. The assay further demonstrates the usefulness of tmRNA/ssrA as a nucleic acid diagnostic target

LP 400-22, A very low-mass and high-velocity white dwarf

We report the identification of LP 400-22 (WD 2234+222) as a very low-mass and high-velocity white dwarf. The ultraviolet GALEX and optical photometric colors and a spectral line analysis of LP 400-22 show this star to have an effective temperature of 11080+/-140 K and a surface gravity of log g = 6.32+/-0.08. Therefore, this is a helium core white dwarf with a mass of 0.17 M_solar. The tangential velocity of this white dwarf is 414+/-43 km/s, making it one of the fastest moving white dwarfs known. We discuss probable evolutionary scenarios for this remarkable object.Comment: accepted for publication in ApJ Letters, made minor correction

Axial Multicentric Osteosarcoma in an English Cocker Spaniel

No abstract available

Ghrelin in the human myometrium

<p>Abstract</p> <p>Background</p> <p>Ghrelin is a 28-amino acid octanolyated peptide, synthesised primarily in the stomach. It stimulates growth hormone release, food intake and exhibits many other diverse effects. Our group have previously determined that ghrelin inhibited human contractility in <it>vitro</it>. The aim of this study therefore, was to investigate the expression of ghrelin, its receptor, the growth hormone secretagogue receptor type 1 (GHS-R1), ghrelin O-acyltransferase (GOAT) which catalyses ghrelin octanoylation, prohormone convertase 1/3 (PC1/3) responsible for pro-ghrelin processing, in human myometrium, during pregnancy prior to labour, during labour and in the non-pregnant state. Modulation of ghrelin and ghrelin receptor expression in cultured myometrial cells was also investigated.</p> <p>Methods</p> <p>mRNA and protein were isolated from human myometrium and the myometrial smooth muscle cell line hTERT-HM; and real-time fluorescence RT-PCR, western blotting and fluorescence microscopy performed. The effects of β-Estradiol and bacterial lipopolysaccharide (LPS) on hTERT-HM gene expression were evaluated by western blotting.</p> <p>Results</p> <p>We have reported for the first time the expression and processing of ghrelin, GHS-R1, GOAT and PC1/3 expression in human myometrium, and also the down-regulation of ghrelin mRNA and protein expression during labour. Furthermore, GHS-R1 protein expression significantly decreased at labour. Myometrial GOAT expression significantly increased during term non-labouring pregnancy in comparison to both non-pregnant and labouring myometrium. Mature PC1/3 protein expression was significantly decreased at term pregnancy and labour in comparison to non-pregnant myometrium. Ghrelin, GHS-R1, GOAT and PC1/3 mRNA and protein expression was also detected in the hTERT-HM cells. Ghrelin protein expression decreased upon LPS treatment in these cells while β-Estradiol treatment increased GHS-R1 expression.</p> <p>Conclusions</p> <p>Ghrelin processing occurred in the human myometrium at term pregnancy and in the non-pregnant state. GOAT expression which increased during term non-labouring pregnancy demonstrating a similar expression pattern to prepro-ghrelin and GHS-R1, decreased at labour, signifying possible myometrial ghrelin acylation. Moreover, the presence of PC1/3 may contribute to pro-ghrelin processing. These results along with the previous <it>in vitro </it>data suggest that myometrially-produced and processed ghrelin plays a significant autocrine or paracrine role in the maintenance of relaxation in this tissue during pregnancy. Furthermore, the significant uterine modulators LPS and β-Estradiol are involved in the regulation of ghrelin and ghrelin receptor expression respectively, in the human myometrium.</p

Coherence and Intermittency of Electron Density in Small-Scale Interstellar Turbulence

Spatial intermittency in decaying kinetic Alfven wave turbulence is investigated to determine if it produces non Gaussian density fluctuations in the interstellar medium. Non Gaussian density fluctuations have been inferred from pulsar scintillation scaling. Kinetic Alfven wave turbulence characterizes density evolution in magnetic turbulence at scales near the ion gyroradius. It is shown that intense localized current filaments in the tail of an initial Gaussian probability distribution function possess a sheared magnetic field that strongly refracts the random kinetic Alfven waves responsible for turbulent decorrelation. The refraction localizes turbulence to the filament periphery, hence it avoids mixing by the turbulence. As the turbulence decays these long-lived filaments create a non Gaussian tail. A condition related to the shear of the filament field determines which fluctuations become coherent and which decay as random fluctuations. The refraction also creates coherent structures in electron density. These structures are not localized. Their spatial envelope maps into a probability distribution that decays as density to the power -3. The spatial envelope of density yields a Levy distribution in the density gradient.Comment: 31 pages, 3 figures. Replacement contains short additions to Secs. 6 and