The Septin-Binding Protein Anillin is Over-Expressed in Diverse Human Tumours

Anillin is an actin-binding protein that can bind septins and is a component of the cytokinetic ring. We assessed the anillin expression in 7,579 human tissue samples and cell lines by DNA micro-array analysis. Anillin is expressed ubiquitously but with variable levels of expression, being highest in the central nervous system. The median level of anillin mRNA expression was higher in tumors than normal tissues (median fold increase 2.58; 95% confidence intervals, 2.19-5.68, P < 0.0001) except in the central nervous system where anillin in RNA levels were lower in tumors. We developed a sensitive reverse transcription-PCR strategy to show that anillin mRNA is expressed in cell lines and in cDNA panels derived from fetal and adult tissues, thus validating the microarray data. We compared anillin with Ki67 in RNA expression and found a significant linear relationship between anillin and Ki67 mRNA expression (Spearmann r similar to 0.6, P < 0.0001). Anillin mRNA expression was analyzed during tumor progression in breast, ovarian, kidney, colorectal, hepatic, lung, endometrial, and pancreatic tumors and in all tissues there was progressive, increase in anillin mRNA expression from normal to benign to malignant to metastatic disease. Finally, we used anti-anillin sera and found nuclear anillin immuncireactivity to be widespread in normal tissues, often not correlating with proliferative compartments. These data provide insight into the existence of non proliferation-associated activities of anillin and roles in interphase nuclei. Thus, anillin is overexpressed in diverse common human tumors, but not simply as a consequence of being a proliferation marker. Anillin may have potential as a novel biomarker.

SEPT9_v4 expression induces morphological change, increased motility and disturbed polarity

Several lines of evidence indicate that altered expression of SEPT9 is seen in human neoplasia. In particular there is evidence of altered expression of the SEPT9_v4 isoform. The functional consequences of this remain unclear. We have studied the expression of wild-type- and GTP-binding mutants (G144V and S148N) of the SEPT9_v4 isoform in the MCF7 cell line as a model for its deregulation in neoplasia. We find that SEPT9_v4 expression induces dramatic actin cytoskeletal reorganization with the formation of processes around the cell periphery. Expression of the SEPT9_v4 isoform and a G144V mutant cause delocalization of endogenous SEPT9 from filamentous structures but the S148N mutant does not have this effect. In addition SEPT9_v4 isoform expression enhances cell motility and is associated with perturbation of directional movement. Expression of SEPT9_v4 GTP binding mutants also has potent effects on morphology and motility and causes loss of normal polarity, as judged by Golgi reorientation assays. The phenotypes induced by expression of the SEPT9_v4 isoform and the GTP mutants provide an insight into possible mechanisms of SEPT9_v4 function and suggest that the GTPase functions have both ras- and rab-like features. We propose a model in which overexpression of the SEPT9_v4 isoform in neoplasia is associated with perturbation of SEPT9 complexes, leading to phenotypes associated with neoplasia. Copyright (c) 2005 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

A peculiar metal-rich star, HD 135485

Local thermodynamic equilibrium (LTE) absolute and differential abundances are presented for a peculiar metal-rich B-type star, HD 135485. These suggest that HD 135485 has a general enrichment of similar to0.5 dex in all the metals observed (C, N, O, Ne, Mg, Al, Si, P, S, Cl, Ar, Sc, Ti, Cr, Mn, Fe and Sr), except for nickel. The helium enhancement and hence hydrogen deficiency can account for less than or equal to 0.2 dex of this enhancement of metals, with the additional enhancement probably being representative of the progenitor gas. However, some of the metals appear to have greater enhancements, which may have occurred during the star's evolution. The significantly larger nitrogen abundance coupled with a modest helium enhancement observed in HD 135485 indicates that carbon- nitrogen (CN) processed material has possibly contaminated the stellar surface. Neon and carbon enhancements may indicate that helium core flashes have also occurred in HD 135485. Some of the iron-group elements (viz. Mn and Ni) appear to have similar abundance patterns to that of silicon Ap stars, but it is uncertain how these abundance patterns formed if they were not present in the progenitor gas. From a kinematical investigation it is unclear whether this star formed in a metal-rich region as implied by its chemical composition. From its position in the Hertzsprung-Russell diagram, HD 135485 would appear to be an evolved star lying close to or on the horizontal branch.

Altered expression of the septin gene, SEPT9, in ovarian neoplasia.

The septin family of genes has been implicated in a variety of cellular processes including cytokinesis, membrane transport and fusion, exocytosis, and apoptosis. One member of the septin family maps to chromosome 17q25.3, a region commonly deleted in sporadic ovarian and breast tumours, and has also been identified as a fusion partner of MLL in acute myeloid leukaemias. The present study demonstrates that the pattern of expression of multiple splice variants of this septin gene is altered in ovarian tumours and cell lines. In particular, expression of the zeta transcript is detectable in the majority of tumours and cell lines, but not in a range of non-malignant adult and fetal tissues. Zeta expression is accompanied by loss of the ubiquitous beta transcript. Somatic mutations of the gene were not detected in ovarian tumours, but it was demonstrated that beta expression in tumour cell lines can be reactivated by 5-azacytidine treatment, suggesting a role for methylation in the control of expression of this gene. Copyright © 2003 John Wiley & Sons, Ltd.

The trade unions membership dynamics and union members’ trajectories

This text presents an analysis of aggregated membership’s dynamics for Spanish trade unions, using ECVT data, as well as union memberships’ trajectories, or members’ decisions about joining the organization, permanency and responsibilities, and subsequent attrition. For the analysis of trajectories we make use of information of the records of actual memberships and the record of quitting of CCOO, and of a survey-questionnaire to a sample of leavers of the same union. This study allows us to confirm a linkage between the decision and motivations to become union member, to participate in union activities, the time of permanency, and the motives to quit the organization. We also identify five types of union members’ trajectories, indicating that, far from views that assert a monolithic structure, unions are complex organizations.

The VLT-FLAMES survey of massive stars: stellar parameters and rotational velocities in NGC 3293, NGC 4755 and NGC 6611

An analysis is presented of VLT-FLAMES spectroscopy for three Galactic clusters, NGC3293, NGC4755 and NGC6611. Non-LTE model atmosphere calculations have been used to estimate effective temperatures (from either the helium spectrum or the silicon ionization equilibrium) and gravities (from the hydrogen spectrum). Projected rotational velocities have been deduced from the helium spectrum (for fast and moderate rotators) or the metal line spectrum (for slow rotators). The origin of the low gravity estimates for apparently near main sequence objects is discussed and is related to the stellar rotational velocity. The atmospheric parameters have been used to estimate cluster distances (which are generally in good agreement with previous determinations) and these have been used to estimate stellar luminosities and evolutionary masses. The observed Hertzsprung-Russell diagrams are compared with theoretical predictions and some discrepancies including differences in the main sequence luminosities are discussed. Cluster ages have been deduced and evidence for non-coeval star formation is found for all three of the clusters. Projected rotational velocities for targets in the older clusters, NGC3293 and NGC4755, have been found to be systematically larger than those for the field, confirming recent results in other similar age clusters. The distribution of projected rotational velocities are consistent with a Gaussian distribution of intrinsic rotational velocities. For the relatively unevolved targets in the older clusters, NGC3293 and NGC4755, the peak of the velocity distribution would be 250 km s(-1) with a full-width-half-maximum of approximately 180 km s(-1). For NGC6611, the sample size is relatively small but implies a lower mean rotational velocity. This may be evidence for the spin-down effect due to angular momentum loss through stellar winds, although our results are consistent with those found for very young high mass stars. For all three clusters we deduce present day mass functions with Gamma-values in the range of -1.5 to -1.8, which are similar to other young stellar clusters in the Milky Way.

The VLT-FLAMES survey of massive stars: mass loss and rotation of early-type stars in the SMC

We have studied the optical spectra of a sample of 31 O- and early B-type stars in the Small Magellanic Cloud, 21 of which are associated with the young massive cluster NGC 346. Stellar parameters are determined using an automated fitting method (Mokiem et al. 2005, A&A, 441, 711), which combines the stellar atmosphere code FASTWIND (Puls et al. 2005, A&A, 435, 669) with the genetic algorithm based optimisation routine PIKAIA (Charbonneau 1995, ApJS, 101, 309). Comparison with predictions of stellar evolution that account for stellar rotation does not result in a unique age, though most stars are best represented by an age of 1-3 Myr. The automated method allows for a detailed determination of the projected rotational velocities. The present day v(r) sin i distribution of the 21 dwarf stars in our sample is consistent with an underlying rotational velocity (v(r)) distribution that can be characterised by a mean velocity of about 160-190 km s(-1) and an effective half width of 100-150 km s(-1). The vr distribution must include a small percentage of slowly rotating stars. If predictions of the time evolution of the equatorial velocity for massive stars within the environment of the SMC are correct (Maeder & Meynet 2001, A&A, 373, 555), the young age of the cluster implies that this underlying distribution is representative for the initial rotational velocity distribution. The location in the Hertzsprung-Russell diagram of the stars showing helium enrichment is in qualitative agreement with evolutionary tracks accounting for rotation, but not for those ignoring vr. The mass loss rates of the SMC objects having luminosities of log L-star/L-circle dot greater than or similar to 5.4 are in excellent agreement with predictions by Vink et al. (2001, A&A, 369, 574). However, for lower luminosity stars the winds are too weak to determine. M accurately from the optical spectrum. Three targets were classified as Vz stars, two of which are located close to the theoretical zero-age main sequence. Three lower luminosity targets that were not classified as Vz stars are also found to lie near the ZAMS. We argue that this is related to a temperature effect inhibiting cooler from displaying the spectral features required for the Vz luminosity class.

Characterisation of two calmodulin-like proteins from the liver fluke, Fasciola hepatica.

Calmodulin is a calcium ion-sensing signalling protein found in eukaryotics. Two calmodulin-like gene sequences were identified in an EST library from adult liver flukes. One codes for a protein (FhCaM1) homologous to mammalian calmodulins (98% identity), whereas the other protein (FhCaM2) has only 41% identity. These genes were cloned into expression vectors and the recombinant proteins were expressed in Escherichia coli. Gel shift assays showed that both proteins bind to calcium, magnesium and zinc ions. Homology models have been built for both proteins. As expected, FhCaM1 has a highly similar structure to other calmodulins. Although FhCaM2 has a similar fold, its surface charge is higher than FhCaM1. One of the potential metal ion-binding sites has lower metal-ion co-ordination capability, while another has an adjacent lysine residue, both of which may decrease the metal-binding affinity. These differences may reflect a specialised role for FhCaM2 in the liver fluke.

Translational control of SEPT9 isoforms is perturbed in disease

A common feature of the mammalian septin gene family is complex genomic architecture with multiple alternate splice variants. Septin 9 has 18 distinct transcripts encoding 15 polypeptides, with two transcripts (SEPT9_v4 and v4*) encoding the same polypeptide. We have previously reported that the ratio of these distinct transcripts is altered in neoplasia, with the v4 transcript being the usual form in normal cells but v4* becoming predominant in tumours. This led us to ask what the functional differences between these two transcripts might be. The 5'-UTRs of v4 and v4* have distinct 5' ends encoded by exons 1 beta (v4) and 1 zeta and 2 (v4*) and a common 3' region and initiating ATG encoded within exon 3. Here we show that the two mRNAs are translated with different efficiencies and that cellular stress can alter this. A putative internal ribosome entry site can be identified in the common region of the v4 and v4* 5'-UTRs and translation is modulated by an upstream open-reading frame in the unique region of the v4 5'-UTR. Germline mutations in hereditary neuralgic amyotrophy (HNA) map to the region which is common to the two UTRs. These mutations dramatically enhance the translational efficiency of the v4 5'-UTR, leading to elevated SEPT9_v4 protein under hypoxic conditions. Our data provide a mechanistic insight into how the HNA mutations can alter the fine control of SEPT9_v4 protein and its regulation under physiologically relevant conditions and are consistent with the episodic and stress-induced nature of the clinical features of HNA.