cut11+: A Gene Required for Cell Cycle-dependent Spindle Pole Body Anchoring in the Nuclear Envelope and Bipolar Spindle Formation in Schizosaccharomyces pombe

The “cut” mutants of Schizosaccharomyces pombe are defective in spindle formation and/or chromosome segregation, but they proceed through the cell cycle, resulting in lethality. Analysis of temperature-sensitive alleles of cut11+ suggests that this gene is required for the formation of a functional bipolar spindle. Defective spindle structure was revealed with fluorescent probes for tubulin and DNA. Three-dimensional reconstruction of mutant spindles by serial sectioning and electron microscopy showed that the spindle pole bodies (SPBs) either failed to complete normal duplication or were free floating in the nucleoplasm. Localization of Cut11p tagged with the green fluorescent protein showed punctate nuclear envelope staining throughout the cell cycle and SPBs staining from early prophase to mid anaphase. This SPB localization correlates with the time in the cell cycle when SPBs are inserted into the nuclear envelope. Immunoelectron microscopy confirmed the localization of Cut11p to mitotic SPBs and nuclear pore complexes. Cloning and sequencing showed that cut11+ encodes a novel protein with seven putative membrane-spanning domains and homology to the Saccharomyces cerevisiae gene NDC1. These data suggest that Cut11p associates with nuclear pore complexes and mitotic SPBs as an anchor in the nuclear envelope; this role is essential for mitosis.

Preselection of retrovirally transduced bone marrow avoids subsequent stem cell gene silencing and age-dependent extinction of expression of human β-globin in engrafted mice

Transcriptional silencing of genes transferred into hematopoietic stem cells poses one of the most significant challenges to the success of gene therapy. If the transferred gene is not completely silenced, a progressive decline in gene expression as the mice age often is encountered. These phenomena were observed to various degrees in mouse transplant experiments using retroviral vectors containing a human β-globin gene, even when cis-linked to locus control region derivatives. Here, we have investigated whether ex vivo preselection of retrovirally transduced stem cells on the basis of expression of the green fluorescent protein driven by the CpG island phosphoglycerate kinase promoter can ensure subsequent long-term expression of a cis-linked β-globin gene in the erythroid lineage of transplanted mice. We observed that 100% of mice (n = 7) engrafted with preselected cells concurrently expressed human β-globin and the green fluorescent protein in 20–95% of their RBC for up to 9.5 mo posttransplantation, the longest time point assessed. This expression pattern was successfully transferred to secondary transplant recipients. In the presence of β-locus control region hypersensitive site 2 alone, human β-globin mRNA expression levels ranged from 0.15% to 20% with human β-globin chains detected by HPLC. Neither the proportion of positive blood cells nor the average expression levels declined with time in transplanted recipients. Although suboptimal expression levels and heterocellular position effects persisted, in vivo stem cell gene silencing and age-dependent extinction of expression were avoided. These findings support the further investigation of this type of vector for the gene therapy of human hemoglobinopathies.

Conserved circadian elements in phylogenetically diverse algae

Circadian expression of the luciferin-binding protein (LBP) from the dinoflagellate Gonyaulax polyedra is regulated at the translational level. A small interval in the lbp 3′-untranslated region, which contains seven UG-repeats, serves as a cis-acting element to which a trans-acting factor (CCTR) binds in a circadian manner. Its binding activity correlates negatively with the circadian expression of LBP. Here I report the identification of a protein in the green alga Chlamydomonas reinhardtii that represents a CCTR analog. It binds both specifically and under control of the circadian clock to the UG-repeat region. The data show for the first time that circadian cis-elements implicated in translational regulation have been conserved during evolution.

Whole-body and intravital optical imaging of angiogenesis in orthotopically implanted tumors

The development of drugs for the control of tumor angiogenesis requires a simple, accurate, and economical assay for tumor-induced vascularization. We have adapted the orthotopic implantation model to angiogenesis measurement by using human tumors labeled with Aequorea victoria green fluorescent protein for grafting into nude mice. The nonluminous induced capillaries are clearly visible against the very bright tumor fluorescence examined either intravitally or by whole-body luminance in real time. The orthotopic implantation model of human cancer has been well characterized, and fluorescence shadowing replaces the laborious histological techniques for determining blood vessel density. Intravital images of orthotopically implanted human pancreatic tumors clearly show angiogenic capillaries at both primary and metastatic sites. A quantitative time course of angiogenesis was determined for an orthotopically growing human prostate tumor periodically imaged intravitally in a single nude mouse over a 19-day period. Whole-body optical imaging of tumor angiogenesis was demonstrated by injecting fluorescent Lewis lung carcinoma cells into the s.c. site of the footpad of nude mice. The footpad is relatively transparent, with comparatively few resident blood vessels, allowing quantitative imaging of tumor angiogenesis in the intact animal. Capillary density increased linearly over a 10-day period as determined by whole-body imaging. Similarly, the green fluorescent protein-expressing human breast tumor MDA-MB-435 was orthotopically transplanted to the mouse fat pad, where whole-body optical imaging showed that blood vessel density increased linearly over a 20-week period. These powerful and clinically relevant angiogenesis mouse models can be used for real-time in vivo evaluation of agents inhibiting or promoting tumor angiogenesis in physiological microenvironments.

A Fruit-Specific Putative Dihydroflavonol 4-Reductase Gene Is Differentially Expressed in Strawberry during the Ripening Process1

A cDNA clone encoding a putative dihydroflavonol 4-reductase gene has been isolated from a strawberry (Fragaria × ananassa cv Chandler) DNA subtractive library. Northern analysis showed that the corresponding gene is predominantly expressed in fruit, where it is first detected during elongation (green stages) and then declines and sharply increases when the initial fruit ripening events occur, at the time of initiation of anthocyanin accumulation. The transcript can be induced in unripe green fruit by removing the achenes, and this induction can be partially inhibited by treatment of de-achened fruit with naphthylacetic acid, indicating that the expression of this gene is under hormonal control. We propose that the putative dihydroflavonol 4-reductase gene in strawberry plays a main role in the biosynthesis of anthocyanin during color development at the late stages of fruit ripening; during the first stages the expression of this gene could be related to the accumulation of condensed tannins.

Misurazione della Sostenibilità in Azienda. Approccio Quantitativo all'Analisi della Letteratura attraverso Bibliografic Coupling.

L'elaborato si impegna ad esporre i risultati derivanti da una revisione estensiva della letteratura riguardante le pratiche di misurazione della sostenibilità in azienda. Si è scelto un approccio al problema di impostazione bibliometrico, consistente in analisi statistiche di dati bibliografici ricavabili osservando le relazioni tra documenti. Si sono utilizzati strumenti di clustering per categorizzare la letteratura ed individuare le aree di maggior interesse potenziali per aziende che desiderino investire con successo nella sostenibilità. Come risultato, si è ottenuta una chiara identificazione delle principali tematiche che devono essere tenute in considerazione nell’affrontare valutazioni relative alla misurazione e valorizzazione dei concetti di sostenibilità e impatto ambientale in ambito aziendale. Il presente lavoro si presenta inoltre come guida metodologica per un approccio quantitativo alla revisione della letteratura, adattabile ed applicabile in maniera iterativa per un continuo monitoraggio del tema o per indagini in altri ambiti. Questa tesi si pone dunque come base per ricerche e analisi successive finalizzate alla comprensione della letteratura.

Do socioeconomic gradients in women's health widen over time and with age

A population-based study was conducted to investigate changes over time in women's well-being and health service use by socio-cconomic status and whether these varied by age. Data from 12,328 mid-age women (aged 45-50 years in 1996) and 10,430 older women (aged 70-75 years) from the Australian Longitudinal Study on Women's Health were analysed. The main outcome measures were changes in the eight dimensions of the Short Form General Health Survey (SF-36) adjusted for baseline scores, lifestyle and behavioural factors; health care utilisation at Survey 2; and rate of deaths (older cohort only). Cross-sectional analyses showed clear socioeconomic differentials in well-being for both cohorts. Differential changes in health across tertiles of socioeconomic status (SES) were more evident in the mid-age cohort than in the older cohort. For the mid-aged women in the low SES tertile, declines in physical functioning (adjusted mean change of -2.4, standard error (SE) 1.1) and general health perceptions (-1.5, SE 1.1) were larger than the high SES group (physical functioning -0.8 SE 1.1, general health perceptions -0.8 SE 1.2). In the older cohort, changes in SF-36 scores over time were similar for all SES groups but women in the high SES group had lower death rates than women in the low SES group (relative risk: 0.79, 95% confidence interval 0.64-0.98). Findings suggest that SES differentials in physical health seem to widen during women's mid-adult years but narrow in older age. Nevertheless, SES remains an important predictor of health, health service use and mortality in older Australian women. (C) 2003 Elsevier Ltd. All rights reserved.

Effect of training on the response of plasma vascular endothelial growth factor to exercise in patients with peripheral arterial disease

Expansion of the capillary network, or angiogenesis, occurs following endurance training. This process, which is reliant on the presence of VEGF (vascular endothelial growth factor), is an adaptation to a chronic mismatch between oxygen demand and supply. Patients with IC (intermittent claudication) experience pain during exercise associated with an inadequate oxygen delivery to the muscles. Therefore the aims of the present study were to examine the plasma VEGF response to acute exercise, and to establish whether exercise training alters this response in patients with IC. In Part A, blood was collected from patients with IC (n = 18) before and after (+ 20 and + 60 min post-exercise) a maximal walking test to determine the plasma VEGF response to acute exercise. VEGF was present in the plasma of patients (45.11 +/- 29.96 pg/ml) and was unchanged in response to acute exercise. Part B was a training study to determine whether exercise training altered the VEGF response to acute exercise. Patients were randomly assigned to a treatment group (TMT; n = 7) that completed 6 weeks of high-intensity treadmill training, or to a control group (CON; n = 6). All patients completed a maximal walking test before and after the intervention, with blood samples drawn as for Part A. Training had no effect on plasma VEGF at rest or in response to acute exercise, despite a significant increase in maximal walking time in the TMT group (915 + 533 to 1206 + 500 s; P = 0.009) following the intervention. The absence of a change in plasma VEGF may reflect altered VEGF binding at the endothelium, although this cannot be confirmed by the present data.

Locating whiteness in the academic production of "I Philosophy" in and of time and place

This review will critically evaluate two recent texts by white academics working across disciplines of cultural studies, history and anthropology and published by UNSW Press, which share a focus on the relationship between Aboriginality, Philosophy, Place and Time in Australia. I write from the position of a queer white academic committed to engaging politically and intellectually with the challenge of Indigenous sovereignties in this place while also aware that my position as a middle class white woman and intellectual imposes limits on what it is possible for me to know about Indigenous epistemologies (see Moreton-Robinson, 2000). In the course of this review I will demonstrate how anthropology's tendency to fix its objects of study within a circumscribed space of 'difference' limits the capacity of texts produced within this discipline to account for racialized struggles over sovereignty. While these struggles are equally embedded in the ethnographic context and the nation's constitution and political institutions, we will see that Muecke and Bird Rose confront problems in analysing the relationship between the intimate space of the 'field', in which one's research subjects quickly become one's 'friends' and/or 'classificatory kin'—on one hand—and the public space of the nation within which statements about Aboriginality by white academics circulate and are vested with an authority that escapes individual intentions and control—on the other.

An inverse methodology for high frequency RF head coil design with pre-emphasized B1 field in MRI

An inverse methodology to assist in the design of radio-frequency (RF) head coils for high field MRI application is described in this work. Free space time-harmonic electromagnetic Green's functions and preemphasized B1 field are used to calculate the current density on the coil cylinder. With B1 field preemphasized and lowered in the middle of the RF transverse plane, the calculated current distribution can generate an internal magnetic field that can reduce the EM field/tissue interactions at high frequencies. The current distribution of a head coil operating at 4 T is calculated using inverse methodology with preemphasized B1 fields. FDTD is employed to calculate B1 field and signal intensity inside a homogenous cylindrical phantom and human head. A comparison with conventional RF birdcage coil is reported here and demonstrated that inverse-method designed coil with preemphasized B1 field can help in decreasing the notorious bright region caused by EM field/tissue interactions in the human head images at 4 T.

Good practice guide in learning and teaching

Once again this publication is produced to celebrate and promote good teaching and learning support and to offer encouragement to those imaginative and innovative staff who continue to wish to challenge students to learn to maximum effect. It is hoped that others will pick up some good ideas from the articles contained in this volume. We have again changed our approach for this 2007/08 edition (our fifth) of the Aston Business School Good Practice Guide. As before, some contributions were selected from those identifying interesting best practice on their Annual Module Reflection Forms in 2006/2007. Brookes? contribution this year is directly from her annual reflection. Other contributors received HELM (Research Centre in Higher Education Learning and Management) small research grants in 2006/2007. Part of the conditions were for them to write an article for this publication. We have also been less tight on the length of the articles this year. Some contributions are, therefore, on the way to being journal articles. HELM will be working with these authors to help develop these for publication. Looking back over the last five years it is brilliant to see how many different people have contributed over the years and, therefore, how much innovative learning and teaching work has been taking place in ABS over this time. In the first edition we were just pleased for people to write a few pages on their teaching. Now things have changed dramatically. The majority of the articles are grounded in empirical research (some funded by HELM small research grants) and Palmer?s article was produced as part of the University?s Postgraduate Certificate in Learning and Teaching. Most encouraging of all, four of this year?s articles have since been developed further and submitted to refereed journals. We await news of publication as we go to press. It is not surprising that how to manage large groups still remains a central theme of the articles, ABS has a large and still growing student body. Essex and Simpson have looked at trying to encourage students to attend taught sessions, on the basis that there is a strong correlation between attendance and higher performance. Their findings are forming the platform of a further study currently being carried out in the Undergraduate Programme. A number of the other articles concentrate on trying to encourage students to engage with study in an innovative way. This is particularly obvious in Shaw?s work. Everyone who has been around campus lately has had evidence that the students on Duncan?s modules have clearly been inspired. I found myself, for example, playing golf in the student dining room as part of this initiative! The articles by Jarzabkowski & Guilietti and Ho involved much larger surveys. This is another first for the Good Practice Guide and marks the first step on what will clearly be larger research efforts for these authors in this area. We look forward to the journal publications which will result from this work. The last articles are the result of HELM?s hosting of the national conference of the Higher Education Academy?s Business, Management, Accounting and Finance (BMAF) Subject Centre Conference in May 2007. Belal and Foster have written about their impressions of the Conference and Andrews has included the paper she gave. The papers on employability and widening participation are the centre of HELM?s current work. In the second volume we mentioned the launch of the School?s Research Centre in Higher Education Learning and Management (HELM). Since then HELM has stimulated a lot of activity across the School (and University) particularly linking research and teaching. A list of the HELM seminars for 2007/2008 is listed as Appendix 1 of this publication. Further details can be obtained from Catherine Foster (c.s.foster@aston.ac.uk), who coordinates the HELM seminars. We have also been working on a list of target journals to guide ABS staff who wish to publish in this area. These are included as Appendix 2 of this publication. May I thank the contributors for taking time out of their busy schedules to write the articles and to Julie Green, the Quality Manager, for putting the varying diverse approaches into a coherent and publishable form and for agreeing to fund the printing of this volume.

In-Motes EYE:a real time application for automobiles in wireless sensor networks

Wireless sensor networks have been identified as one of the key technologies for the 21st century. In order to overcome their limitations such as fault tolerance and conservation of energy, we propose a middleware solution, In-Motes. In-Motes stands as a fault tolerant platform for deploying and monitoring applications in real time offers a number of possibilities for the end user giving him in parallel the freedom to experiment with various parameters, in an effort the deployed applications to run in an energy efficient manner inside the network. The proposed scheme is evaluated through the In-Motes EYE application, aiming to test its merits under real time conditions. In-Motes EYE application which is an agent based real time In-Motes application developed for sensing acceleration variations in an environment. The application was tested in a prototype area, road alike, for a period of four months.

In-Motes Bins:a real time application for environmental monitoring in wireless sensor networks

In-Motes Bins is an agent based real time In-Motes application developed for sensing light and temperature variations in an environment. In-Motes is a mobile agent middleware that facilitates the rapid deployment of adaptive applications in Wireless Sensor Networks (WSN's). In-Motes Bins is based on the injection of mobile agents into the WSN that can migrate or clone following specific rules and performing application specific tasks. Using In-Motes we were able to create and rapidly deploy our application on a WSN consisting of 10 MICA2 motes. Our application was tested in a wine store for a period of four months. In this paper we present the In-Motes Bins application and provide a detailed evaluation of its implementation. © 2007 IEEE.

On realizing the principles of Gaudium et spes in a post-communist country: Searching for christian entrepreneurs and roots of individuals in Hungary at the time of globalization

Due to the communist regime in Hungary the values and principles of the Second Vatican Counsil could hardly achieve their goal in the region and the situation is almost the same even today. This paper examines two levels of society where the thoughts of Gaudium et spes might have appeared: we have explored that there are Christian companies existing about 15 years since the political transition in 1990 and we made a research among individuals in rural environment, how could they preserve their human wholeness described in GS, in other words, how could they keep their social, cultural, natural, religiuos and local roots amongst the consumer society that has been developped in Hungary at the time of capitalism. Regarding the Christian companies our research could produce a positive result: we have explored that although the Christian companies survayed hardly know the Church’s social doctrine, they live and operate according to it. At the same time in the realm of individuals we cannot tell good news of this kind. Most of the persons interviewed have already lost or are near to loose their roots, that is their human wholeness. Our final conclusion is that our hope for preserving even strenghtening the values of GS in the Hungarian society is in the communities, be it work communities, as John Paul II. mentioned in his encyclical Sollicitudo rei Socialis. The paper presents the details and conclusions of our researches.

Physiological response measures (respiration, clearance, byssus production, survival) of Perna viridis during a 91 day exposure experiment to microplastics in Bogor, Indonesia, 2014

Perna viridis from the Bay of Jakarta was exposed to different concentrations (0, 21.6, 216 and 2160 mg/l) of PVC microplastic particles for 91 days in a controlled laboratory experiment. Particles were negatively buoyant, but were regularly resuspended from the sediment, mimicking tidal events. The particles were contaminated with the organic pollutant fluoranthene, except for one control group, which was exposed to the highest plastic concentration (2160 mg/l) but with clean particles. Within the 91 days survival was monitored. After 40 - 44 days of the exposure, physiological responses of all mussel individuals were measured. Respiration rates were measured as the decrease of oxygen in a sealed container in 20 minutes. Clearance rates were determined by measuring the depletion of algal cells in the water in 30 minutes. Byssus production was assessed by counting the number of newly formed byssus discs within 24 hours.