bluuebunny/crossref_metadata_2025 huggingface dataset:
Virological diagnosis in community-acquired pneumonia in immunocompromised patients
Authors: Camps Serra M. , Cervera C. , Pumarola T. , Moreno A. , Perelló R. , Torres A. , Jiménez de Anta M. T. , Marcos M. A.
Published: October 2007

Community-acquired pneumonia (CAP) is a serious lower respiratory tract infection associated with significant morbidity and mortality in immunocompromised patients. The present study evaluated the clinical spectrum of CAP in immunocompromised hosts and the role of respiratory viruses, as well as the yield of viral diagnostic methods.

Conventional microbiological tests were routinely performed in immunocompromised patients with CAP. Nasopharyngeal swabs were processed for respiratory viruses by indirect immunofluorescence assay, cell culture and PCR. Four groups were defined according to aetiology of CAP, as follows: group 1 (nonviral), group 2 (mixed, nonviral and viral), group 3 (only viral) and group 4 (unknown aetiology).

Over a 1-yr period, 92 patients were included. An aetiological diagnosis was achieved in 61 (66%) patients: 38 (41%), group 1; 12 (13%), group 2; and 11 (12%), group 3. The most frequent pathogen detected wasStreptococcus pneumoniae(n = 29, 48%), followed by rhinovirus (n = 11, 18%). PCR identified 95% of respiratory viruses. Clinical characteristics could not reliably distinguish among the different aetiological groups.

Respiratory viruses represent a substantial part of the aetiologies of community-acquired pneumonia in immunocompromised patients and its routine assessment through PCR in nasopharyngeal swabs should be considered in the clinical care of these patients.
External validation of a COPD diagnostic questionnaire
Authors: Kotz D. , Nelemans P. , van Schayck C. P. , Wesseling G. J.
Published: October 2007

The aim of the present study was to determine the external validity of a recently developed questionnaire for the identification of patients at increased risk of airflow limitation in smokers from the general population in the provinces of Dutch and Belgian Limburg (regions surrounding Maastricht, the Netherlands).

As part of a study on the early detection of airflow limitation and subsequent smoking cessation treatment (International Standard Randomised Controlled Trial Number: 64481813), the recently developed chronic obstructive pulmonary disease (COPD) diagnostic questionnaire was used in current smokers aged 40–70 yrs, with a smoking history of ≥10 pack-yrs, who reported one or more respiratory symptom (cough, sputum production or dyspnoea), but who had no diagnosis of a respiratory disease (COPD or asthma). Spirometry performed according to American Thoracic Society/European Respiratory Society criteria served as a reference test.

Of the 676 subjects who entered the analyses, 398 showed normal lung function and 278 had a diagnosis of COPD (post-bronchodilator forced expiratory volume in one second/forced vital capacity of <0.70). The ability of the COPD diagnostic questionnaire to discriminate between subjects with and without COPD was poor (area under the receiver operating characteristic curve of 0.65).

In a high-risk population consisting of middle-aged current smokers with a smoking history of ≥10 pack-yrs, the chronic obstructive pulmonary disease diagnostic questionnaire is probably not useful as a diagnostic tool for the identification of patients with an increased risk of airflow limitation.
Sequential measurements of procalcitonin levels in diagnosing ventilator-associated pneumonia
Authors: Ramirez P. , Garcia M. A. , Ferrer M. , Aznar J. , Valencia M. , Sahuquillo J. M. , Menéndez R. , Asenjo M. A. , Torres A.
Published: October 2007

The utility of procalcitonin levels to improve the accuracy of clinical and microbiological parameters in diagnosing ventilator-associated pneumonia (VAP) was evaluated.

Sequential measurement of procalcitonin and C-reactive protein levels and the calculation of the simplified Clinical Pulmonary Infection Scores (CPIS) were performed in 44 patients mechanically-ventilated for >48 h with neither active infection for the duration or suspicion of VAP. Patients who developed extrapulmonary infection were excluded.

In total, 20 cases were suspected of having VAP and diagnosis was microbiologically confirmed in nine. In patients with confirmed VAP, procalcitonin levels were higher than in those without VAP. C-reactive protein levels and CPIS were lower in patients without suspected VAP, but could not discriminate confirmed and nonconfirmed suspicion of VAP. The best sensitivity and specificity (78 and 97%, respectively) corresponded to procalcitonin. The CPIS resulted in the same sensitivity, but had a lower specificity (80%). C-reactive protein had the worst sensitivity (56%), but a good specificity (91%). A CPIS ≥6 combined with serum levels of procalcitonin ≥2.99 ng·mL−1did not improve the sensitivity (67%), but resulted in 100% specificity.

Procalcitonin might be useful in the diagnosis of ventilator-associated pneumonia. Combined values of Clinical Pulmonary Infection Scores and procalcitonin below the cut-off points excluded false-positive diagnoses of ventilator-associated pneumonia.
Polymorphisms in the epidermal growth factor receptor gene and the risk of primary lung cancer: a case-control study
Authors: Choi Jin Eun , Ha Park Sun , Kim Kyung Mee , Lee Won Kee , Kam Sin , Cha Sung Ick , Ho Kim Chang , Mo Kang Young , Kim Young-Chul , Han Sung Beom , Jung Tae Hoon , Park Jae Yong
Published: October 2007

Background

Polymorphisms in Epidermal Growth Factor Receptor (EGFR) gene may influence EGFR production and/or activity, thereby modulating susceptibility to lung cancer. To test this hypothesis, we investigated the association between polymorphisms in theEGFRgene and the risk of lung cancer in a Korean population.

Methods

We first examined the frequencies of 39 candidate polymorphisms in theEGFRgene in 27 healthy Korean individuals. After then, we genotyped five polymorphisms (127378C>T, 142285G>A, 162093G>A, 181946C>T and 187114T>C) that have variant allele frequencies greater than 10%, in 582 lung cancer patients and in 582 healthy controls.

Results

Of the 5 polymorphisms, the 181946C>T genotype distribution was significantly different between the cases and controls (P= 0.04). Compared with the 181946 CC + CT genotype, the 181946 TT genotype was associated with a significantly decreased risk of lung cancer (adjusted OR = 0.63, 95% CI = 0.45–0.88,P= 0.007). When the analyses were stratified by smoking status, the protective effect of the TT genotype was statistically significant in ever-smokers (adjusted OR = 0.59, 95% CI = 0.41–0.86,P= 0.007), but not in never-smokers (adjusted OR = 0.89, 95% CI = 0.45–1.75,P= 0.73;P= 0.08, test for homogeneity). Consistent with the results of the genotyping analysis, the CGGCT haplotype with the 181946C allele was associated with a significantly increased risk of lung cancer compared to the CGGTT haplotype carrying the 181946T allele (adjusted OR = 1.50, 95% CI = 1.09–2.07,P= 0.012 and Bonferroni correctedP-value = 0.048).

Conclusion

These results suggest that theEGFRpolymorphisms, particularly the 181945C>T polymorphism, could be used as markers for the genetic susceptibility to lung cancer.
An integrative expression vector for Actinosynnema pretiosum
Authors: Goh Shan , Camattari Andrea , Ng Daniel , Song Ruth , Madden Kevin , Westpheling Janet , Wong Victor VT
Published: October 2007

Background

The Actinomycete Actinosynnema pretiosum ssp. auranticum has commercial importance due to its production of ansamitocin P-3 (AP-3), a potent antitumor agent. One way to increase AP-3 production would be to constitutively express selected genes so as to relieve bottlenecks in the biosynthetic pathway; however, an integrative expression vector for A. pretiosum is lacking. The aim of this study was to construct a vector for heterologous gene expression in A. pretiosum.

Results

A series of integrative expression vectors have been made with the following features: the IS117 transposase from Streptomyces coelicolor, the constitutive ermE* promoter from Saccharopolyspora erythraea, different ribosome-binding site (RBS) sequences and xylE as a translational reporter. Positive E. coli clones and A. pretiosum transconjugants were assayed by catechol. pAP42, containing an E. coli consensus RBS, and pAP43, containing an asm19 RBS, gave strong and moderate gene expression, respectively. In addition, an operon construct capable of multi-gene expression was created. Plasmid integration sites in transconjugants were investigated and four different sites were observed. Although the most common integration site was within a putative ORF with sequence similarity to NADH-flavin reductase, AP-3 levels and cell growth of transconjugants were unaffected.

Conclusion

A set of integrative vectors for constitutive gene expression in A. pretiosum has been constructed. Gene translation is easily determined by colorimetric assay on an agar plate. The vectors are suitable for studies relating to AP-3 biosynthesis as they do not affect AP-3 production.
Susceptibility of Anopheles gambiae and Anopheles stephensi to tropical isolates of Plasmodium falciparum
Authors: Hume Jennifer CC , Tunnicliff Mark , Ranford-Cartwright Lisa C , Day Karen P
Published: October 2007

Background

The susceptibility of anopheline mosquito species to Plasmodium infection is known to be variable with some mosquitoes more permissive to infection than others. Little work, however, has been carried out investigating the susceptibility of major malaria vectors to geographically diverse tropical isolates of Plasmodium falciparum aside from examining the possibility of infection extending its range from tropical regions into more temperate zones.

Methods

This study investigates the susceptibility of two major tropical mosquito hosts (Anopheles gambiae and Anopheles stephensi) to P. falciparum isolates of different tropical geographical origins. Cultured parasite isolates were fed via membrane feeders simultaneously to both mosquito species and the resulting mosquito infections were compared.

Results

Infection prevalence was variable with African parasites equally successful in both mosquito species, Thai parasites significantly more successful in An. stephensi, and PNG parasites largely unsuccessful in both species.

Conclusion

Infection success of P. falciparum was variable according to geographical origin of both the parasite and the mosquito. Data presented raise the possibility that local adaptation of tropical parasites and mosquitoes has a role to play in limiting gene flow between allopatric parasite populations.
Single-walled carbon nanotube interactions with HeLa cells
Authors: Yehia Hadi N , Draper Rockford K , Mikoryak Carole , Walker Erin Kate , Bajaj Pooja , Musselman Inga H , Daigrepont Meredith C , Dieckmann Gregg R , Pantano Paul
Published: October 2007

This work concerns exposing cultured human epithelial-like HeLa cells to single-walled carbon nanotubes (SWNTs) dispersed in cell culture media supplemented with serum. First, the as-received CoMoCAT SWNT-containing powder was characterized using scanning electron microscopy and thermal gravimetric analyses. Characterizations of the purified dispersions, termed DM-SWNTs, involved atomic force microscopy, inductively coupled plasma – mass spectrometry, and absorption and Raman spectroscopies. Confocal microRaman spectroscopy was used to demonstrate that DM-SWNTs were taken up by HeLa cells in a time- and temperature-dependent fashion. Transmission electron microscopy revealed SWNT-like material in intracellular vacuoles. The morphologies and growth rates of HeLa cells exposed to DM-SWNTs were statistically similar to control cells over the course of 4 d. Finally, flow cytometry was used to show that the fluorescence from MitoSOX™ Red, a selective indicator of superoxide in mitochondria, was statistically similar in both control cells and cells incubated in DM-SWNTs. The combined results indicate that under our sample preparation protocols and assay conditions, CoMoCAT DM-SWNT dispersions are not inherently cytotoxic to HeLa cells. We conclude with recommendations for improving the accuracy and comparability of carbon nanotube (CNT) cytotoxicity reports.
Isolated HIV-1 core is active for reverse transcription
Authors: Warrilow David , Stenzel Deborah , Harrich David
Published: October 2007

Whether purified HIV-1 virion cores are capable of reverse transcription or require uncoating to be activated is currently controversial. To address this question we purified cores from a virus culture and tested for the ability to generate authentic reverse transcription products. A dense fraction (approximately 1.28 g/ml) prepared without detergent, possibly derived from disrupted virions, was found to naturally occur as a minor sub-fraction in our preparations. Core-like particles were identified in this active fraction by electron microscopy. We are the first to report the detection of authentic strong-stop, first-strand transfer and full-length minus strand products in this core fraction without requirement for an uncoating activity.
Metamorphosis of an identified serotonergic neuron in the Drosophila olfactory system
Authors: Roy Bidisha , Singh Ajeet P , Shetty Chetak , Chaudhary Varun , North Annemarie , Landgraf Matthias , VijayRaghavan K , Rodrigues Veronica
Published: October 2007

Background

Odors are detected by sensory neurons that carry information to the olfactory lobe where they connect to projection neurons and local interneurons in glomeruli: anatomically well-characterized structures that collect, integrate and relay information to higher centers. Recent studies have revealed that the sensitivity of such networks can be modulated by wide-field feedback neurons. The connectivity and function of such feedback neurons are themselves subject to alteration by external cues, such as hormones, stress, or experience. Very little is known about how this class of central neurons changes its anatomical properties to perform functions in altered developmental contexts. A mechanistic understanding of how central neurons change their anatomy to meet new functional requirements will benefit greatly from the establishment of a model preparation where cellular and molecular changes can be examined in an identified central neuron.

Results

In this study, we examine a wide-field serotonergic neuron in the Drosophila olfactory pathway and map the dramatic changes that it undergoes from larva to adult. We show that expression of a dominant-negative form of the ecdysterone receptor prevents remodeling. We further use different transgenic constructs to silence neuronal activity and report defects in the morphology of the adult-specific dendritic trees. The branching of the presynaptic axonal arbors is regulated by mechanisms that affect axon growth and retrograde transport. The neuron develops its normal morphology in the absence of sensory input to the antennal lobe, or of the mushroom bodies. However, ablation of its presumptive postsynaptic partners, the projection neurons and/or local interneurons, affects the growth and branching of terminal arbors.

Conclusion

Our studies establish a cellular system for studying remodeling of a central neuromodulatory feedback neuron and also identify key elements in this process. Understanding the morphogenesis of such neurons, which have been shown in other systems to modulate the sensitivity and directionality of response to odors, links anatomy to the development of olfactory behavior.
Development of musculoskeletal toxicity without clear benefit after administration of PG-116800, a matrix metalloproteinase inhibitor, to patients with knee osteoarthritis: a randomized, 12-month, double-blind, placebo-controlled study
Authors: Krzeski Piotr , Buckland-Wright Chris , Bálint Géza , Cline Gary A , Stoner Karen , Lyon Robert , Beary John , Aronstein William S , Spector Tim D
Published: October 2007

We performed a randomized, double-blind, placebo-controlled, multicenter, parallel-group, dose-response study of the efficacy and safety of the oral administration of PG-116800, a matrix metalloproteinase (MMP) inhibitor, in patients with mild to moderate knee osteoarthritis. The primary efficacy endpoints included the progression of joint space narrowing in the osteoarthritic knee, as measured by microfocal radiography with fluoroscopic positioning, and the reduction of symptoms (pain and stiffness) and/or the improvement of function, as measured by the Western Ontario and McMaster Universities osteoarthritis index (WOMAC). Four hundred and one patients were randomly assigned to either placebo (n= 80) or one of fourdoses of PG-116800: 25 mg (n= 81), 50 mg (n= 80), 100 mg (n= 80), or 200 mg (n= 80) taken twice daily for 12 months. During the study, the 200-mg dose was discontinued based on an increased frequency of musculoskeletal adverse effects. After 1 year of treatment, no statistically significant difference was observed between placebo and PG-116800 with regard to mean changes in minimum joint space width of the knee or to WOMAC scores. The most frequent adverse effect was arthralgia (35%). Twenty-three percent of evaluable patients had at least a 30% decrease from baseline of at least onerange-of-motion measurement of either shoulder at a follow-up visit. The percentage of patients with reduction in range of motion was significantly greater in the twohighest dose groups relative to placebo. Thirteen percent of patients, half of whom were in the 200-mg group, reported hand adverse events (oedema, palmar fibrosis, Dupuytren contracture, or persistent tendon thickness or nodules). The threemost frequent shoulder adverse events were reversible arthralgia, stiffness, and myalgia, which mostly affected the twohighest dose groups. The unfavorable risk-benefit balance of the MMP inhibitor PG-116800 in patients with knee osteoarthritis precludes further development of the compound for this indication. This study adds to the weight of evidence suggesting that side effect profiles of MMP inhibitors in general make them unsuitable for use in osteoarthritis.

Trial Registration

ClinicalTrials.gov NCT00041756.
Placental Productions and Expressions of Soluble Endoglin, Soluble fms-Like Tyrosine Kinase Receptor-1, and Placental Growth Factor in Normal and Preeclamptic Pregnancies
Authors: Gu Yang , Lewis David F. , Wang Yuping
Published: October 2007

Context: Increased production of antiangiogenic factors soluble endoglin (sEng) and soluble fms-like tyrosine kinase receptor-1 (sFlt-1) by the placenta contributes to the pathophysiology in preeclampsia (PE).

Objective: Our objective was to determine the differences in endoglin (Eng), fms-like tyrosine kinase receptor-1 (Flt-1), and placental growth factor (PlGF) expressions between normal and PE placentas and sEng, sFlt-1, and PlGF production by trophoblast cells (TC) cultured under lowered oxygen conditions.

Methods: TCs isolated from normal and PE placentas were cultured under regular (5% CO2/air) and lowered (2% O2/5% CO2/93% N2) oxygen conditions. sEng, sFlt-1, and PlGF productions were determined by ELISA. Protein expressions for Eng, Flt-1, and PlGF in the placental tissues were accessed by immunohistochemical staining and Western blot analysis. Deglycosylated Eng, Flt-1, and PlGF protein expressions in placental tissues were also examined.

Results: PE TCs produced significantly more sEng, sFlt-1, and PlGF compared with those from normal TCs (P < 0.05). Under lowered oxygen conditions, PE TCs, but not normal TCs, released more sEng and sFlt-1. In contrast, both normal and PE TCs released less PlGF (P < 0.05). Enhanced expressions of Eng and Flt-1, as well as glycosylated Eng and Flt-1, were observed in PE placentas. Immunoblot also revealed that TCs released glycosylated sFlt-1, but not sEng, in culture.

Conclusions: PE TCs produce more sEng, sFlt-1, and PlGF than normal TCs. Lowered oxygen conditions promote sEng and sFlt-1, but reduce PlGF, productions by PE TCs. More glycosylated sEng and sFlt-1 are present in PE placentas. Trophoblasts release glycosylated sFlt-1, but unglycosylated sEng, in culture.
MEF2: a central regulator of diverse developmental programs
Authors: Potthoff Matthew J. , Olson Eric N.
Published: October 2007

The myocyte enhancer factor 2 (MEF2) transcription factor acts as a lynchpin in the transcriptional circuits that control cell differentiation and organogenesis. The spectrum of genes activated by MEF2 in different cell types depends on extracellular signaling and on co-factor interactions that modulate MEF2 activity. Recent studies have revealed MEF2 to form an intimate partnership with class IIa histone deacetylases, which together function as a point of convergence of multiple epigenetic regulatory mechanisms. We review the myriad roles of MEF2 in development and the mechanisms through which it couples developmental, physiological and pathological signals with programs of cell-specific transcription.
Nuclear accumulation of Smad complexes occurs only after the midblastula transition inXenopus
Authors: Saka Yasushi , Hagemann Anja I. , Piepenburg Olaf , Smith James C.
Published: October 2007

Activin and the Nodal-related proteins induce mesendodermal tissues during Xenopus development. These signals act through specific receptors to cause the phosphorylation, at their carboxyl termini, of Smad2 and Smad3. The phosphorylated Smad proteins form heteromeric complexes with Smad4 and translocate into the nucleus to activate the transcription, after the midblastula transition, of target genes such as Xbra and goosecoid(gsc). In this paper we use bimolecular fluorescence complementation(BiFC) to study complex formation between Smad proteins both in vivo and in response to exogenous proteins. The technique has allowed us to detect Smad2-Smad4 heteromeric interactions during normal Xenopusdevelopment and Smad2 and Smad4 homo- and heteromers in isolated Xenopus blastomeres. Smad2-Smad2 and Smad2-Smad4 complexes accumulate rapidly in the nuclei of responding cells following Activin treatment, whereas Smad4 homomeric complexes remain cytoplasmic. When cells divide, Smad2-Smad4 complexes associate with chromatin, even in the absence of ligand. Our observation that Smad2-Smad4 complexes accumulate in the nucleus only after the midblastula transition, irrespective of the stage at which cells were treated with Activin, may shed light on the mechanisms of developmental timing.
Synaptic differentiation is defective in mice lacking acetylcholine receptor β-subunit tyrosine phosphorylation
Authors: Friese Matthew B. , Blagden Chris S. , Burden Steven J.
Published: October 2007

Agrin activates MuSK, a receptor tyrosine kinase expressed in skeletal muscle, leading to tyrosine phosphorylation of the acetylcholine receptor(AChR) β-subunit and clustering of AChRs. The importance of AChRβ-subunit tyrosine phosphorylation in clustering AChRs and regulating synaptic differentiation is poorly understood. We generated mice with targeted mutations in the three intracellular tyrosines of the AChR β-subunit(AChR-β3F/3F). Mice lacking AChRβ-subunit tyrosine phosphorylation thrive postnatally and have no overt behavioral defects, indicating that AChR β-subunit tyrosine phosphorylation is not essential for the formation of neuromuscular synapses. Nonetheless, the size of synapses and the density of synaptic AChRs are reduced in AChR- β3F/3F mutant mice. Moreover, synapses are structurally simplified and the organization of postjunctional folds is aberrant in mice lacking tyrosine phosphorylation of the AChR β-subunit. Furthermore, mutant AChRs cluster poorly in response to agrin and are readily extracted from the cell surface of cultured myotubes by non-ionic detergent. These data indicate that tyrosine phosphorylation of the AChR β-subunit has an important role in organizing AChRs and regulating synaptic differentiation.
SH4-domain-induced plasma membrane dynamization promotes bleb-associated cell motility
Authors: Tournaviti Stella , Hannemann Sebastian , Terjung Stefan , Kitzing Thomas M. , Stegmayer Carolin , Ritzerfeld Julia , Walther Paul , Grosse Robert , Nickel Walter , Fackler Oliver T.
Published: October 2007

SH4 domains provide bipartite membrane-targeting signals for oncogenic Src family kinases. Here we report the induction of non-apoptotic plasma membrane (PM) blebbing as a novel and conserved activity of SH4 domains derived from the prototypic Src kinases Src, Fyn, Yes and Lck as well as the HASPB protein of Leishmania parasites. SH4-domain-induced blebbing is highly dynamic, with bleb formation and collapse displaying distinct kinetics. These reorganizations of the PM are controlled by Rho but not Rac or Cdc42 GTPase signalling pathways. SH4-induced membrane blebbing requires the membrane association of the SH4 domain, is regulated by the activities of Rock kinase and myosin II ATPase, and depends on the integrity of F-actin as well as microtubules. Endogenous Src kinase activity is crucial for PM blebbing in SH4-domain-expressing cells, active Src and Rock kinases are enriched in SH4-domain-induced PM blebs, and PM blebbing correlates with enhanced cell invasion in 3D matrices. These results establish a novel link between SH4 domains, Src activity and Rho signalling, and implicate SH4-domain-mediated PM dynamization as a mechanism that influences invasiveness of cells transformed by SH4-domain-containing oncoproteins.
Depletion of topoisomerase IIα leads to shortening of the metaphase interkinetochore distance and abnormal persistence of PICH-coated anaphase threads
Authors: Spence Jennifer M. , Phua Hui Hui , Mills Walter , Carpenter Adam J. , Porter Andrew C. G. , Farr Christine J.
Published: October 2007

Topoisomerase II (topo II) is a major component of mitotic chromosomes, and its unique decatenating activity has been implicated in many aspects of chromosome dynamics, of which chromosome segregation is the most seriously affected by loss of topo II activity in living cells. There is considerable evidence that topo II plays a role at the centromere including: the centromere-specific accumulation of topo II protein; cytogenetic/molecular mapping of the catalytic activity of topo II to active centromeres; the influence of sumoylated topo II on sister centromere cohesion; and its involvement in the activation of a Mad2-dependent spindle checkpoint. By using a human cell line with a conditional-lethal mutation in the gene encoding DNA topoisomerase IIα, we find that depletion of topo IIα, while leading to a disorganised metaphase plate, does not have any overt effect on general assembly of kinetochores. Fluorescence in situ hybridisation suggested that centromeres segregate normally, most segregation errors being chromatin bridges involving longer chromosome arms. Strikingly, a linear human X centromere-based minichromosome also displayed a significantly increased rate of missegregation. This sensitivity to depletion of topo IIα might be linked to structural alterations within the centromere domain, as indicated by a significant shortening of the distance across metaphase sister centromeres and the abnormal persistence of PICH-coated connections between segregating chromatids.
Targeting of the type II inositol polyphosphate 5-phosphatase INPP5B to the early secretory pathway
Authors: Williams Catrin , Choudhury Rawshan , McKenzie Eddie , Lowe Martin
Published: October 2007

The inositol polyphosphate 5-phosphatase INPP5B is closely related to the Lowe syndrome protein OCRL1, sharing a similar substrate specificity, domain organisation and an ability to compensate for loss of OCRL1 in knockout mice. The cellular localisation and functions of INPP5B have remained poorly defined until recently, when a role within the endocytic pathway was suggested. Here, we report that INPP5B is also localised to the early secretory pathway including the Golgi apparatus and ER-to-Golgi intermediate compartment (ERGIC). Consistent with this localisation, INPP5B binds to specific RAB proteins within the secretory pathway, and mutational analysis indicates that RAB binding is required for efficient Golgi targeting of INPP5B. Unlike OCRL1, INPP5B interacts with neither clathrin nor α-adaptin and is largely absent from clathrin-coated intermediates. Expression of INPP5B but not OCRL1 alters the distribution of the cycling protein ERGIC53 when cells are incubated at low temperature (15°C) or in the presence of brefeldin A, causing ERGIC53 to accumulate in the ERGIC, with a concomitant loss from the ER. Our data suggest a role for INPP5B in retrograde ERGIC-to-ER transport and imply that it has functions distinct from those of OCRL1 within both the secretory and endocytic pathways.
The mechanical effects of deposition patterns in welding-based layered manufacturing
Authors: Mughal M P , Mufti R A , Fawad H
Published: October 2007

This paper presents a finite element (FE)-based three-dimensional analysis to study the structural effects of deposition patterns in welding-based layered manufacturing (LM). A commercial finite element software ANSYS is used to simulate the deposition incorporating a double ellipsoidal heat source, material addition, and temperature-dependent material properties. Simulations carried out with various deposition sequences revealed that the thermal and structural effects on the workpiece are different for different patterns. The sequence starting from outside and ending at the centre is identified as the one which produces minimum warpage.
Towards an integrated CADCAM process for the production of forming shoulders with exact geometry
Authors: Hicks B , Mullineux G , Matthews J , Medland T
Published: October 2007

This paper considers the application of advanced CADCAM techniques for the production of the complex machine tooling used in packaging fast-moving consumer goods. In particular, the paper discusses the practical considerations and design issues that need to be addressed in order to construct CNC machined tooling with exact geometry. The paper considers in detail the design of ‘forming shoulders’, a common means to form bags, and begins with a mathematical model for defining its complex geometry. A CADCAM method for producing a forming shoulder with exact geometry is then summarized. In practice, it is necessary to modify the geometry of the forming shoulder in order to produce an overlap of material for the creation of a bag. This process also demands that the geometry of the forming shoulder ‘overlaps‘, which has certain implications for the CNC machining of the surface. These considerations and the means by which they are overcome are discussed with respect to a series of prototypes and experimental trials. The performance of a forming shoulder, produced by way of the integrated CADCAM process, during extended production trials is then discussed.
An Hfq-like protein in archaea: Crystal structure and functional characterization of the Sm protein from Methanococcus jannaschii
Authors: Nielsen Jesper S. , Bøggild Andreas , Andersen Christian B.F. , Nielsen Gorm , Boysen Anders , Brodersen Ditlev E. , Valentin-Hansen Poul
Published: October 2007

The Sm and Sm-like proteins are conserved in all three domains of life and have emerged as important players in many different RNA-processing reactions. Their proposed role is to mediate RNA–RNA and/or RNA–protein interactions. In marked contrast to eukaryotes, bacteria appear to contain only one distinct Sm-like protein belonging to the Hfq family of proteins. Similarly, there are generally only one or two subtypes of Sm-related proteins in archaea, but at least one archaeon, Methanococcus jannaschii, encodes a protein that is related to Hfq. This archaeon does not contain any gene encoding a conventional archaeal Sm-type protein, suggesting that Hfq proteins and archaeal Sm-homologs can complement each other functionally. Here, we report the functional characterization of M. jannaschii Hfq and its crystal structure at 2.5 Å resolution. The protein forms a hexameric ring. The monomer fold, as well as the overall structure of the complex is similar to that found for the bacterial Hfq proteins. However, clear differences are seen in the charge distribution on the distal face of the ring, which is unusually negative in M. jannaschii Hfq. Moreover, owing to a very short N-terminal α-helix, the overall diameter of the archaeal Hfq hexamer is significantly smaller than its bacterial counterparts. Functional analysis reveals that Escherichia coli and M. jannaschii Hfqs display very similar biochemical and biological properties. It thus appears that the archaeal and bacterial Hfq proteins are largely functionally interchangeable.
The RNA binding protein RNPS1 alleviates ASF/SF2 depletion-induced genomic instability
Authors: Li Xialu , Niu Tianhui , Manley James L.
Published: October 2007

Formation of transcription-induced R-loops poses a critical threat to genomic integrity throughout evolution. We have recently shown that the SR protein ASF/SF2 prevents R-loop formation in vertebrates by cotranscriptionally binding to nascent mRNA precursors to prevent their reassociation with template DNA. Here, we identify another RNA binding protein, RNPS1, that when overexpressed strongly suppresses the high molecular weight (HMW) DNA fragmentation, hypermutation, and G2 cell cycle arrest phenotypes of ASF/SF2-depleted cells. Furthermore, ablation of RNPS1 by RNA interference in HeLa cells leads to accumulation of HMW DNA fragments. As ASF/SF2 depletion does not influence RNPS1 expression, and RNPS1 cannot compensate for ASF/SF2 function in splicing, our data suggest that RNPS1 is able to function together with ASF/SF2 to form RNP complexes on nascent transcripts, and thereby prevent formation of transcriptional R-loops.
Nop9 is an RNA binding protein present in pre-40S ribosomes and required for 18S rRNA synthesis in yeast
Authors: Thomson Emma , Rappsilber Juri , Tollervey David
Published: October 2007

Proteomic analyses in yeast have identified a large number of proteins that are associated with preribosomal particles. However, the product of the yeast ORF YJL010C, herein designated as Nop9, failed to be identified in any previous physical or genetic analysis of preribosomes. Here we report that Nop9 is a nucleolar protein, which is associated with 90S and 40S preribosomes. In cells depleted of Nop9p, early cleavages of the 35S pre-rRNA are inhibited, resulting in the nucleolar retention of accumulated precursors and a failure to synthesize 18S rRNA. Nop9 contains multiple pumilio-like putative RNA binding repeats and displays robust in vitro RNA binding activity. The identification of Nop9p as a novel, essential factor in the nuclear maturation of 90S and pre-40S ribosomal subunits shows that the complement of ribosome synthesis factors remains incomplete.
Evolution of small nuclear RNAs in S. cerevisiae, C. albicans, and other hemiascomycetous yeasts
Authors: Mitrovich Quinn M. , Guthrie Christine
Published: October 2007

The spliceosome is a large, dynamic ribonuclear protein complex, required for the removal of intron sequences from newly synthesized eukaryotic RNAs. The spliceosome contains five essential small nuclear RNAs (snRNAs): U1, U2, U4, U5, and U6. Phylogenetic comparisons of snRNAs from protists to mammals have long demonstrated remarkable conservation in both primary sequence and secondary structure. In contrast, the snRNAs of the hemiascomycetous yeast Saccharomyces cerevisiae have highly unusual features that set them apart from the snRNAs of other eukaryotes. With an emphasis on the pathogenic yeast Candida albicans, we have now identified and compared snRNAs from newly sequenced yeast genomes, providing a perspective on spliceosome evolution within the hemiascomycetes. In addition to tracing the origins of previously identified snRNA variations present in Saccharomyces cerevisiae, we have found numerous unexpected changes occurring throughout the hemiascomycetous lineages. Our observations reveal interesting examples of RNA and protein coevolution, giving rise to altered interaction domains, losses of deeply conserved snRNA-binding proteins, and unique snRNA sequence changes within the catalytic center of the spliceosome. These same yeast lineages have experienced exceptionally high rates of intron loss, such that modern hemiascomycetous genomes contain introns in only ∼5% of their genes. Also, the splice site sequences of those introns that remain adhere to an unusually strict consensus. Some of the snRNA variations we observe may thus reflect the altered intron landscape with which the hemiascomycetous spliceosome must contend.
Wild-type is the optimal sequence of the HDV ribozyme under cotranscriptional conditions
Authors: Chadalavada Durga M. , Cerrone-Szakal Andrea L. , Bevilacqua Philip C.
Published: October 2007

RNA viruses are responsible for a variety of human diseases, and the pathogenicity of RNA viruses is often attributed to a high rate of mutation. Self-cleavage activity of the wild-type hepatitis delta virus (HDV) ribozyme as measured in standard divalent ion renaturation assays is biphasic and mostly slow and can be improved by multiple rational changes to ribozyme sequence or by addition of chemical denaturants. This is unusual in the sense that wild type is the most catalytically active sequence for the majority of protein enzymes, and RNA viruses are highly mutable. To see whether the ribozyme takes advantage of fast-reacting sequence changes in vivo, we performed alignment of 76 genomic and 269 antigenomic HDV isolates. Paradoxically, the sequence for the ribozyme was found to be essentially invariant in nature. We therefore tested whether three ribozyme sequence changes that improve self-cleavage under standard divalent ion renaturation assays also improve self-cleavage during transcription. Remarkably, wild type was as fast, or faster, than these mutants under cotranscriptional conditions. Slowing the rate of transcription or adding the hepatitis delta antigen protein only further stimulated cotranscriptional self-cleavage activity. Thus, the relative activity of HDV ribozyme mutants depends critically on whether the reaction is assayed under in vivo-like conditions. A model is presented for how wild-type ribozyme sequence and flanking sequence work in concert to promote efficient self-cleavage during transcription. Wild type being the optimal ribozyme sequence under in vivo-like conditions parallels the behavior of most protein enzymes.
Should β-Blockers Be Used in the Treatment of Cocaine-Associated Acute Coronary Syndrome?
Authors: Page Robert Lee , Utz Kenneth J , Wolfel Eugene E
Published: October 2007

Objective:

To critically evaluate the 30 year debate of β-blocker use in cocaine-induced acute coronary syndrome (CIACS).

Data Sources:

An Ovid MEDLINE In-Process and Other Non-Indexed Citations, Ovid MEDLINE Daily, and Ovid MEDLINE (1966–August 21, 2007) search of the medical literature was conducted using the key terms cocaine, myocardial infarction, acute coronary syndrome, and adrenergic β-antagonists.

Study Selection and Data Extraction:

All clinical trials, case reports, national cardiovascular guidelines, and reviews published in English were evaluated. Case reports were included based on whether (1) acute coronary syndrome was suspected, (2) a β-blocker was used during the treatment course, and (3) objective and subjective patient-specific information was documented.

Data Synthesis:

Three case reports and 2 placebo-controlled trials were identified that used 4 β-blockers (atenolol, labetalol, metoprolol, propranolol). Three national guidelines addressed β-blocker use. Although published data are limited, propranolol and labetalol exert minimal to no effect on alleviating cocaine-induced coronary vasoconstriction. None of the evaluated national guidelines recommends β-blockers as first-line agents in CIACS management.

Conclusion:

β-Blockers should not be considered first-line agents for controlling chest pain in patients with documented CIACS. If long-term β-blockade is warranted, its benefits should be weighed against recurrent use of cocaine and possible exacerbation of acute coronary syndrome. Given that carvedilol exhibits ancillary pharmacologic proprieties beneficial in CIACS, and post-myocardial infarction mortality data are available regarding its use, this agent could be considered to be appropriate therapy.
Emergence and Evolution of Pharmacogenetics and Pharmacogenomics in Clinical Pharmacy over the Past 40 Years
Authors: Streetman Daniel S
Published: October 2007

2006 marked the 40th year of publication for The Annals. Throughout its history, The Annals has provided important contributions to the development of clinical pharmacy. In 2007, we are continuing to publish articles reflecting on the history of clinical pharmacy through the eyes of practitioners, including those pioneering clinical pharmacy, as well as those whohave more recently entered the profession and a well-established specialty. In addition, we are presenting articles and editorials from the early history of The Annals that have given direction and shape to the practice of clinical pharmacy (see page 2042).
Reviews
Authors: Bailey Christopher , Rust Chris
Published: October 2007

Improving Teaching and Learning: A Whole Institution Approach, Vaneeta-Marie D'Andrea and David Gosling (2005) Society for Research into Higher Education and Open University Press, 245 pp., ISBN 13-978-0335-21068-8 (pbk), 978-0335-21069-5 (hbk), No cover price given

Effective
Postgraduate Supervision: Improving the Student/Supervisor Relationship, Adrian R. Eley and Roy Jennings (2005) Maidenhead: Open University Press, 184 pp., ISBN 13-978-0335-217076 (pbk), 19.99; 978-0335-217083 (hbk), 60.00
Phoenix from the Ashes. The PSOE Government under Rodrguez Zapatero 20042007: A new model for social democracy?
Authors: Kennedy Paul
Published: October 2007

With the Spanish general election approaching, this article considers the political lessons for social democratic parties to be drawn from the PSOE's experience in government and argues that its strategy has been based on contrasting approaches in the economic and non-economic policy areas. Continuity has characterised the PSOE's economic policy orientation, as it has broadly maintained the policy inherited from the previous PP government. In contrast, the PSOE has put in place a number of innovative reforms in areas beyond the economic policy arena, particularly with respect to policy on civil and gender rights. This combination has allowed the government under Rodrguez Zapatero to prioritise macroeconomic stability whilst at the same time distinguishing itself from previous PSOE and PP governments via a package of reforms which, it argues, has formed the basis of a distinctive national social democratic agenda founded on economic efficiency, social justice and individual freedoms.
Memory and trauma of the Portuguese agrarian reform: A case study
Authors: de Almeida Maria Antnia Pires
Published: October 2007

The author analyses the agrarian reform in the southern part of Portugal, which took place as a result of radical legislation issued right after the 25 April 1974 revolution. The municipality of Avis is presented as an example of this movement, because of its charismatic leaders and
the huge adhesion of their followers, who set in motion land occupations throughout the entire region. Personal and group motivations are described, using both written (institutional and literary) and oral sources. Thirty years later, consequences of the movement can still be found in the
region and new ways of life were established in order to cope with the changing rural world.
“You Feed Us So that We Can Fight Against You”. Concepts of the Art and State in the Hungarian Avant-Garde
Authors: Forgács Éva
Published: October 2007

Although the Hungarian avant-garde (1909-late 1930s) was radically innovative, it contains a classicist element that has not been adequately studied, probably because it contradicts quintessential avant-garde features. The young György Lukács supported the avant-garde painters because he preferred their newfound order and control to the ephemeral and the subjective in Hungarian Impressionism. – The classicist vein of the Hungarian avant-garde related to its political program. Poet, writer, painter, editor and publisher Lajos Kassák, who became the leader of the avant-garde, articulated several times his position with regard to the state: as a social democrat and anarchist anti-war activist during World War I.; as an oppositional artist during the 1919 Hungarian Commune who nevertheless claimed state support; as a leading avant-garde authority in exile (1919–1926); and as an editor and poet with increasingly classicist leanings after his return to Budapest. Between 1919 and the 1930s, he aspired both to the position of an anti-establishment artist and to that of a rival to political leadership. – Kassák adopted Dada in 1921 but categorically rejected it a year later in favor of the austere order of constructivist geometry. His choice indicated not only his preference of clarity and authority but also a Central-European (mis)understanding of Russian Constructivism.
L'avant-garde littéraire roumaine et la politique
Authors: Pop Ion
Published: October 2007

Voicing initially a rather general critique of bourgeois culture and society – which was characteristic also of other European vanguard movements such as futurism, Dada, constructivism and surrealism – many militants of the Romanian avant-garde groups moved in the 1930s and 1940s towards an outspoken Marxist engagement, as evinced by the publications of Stephan Roll (pseudonym of Gheorghe Dinu) in the last issues of the review unu. As a result of this engagement, Marxist avant-gardists came to regard the avant-garde itself as a product of a decadent, bourgeois mindset, and called for a “true” aesthetic engagement in proletarian struggles, following the example of the USSR. Signs of an “engagement” may also be discerned in the surrealists Gherasim Luca and Gellu Naum in the 1940s. In line with André Breton they tried to reconcile the surrealist programme with the “total liberation of mankind.” Ironically, however, the Romanian communist regime muzzled the avant-garde: the old avant-garde militants had to submit to the ideological commands of “social realism” or leave their country.
Regarding Paintings. Works of El Greco in Simon Vestdijk's Het vijfde zegel
Authors: Heynders Odile
Published: October 2007

Het vijfde zegel (The Fifth Seal), published by the Dutch writer Simon Vestdijk in 1937, follows the nineteenth-century tradition of literary novels on painting, practised by such writers as Stendhal, Gautier, and Zola. But Vestdijk also offers a more modern case of intermediality, for the novel can be understood as a psychologically motivated reading of El Greco's life and final phase of work. Protagonist and paintings develop through apollinic (orderly) and dionysic (creative) conventions. The novel can also be read as critique of avant-garde conceptions of representation. The interpretation of this fascinating text is accompanied by some general reflections on ekphrasis.
Vulnerable Adults in Research: From Consent to Assent?
Authors: Edwards Sarah J.L , Guest Stephen
Published: October 2007

Some scientific advances can become realities only with the help of vulnerable people who do not have legal capacity to consent to research. When dealing with legally non-competent adults, practitioners now consider the patient's own wishes more than ever before, and so they have pioneered the practice of seeking what has become known as their ‘assent’ before treating them. Assent has been defined as a person's ‘permission or affirmative agreement to something’. People who do not possess sufficient capacity to make a