bluuebunny/crossref_metadata_2025 huggingface dataset:
The role of lipid composition for insertion and stabilization of amino acids in membranes
Authors: Johansson Anna C. V. , Lindahl Erik
Published: May 2009

While most membrane protein helices are clearly hydrophobic, recent experiments have indicated that it is possible to insert marginally hydrophobic helices into bilayers and have suggested apparent in vivo free energies of insertion for charged residues that are low, e.g., a few kcals for arginine. In contrast, a number of biophysical simulation studies have predicted that the bilayer interior is close to a pure hydrophobic environment with large penalties for hydrophilic amino acids—and yet the experimental scales do significantly better at predicting actual membrane proteins from sequence. Here, we have systematically studied the dependence of the free energy profiles on lipid properties, including tail length, saturation, headgroup hydrogen bond strength, and charge, both to see to whether the in vivo insertion can be explained in whole or part from lipid composition of the endoplasmic reticulum (ER) membranes, and if the solvation properties can help interpret how protein function depends on the lipids. We find that lipid charge is important to stabilize charged amino acids inside the bilayer (with implications, e.g., for ion channels), that thicker bilayers have higher solvation costs for hydrophilic side chains, and that headgroup hydrogen bond strength determines how adaptive the lipids are as a hydrophobic/hydrophilic solvent. None of the different free energy profiles are even close to the low apparent in vivo insertion cost, which suggests that regardless of the specific ER membrane composition the current experimental results cannot be explained by normal lipid-type variation.
Electromagnetic source transformations using superellipse equations
Authors: Allen Jeffery , Kundtz Nathan , Roberts Daniel A. , Cummer Steven A. , Smith David R.
Published: May 2009

Transformation optics can be used to design media with unique properties that alter the behavior of electromagnetic waves in passive space and recently in space containing source distributions. We present source transformations where current from a linear radiator is spread over a cylindrical shell with various cross sections. The semianalytic transformations are based on superellipse equations. Finite-element full-wave simulations of transformations from a dipole to a cylinder, diamond-shaped cylinder, and flattened cylinder are presented. The radiation pattern of the dipole seen by an outside observer is replicated in all cases demonstrating the potential applicability of source transformations to conformal antenna design.
Comment on “Rewiring networks for synchronization” [Chaos 18, 037105 (2008)]
Authors: Jalili Mahdi , Ajdari Rad Ali
Published: May 2009

This is a comment on a recent paper by A. Hagberg and D. A. Schult [Chaos 18, 037105 (2008)]. By taking the eigenratio of the Laplacian of an undirected and unweighted network as its synchronizability measure, they proposed a greedy rewiring algorithm for enhancing the synchronizability of the network. The algorithm is not capable of avoiding local minima, and as a consequence, for each initial network, different optimized networks with different synchronizabilities are obtained. Here, we show that by employing a simulated annealing based optimization method, it is possible to further enhance the synchronizability of the network. Moreover, using this approach, the optimized network is not biased by the initial network and regardless of the initial networks, the final optimized networks have similar synchronization properties.
Adjustable fetal phantom for pulse oximetry
Authors: Stubán Norbert , Niwayama Masatsugu
Published: May 2009

As the measuring head of a fetal pulse oximeter must be attached to the head of the fetus inside the mother’s uterus during labor, testing, and developing of fetal pulse oximeters in real environment have several difficulties. A fetal phantom could enable evaluation of pulse oximeters in a simulated environment without the restrictions and difficultness of medical experiments in the labor room. Based on anatomic data we developed an adjustable fetal head phantom with three different tissue layers and artificial arteries. The phantom consisted of two arteries with an inner diameter of 0.2 and 0.4 mm. An electronically controlled pump produced pulse waves in the arteries. With the phantom we investigated the sensitivity of a custom-designed wireless pulse oximeter at different pulsation intensity and artery diameters. The results showed that the oximeter was capable of identifying 4% and 2% changes in diameter between the diastolic and systolic point in arteries of over 0.2 and 0.4 mm inner diameter, respectively. As the structure of the phantom is based on reported anatomic values, the results predict that the investigated custom-designed wireless pulse oximeter has sufficient sensitivity to detect the pulse waves and to calculate the R rate on the fetal head.
Mass- and energy-analyses of ions from plasma by means of a miniature Thomson spectrometer
Authors: Sadowski M. J. , Czaus K. , Malinowski K. , Skladnik-Sadowska E. , Zebrowski J.
Published: May 2009

The paper presents an improved version of a miniature mass-spectrometer of the Thomson-type, which has been adopted for ion analysis near the dense plasma region inside a vacuum chamber. Problems connected with the separation of ions from plasma streams are considered. Input diaphragms and pumping systems, needed to ensure good vacuum inside the analyzing region, are described. The application of the miniature Thomson-type analyzer is illustrated by ion parabolas recorded in plasma-focus facility and rod plasma injector experiment. A quantitative analysis of the recorded ion parabolas is presented. Factors influencing accuracy of the ion analysis are discussed and methods of the spectrometer calibration are described.
Sheath and boundary conditions for plasma simulations of a Hall thruster discharge with magnetic lenses
Authors: Keidar Michael , Beilis Isak I.
Published: May 2009

The effect of magnetic lens configuration on sheath in a Hall thruster discharge channels is considered. A model of the plasma-wall transition is developed in the case of large magnetic field incidence angle with respect to the wall. Peculiarity of such case consists in that the potential drop across the magnetic field is set externally. In this case, standard boundary conditions at the sheath edge for plasma simulations fail and a new formulation for those boundary conditions is proposed. The results obtained demonstrate importance of the effect of the magnetic field incidence angle on the sheath boundary conditions for plasma simulations as well as on the energy balance in the Hall thruster discharge.
Nested multilayered X pinches for generators with mega-ampere current level
Authors: Shelkovenko T. A. , Pikuz S. A. , McBride R. D. , Knapp P. F. , Wilhelm H. , Hammer D. A. , Sinars D. B.
Published: May 2009

A symmetric X pinch configuration that is conducive to using large numbers of wires on ≥1 MA pulsed power generators has been tested at 1 MA. Using an initial configuration of wires before their twisting, similar to nested cylindrical wire arrays, enables a geometrically simple, compact, multilayer wire configuration at the X pinch crossing region. Multilayer X pinches with the same or different materials in the inner and outer wire layers were tested. Optimization resulted in X pinch radiation sources with peak power comparable to the most successful single layer X pinch, but with a compact, single bright X radiation source more reliably obtained using the nested configuration.
Photoemission study of fully silicided Pd2Si gates with interface modification induced by dopants
Authors: Hosoi Takuji , Ohta Akio , Miyazaki Seiichi , Shiraishi Hiroyuki , Shibahara Kentaro
Published: May 2009

Work function shifts for fully silicided (FUSI) Pd2Si gates with BF2 and P predoping, originally observed as the flatband voltage shifts in metal-oxide-semiconductor (MOS) diodes, were also reproducible for backside x-ray photoelectron spectroscopy measurements. The Pd composition ratios of FUSI layers were reduced in the vicinity of the gate SiO2 layer, which directly affects apparent work function in the MOS system. However, other non-negligible changes, such as pileup of predoped elements and Pd precipitation, were also found for the predoped cases. The work function shift by predoping should be considered to be a result of interaction of such multiple factors.
Embryonic pig pancreatic tissue for the treatment of diabetes in a nonhuman primate model
Authors: Hecht Gil , Eventov-Friedman Smadar , Rosen Chava , Shezen Elias , Tchorsh Dalit , Aronovich Anna , Freud Enrique , Golan Hana , El-Hasid Ronit , Katchman Helena , Hering Bernhard J. , Zung Amnon , Kra-Oz Zipi , Shaked-Mishan Pninit , Yusim Alex , Shtabsky Alex , Idelevitch Pavel , Tobar Ana , Harmelin Alon , Bachar-Lustig Esther , Reisner Yair
Published: May 2009

Xenotransplantation of pig tissues has great potential to overcome the shortage of organ donors. One approach to address the vigorous immune rejection associated with xenotransplants is the use of embryonic precursor tissue, which induces and utilizes host vasculature upon its growth and development. Recently, we showed in mice that embryonic pig pancreatic tissue from embryonic day 42 (E42) exhibits optimal properties as a β cell replacement therapy. We now demonstrate the proof of concept in 2 diabetic Cynomolgus monkeys, followed for 393 and 280 days, respectively. A marked reduction of exogenous insulin requirement was noted by the fourth month after transplantation, reaching complete independence from exogenous insulin during the fifth month after transplantation, with full physiological control of blood glucose levels. The porcine origin of insulin was documented by a radioimmunoassay specific for porcine C-peptide. Furthermore, the growing tissue was found to be predominantly vascularized with host blood vessels, thereby evading hyperacute or acute rejection, which could potentially be mediated by preexisting anti-pig antibodies. Durable graft protection was achieved, and most of the late complications could be attributed to the immunosuppressive protocol. While fine tuning of immune suppression, tissue dose, and implantation techniques are still required, our results demonstrate that porcine E-42 embryonic pancreatic tissue can normalize blood glucose levels in primates. Its long-term proliferative capacity, its revascularization by host endothelium, and its reduced immunogenicity, strongly suggest that this approach could offer an attractive replacement therapy for diabetes.
Structure of bacteriophage SPP1 head-to-tail connection reveals mechanism for viral DNA gating
Authors: Lhuillier Sophie , Gallopin Matthieu , Gilquin Bernard , Brasilès Sandrine , Lancelot Nathalie , Letellier Guillaume , Gilles Mathilde , Dethan Guillaume , Orlova Elena V. , Couprie Joël , Tavares Paulo , Zinn-Justin Sophie
Published: May 2009

In many bacterial viruses and in certain animal viruses, the double-stranded DNA genome enters and exits the capsid through a portal gatekeeper. We report a pseudoatomic structure of a complete portal system. The bacteriophage SPP1 gatekeeper is composed of dodecamers of the portal protein gp6, the adaptor gp15, and the stopper gp16. The solution structures of gp15 and gp16 were determined by NMR. They were then docked together with the X-ray structure of gp6 into the electron density of the ≈1-MDa SPP1 portal complex purified from DNA-filled capsids. The resulting structure reveals that gatekeeper assembly is accompanied by a large rearrangement of the gp15 structure and by folding of a flexible loop of gp16 to form an intersubunit parallel β-sheet that closes the portal channel. This stopper system prevents release of packaged DNA. Disulfide cross-linking between β-strands of the stopper blocks the key conformational changes that control genome ejection from the virus at the beginning of host infection.
Location of chloride and its possible functions in oxygen-evolving photosystem II revealed by X-ray crystallography
Authors: Kawakami Keisuke , Umena Yasufumi , Kamiya Nobuo , Shen Jian-Ren
Published: May 2009

The chloride ion, Cl−, is an essential cofactor for oxygen evolution of photosystem II (PSII) and is closely associated with the Mn4Ca cluster. Its detailed location and function have not been identified, however. We substituted Cl−with a bromide ion (Br−) or an iodide ion (I−) in PSII and analyzed the crystal structures of PSII with Br−and I−substitutions. Substitution of Cl−with Br−did not inhibit oxygen evolution, whereas substitution of Cl−with I−completely inhibited oxygen evolution, indicating the efficient replacement of Cl−by I−. PSII with Br−and I−substitutions were crystallized, and their structures were analyzed. The results showed that there are 2 anion-binding sites in each PSII monomer; they are located on 2 sides of the Mn4Ca cluster at equal distances from the metal cluster. Anion-binding site 1 is close to the main chain of D1-Glu-333, and site 2 is close to the main chain of CP43-Glu-354; these 2 residues are coordinated directly with the Mn4Ca cluster. In addition, site 1 is located in the entrance of a proton exit channel. These results indicate that these 2 Cl−anions are required to maintain the coordination structure of the Mn4Ca cluster as well as the proposed proton channel, thereby keeping the oxygen-evolving complex fully active.
A tumor suppressive coactivator complex of p53 containing ASC-2 and histone H3-lysine-4 methyltransferase MLL3 or its paralogue MLL4
Authors: Lee Jeongkyung , Kim Dae-Hwan , Lee Seunghee , Yang Qi-Heng , Lee Dong Kee , Lee Soo-Kyung , Roeder Robert G. , Lee Jae W.
Published: May 2009

ASC-2, a multifunctional coactivator, forms a steady-state complex, named ASCOM (for
AS
C-2
COM
plex), that contains the histone H3-lysine-4 (H3K4)-methyltransferase MLL3 or its paralogue MLL4. Somewhat surprisingly, given prior indications of redundancy between MLL3 and MLL4, targeted inactivation of the MLL3 H3K4-methylation activity in mice is found to result in ureter epithelial tumors. Interestingly, this phenotype is exacerbated in a
p53
+/−
background and the tumorigenic cells are heavily immunostained for γH2AX, indicating a contribution of MLL3 to the DNA damage response pathway through p53. Consistent with the in vivo observations, and the demonstration of a direct interaction between p53 and ASCOM, cell-based assays have revealed that ASCOM, through ASC-2 and MLL3/4, acts as a p53 coactivator and is required for H3K4-trimethyation and expression of endogenous p53-target genes in response to the DNA damaging agent doxorubicin. In support of redundant functions for MLL3 and MLL4 for some events, siRNA-mediated down-regulation of both MLL3 and MLL4 is required to suppress doxorubicin-inducible expression of several p53-target genes. Importantly, this study identifies a specific H3K4 methytransferase complex, ASCOM, as a physiologically relevant coactivator for p53 and implicates ASCOM in the p53 tumor suppression pathway in vivo.
Conserved waters mediate structural and functional activation of family A (rhodopsin-like) G protein-coupled receptors
Authors: Angel Thomas E. , Chance Mark R. , Palczewski Krzysztof
Published: May 2009

G protein-coupled receptors with seven transmembrane α-helices (GPCRs) comprise the largest receptor superfamily and are involved in detecting a wide variety of extracellular stimuli. The availability of high-resolution crystal structures of five prototypical GPCRs, bovine and squid rhodopsin, engineered A
2A
-adenosine, β
1
- and β
2
-adrenergic receptors, permits comparative analysis of features common to these and likely all GPCRs. We provide an analysis of the distribution of water molecules in the transmembrane region of these GPCR structures and find conserved contacts with microdomains demonstrated to be involved in receptor activation. Colocalization of water molecules associating with highly conserved and functionally important residues in several of these GPCR crystal structures supports the notion that these waters are likely to be as important to proper receptor function as the conserved residues. Moreover, in the absence of large conformational changes in rhodopsin after photoactivation, we propose that ordered waters contribute to the functional plasticity needed to transmit activation signals from the retinal-binding pocket to the cytoplasmic face of rhodopsin and that fundamental features of the mechanism of activation, involving these conserved waters, are shared by many if not all family A receptors.
The Yb protein defines a novel organelle and regulates male germline stem cell self-renewal in Drosophila melanogaster
Authors: Szakmary Akos , Reedy Mary , Qi Hongying , Lin Haifan
Published: May 2009

Yb regulates the proliferation of both germline and somatic stem cells in the Drosophila melanogaster ovary by activating piwi and hh expression in niche cells. In this study, we show that Yb protein is localized as discrete cytoplasmic spots exclusively in the somatic cells of the ovary and testis. These spots, which are different from all known cytoplasmic structures in D. melanogaster, are evenly electron-dense spheres 1.5 µm in diameter (herein termed the Yb body). The Yb body is frequently associated with mitochondria and a less electron-dense sphere of similar size that appears to be RNA rich. There are one to two Yb bodies/cell, often located close to germline cells. The N-terminal region of Yb is required for hh expression in niche cells, whereas the C-terminal region is required for localization to Yb bodies. The entire Yb protein is necessary for piwi expression in niche cells. A double mutant of Yb and a novel locus show male germline loss, revealing a function for Yb in male germline stem cell maintenance.
Mechanism of olfactory masking in the sensory cilia
Authors: Takeuchi Hiroko , Ishida Hirohiko , Hikichi Satoshi , Kurahashi Takashi
Published: May 2009

Olfactory masking has been used to erase the unpleasant sensation in human cultures for a long period of history. Here, we show a positive correlation between the human masking and the odorant suppression of the transduction current through the cyclic nucleotide–gated (CNG) and Ca2+-activated Cl− (Cl(Ca)) channels. Channels in the olfactory cilia were activated with the cytoplasmic photolysis of caged compounds, and their sensitiveness to odorant suppression was measured with the whole cell patch clamp. When 16 different types of chemicals were applied to cells, cyclic AMP (cAMP)-induced responses (a mixture of CNG and Cl(Ca) currents) were suppressed widely with these substances, but with different sensitivities. Using the same chemicals, in parallel, we measured human olfactory masking with 6-rate scoring tests and saw a correlation coefficient of 0.81 with the channel block. Ringer's solution that was just preexposed to the odorant-containing air affected the cAMP-induced current of the single cell, suggesting that odorant suppression occurs after the evaporation and air/water partition of the odorant chemicals at the olfactory mucus. To investigate the contribution of Cl(Ca), the current was exclusively activated by using the ultraviolet photolysis of caged Ca, DM-nitrophen. With chemical stimuli, it was confirmed that Cl(Ca) channels were less sensitive to the odorant suppression. It is interpreted, however, that in the natural odorant response the Cl(Ca) is affected by the reduction of Ca2+ influx through the CNG channels as a secondary effect. Because the signal transmission between CNG and Cl(Ca) channels includes nonlinear signal-boosting process, CNG channel blockage leads to an amplified reduction in the net current. In addition, we mapped the distribution of the Cl(Ca) channel in living olfactory single cilium using a submicron local [Ca2+]i elevation with the laser photolysis. Cl(Ca) channels are expressed broadly along the cilia. We conclude that odorants regulate CNG level to express masking, and Cl(Ca) in the cilia carries out the signal amplification and reduction evenly spanning the entire cilia. The present findings may serve possible molecular architectures to design effective masking agents, targeting olfactory manipulation at the nano-scale ciliary membrane.
Protein kinase A–dependent modulation of Ca2+ sensitivity in cardiac and fast skeletal muscles after reconstitution with cardiac troponin
Authors: Matsuba Douchi , Terui Takako , O-Uchi Jin , Tanaka Hiroyuki , Ojima Takao , Ohtsuki Iwao , Ishiwata Shin'ichi , Kurihara Satoshi , Fukuda Norio
Published: May 2009

Protein kinase A (PKA)-dependent phosphorylation of troponin (Tn)I represents a major physiological mechanism during β-adrenergic stimulation in myocardium for the reduction of myofibrillar Ca2+ sensitivity via weakening of the interaction with TnC. By taking advantage of thin filament reconstitution, we directly investigated whether or not PKA-dependent phosphorylation of cardiac TnI (cTnI) decreases Ca2+ sensitivity in different types of muscle: cardiac (porcine ventricular) and fast skeletal (rabbit psoas) muscles. PKA enhanced phosphorylation of cTnI at Ser23/24 in skinned cardiac muscle and decreased Ca2+ sensitivity, of which the effects were confirmed after reconstitution with the cardiac Tn complex (cTn) or the hybrid Tn complex (designated as PCRF; fast skeletal TnT with cTnI and cTnC). Reconstitution of cardiac muscle with the fast skeletal Tn complex (sTn) not only increased Ca2+ sensitivity, but also abolished the Ca2+-desensitizing effect of PKA, supporting the view that the phosphorylation of cTnI, but not that of other myofibrillar proteins, such as myosin-binding protein C, primarily underlies the PKA-induced Ca2+ desensitization in cardiac muscle. Reconstitution of fast skeletal muscle with cTn decreased Ca2+ sensitivity, and PKA further decreased Ca2+ sensitivity, which was almost completely restored to the original level upon subsequent reconstitution with sTn. The essentially same result was obtained when fast skeletal muscle was reconstituted with PCRF. It is therefore suggested that the PKA-dependent phosphorylation or dephosphorylation of cTnI universally modulates Ca2+ sensitivity associated with cTnC in the striated muscle sarcomere, independent of the TnT isoform.
Substrate‐dependent bidirectional modulation of P‐glycoprotein‐mediated drug resistance by erlotinib
Authors: Noguchi Kohji , Kawahara Haruka , Kaji Airi , Katayama Kazuhiro , Mitsuhashi Junko , Sugimoto Yoshikazu
Published: May 2009

Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR‐TKIs) inhibit the function of certain adenosine triphosphate (ATP)‐binding cassette transporters, including P‐glycoprotein/ABCB1 and breast cancer resistance protein (BCRP)/ABCG2. We previously reported an antagonistic activity of gefitinib towards BCRP. We have now analyzed the effects of erlotinib, another EGFR‐TKI, on P‐glycoprotein and BCRP. As with gefitinib, erlotinib effectively reversed BCRP‐mediated resistance to SN‐38 (7‐ethyl‐10‐hydroxycamptothecin) and mitoxantrone. In contrast, we found that erlotinib effectively suppressed P‐glycoprotein‐mediated resistance to vincristine and paclitaxel, but did not suppress resistance to mitoxantrone and doxorubicin. Conversely, erlotinib appeared to enhance P‐glycoprotein‐mediated resistance to mitoxantrone in K562/MDR cells. This bidirectional activity of erlotinib was not observed with verapamil, a typical P‐glycoprotein inhibitor. Flow cytometric analysis showed that erlotinib co‐treatment restored intracellular accumulation of mitoxantrone in K562 cells expressing BCRP, but not in cells expressing P‐glycoprotein. Consistently, erlotinib did not inhibit mitoxantrone efflux in K562/MDR cells although it did vincristine efflux in K562/MDR cells and mitoxantrone efflux in K562/BCRP cells. Intravesicular transport assay showed that erlotinib inhibited both P‐glycoprotein‐mediated vincristine transport and BCRP‐mediated estrone 3‐sulfate transport. Intriguingly, Lineweaver‐Burk plot suggested that the inhibitory mode of erlotinib was a mixed type for P‐glycoprotein‐mediated vincristine transport whereas it was a competitive type for BCRP‐mediated estrone 3‐sulfate transport. Collectively, these observations indicate that the pharmacological activity of erlotinib on P‐glycoprotein‐mediated drug resistance is dependent upon the transporter substrate. These findings will be useful in understanding the pharmacological interactions of erlotinib used in combinational chemotherapy. (Cancer Sci 2009; 100: 1701–1707)
Sprouty4 deficiency potentiates Ras‐independent angiogenic signals and tumor growth
Authors: Taniguchi Koji , Ishizaki Takuma , Ayada Toranoshin , Sugiyama Yuki , Wakabayashi Yu , Sekiya Takashi , Nakagawa Ryusuke , Yoshimura Akihiko
Published: May 2009

Sprouty proteins have been shown to negatively regulate a variety of receptor tyrosine kinase (RTK) signaling pathways and are considered to be tumor suppressor proteins. The pathophysiological functions of Sproutys in vivo remain to be investigated. In this study, we examined the physiological function of Sprouty4 as an angiogenic regulator, using Sprouty4 knockout (KO) mice and cells. We found that transplanted tumor cells grow much faster in Sprouty4 KO mice than in wild type (WT) mice, which we associate with enhanced neovascularization in the tumors transplanted into Sprouty4 KO mice. Moreover, vascular endothelial growth factor (VEGF)‐A‐induced angiogenesis and vascular permeability in vivo were enhanced in Sprouty4 KO mice compared with WT mice. Ex vivo angiogenesis, which we induced by VEGF‐A, basic fibroblast growth factor (bFGF), and sphingosine‐1‐phosphate (S1P), was also enhanced in the aortas of Sprouty4 KO mice. We demonstrated that Sprouty4 suppresses Ras‐independent VEGF‐A and S1P signaling, while it does not affect Ras‐dependent VEGF‐C signaling. These data indicate that Sprouty4 selectively suppresses Ras‐independent angiogenic factor signals and is an important negative regulator of pathophysiological angiogenesis. (Cancer Sci 2009; 100: 1648–1654)
Exposure of children and adolescents to alcohol advertising on Australian metropolitan free‐to‐air television
Authors: Fielder Lynda , Donovan Robert J. , Ouschan Robyn
Published: May 2009

Aim  This study investigated the exposure of underage youth to alcohol television advertising on metropolitan free‐to‐air television in the five mainland capital city markets of Australia.

Design  Exposure levels (target audience rating points; TARPs) were obtained for all alcohol advertisements screened from November 2005 to October 2006 in each capital city market for: children 0–12 years; underage teens 13–17 years; young adults 18–24 years; and mature adults 25+ years. The 30 most exposed advertisements across age groups were then content‐analysed for elements appealing to children and underage youth.

Results  In each of the five metropolitan markets, mature adults were most exposed to alcohol advertising. Children were exposed to one‐third the level of mature adults and underage teens to approximately the same level as young adults. However, there was considerable variation in media weight between markets, such that underage teens in two markets had higher advertising TARPs than young adults in other markets.

All 30 highest exposed advertisements contained at least one element known to appeal to children and underage youth, with 23 containing two or more such elements. Fifteen of the 30 advertisements featured an animal.

Conclusions  The self‐regulation system in Australia does not protect children and youth from exposure to alcohol advertising, much of which contains elements appealing to these groups.
Sexual orientation, substance use behaviors and substance dependence in the United States
Authors: McCabe Sean Esteban , Hughes Tonda L. , Bostwick Wendy B. , West Brady T. , Boyd Carol J.
Published: May 2009

Aims  To assess past‐year prevalence rates of substance use behaviors and substance dependence across three major dimensions of sexual orientation (identity, attraction and behavior) in a large national sample of adult women and men in the United States.

Design  Data were collected from structured diagnostic face‐to‐face interviews using the Alcohol Use Disorder and Associated Disabilities Interview Schedule DSM‐IV version IV (AUDADIS‐IV).

Setting  Prevalence estimates were based on data collected from the 2004–2005 (wave 2) National Epidemiologic Survey on Alcohol and Related Conditions (NESARC).

Participants  A large national sample of 34 653 adults aged 20 years and older: 52% female, 71% white, 12% Hispanic, 11% African American, 4% Asian and 2% Native American.

Findings  Approximately 2% of the population self‐identified as lesbian, gay or bisexual; 4% reported at least one life‐time same‐sex sexual partner and 6% reported same‐sex sexual attraction. Although non‐heterosexual orientation was generally associated with a higher risk of substance use and substance dependence, the majority of sexual minority respondents did not report substance use or meet criteria for DSM‐IV substance dependence. There was considerable variation in substance use outcomes across sexual orientation dimensions; these variations were more pronounced among women than among men.

Conclusions  Results support previous research findings of heightened risk of substance use and substance dependence among some sexual minority groups and point to the need for research that examines the reasons for such differences. Results also highlight important gender differences and question previous findings indicating uniformly higher risk for substance dependence among sexual minorities. Risks appear to vary based on gender and how sexual orientation is defined. Findings have implications for more effective prevention and intervention efforts that target subgroups at greatest risk.
Recanting of life‐time inhalant use: how big a problem and what to make of it
Authors: Martino Steven C. , McCaffrey Daniel F. , Klein David J. , Ellickson Phyllis L.
Published: May 2009

Aims  To establish the prevalence of recanting of life‐time inhalant use, identify correlates of recanting to gain insight to its causes and develop a method for distinguishing recanters who truly are versus are not life‐time users of inhalants.

Design and setting  Longitudinal survey data from students in 62 South Dakota middle schools who were participating in a field trial to evaluate a school‐based drug prevention program.

Measurements  At grades 7–8, participants reported on their life‐time inhalant use, other drug use and drug‐related beliefs, attitudes and behaviors.

Findings  Forty‐nine per cent of students who reported life‐time inhalant use at grade 7 recanted their reports a year later. Comparison of students who recanted inhalant use with those who did or did not report inhalant use consistently on drug‐related beliefs, attitudes and behaviors at grades 7 and 8 suggested that, whereas some inhalant use recanting reflects denial of past behavior, some reflects erroneous initial reporting. Based on a latent mixture model fitted to the multivariate distribution of grade 7 and grade 8 responses of recanters and consistent reporters, we calculated the probability that each recanter was, in fact, a life‐time inhalant user. An estimated 67% of the recanters in our sample appear to be life‐time inhalant users who admitted use in grade 7 and then denied that use at grade 8; 33% appear to be students who reported use incorrectly at grade 7 and then corrected that error at grade 8.

Conclusions  Inhalant use recanting is a significant problem that, if not handled carefully, is likely to have a considerable impact on our understanding of the etiology of inhalant use and efforts to prevent it.
Older adults' alcohol consumption and late‐life drinking problems: a 20‐year perspective
Authors: Moos Rudolf H. , Schutte Kathleen K. , Brennan Penny L. , Moos Bernice S.
Published: May 2009

Aims  The aim of this study was to identify changes in patterns of alcohol consumption over a 20‐year interval among older women and men, and to examine the associations between guideline‐defined excessive drinking and late‐life drinking problems.

Design, participants and measures  A community sample of 719 adults between 55 and 65 years of age who consumed alcohol at or prior to baseline participated in a survey of alcohol consumption and drinking problems and was followed 10 years and 20 years later.

Findings  The likelihood of excessive drinking declined over the 20‐year interval as adults matured into their 70s and 80s. However, at ages 75–85, 27.1% of women and 48.6% of men consumed more than two drinks per day or seven drinks per week. At comparable guideline levels of alcohol consumption, older men were more likely to have drinking problems than were older women. Consumption of more than two drinks per day or seven drinks per week was identified as a potential conservative guideline for identifying excessive drinking associated with an elevated likelihood of drinking problems.

Conclusions  A substantial percentage of older adults who consume alcohol engage in guideline‐defined excessive drinking and incur drinking problems. The finding that older men may be more likely than older women to experience problems when they drink beyond guideline levels suggests that alcohol guidelines for men should not be set higher than those for women.
Anti‐tissue transglutaminase antibodies in the follow‐up of adult coeliac disease
Authors: DIPPER C. R. , MAITRA S. , THOMAS R. , LAMB C. A. , MCLEAN‐TOOKE A. P. C. , WARD R. , SMITH D. , SPICKETT G. , MANSFIELD J. C.
Published: May 2009

Background  The detection of auto antibodies directed against tissue transglutaminase (anti‐tTG antibodies) has a well‐established role in the diagnosis of coeliac disease, but the value of these antibodies in long‐term follow‐up is controversial.

Aims  To determine if serial anti‐tTG antibody measurements could confirm adherence to a gluten‐free diet (GFD) and identify patients at risk of disease complications.

Methods  In a 54‐month cohort follow‐up study, 182 adult patients were assessed. Data recorded included self‐assessment of GFD adherence; anti‐tTG antibody concentration and serum ferritin, vitamin B12 and folate. Where available, bone mineral density (BMD) and duodenal histology data were retrieved.

Results  Persistently elevated anti‐tTG antibody levels were significantly associated with abnormal duodenal histology (P < 0.001), low ferritin (P < 0.01) and poor adherence to the GFD (P < 0.001). The specificity was >85% while the sensitivity was 39–60%. Anti‐tTG antibody concentrations fell rapidly following successful initiation of a GFD, and maintenance of normalization identified those who continued to be adherent to the diet.

Conclusions  This study supports a strategy of using anti‐tTG antibody concentrations to monitor newly diagnosed and established patients with coeliac disease, and to target dietetic intervention to reduce the risk of complication.
Clinical Trial: High‐dose furosemide plus small‐volume hypertonic saline solutions vs. repeated paracentesis as treatment of refractory ascites
Authors: LICATA G. , TUTTOLOMONDO A. , LICATA A. , PARRINELLO G. , DI RAIMONDO D. , DI SCIACCA R. , CAMMÀ C. , CRAXÌ A. , PATERNA S. , PINTO A.
Published: May 2009

Background  In patients with cirrhosis, ascites is defined as refractory when it cannot be mobilized or recurs early in standard diuretic therapy.

Aim  To compare the safety and efficacy of intravenous high‐dose furosemide + hypertonic saline solutions (HSS) with repeated paracentesis in patients with cirrhosis and refractory ascites.

Patients and methods  Eighty‐four subjects (59/25 M/F) with cirrhosis, mostly of viral aetiology, admitted for refractory ascites, were randomly assigned to receive furosemide (250–1000 mg/bid i.v.) plus HSS (150 mL H2O with NaCl 1.4–4.6% or 239–187 mEq/L) (60 patients, Group A) or to repeated paracentesis and a standard diuretic schedule (24 patients, Group B).

Results  During hospitalization, Group A patients had more diuresis (1605 ± 131 mL vs. 532 ± 124 mL than Group B patients; P < 0.001) and a greater loss of weight at discharge (−8.8 ± 4.8 kg vs. −4.5 ± 3.8 kg, P < 0.00). Control of ascites, pleural effusions and/or leg oedema was deemed significantly better in Group A.

Conclusions  This randomized pilot study suggests that HHS plus high‐dose furosemide is a safe and effective alternative to repeated paracentesis when treating hospitalized patients with cirrhosis and refractory ascites. Larger studies will be needed to evaluate long‐term outcomes such as readmission and mortality.
Effect of mesalazine on mucosal immune biomarkers in irritable bowel syndrome: a randomized controlled proof‐of‐concept study
Authors: CORINALDESI R. , STANGHELLINI V. , CREMON C. , GARGANO L. , COGLIANDRO R. F. , DE GIORGIO R. , BARTESAGHI G. , CANOVI B. , BARBARA G.
Published: May 2009

Background  Intestinal immune infiltration contributes to symptoms in patients with irritable bowel syndrome (IBS).

Aim  To assesses the effect of mesalazine (mesalamine) on mucosal immune cells in patients with IBS, through a pilot study.

Methods  A randomized, double‐blind, placebo‐controlled trial in 20 patients with IBS in tertiary care setting. Patients were randomized to receive placebo or 800 mg mesalazine three times daily for 8 weeks. The primary endpoint was a significant reduction in total colonic immune cells on biopsies obtained at the end of treatment compared to baseline. Secondary endpoints included effects on subsets of immune cells, inflammatory mediators and symptom severity. Intention‐to‐treat analysis was performed.

Results  Mesalazine markedly reduced immune cells as compared with placebo (P = 0.0082); this effect was ascribed to a marked inhibition of mast cells (P = 0.0014). Mesalazine significantly increased general well‐being (P = 0.038), but had no significant effects on abdominal pain (P = 0.084), bloating (P = 0.177) or bowel habits. No serious drug‐related adverse events were reported during the study.

Conclusions  Mesalazine is an effective and safe approach to reduce mast cell infiltration and may improve general well‐being in patients with IBS. These results support the hypothesis that immune mechanisms represent potential therapeutic targets in IBS.
A role of Gab2 association in Flt3 ITD mediated Stat5 phosphorylation and cell survival
Authors: Masson Kristina , Liu Tao , Khan Rasheed , Sun Jianmin , Rönnstrand Lars
Published: May 2009

The haematopoietic growth factor receptor Flt3 has been implicated as major cause of transformation in acute myeloid leukaemia. Intracellular signals mediated by wild‐type Flt3 are involved in cell differentiation and survival whereas signalling via the mutant Flt3 ITD (internal tandem duplication) promotes enhanced cell growth. In this study, we identified tyrosines 768, 955 and 969 of Flt3 as phosphorylation sites and mediators of growth factor receptor binding protein 2 (Grb2) interaction, leading to the association of Grb2 associated binder 2 (Gab2) and contributing to proliferation and survival. Ba/F3 cells were transfected with either the wild‐type Flt3 or the ITD, with or without a triple mutation of the Grb2 binding sites, and characterised in terms of proliferation and viability. Interestingly, the Flt3 ITD promoted increased survival but after introducing the triple mutation, this phenotype was lost. When looking into different downstream pathways, this effect was mainly caused by decreased phosphoinositide 3‐kinase and Stat5 signalling, and the Flt3 ITD carrying the Grb2 binding mutations showed less Akt and Stat5 activation compared to the regular Flt3 ITD receptor. These findings not only reveal novel phosphorylation sites in Flt3 but contribute to the understanding of the molecular mechanism by which Flt3 ITD functions in pathological conditions.
Outcomes for patients who fail high dose chemoradiotherapy and autologous stem cell rescue for relapsed and primary refractory Hodgkin lymphoma
Authors: Moskowitz Alison J. , Perales Miguel‐Angel , Kewalramani Tarun , Yahalom Joachim , Castro‐Malaspina Hugo , Zhang Zhigang , Vanak Jill , Zelenetz Andrew D. , Moskowitz Craig H.
Published: May 2009

Most patients with Hodgkin lymphoma (HL) are cured with first and second‐line treatment; however, the outcome is unknown for those who fail high dose chemoradiotherapy with autologous stem cell transplant (HDT‐ASCT). This report is an analysis of patients with relapsed and primary refractory HL who were treated with HDT‐ASCT and failed due to progression of disease (POD). Two hundred and two patients received HDT‐ASCT at Memorial Sloan Kettering Cancer Center for relapsed or refractory HL between December 1994 and 2005 and 71 failed due to POD. The median survival following HDT‐ASCT failure was 25 months. Only 16 (23%) of the 71 patients are currently alive, nine of whom are in remission. Multivariate analysis revealed two factors associated with poor outcome: relapse within 6 months of HDT‐ASCT and primary refractory disease. The only factor associated with improved survival was the ability to receive a second transplant, in particular, reduced intensity allogeneic transplant (RIT). Novel therapies are needed for patients who fail HDT‐ASCT, particularly those with primary refractory disease and those who relapse within 6 months of HDT‐ASCT. Future studies should focus on prospectively evaluating RIT following HDT‐ASCT failure in patients with remission duration from HDT‐ASCT of >6 months.
Low plasma HDL‐c, a vascular risk factor in high risk patients independent of LDL‐c
Authors: Hajer G. R. , Van Der Graaf Y. , Bots M. L. , Algra A. , Visseren F. L. J. ,   on behalf of the SMART Study Group
Published: May 2009

Background  High concentrations of high density lipoprotein‐cholesterol (HDL‐c) are associated with lower cardiovascular risk, but it is not known whether this is also the case in the presence of intensive low density lipoprotein‐cholesterol (LDL‐C) therapy. In this study, we determined the relationship between HDL‐c and new non‐fatal or fatal vascular events in patients with various manifestations of clinical evident vascular disease and evaluated whether this relationship is modified by LDL‐c levels.

Materials and methods  Prospective single centre, cohort study of 3837 patients with a history or recent diagnosis of clinical manifest vascular disease (coronary, cerebrovascular, peripheral arterial disease or abdominal aortic aneurysm) The relationship between HDL‐c quintiles and time to a new event (myocardial infarction, ischaemic stroke, vascular death) was quantified with Cox‐regression models and adjusted for potential confounders (age, gender, body mass index, type 2 diabetes, triglycerides, smoking, use of alcohol and lipid‐lowering therapy). Effect modification of LDL‐c was assessed with interaction terms.

Results  During a median follow up of 3·3 (range 0·1–9·5) years, a total of 465 first new events occurred. Compared with the lowest quintile, the upper quintile of HDL‐c levels was associated with a lower risk for new events; Hazard Ratio 0·61 (95% CI 0·43–0·86) irrespective of the localisation of vascular disease and use of lipid‐lowering medication. Higher HDL‐c levels were associated with comparably lower risks for vascular events in patients with LDL‐c levels above and below 2·5 mmol L−1 (P‐values for interaction > 0·05).

Conclusions  Patients with various clinical manifestations of vascular diseases in the highest HDL‐c quintile have a lower risk for vascular events compared with patients in the lowest HDL‐c quintile. Further, the current results expand the evidence by showing that also in a cohort of patients with various localisations of clinical evident vascular disease, in which statins were widely used, higher HDL‐c levels confer a lower risk for developing new vascular events, irrespective of the localisation of vascular disease, use of lipid‐lowering medication and plasma LDL‐c concentration.
Toward a full multiscale approach to interpret potential fields
Authors: Cella Federico , Fedi Maurizio , Florio Giovanni
Published: May 2009

The way potential fields convey source information depends on the scale at which the field is analysed. In this sense a multiscale analysis is a useful method to study potential fields particularly when the main field contributions are caused by sources with different depths and extents. Our multiscale approach is built with a stable transformation, such as depth from extreme points. Its stability results from mixing, in a single operator, the wavenumber low‐pass behaviour of the upward continuation transformation of the field with the enhancement high‐pass properties of n‐order derivative transformations. So, the complex reciprocal interference of several field components may be efficiently faced at several scales of the analysis and the depth to the sources may be estimated together with the homogeneity degrees of the field. In order to estimate the source boundaries we use another multiscale method, the multiscale derivative analysis, which utilizes a generalized concept of horizontal derivative and produces a set of boundary maps at different scales. We show through synthetic examples and application to the gravity field of Southern Italy that this multiscale behaviour makes this technique quite different from other source boundary estimators.

The main result obtained by integrating multiscale derivative analysis with depth from extreme points is the retrieval of rather effective information of the field sources (horizontal boundaries, depth, structural index). This interpretative approach has been used along a specific transect for the analysis of the Bouguer anomaly field of Southern Apennines. It was set at such scales, so to emphasize either regional or local features along the transect. Two different classes of sources were individuated. The first one includes a broad, deep source with lateral size of 45∼50 km, at a depth of 13 km and having a 0.5 structural index. The second class includes several narrower sources located at shallowest depths, ranging from 3–6 km, with lateral size not larger than 5 km and structural indexes ranging from 1–1.5. Within a large‐scale geological framework, these results could help to outline the mean structural features at crustal depths.
The utility of rapid application development in large‐scale, complex projects
Authors: Berger Hilary , Beynon‐Davies Paul
Published: May 2009

This paper describes an investigation into the utility of an agile development approach known as rapid application development (RAD) within a large‐scale development project conducted within the public sector in the UK. Features of RAD as an ‘agile’ information systems development method (ISDM) are discussed and previous research identifying properties of development environments most conducive to its application are described. A case study is then presented based upon a long‐term qualitative investigation of the ‘unbundling’ of a commercial RAD‐like ISDM. The evidence from this case leads us to question established wisdom in relation to appropriate adoption and application of agile development within large‐scale and complex development environments.