bluuebunny/crossref_metadata_2025 huggingface dataset:
Voltage-dependent Gating Rearrangements in the Intracellular T1–T1 Interface of a K+ Channel
Authors: Wang Guangyu , Covarrubias Manuel
Published: March 2006

The intracellular tetramerization domain (T1) of most eukaryotic voltage-gated potassium channels (Kv channels) exists as a “hanging gondola” below the transmembrane regions that directly control activation gating via the electromechanical coupling between the S4 voltage sensor and the main S6 gate. However, much less is known about the putative contribution of the T1 domain to Kv channel gating. This possibility is mechanistically intriguing because the T1–S1 linker connects the T1 domain to the voltage-sensing domain. Previously, we demonstrated that thiol-specific reagents inhibit Kv4.1 channels by reacting in a state-dependent manner with native Zn2+ site thiolate groups in the T1–T1 interface; therefore, we concluded that the T1–T1 interface is functionally active and not protected by Zn2+ (Wang, G., M. Shahidullah, C.A. Rocha, C. Strang, P.J. Pfaffinger, and M. Covarrubias. 2005. J. Gen. Physiol. 126:55–69). Here, we co-expressed Kv4.1 channels and auxiliary subunits (KChIP-1 and DPPX-S) to investigate the state and voltage dependence of the accessibility of MTSET to the three interfacial cysteines in the T1 domain. The results showed that the average MTSET modification rate constant (kMTSET) is dramatically enhanced in the activated state relative to the resting and inactivated states (∼260- and ∼47-fold, respectively). Crucially, under three separate conditions that produce distinct activation profiles, kMTSET is steeply voltage dependent in a manner that is precisely correlated with the peak conductance–voltage relations. These observations strongly suggest that Kv4 channel gating is tightly coupled to voltage-dependent accessibility changes of native T1 cysteines in the intersubunit Zn2+ site. Furthermore, cross-linking of cysteine pairs across the T1–T1 interface induced substantial inhibition of the channel, which supports the functionally dynamic role of T1 in channel gating. Therefore, we conclude that the complex voltage-dependent gating rearrangements of eukaryotic Kv channels are not limited to the membrane-spanning core but must include the intracellular T1–T1 interface. Oxidative stress in excitable tissues may perturb this interface to modulate Kv4 channel function.
Experiences of the First 16 Hospitals Using Copper-Silver Ionization forLegionellaControl: Implications for the Evaluation of Other Disinfection Modalities
Authors: Stout Janet E. , Yu Victor L.
Published: March 2006

Background and Objectives:

Hospital-acquired legionnaires' disease can be prevented by disinfection of hospital water systems. This study assessed the long-term efficacy of copper-silver ionization as a disinfection method in controllingLegionellain hospital water systems and reducing the incidence of hospital-acquired legionnaires' disease. A standardized, evidence-based approach to assist hospitals with decision making concerning the possible purchase of a disinfection system is presented.

Design:

The first 16 hospitals to install copper-silver ionization systems forLegionelladisinfection were surveyed. Surveys conducted in 1995 and 2000 documented the experiences of the hospitals with maintenance of the system, contamination of water withLegionella, and occurrence of hospital-acquired legionnaires' disease. All were acute care hospitals with a mean of 435 beds.

Results:

All 16 hospitals reported cases of hospital-acquired legionnaires' disease prior to installing the copper-silver ionization system. Seventy-five percent had previously attempted other disinfection methods including superheat and flush, ultraviolet light, and hyperchlorination. By 2000, the ionization systems had been operational from 5 to 11 years. Prior to installation, 47% of the hospitals reported that more than 30% of distal water sites yieldedLegionella. In 1995, after installation, 50% of the hospitals reported 0% positivity, and 43% still reported 0% in 2000. Moreover, no cases of hospital-acquired legionnaires' disease have occurred in any hospital since 1995.

Conclusions:

This study represents the final step in a proposed 4-step evaluation process of disinfection systems that includes (1) demonstrated efficacy ofLegionellaeradication in vitro using laboratory assays, (2) anecdotal experiences in preventing legionnaires' disease in individual hospitals, (3) controlled studies in individual hospitals, and (4) validation in confirmatory reports from multiple hospitals during a prolonged time (5 to 11 years in this study). Copper-silver ionization is now the only disinfection modality to have fulfilled all four evaluation criteria.
A Nosocomial Outbreak ofLegionella pneumophilaCaused by Contaminated Transesophageal Echocardiography Probes
Authors: Levy Pierre-Yves , Teysseire Nadine , Etienne Jérôme , Raoult Didier
Published: March 2006

A case-control study of three cases ofLegionella pneumophilapneumonia identified transesophageal echocardiography (TEE) as a risk factor. Patient isolates and environmental strains from water used for rinsing TEE probes were identical by pulsed-fleld gel electrophoresis. This is the first report of endoscopy as a potential source of legionellosis.
Epidemiologic Investigation ofBurkholderia cepaciaAcquisition in Two Pediatric Intensive Care Units
Authors: Loukil Chawki , Saizou Carole , Doit Catherine , Bidet Philippe , Mariani-Kurkdjian Patricia , Aujard Yannick , Beaufils François , Bingen Edouard
Published: March 2006

Objectives:

To investigate and describe an outbreak ofBurkholderia cepaciain a neonatal intensive care unit (NICU) and a pediatric intensive care unit (PICU), and to report the interventions leading to the cessation of the outbreak.

Design:

We conducted an epidemiologic investigation of an outbreak ofB. cepaciacolonization or infection in two clinical wards during a 35-month period (December 1998 to October 2001).

Setting:

A 500-bed, university hospital-affiliated, tertiary-care pediatric institution in Paris, France, with a 22-bed PICU and 31-bed NICU.

Methods:

Ribotyping was used to determine the genotypes ofB. cepaciaisolates. Procedures for the maintenance and disinfection of respiratory therapy devices were reviewed.

Results:

Thirty-two children were colonized (n = 14) or infected (n = 18) byB. cepaciain 2 wards (28 in the PICU and 4 in the NICU). In the PICU, a single ribotype was found among the isolates obtained from all of the patients except 1, and from the 6 isolates obtained from respiratory therapy devices (ie, heated humidifier water). In the NICU, the isolates obtained from the patients harbored a single ribotype unrelated to that of the epidemic strain isolated in the PICU; no environmental source of infection was found.

Conclusion:

Two different outbreaks appeared to be associated with 2 ribotypes, 1 of which was linked to patient-to-patient transmission via respiratory therapy devices. Complete elimination of the outbreak was achieved only when disposable, sterilizable, or easy-to-disinfect materials were used in the PICU. The source of infection in the NICU was not found.
Epidemiology of Methicillin-ResistantStaphylococcus aureusat a University Hospital in the Canary Islands
Authors: Montesinos Isabel , Salido Eduardo , Delgado Teresa , Lecuona Maria , Sierra Antonio
Published: March 2006

Objectives:

To describe the epidemiology of methicillin-resistantStaphylococcus aureus(MRSA) at a university hospital in Tenerife, Canary Islands, during a 40-month period and to evaluate the effectiveness of the application of control measures.

Design:

Laboratory-based surveillance, medical charts and microbiological records review, and characterization of strains by pulsed-field gel electrophoresis (PFGE) were used to describe the epidemiology. Infection control practices were introduced as an intervention.

Setting:

A 650-bed, tertiary-care university hospital.

Subjects:

Patients with clinical and nasal isolates of MRSA and colonized staff members.

Results:

The rate of nosocomial MRSA infections was 32.5% for 1997, 17.9% for 1998, 14.5% for 1999, and 25.6% during the first 4 months of 2000. The major sites of isolation for nosocomial MRSA infection included surgical wounds (25%) and the lower respiratory tract (24%). Intensive care units and surgical specialties had more frequent MRSA cases. Characteristics associated with nosocomial MRSA isolates included prior use of intensive antibiotic therapy, prolonged hospital stays, major underlying illness, invasive procedures, and older age. PFGE type A (subtype A1) was the strain most frequently found and the only PFGE type involved in clusters.

Conclusions:

Surveillance cultures and contact droplet precautions were followed by decreased rates for 2 years. Nevertheless, the spread of PFGE subtype Al to many different areas of the hospital and the increase in incidence during the first third of 2000 indicates either that surveillance cultures were not used widely enough or that compliance with isolation measures was suboptimal.
The Cadherin-11 Cytoplasmic Juxtamembrane Domain Promotes α-Catenin Turnover at Adherens Junctions and Intercellular Motility
Authors: Kiener Hans P. , Stipp Christopher S. , Allen Philip G. , Higgins Jonathan M.G. , Brenner Michael B.
Published: March 2006

Cadherins mediate homophilic cell adhesion and contribute to tissue morphogenesis and architecture. Cadherin cell adhesion contacts are actively remodeled and impact cell movement and migration over other cells. We found that expression of a mutant cadherin-11 lacking the cytoplasmic juxtamembrane domain (JMD) diminished the turnover of α-catenin at adherens junctions as measured by fluorescence recovery after photobleaching. This resulted in markedly diminished cell intercalation into monolayers reflecting reduced cadherin-11-dependent cell motility on other cells. Furthermore, the actin cytoskeleton in cadherin-11 ΔJMD cells revealed a more extensive cortical F-actin ring that correlated with significantly higher levels of activated Rac1. Together, these data implicate the cadherin-11 cytoplasmic JMD as a regulator of α-catenin turnover at adherens junctions and actin-cytoskeletal organization that is critical for intercellular motility and rearrangement in multicellular clusters.
JRAB/MICAL-L2 Is a Junctional Rab13-binding Protein Mediating the Endocytic Recycling of Occludin
Authors: Terai Tomoya , Nishimura Noriyuki , Kanda Ikuno , Yasui Natsuo , Sasaki Takuya
Published: March 2006

The dynamic turnover of tight junctions (TJs) is essential for epithelial-mesenchymal transitions and/or mesenchymal-epithelial transitions during epithelial morphogenesis. We previously demonstrated that Rab13 specifically mediates the endocytic recycling of occludin. Here, we identified MICAL-L2 (molecule interacting with CasL-like 2) as a novel Rab13-binding protein. Immunoprecipitation and immunofluorescence microscopy showed that MICAL-L2 specifically bound to the GTP-bound form of Rab13 via its C terminus, which contained a coiled-coil domain, and localized at TJs in epithelial MTD-1A cells. Recycling assay demonstrated that a MICAL-L2 mutant lacking the Rab13-binding domain (MICAL-L2-N) specifically inhibited the endocytic recycling of occludin but not transferrin receptor. Ca2+switch assay further revealed that MICAL-L2-N as well as Rab13 Q67L inhibited the recruitment of occludin to the plasma membrane, the development of transepithelial electrical resistance, and the formation of a paracellular diffusion barrier. MICAL-L2 was displaced from TJs upon actin depolymerization and was distributed along radiating actin cables and stress fibers in Ca2+-depleted MTD-1A and fibroblastic NIH3T3 cells, respectively. These results suggest that MICAL-L2 mediates the endocytic recycling of occludin and the formation of functional TJs by linking Rab13 to actin cytoskeleton. We rename MICAL-L2 as JRAB (junctional Rab13-binding protein).
Enhancement of Actin-depolymerizing Factor/Cofilin-dependent Actin Disassembly by Actin-interacting Protein 1 Is Required for Organized Actin Filament Assembly in theCaenorhabditis elegansBody Wall Muscle
Authors: Mohri Kurato , Ono Kanako , Yu Robinson , Yamashiro Sawako , Ono Shoichiro
Published: March 2006

Regulated disassembly of actin filaments is involved in several cellular processes that require dynamic rearrangement of the actin cytoskeleton. Actin-interacting protein (AIP) 1 specifically enhances disassembly of actin-depolymerizing factor (ADF)/cofilin-bound actin filaments. In vitro, AIP1 actively disassembles filaments, caps barbed ends, and binds to the side of filaments. However, how AIP1 functions in the cellular actin cytoskeletal dynamics is not understood. We compared biochemical and in vivo activities of mutant UNC-78 proteins and found that impaired activity of mutant UNC-78 proteins to enhance disassembly of ADF/cofilin-bound actin filaments is associated with inability to regulate striated organization of actin filaments in muscle cells. Six functionally important residues are present in the N-terminal β-propeller, whereas one residue is located in the C-terminal β-propeller, suggesting the presence of two separate sites for interaction with ADF/cofilin and actin. In vitro, these mutant UNC-78 proteins exhibited variable alterations in actin disassembly and/or barbed end-capping activities, suggesting that both activities are important for its in vivo function. These results indicate that the actin-regulating activity of AIP1 in cooperation with ADF/cofilin is essential for its in vivo function to regulate actin filament organization in muscle cells.
Type I Collagen in Hsp47-null Cells Is Aggregated in Endoplasmic Reticulum and Deficient in N-Propeptide Processing and Fibrillogenesis
Authors: Ishida Yoshihito , Kubota Hiroshi , Yamamoto Akitsugu , Kitamura Akira , Bächinger Hans Peter , Nagata Kazuhiro
Published: March 2006

Heat-shock protein of 47 kDa (Hsp47) is a molecular chaperone that recognizes collagen triple helices in the endoplasmic reticulum (ER). Hsp47-knockout mouse embryos are deficient in the maturation of collagen types I and IV, and collagen triple helices formed in the absence of Hsp47 show increased susceptibility to protease digestion. We show here that the fibrils of type I collagen produced by Hsp47-/-cells are abnormally thin and frequently branched. Type I collagen was highly accumulated in the ER of Hsp47-/-cells, and its secretion rate was much slower than that of Hsp47+/+cells, leading to accumulation of the insoluble aggregate of type I collagen within the cells. Transient expression of Hsp47 in the Hsp47-/-cells restored normal extracellular fibril formation and intracellular localization of type I collagen. Intriguingly, type I collagen with unprocessed N-terminal propeptide (N-propeptide) was secreted from Hsp47-/-cells and accumulated in the extracellular matrix. These results indicate that Hsp47 is required for correct folding and prevention of aggregation of type I collagen in the ER and that this function is indispensable for efficient secretion, processing, and fibril formation of collagen.
Tobacco mosaic virus (TMV) and potato virus X (PVX) coat proteins confer heterologous interference to PVX and TMV infection, respectively
Authors: Bazzini A. A. , Asurmendi S. , Hopp H. E. , Beachy R. N.
Published: March 2006

Replication ofPotato virus X(PVX) was reduced in transgenic protoplasts that accumulated wild-type coat protein (CPWT) ofTobacco mosaic virus(TMV) or a mutant CP, CPT42W, that produced highly ordered states of aggregation, including pseudovirions. This reaction is referred to as heterologous CP-mediated resistance. However, protoplasts expressing a CP mutant that abolished aggregation and did not produce pseudovirions, CPT28W, did not reduce PVX replication. Similarly, in transgenic tobacco plants producing TMV CPWTor CPT42W, there was a delay in local cell-to-cell spread of PVX infection that was not observed in CPT28Wplants or in non-transgenic plants. The results suggest that the quaternary structure of the TMV CP regulates the mechanism(s) of heterologous CP-mediated resistance. Similarly, transgenic protoplasts that produced PVX CP conferred transient protection against infection by TMV RNA. Transgenic plants that accumulated PVX CP reduced the cell-to-cell spread of infection and resulted in a delay in systemic infection following inoculation with TMV or TMV RNA. Heterologous CP-mediated resistance was characterized by a brief delay in systemic infection, whilst homologous CP-mediated resistance conferred reduced or no systemic infection.
PrPSc accumulation in fetal cotyledons of scrapie-resistant lambs is influenced by fetus location in the uterus
Authors: Alverson Janet , O'Rourke Katherine I. , Baszler Timothy V.
Published: March 2006

Placentae from scrapie-infected ewes have been shown to accumulate PrPSc when the genotype of the fetus is of a susceptible genotype (VRQ/VRQ, ARQ/VRQ or ARQ/ARQ). Cotyledons from fetuses of genotypes ARR/ARR, ARQ/ARR and ARQ/VRR have previously been shown to be resistant to PrPSc accumulation. By using ewes from a naturally infected scrapie flock, cotyledons from fetuses of multiple births of different genotypes were examined. PrPSc was detected in fetal cotyledons of genotype ARQ/ARQ, but not in cotyledons from their dizygotic twin of genotype ARQ/ARR. This confirms earlier reports of single fetuses of these genotypes, but is the first description of such a finding in twin fetuses, one of each genotype. It is also demonstrated that cotyledons from sibling fetuses of genotypes ARQ/VRQ and ARQ/ARQ have different patterns of PrPSc accumulation depending on whether the dam is of genotype ARQ/ARQ or ARQ/VRQ. Lastly, it is shown that cotyledons from fetuses with resistant genotypes are weakly positive for PrPSc when they have shared the same pregnant uterine horn with a fetus of a susceptible genotype with cotyledons positive for the detection of PrPSc. Additionally, a PCR product for the Sry gene, a product specific to males, was found in cotyledons from female fetuses that had shared a uterine horn with a male fetus. This indicates that some sharing of fetal blood occurs between placentomes and fetuses residing in the same uterine horn, which can result in PrPSc accumulation in cotyledons with resistant genotypes.
Site-directed mutagenesis of the VP2 gene of Chicken anemia virus affects virus replication, cytopathology and host-cell MHC class I expression
Authors: Peters Michelle A. , Crabb Brendan S. , Washington Elizabeth A. , Browning Glenn F.
Published: March 2006

Chicken anemia virus (CAV) is an immunosuppressive pathogen of chickens. To further examine the role of viral protein 2 (VP2), which possesses dual-specificity protein phosphatase (DSP) activity, in viral cytopathogenicity and its influence on viral growth and virulence, an infectious genomic clone of CAV was subjected to site-directed mutagenesis. Substitution mutations C87R, R101G, K102D and H103Y were introduced into the DSP catalytic motif and R129G, Q131P, R/K/K150/151/152G/A/A, D/E161/162G/G, L163P, D169G and E186G into a region predicted to have a high degree of secondary structure. All mutant constructs were infectious, but their growth curves differed. The growth curve for mutant virus R/K/K150/151/152G/A/A was similar to that for wild-type virus, a second cluster of mutant viruses had an extended latent period and a third cluster of mutant viruses had extended latent and eclipse periods. All mutants had a reduced cytopathogenic effect in infected cells and VP3 was restricted to the cytoplasm. Mutation of the second basic residue (K102D) in the atypical DSP signature motif resulted in a marked reduction in virus replication efficiency, whereas mutation of the first basic residue (R101G) attenuated cytopathogenicity, but did not reduce replication efficiency. Expression of major histocompatibility complex (MHC) class I was markedly downregulated in cells infected with wild-type CAV, but not in those infected with mutants. This study further demonstrates the significance of VP2 in CAV replication and shows that specific mutations introduced into the gene encoding this protein can reduce virus replication, cytopathogenicity and downregulation of MHC I in infected cells.
Herpesvirus of turkey reconstituted from bacterial artificial chromosome clones induces protection against Marek's disease
Authors: Baigent Susan J. , Petherbridge Lawrence J. , Smith Lorraine P. , Zhao Yuguang , Chesters Peter M. , Nair Venugopal K.
Published: March 2006

Herpesvirus of turkey (HVT) is an alphaherpesvirus that is widely used as a live vaccine against Marek's disease because of its antigenic relationship with Marek's disease virus (MDV). In spite of a similar genome structure, HVT has several unique genes, the functions of which are not completely understood. As a first step in carrying out detailed analysis of the functions of the HVT genes, a full-length infectious bacterial artificial chromosome (BAC) clone of HVT was constructed. DNA from two independent BAC clones, upon transfection into chicken embryo fibroblasts, produced plaques similar to those produced by the wild-type virus. Viruses derived from the BAC clones were stable during in vitro passage, but showed differences in in vitro growth kinetics compared with the wild-type virus. Using a one-step mutagenesis protocol to delete the essential glycoprotein B gene from the HVT genome, followed by construction of the revertant virus, BAC clones of HVT were shown to be amenable to standard mutagenesis techniques. In spite of the difference in in vitro growth, viruses from both clones induced 100 % protection against infection by the virulent MDV strain RB-1B, indicating that the BAC-derived viruses could be used as vaccines with efficacies similar to that of the parental virus. The construction of HVT BAC is a major step in understanding the functions of HVT genes by exploiting the power of BAC technology. Furthermore, the availability of the BAC clones enables use of HVT as a vector for expressing foreign genes.
Recombination of Feline calicivirus within an endemically infected cat colony
Authors: Coyne K. P. , Reed F. C. , Porter C. J. , Dawson S. , Gaskell R. M. , Radford A. D.
Published: March 2006

To understand the evolution of the family Caliciviridae, the persistence of Feline calicivirus (FCV) was studied within an endemically infected cat colony. Polymerase and capsid sequences were analysed for 34 FCV isolates obtained over a 4 year period. Initially, the colony was infected with one strain of virus, but a second distinct strain was later identified. Subsequently, the emergence of a recombinant virus was observed, containing elements of both of the strains circulating within the colony. The recombination event mapped close to the ORF1/ORF2 junction. This is consistent with recombination in other caliciviruses, suggesting a common mechanism within this family. This is the first report of recombination within the genus Vesivirus in the family Caliciviridae and the first time that a recombination event has been observed where the parental strains have also been identified.
Herpes simplex virus type 1 glycoprotein L mutants that fail to promote trafficking of glycoprotein H and fail to function in fusion can induce binding of glycoprotein L-dependent anti-glycoprotein H antibodies
Authors: Klyachkin Yuri M. , Stoops Krista D. , Geraghty Robert J.
Published: March 2006

The herpes simplex virus type 1 (HSV-1) glycoproteins H (gH) and L (gL) form a heterodimer and efficient expression of gH at the virion or cell surface is dependent upon gL. Five carboxy-terminal deletion mutants of gL were created and their ability to interact with and mediate cell-surface expression of gH, to promote binding of gL-dependent anti-gH antibodies and to contribute to cell fusion was analysed. All of the gL mutants bound gH, but only two mutants, containing the amino-terminal 161 or 168 aa of gL, mediated cell-surface expression of gH, and only gL161 and gL168 functioned in cell fusion. The binding of gL to gH, therefore, was not sufficient to ensure gH cell-surface expression and it was not possible to separate the gH-trafficking role of gL from gL function in fusion. Co-expression of gH with any gL mutant conferred binding of the anti-gH mAbs 53S and LP11. If the acquisition of 53S and LP11 binding to gH reflects a gL-induced conformational change, such a change is not sufficient to mediate trafficking of the gH–gL heterodimer.
Varicella-zoster virus influences the activities of components and targets of the ERK signalling pathway
Authors: Rahaus Markus , Desloges Nathalie , Wolff Manfred H.
Published: March 2006

Varicella-zoster virus (VZV) is ultimately dependent upon its host cell for replication. To ensure its reproduction, VZV reorganizes various cellular functions by taking advantage of pre-existing signalling pathways. Recently, it was demonstrated that the activation of stress-related mitogen-activated protein kinase pathways following infection led to increased phosphorylation of cellular transcription factors involved in VZV gene expression. Here, it was shown that members of the extracellular signal-regulated kinase (ERK) pathway are also influenced following VZV infection: c-Raf remained inactive in infected MeWo cells, whereas MEK1/2 and ERK1/2 were phosphorylated transiently, reaching their highest level of phosphorylation at between 10 and 12 h post-infection. Inhibition of this pathway resulted in a severe reduction in viral progeny and in an increased apoptotic response, indicating that the functionality of this cascade is essential for successful high-rate replication. In addition, the activities of Bad, a cytoplasmic target of ERK via ribosomal S6 kinase, and the nuclear-localized target c-Myc were analysed. Bad is a member of the Bcl-2 family and has a key function in regulating apoptosis. Pro-apoptotic functions of Bad are repressed by phosphorylation. A 10-fold increase in Bad phosphorylation at Ser-112 was detected following infection, which was suppressed after inhibition of ERK. The transcription factor c-Myc is involved in the regulation of cell growth and apoptosis. By performing immunoblots and quantitative RT-PCR, suppression of c-Myc expression was demonstrated at both the transcriptional and translational levels in VZV-infected cells. These results suggest that VZV optimizes the conditions for its replication in different ways: upregulation of proviral-acting systems and suppression of potentially antiviral-acting systems.
Prevalence of swine Torque teno virus in post-weaning multisystemic wasting syndrome (PMWS)-affected and non-PMWS-affected pigs in Spain
Authors: Kekarainen Tuija , Sibila Marina , Segalés Joaquim
Published: March 2006

The present study was designed to investigate the prevalence of swineTorque teno virus(TTV) in post-weaning multisystemic wasting syndrome (PMWS)-affected and non-affected Spanish swine. Nested PCR (nPCR) assays to detect two distinct TTV genogroups were applied. A significantly higher prevalence of TTV infection was found in sera from PMWS-affected animals (97 %) than in sera from non-PMWS-affected animals (78 %). Whilst PMWS-affected pigs (91 %) were more likely to be infected with TTV from genogroup 2 than non-PMWS-affected swine (72 %), no such difference was observed with genogroup 1. Nucleotide sequences of nPCR products were 91–99 % identical between strains within a genogroup. In contrast, inter-genogroup sequence identities were 49–58 %. Phylogenetic analyses demonstrated that genogroups form different clusters without association with PMWS or porcine circovirus type 2 infection status of the animals. These results indicate a high prevalence of both swine TTV genogroups in Spain, being present more frequently in PMWS-affected animals than in non-PMWS-affected animals.
Disentangling information flow in the Ras-cAMP signaling network
Authors: Carter Gregory W. , Rupp Steffen , Fink Gerald R. , Galitski Timothy
Published: March 2006

The perturbation of signal-transduction molecules elicits genomic-expression effects that are typically neither restricted to a small set of genes nor uniform. Instead there are broad, varied, and complex changes in expression across the genome. These observations suggest that signal transduction is not mediated by isolated pathways of information flow to distinct groups of genes in the genome. Rather, multiple entangled paths of information flow influence overlapping sets of genes. Using the Ras-cAMP pathway in Saccharomyces cerevisiae as a model system, we perturbed key pathway elements and collected genomic-expression data. Singular value decomposition was applied to separate the genome-wide transcriptional response into weighted expression components exhibited by overlapping groups of genes. Molecular interaction data were integrated to connect gene groups to perturbed signaling elements. The resulting series of linked subnetworks maps multiple putative pathways of information flow through a dense signaling network, and provides a set of testable hypotheses for complex gene-expression effects across the genome.
Post‐1990 Screen Memories: How East and West German Cinema Remembers the Third Reich and the Holocaust
Authors: Berghahn Daniela
Published: March 2006

The following article examines the contribution of German feature films about the Third Reich and the Holocaust to memory discourse in the wake of German unification. A comparison between East and West German films made since the 1990s reveals some startling asymmetries and polarities. While East German film‐makers, if they continued to work in Germany's reunified film industry at all, made very few films about the Third Reich, West German directors took advantage of the recent memory boom. Whereas films made by East German directors, such as Erster Verlust and Der Fall Ö, suggest, in liberating contradiction to the anti‐fascist interpretation of history, that East Germany shared the burden of guilt, West German productions subscribe to the normalisation discourse that has gained ideological hegemony in the East‐West‐German memory contest since unification. Films such as Aimée & Jaguar and Rosenstraße construct a memory of the past that is no longer encumbered by guilt, principally because the relationship between Germans and Jews is re‐imagined as one of solidarity. As post‐memory films, they take liberties with the traumatic memory of the past and, by following the generic conventions of melodrama, family saga and European heritage cinema, even lend it popular appeal.
An Unravelling Development Strategy? Garment Assembly in the Caribbean Basin after the Multifibre Arrangement
Authors: Heron Tony
Published: March 2006

The purpose of this article is to assess the likely impact of the ending of the Multifibre Arrangement (MFA) on garment assembly in the Caribbean Basin. Over the last fifteen years or so garment assembly has come to represent one of the region's few economic ‘success stories’ and has accordingly helped to offset the loss of export revenue from traditional commodities such as coffee and sugar. With the ending of the MFA on 1 January 2005, however, the long‐term viability of this trade has been thrown into doubt. This article thus seeks to assess the prospects for continued garment assembly in the Caribbean Basin against the backdrop of these developments.
Survivin expression, apoptosis and proliferation in chronic myelomonocytic leukemia
Authors: Invernizzi Rosangela , Travaglino Erica , Benatti Chiara , Malcovati Luca , Porta Matteo Della , Cazzola Mario , Ascari Edoardo
Published: March 2006

Abstract:  We analyzed the expression of the inhibitor of apoptosis survivin by immunocytochemistry in bone marrow cells from patients with chronic myelomonocytic leukemia (CMML) to evaluate possible abnormalities in comparison with other myelodysplastic (MDS) and myeloproliferative syndromes, and to investigate a possible correlation between survivin expression and altered apoptosis or proliferation, or relevant laboratory and clinical findings. Thirty‐four patients with CMML [18 MDS‐CMML and 16 myeloproliferative disorder (MPD)‐CMML], 90 with MDS, 41 with acute myeloid leukemia (AML), 19 with chronic MPD and 25 control subjects were studied. In normal samples survivin was never detectable. In CMML survivin levels higher than in MDS and AML (P < 0.0001), but similar to those found in MPD were observed. In CMML and MDS apoptosis was significantly higher compared to normal controls and all other subtypes of leukemias (P < 0.0001). Proliferation did not differ significantly in normal controls, MDS and CMML; the lowest levels were observed in AML and MPD (P < 0.0001). In CMML there was no correlation between survivin expression and blast cell percentage, apoptosis or proliferation, FAB or WHO subgroup. Proliferation was higher in MDS‐CMML and tended to correlate with overall survival. CMML‐2 cases with higher survivin expression showed higher evolution rate and shorter survival. In conclusion, CMML is characterized by high proliferation and apoptosis. Survivin overexpression, by disrupting the balance between cell proliferation/differentiation and apoptosis, may play an important role in its pathophysiology. The detection of survivin‐deregulated expression may provide a useful tool for diagnosis, prognosis and a possible target for experimental treatments.
Mania as a presenting symptom of Wilson's disease
Authors: Chand Prabhat Kumar , Murthy Pratima
Published: March 2006

Objective:

Wilson's disease commonly presents with neurological or hepatic manifestations. When it presents with only psychiatric symptoms, or with extrapyramidal symptoms secondary to neuroleptic exposure, the diagnosis of underlying Wilson's disease may be missed.

Methods:

An 18-year-old boy presented to the psychiatric clinic with a manic syndrome and high propensity for extrapyramidal symptoms to neuroleptic. Initial examination revealed splenomegaly and pancytopenia. Subsequent detection of Kayser–Fleischer ring and typical biochemical findings confirmed the diagnosis of Wilson's disease.

Results:

While the psychiatric symptoms came under control with lithium carbonate, extrapyramidal symptoms continue to persist even after neuroleptic withdrawal. Pancytopenia thought to be due to hypersplenism persists, and patient has developed features of liver cirrhosis. Treatment with zinc and folic acid has been started, and the patient is under evaluation for treatment with penicillamine.

Conclusion:

The psychiatrist needs to recognize that Wilson's disease can uncommonly present with isolated psychiatric symptoms, including mania. Early and severe extrapyramidal symptoms secondary to neuroleptic exposure in an adolescent age group warrants a detailed evaluation to rule out underlying neuropsychiatric conditions.
Cognitive performance following fluoxetine treatment in depressed patients post myocardial infarction
Authors: Strik Jacqueline J. M. H. , Honig Adriaan , Klinkenberg Edwin , Dijkstra Jeanette , Jolles Jelle
Published: March 2006

Background:

As depression is a considerable risk factor for an unfavourable course of myocardial infarction (MI), antidepressant treatment of post-MI depression and, inherent to MI status, polypharmacy has become an important issue.

Objective:

The present study is the first to evaluate cognitive side effects of fluoxetine, as part of a placebo-controlled double-blind trial, in patients with post-first MI depression.

Methods:

Cognitive performance of 54 depressed patients post first-MI, treated with fluoxetine or placebo was compared. Cognitive performance was tested before and after 9 weeks of treatment using the Visual Verbal Learning Test, Concept Shifting Task, Stroop Colour-Word Test and Letter-Digit-Substitution Test.

Results:

The median number of cardiovascular drugs taken by MI patients was 4.9. There were no differences between the fluoxetine and the placebo group on cognitive performance.

Conclusion:

In sum, there were no negative side effects of fluoxetine compared with placebo on cognition in depressed MI patients, simultaneously treated with cardiac drugs.
Changes in Canadian heroin supply coinciding with the Australian heroin shortage
Authors: Wood Evan , Stoltz Jo‐Anne , Li Kathy , Montaner Julio S. G. , Kerr Thomas
Published: March 2006

Aims  Previous studies have largely attributed the Australian heroin shortage to increases in local law enforcement efforts. Because western Canada receives heroin from similar source nations, but has not measurably increased enforcement practices or funding levels, we sought to examine trends in Canadian heroin‐related indices before and after the Australian heroin shortage, which began in approximately January 2001.

Methods  During periods before and after January 2001, we examined the number of fatal overdoses and ambulance responses to heroin‐related overdoses that required the use of naloxone in British Columbia, Canada. As an overall marker of Canadian supply reduction, we also examined the quantity of heroin seized during this period. Lastly, we examined trends in daily heroin use among injection drug users enrolled in the Vancouver Injection Drug Users Study (VIDUS).

Results  There was a 35% reduction in overdose deaths, from an annual average of 297 deaths during the years 1998–2000 in comparison to an average of 192 deaths during 2001–03. Similarly, use of naloxone declined 45% in the period coinciding with the Australian heroin shortage. Interestingly, the weight of Canadian heroin seized declined 64% coinciding with the Australian heroin shortage, from an average of 184 kg during 1998–2000 to 67 kg on average during 2001–03. Among 1587 VIDUS participants, the period coinciding with the Australian heroin shortage was associated independently with reduced daily injection of heroin [adjusted odds ratio: 0.55 (95% CI: 0.50–0.61); P < 0.001].

Conclusions  Massive decreases in three independent markers of heroin use have been observed in western Canada coinciding with the Australian heroin shortage, despite no increases in funding to Canadian enforcement efforts. Markedly reduced Canadian seizure activity also coincided with the Australian heroin shortage. These findings suggest that external global heroin supply forces deserve greater investigation and credence as a potential explanation for the Australian heroin shortage.
Evaluating the effectiveness of proactive telephone counselling for smoking cessation in a randomized controlled trial
Authors: Gilbert Hazel , Sutton Stephen
Published: March 2006

Aim  To evaluate the effectiveness of repeated‐contact proactive telephone counselling for smoking cessation in a UK setting.

Design  Randomized controlled trial.

Setting  The Quitline®, an established national telephone counselling service available throughout the UK.

Participants and intervention  A total of 1457 callers to the Quitline in 2000 and 2001 were allocated randomly to a Control group to receive usual care or to a Repeated Contact group to be offered five proactive calls in addition to usual care. 

Measurements  Prolonged abstinence and 24‐hour point‐prevalent abstinence 6 and 12 months after recruitment, quit attempts and 24‐hour periods of abstinence in non‐quitters.

Findings  No significant differences were found between the Repeated Contact and Control groups on prolonged or point‐prevalent abstinence. On an intention‐to‐treat basis, 9.5% of the Control group were abstinent for longer than 6 months at the 12‐month follow‐up, compared with 9.3% of the Repeated Contact group; 18.9% and 20.2%, respectively, were point‐prevalent abstinent at the 6‐month follow‐up. Significantly more non‐quitters in the Control group made a quit attempt in the first 6 months following recruitment than in the Repeated Contact group (62.6%/56.1%, P < 0.05).

Conclusions  Proactive telephone counselling did not significantly increase abstinence rates, and appeared to decrease quit attempts, in callers to the Quitline. A non‐structured, client‐led counselling protocol and insufficient pre‐quit motivational counselling could account for the lack of effect.
Differential regulation of platelet aggregation and aminophospholipid exposure by calpain
Authors: Bachelot‐Loza Christilla , Badol Perrine , Brohard‐Bohn Brigitte , Fraiz Nuria , Cano Ernesto , Rendu Francine
Published: March 2006

Aggregation, exposure of procoagulant phospholipids and shedding of microparticles are platelet responses that depend on activating conditions. To determine how these different responses are interconnected, we simultaneously measured fibrinogen (Fg) binding and aminophospholipid exposure on activated platelets by means of flow cytometry. Low calcium ionophore (A23187) concentrations induced Fg binding but not annexin V binding. In contrast, high A23187 concentrations induced annexin V binding but not Fg binding. Collagen, both alone and in the presence of thrombin, induced both Fg and annexin V binding. Dual labelling was found on 38 ± 9% of platelets stimulated by thrombin plus collagen. The regulatory role of calpain in these platelet functions was investigated. When calpain was partially inhibited by 2 μg/ml calpeptin, Fg binding still occurred but aminophospholipid exposure was limited. By contrast, complete inhibition of calpain by 100 μg/ml calpeptin or E64d decreased Fg binding but enhanced aminophospholipid exposure. In these latter conditions, cytosolic calcium‐extruding systems were inhibited. The results suggest that (i) conditions that favour aminophospholipid exposure tend to decrease the aggregation process and (ii) calpain determines the switch to either aggregation or aminophospholipid exposure by controlling intracellular calcium.
Monocyte‐derived dendritic cells from horses differ from dendritic cells of humans and mice
Authors: Mauel Susanne , Steinbach Falko , Ludwig Hanns
Published: March 2006

Dendritic cells (DC) are the initiators of immune responses and are present in most tissues in vivo. To generate myeloid DC from monocytes (MoDC) in vitro the necessary cytokines are granulocyte–macrophage colony‐stimulating factor (GM‐CSF) and interleukin‐4 (IL‐4). Using degenerated primers delineated from other species and rapid amplification of cDNA ends reverse transcription–polymerase chain reaction (RACE RT‐PCR), the cDNA of equine (eq.) GM‐CSF was cloned and found to have a point deletion at the 3′‐end of eq.GM‐CSF, resulting in a 24‐nucleotide extended open reading frame not described in any species thus far. For differentiating eq.MoDC, monocytes were stimulated with eq.GM‐CSF and eq.IL‐4. The eq.MoDC was analysed by both light and electron microscopy and by flow cytometry and mixed lymphocyte reaction. The eq.MoDC obtained had the typical morphology and function of DC, including the ability to stimulate allogeneic T cells in a mixed lymphocyte reaction. In contrast to the human system, however, monocytes had to be differentiated for 6–7 days before immature DC were obtained. Our data also indicate that lipopolysaccharide or poly(I:C) alone are not sufficient to confer the full phenotypic transition into mature DC. Thus our study contributes to understanding the heterogeneity of immunity and adds important information on the equine immune system, which is clearly distinct from those of mice or man.
Interleukin‐4‐dependent innate collaboration between iNKT cells and B‐1 B cells controls adaptative contact sensitivity
Authors: Campos Regis A. , Szczepanik Marian , Itakura Atsuko , Lisbonne Mariette , Dey Neelendu , Leite‐de‐Moraes Maria C. , Askenase Philip W.
Published: March 2006

We showed that hepatic Vα14+ invariant natural killer T (iNKT) cells, via their rapid interleukin (IL)‐4 production, activate B‐1 cells to initiate contact sensitivity (CS). This innate collaboration was absent in IL‐4–/– and signal transducer and activator of transcription (STAT)‐6–/– mice and was inhibited by anti‐IL‐4 treatment. These mice have defective CS because they fail to locally recruit the sensitized effector T cells of acquired immunity. Their CS is reconstituted by transfer of downstream‐acting 1‐day immune B‐1 cells from wild‐type mice. Responses were not reconstituted with B‐1 cells from IL‐4 receptor‐α–/– or STAT‐6–/– mice, nor by IL‐4 treatment of B cell‐deficient mice at immunization. Finally, IL‐4 was preferentially and transiently produced by hepatic iNKT cells within 7 min after sensitization to mediate collaboration between innate‐like iNKT cells and the B‐1 B cells that participate in the recruitment of effector T cells in vivo.
Drinking a lot is good for dendritic cells
Authors: Norbury Christopher C.
Published: March 2006

Macropinocytosis is the actin‐dependent formation of large vesicles, which allow the internalization of large quantities of fluid‐phase solute. In the majority of cells examined, an exogenous stimulus is required to induce the initiation of this endocytic pathway. However, dendritic cells are thought to constitutively macropinocytose large quantities of exogenous solute as part of their sentinel function. In this review we discuss the evidence that dendritic cells macropinocytose exogenous solute and subsequently present antigenic peptides derived from internalized material to T cells. In addition, we put these data into the context of immune surveillance in vivo.
Long‐term fire exclusion and ant community structure in an Australian tropical savanna: congruence with vegetation succession
Authors: Andersen Alan N. , Hertog Tony , Woinarski John C. Z.
Published: March 2006

Aim  To examine the extent to which succession from tropical savanna to rain forest in the long‐term absence of fire is matched by successional changes in ant communities. This is done by describing ant community responses to 23 years of fire exclusion in a northern Australian tropical savanna, with a particular focus on the extent of colonization by specialist rain forest taxa.

Location  Solar Village, near Darwin in Australia's Northern Territory.

Methods  Ants were sampled within 12 plots located inside (‘unburnt’– protected from fire for 23 years) and outside (burnt every 1–2 years) Solar Village in ridge and slope habitat dominated by Eucalyptus spp. The litter, ground‐foraging and arboreal faunas were sampled separately, using Berlese funnels, unbaited pitfall traps and baited pitfall traps attached to tree trunks, respectively. Each species was assigned a forest‐association score ranging from 0 (open savanna species) to 3 (specialist forest species) based on their known habitat preferences in the region.

Results  A total of 85 ant species from 35 genera were recorded, with multivariate analysis demonstrating distinct litter, ground and arboreal communities. Ant communities also varied substantially with topographic position, which interacted strongly with fire exclusion. A total of 72 species were recorded in burnt habitat, compared with only 45 in unburnt, and the number of ant species records was also about twice as high in burnt compared with unburnt habitat. Fire exclusion has resulted in a dramatic increase in forest‐associated taxa (those occurring in forest and denser, but rarely open, savanna), with such species representing 51% of species records in unburnt habitat compared with 19% in burnt. However, only five specialist forest species were recorded, representing < 1% of total ant records.

Main conclusions  Fire exclusion at Solar Village has markedly increased the prevalence of forest‐associated ant species, but has led to only very minor incursions by specialist rain forest ant taxa. These responses match very closely those of the vegetation.