bluuebunny/crossref_metadata_2025 huggingface dataset:
Alanine Scanning of Poliovirus 2C
ATPase
Reveals New Genetic Evidence that Capsid Protein/2C
ATPase
Interactions Are Essential for Morphogenesis
Authors: Wang Chunling , Jiang Ping , Sand Claire , Paul Aniko V. , Wimmer Eckard
Published: July 2012

Polypeptide 2C
ATPase
is one of the most thoroughly studied but least understood proteins in the life cycle of poliovirus. Within the protein, multiple functional domains important for uncoating, host cell membrane alterations, and RNA replication and encapsidation have previously been identified. In this study, charged to alanine-scanning mutagenesis was used to generate conditional-lethal mutations in hitherto-uncharacterized domains of the 2C
ATPase
polypeptide, particularly those involved in morphogenesis. Adjacent or clustered charged amino acids (2 to 4), scattered along the 2C
ATPase
coding sequence, were replaced with alanines. RNA transcripts of mutant poliovirus cDNA clones were transfected into HeLa cells. Subsequently, 10 lethal, 1 severely temperature-sensitive, 2 quasi-infectious, and 3 wild type-like mutants were identified. Using a luciferase-containing reporter virus, we demonstrated RNA replication defects in all lethal and quasi-infectious mutants. Temperature-sensitive mutants were defective in RNA replication only at the restricted temperatures. Furthermore, we characterized a quasi-infectious mutant (K
6
A/K
7
A) that produced a suppressor mutation (G
1
R) and a novel 2B∧2C
ATPase
cleavage site (Q∧R). Surprisingly, this cleavage site mutation did not interfere with normal processing of the polyprotein. These mutants have led to the identification of several new sites within the 2C
ATPase
polypeptide that are required for RNA replication. In addition, analysis of the suppressor mutants has revealed a new domain near the C terminus of 2C
ATPase
that is involved in encapsidation, possibly achieved through interaction with an amino acid sequence between NTP binding motifs A and B of 2C
ATPase
. Most importantly, the identification of suppressor mutations in both 2C
ATPase
and the capsid domains (VP1 and VP3) of poliovirus has confirmed that an interaction between 2C
ATPase
and capsid proteins is involved in viral morphogenesis.
Interferon Response Factors 3 and 7 Protect against Chikungunya Virus Hemorrhagic Fever and Shock
Authors: Rudd Penny A. , Wilson Jane , Gardner Joy , Larcher Thibaut , Babarit Candice , Le Thuy T. , Anraku Itaru , Kumagai Yutaro , Loo Yueh-Ming , Gale Michael , Akira Shizuo , Khromykh Alexander A. , Suhrbier Andreas
Published: July 2012

Chikungunya virus (CHIKV) infections can produce severe disease and mortality. Here we show that CHIKV infection of adult mice deficient in interferon response factors 3 and 7 (IRF3/7
−/−
) is lethal. Mortality was associated with undetectable levels of alpha/beta interferon (IFN-α/β) in serum, ∼50- and ∼10-fold increases in levels of IFN-γ and tumor necrosis factor (TNF), respectively, increased virus replication, edema, vasculitis, hemorrhage, fever followed by hypothermia, oliguria, thrombocytopenia, and raised hematocrits. These features are consistent with hemorrhagic shock and were also evident in infected IFN-α/β receptor-deficient mice.
In situ
hybridization suggested CHIKV infection of endothelium, fibroblasts, skeletal muscle, mononuclear cells, chondrocytes, and keratinocytes in IRF3/7
−/−
mice; all but the latter two stained positive in wild-type mice. Vaccination protected IRF3/7
−/−
mice, suggesting that defective antibody responses were not responsible for mortality. IPS-1- and TRIF-dependent pathways were primarily responsible for IFN-α/β induction, with IRF7 being upregulated >100-fold in infected wild-type mice. These studies suggest that inadequate IFN-α/β responses following virus infection can be sufficient to induce hemorrhagic fever and shock, a finding with implications for understanding severe CHIKV disease and dengue hemorrhagic fever/dengue shock syndrome.
Rotavirus VP8*: Phylogeny, Host Range, and Interaction with Histo-Blood Group Antigens
Authors: Liu Yang , Huang Pengwei , Tan Ming , Liu Yiliu , Biesiada Jacek , Meller Jarek , Castello Alejandro A. , Jiang Baoming , Jiang Xi
Published: July 2012

The distal portion of rotavirus (RV) VP4 spike protein (VP8*) is implicated in binding to cellular receptors, thereby facilitating viral attachment and entry. While VP8* of some animal RVs engage sialic acid, human RVs often attach to and enter cells in a sialic acid-independent manner. A recent study demonstrated that the major human RVs (P[4], P[6], and P[8]) recognize human histo-blood group antigens (HBGAs). In this study, we performed a phylogenetic analysis of RVs and showed further variations of RV interaction with HBGAs. On the basis of the VP8* sequences, RVs are grouped into five P genogroups (P[I] to P[V]), of which P[I], P[IV], and P[V] mainly infect animals, P[II] infects humans, and P[III] infects both animals and humans. The sialic acid-dependent RVs (P[1], P[2], P[3], and P[7]) form a subcluster within P[I], while all three major P genotypes of human RVs (P[4], P[6], and P[8]) are clustered in P[II]. We then characterized three human RVs (P[9], P[14], and P[25]) in P[III] and observed a new pattern of binding to the type A antigen which is distinct from that of the P[II] RVs. The binding was demonstrated by hemagglutination and saliva binding assay using recombinant VP8* and native RVs. Homology modeling and mutagenesis study showed that the locations of the carbohydrate binding interfaces are shared with the sialic acid-dependent RVs, although different amino acids are involved. The P[III] VP8* proteins also bind the A antigens of the porcine and bovine mucins, suggesting the A antigen as a possible factor for cross-species transmission of RVs. Our study suggests that HBGAs play an important role in RV infection and evolution.
Cyclophilins Facilitate Dissociation of the Human Papillomavirus Type 16 Capsid Protein L1 from the L2/DNA Complex following Virus Entry
Authors: Bienkowska-Haba Malgorzata , Williams Carlyn , Kim Seong Man , Garcea Robert L. , Sapp Martin
Published: July 2012

Human papillomaviruses (HPV) are composed of the major and minor capsid proteins, L1 and L2, that encapsidate a chromatinized, circular double-stranded DNA genome. At the outset of infection, the interaction of HPV type 16 (HPV16) (pseudo)virions with heparan sulfate proteoglycans triggers a conformational change in L2 that is facilitated by the host cell chaperone cyclophilin B (CyPB). This conformational change results in exposure of the L2 N terminus, which is required for infectious internalization. Following internalization, L2 facilitates egress of the viral genome from acidified endosomes, and the L2/DNA complex accumulates at PML nuclear bodies. We recently described a mutant virus that bypasses the requirement for cell surface CyPB but remains sensitive to cyclosporine for infection, indicating an additional role for CyP following endocytic uptake of virions. We now report that the L1 protein dissociates from the L2/DNA complex following infectious internalization. Inhibition and small interfering RNA (siRNA)-mediated knockdown of CyPs blocked dissociation of L1 from the L2/DNA complex.
In vitro
, purified CyPs facilitated the dissociation of L1 pentamers from recombinant HPV11 L1/L2 complexes in a pH-dependent manner. Furthermore, CyPs released L1 capsomeres from partially disassembled HPV16 pseudovirions at slightly acidic pH. Taken together, these data suggest that CyPs mediate the dissociation of HPV L1 and L2 capsid proteins following acidification of endocytic vesicles.
Gag-Specific Cellular Immunity Determines
In Vitro
Viral Inhibition and
In Vivo
Virologic Control following Simian Immunodeficiency Virus Challenges of Vaccinated Rhesus Monkeys
Authors: Stephenson Kathryn E. , Li Hualin , Walker Bruce D. , Michael Nelson L. , Barouch Dan H.
Published: July 2012

A comprehensive vaccine for human immunodeficiency virus type 1 (HIV-1) would block HIV-1 acquisition as well as durably control viral replication in breakthrough infections. Recent studies have demonstrated that Env is required for a vaccine to protect against acquisition of simian immunodeficiency virus (SIV) in vaccinated rhesus monkeys, but the antigen requirements for virologic control remain unclear. Here, we investigate whether CD8
+
T lymphocytes from vaccinated rhesus monkeys mediate viral inhibition
in vitro
and whether these responses predict virologic control following SIV challenge. We observed that CD8
+
lymphocytes from 23 vaccinated rhesus monkeys inhibited replication of SIV
in vitro
. Moreover, the magnitude of inhibition prior to challenge was inversely correlated with set point SIV plasma viral loads after challenge. In addition, CD8 cell-mediated viral inhibition in vaccinated rhesus monkeys correlated significantly with Gag-specific, but not Pol- or Env-specific, CD4
+
and CD8
+
T lymphocyte responses. These findings demonstrate that
in vitro
viral inhibition following vaccination largely reflects Gag-specific cellular immune responses and correlates with
in vivo
virologic control following infection. These data suggest the importance of including Gag in an HIV-1 vaccine in which virologic control is desired.
Interaction between the Human Cytomegalovirus Tegument Proteins UL94 and UL99 Is Essential for Virus Replication
Authors: Phillips Stacia L. , Cygnar Daniel , Thomas Alexandra , Bresnahan Wade A.
Published: July 2012

Human cytomegalovirus (HCMV) virions are structurally complex, and the mechanisms by which they are assembled are poorly understood, especially with respect to the cytoplasmic phase of assembly, during which the majority of the tegument is acquired and final envelopment occurs. These processes occur at a unique cytoplasmic structure called the assembly complex, which is formed through a reorganization of the cellular secretory apparatus. The HCMV tegument protein UL99 (pp28) is essential for viral replication at the stage of secondary envelopment. We previously demonstrated that UL99 interacts with the essential tegument protein UL94 in infected cells as well as in the absence of other viral proteins. Here we show that UL94 and UL99 alter each other's localization and that UL99 stabilizes UL94 in a binding-dependent manner. We have mapped the interaction between UL94 and UL99 to identify the amino acids of each protein that are required for their interaction. Mutation of these amino acids in the context of the viral genome demonstrates that HCMV is completely defective for replication in the absence of the interaction between UL94 and UL99. Further, we demonstrate that in the absence of their interaction, both UL94 and UL99 exhibit aberrant localization and do not accumulate at the assembly complex during infection. Taken together, our data suggest that the interaction between UL94 and UL99 is essential for the proper localization of each protein to the assembly complex and thus for the production of infectious virus.
Dissecting Quasi-Equivalence in Nonenveloped Viruses: Membrane Disruption Is Promoted by Lytic Peptides Released from Subunit Pentamers, Not Hexamers
Authors: Domitrovic Tatiana , Matsui Tsutomu , Johnson John E.
Published: July 2012

Nonenveloped viruses often invade membranes by exposing hydrophobic or amphipathic peptides generated by a proteolytic maturation step that leaves a lytic peptide noncovalently associated with the viral capsid. Since multiple copies of the same protein form many nonenveloped virus capsids, it is unclear if lytic peptides derived from subunits occupying different positions in a quasi-equivalent icosahedral capsid play different roles in host infection. We addressed this question with
Nudaurelia capensis omega virus
(NωV), an insect RNA virus with an icosahedral capsid formed by protein α, which undergoes autocleavage during maturation, producing the lytic γ peptide. NωV is a unique model because autocatalysis can be precisely initiated
in vitro
and is sufficiently slow to correlate lytic activity with γ peptide production. Using liposome-based assays, we observed that autocatalysis is essential for the potent membrane disruption caused by NωV. We observed that lytic activity is acquired rapidly during the maturation program, reaching 100% activity with less than 50% of the subunits cleaved. Previous time-resolved structural studies of partially mature NωV particles showed that, during this time frame, γ peptides derived from the pentamer subunits are produced and are organized in a vertical helical bundle that is projected toward the particle surface, while identical polypeptides in quasi-equivalent subunits are produced later or are in positions inappropriate for release. Our functional data provide experimental support for the hypothesis that pentamers containing a central helical bundle, observed in different nonenveloped virus families, are a specialized lytic motif.
Adaptive Changes in Alphavirus mRNA Translation Allowed Colonization of Vertebrate Hosts
Authors: Ventoso Iván
Published: July 2012

Members of the
Alphavirus
genus are arboviruses that alternate replication in mosquitoes and vertebrate hosts. In vertebrate cells, the alphavirus resists the activation of antiviral RNA-activated protein kinase (PKR) by the presence of a prominent RNA structure (downstream loop [DLP]) located in viral 26S transcripts, which allows an eIF2-independent translation initiation of these mRNAs. This article shows that DLP structure is essential for replication of Sindbis virus (SINV) in vertebrate cell lines and animals but is dispensable for replication in insect cells, where no ortholog of the vertebrate
PKR
gene has been found. Sequence comparisons and structural RNA analysis revealed the evolutionary conservation of DLP in SINV and predicted the existence of equivalent DLP structures in many members of the
Alphavirus
genus. A mutant SINV lacking the DLP structure evolved in murine cells to recover a wild-type phenotype by creating an alternative structure in the RNA that restored the translational independence for eIF2. Genetic, phylogenetic, and biochemical data presented here support an evolutionary scenario for the natural history of alphaviruses, in which the acquisition of DLP structure in their mRNAs probably allowed the colonization of vertebrate host and the consequent geographic expansion of some of these viruses worldwide.
Conformational Switch-Defective ϕX174 Internal Scaffolding Proteins Kinetically Trap Assembly Intermediates before Procapsid Formation
Authors: Gordon Emile B. , Knuff Christopher J. , Fane Bentley A.
Published: July 2012

Conformational switching is an overarching paradigm in which to describe scaffolding protein-mediated virus assembly. However, rapid morphogenesis with small assembly subunits hinders the isolation of early morphogenetic intermediates in most model systems. Consequently, conformational switches are often defined by comparing the structures of virions, procapsids and aberrantly assembled particles. In contrast, ϕX174 morphogenesis proceeds through at least three preprocapsid intermediates, which can be biochemically isolated. This affords a detailed analysis of early morphogenesis and internal scaffolding protein function. Amino acid substitutions were generated for the six C-terminal, aromatic amino acids that mediate most coat-internal scaffolding protein contacts. The biochemical characterization of mutant assembly pathways revealed two classes of molecular defects, protein binding and conformational switching, a novel phenotype. The conformational switch mutations kinetically trapped assembly intermediates before procapsid formation. Although mutations trapped different particles, they shared common second-site suppressors located in the viral coat protein. This suggests a fluid assembly pathway, one in which the scaffolding protein induces a single, coat protein conformational switch and not a series of sequential reactions. In this model, an incomplete or improper switch would kinetically trap intermediates.
Virion-Associated Complement Regulator CD55 Is More Potent than CD46 in Mediating Resistance of Mumps Virus and Vesicular Stomatitis Virus to Neutralization
Authors: Johnson John B. , Lyles Douglas S. , Alexander-Miller Martha A. , Parks Griffith D.
Published: July 2012

Enveloped viruses can incorporate host cell membrane proteins during the budding process. Here we demonstrate that mumps virus (MuV) and vesicular stomatitis virus (VSV) assemble to include CD46 and CD55, two host cell regulators which inhibit propagation of complement pathways through distinct mechanisms. Using viruses which incorporated CD46 alone, CD55 alone, or both CD46 and CD55, we have tested the relative contribution of these regulators in resistance to complement-mediated neutralization. Virion-associated CD46 and CD55 were biologically active, with VSV showing higher levels of activity of both cofactors, which promoted factor I-mediated cleavage of C3b into iC3b as well as decay-accelerating factor (DAF) activity against the C3 convertase, than MuV. Time courses ofin vitroneutralization with normal human serum (NHS) showed that both regulators could delay neutralization, but viruses containing CD46 alone were neutralized faster and more completely than viruses containing CD55 alone. A dominant inhibitory role for CD55 was most evident for VSV, where virus containing CD55 alone was not substantially different in neutralization kinetics from virus harboring both regulators. Electron microscopy showed that VSV neutralization proceeded through virion aggregation followed by lysis, with virion-associated CD55 providing a delay in both aggregation and lysis more substantial than that conferred by CD46. Our results demonstrate the functional significance of incorporation of host cell factors during virion envelope assembly. They also define pathways of virus complement-mediated neutralization and suggest the design of more effective viral vectors.
A Myeloid Progenitor Cell Line Capable of Supporting Human Cytomegalovirus Latency and Reactivation, Resulting in Infectious Progeny
Authors: O'Connor Christine M. , Murphy Eain A.
Published: July 2012

Human cytomegalovirus (HCMV) is a herpesvirus that establishes a lifelong, latent infection within a host. At times when the immune system is compromised, the virus undergoes a lytic reactivation producing infectious progeny. The identification and understanding of the biological mechanisms underlying HCMV latency and reactivation are not completely defined. To this end, we have developed a tractablein vitromodel system to investigate these phases of viral infection using a clonal population of myeloid progenitor cells (Kasumi-3 cells). Infection of these cells results in maintenance of the viral genome with restricted viral RNA expression that is reversed with the addition of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA, also known as PMA). Additionally, a latent viral transcript (LUNA) is expressed at times where viral lytic transcription is suppressed. Infected Kasumi-3 cells initiate production of infectious virus following TPA treatment, which requires cell-to-cell contact for efficient transfer of virus to other cell types. Importantly, lytically infected fibroblast, endothelial, or epithelial cells can transfer virus to Kasumi-3 cells, which fail to initiate lytic replication until stimulated with TPA. Finally, inflammatory cytokines, in addition to the pharmacological agent TPA, are sufficient for transcription of immediate-early (IE) genes following latent infection. Taken together, our findings argue that the Kasumi-3 cell line is a tractablein vitromodel system with which to study HCMV latency and reactivation.
Stem Cell-Derived Human Intestinal Organoids as an Infection Model for Rotaviruses
Authors: Finkbeiner Stacy R. , Zeng Xi-Lei , Utama Budi , Atmar Robert L. , Shroyer Noah F. , Estes Mary K.
Published: July 2012

Directed differentiation of stem cell lines into intestine-like tissue called induced human intestinal organoids (iHIOs) is now possible (J. R. Spence, C. N. Mayhew, S. A. Rankin, M. F. Kuhar, J. E. Vallance, K. Tolle, E. E. Hoskins, V. V. Kalinichenko, S. I. Wells, A. M. Zorn, N. F. Shroyer, and J. M. Wells, Nature 470:105-109, 2011). We tested iHIOs as a new model to cultivate and study fecal viruses. Protocols for infection of iHIOs with a laboratory strain of rotavirus, simian SA11, were developed. Proof-of-principle analyses showed that iHIOs support replication of a gastrointestinal virus, rotavirus, on the basis of detection of nonstructural viral proteins (nonstructural protein 4 [NSP4] and NSP2) by immunofluorescence, increased levels of viral RNA by quantitative reverse transcription-PCR (qRT-PCR), and production of infectious progeny virus. iHIOs were also shown to support replication of 12/13 clinical rotavirus isolates directly from stool samples. An unexpected finding was the detection of rotavirus infection not only in the epithelial cells but also in the mesenchymal cell population of the iHIOs. This work demonstrates that iHIOs offer a promising new model to study rotaviruses and other gastrointestinal viruses.

IMPORTANCE
Gastrointestinal viral infections are a major cause of illness and death in children and adults. The ability to fully understand how viruses interact with human intestinal cells in order to cause disease has been hampered by insufficient methods for growing many gastrointestinal viruses in the laboratory. Induced human intestinal organoids (iHIOs) are a promising new model for generating intestine-like tissue. This is the first report of a study using iHIOs to cultivate any microorganism, in this case, an enteric virus. The evidence that both laboratory and clinical rotavirus isolates can replicate in iHIOs suggests that this model would be useful not only for studies of rotaviruses but also potentially of other infectious agents. Furthermore, detection of rotavirus proteins in unexpected cell types highlights the promise of this system to reveal new questions about pathogenesis that have not been previously recognized or investigated in other intestinal cell culture models.
Ethanolamine: A Signal to Commence a Host-Associated Lifestyle?
Authors: Garsin Danielle A.
Published: July 2012

Ethanolamine (EA) is a compound prevalent in the gastrointestinal (GI) environment. The ability to catabolize this compound has been associated with important GI pathogens, including enterohemorrhagic
Escherichia coli
O157:H7 (EHEC). It has been hypothesized that the ability of EHEC to utilize EA as a source of nitrogen provides EHEC with an important competitive advantage in the gut. However, new work by Kendall et al. (mBio 3:e00050-12, 2012) suggests that the role of EA in EHEC pathogenesis may be more fundamental; EA appears to be a signal for EHEC to commence its virulence program. In this commentary, I review the previously described connections of EA to bacterial pathogenesis in the GI tract, highlight the important findings of this new study, and note areas where further research is needed to fully comprehend the connection of EA utilization to bacterial pathogenesis.
Inconsistencies in Nice Guidance for Acupuncture: Reanalysis and Discussion
Authors: Latimer Nicholas R , Bhanu Aathish Chandrika , Whitehurst David G T
Published: July 2012

Background

Acupuncture received a positive recommendation in the National Institute for Health and Clinical Excellence (NICE) clinical guideline for low back pain (LBP). However, no such recommendation was forthcoming in the NICE clinical guideline for osteoarthritis (OA). Importantly, the two guidelines adopted different treatment comparators in their economic analyses of acupuncture; in the LBP guideline ‘usual care’ was used (with no consideration of placebo/sham interventions), whereas ‘sham acupuncture’ was the comparator in the OA guideline.

Objective

To analyse the implications of using different control group comparators when estimating the cost-effectiveness of acupuncture therapy.

Methods

The NICE OA economic analysis for acupuncture was replicated using ‘usual care’ (ie, no placebo/sham component) as the treatment comparator. A ‘transfer-to-utility’ technique was used to transform Western Ontario and McMaster Osteoarthritis scores into EQ-5D utility scores to allow quality-adjusted life year (QALY) gains to be estimated. QALY estimates were combined with direct incremental cost estimates of acupuncture treatment to determine incremental cost-effectiveness ratios (ICERs).

Results

When ‘usual care’ was used as the treatment comparator, ICER point estimates were below £20 000 per QALY gained for each acupuncture trial analysed in the OA clinical guideline. In the original analysis, using placebo/sham acupuncture as the treatment comparator, ICERs were generally above £20 000 per QALY gained.

Conclusion

The treatment comparator chosen in economic evaluations of acupuncture therapy is likely to be a strong determinant of the cost-effectiveness results. Different comparators used in the OA and LBP NICE guidelines may have led to the divergent recommendations in the guidelines.
An ecological study on the geographic patterns of ischaemic heart disease in Portugal and its association with demography, economic factors and health resources distribution
Authors: Ferreira-Pinto Luís Manuel , Rocha-Gonçalves Francisco , Teixeira-Pinto Armando
Published: July 2012

Objectives

Being one of the main causes of morbidity and mortality in developed countries, ischaemic heart disease's (IHD) incidence and mortality present clear differences between and within countries. Several authors already proposed possible explanations based on the demography, environmental factors, diet and level of urbanisation. This study reflects the Portuguese reality concerning IHD, by analysing the geographical distribution of hospital admissions and mortality due to this condition, in Portugal, and its association with demography, economical factors and the distribution of healthcare resources at the regional level.

Design

Ecological study.

Setting

Data from all Portuguese Public Hospitals were obtained using the National Registry of Hospital Admissions, between 2000 and 2007, and data on demography, economical factors and health resources distribution were obtained from the National Institute of Statistics.

Participants

Aggregated statistics on hospital admissions and mortality were computed for 278 counties based on almost 200 000 admissions.

Primary and secondary outcome measures

Mortality rate; hospital admissions rate.

Results

The geographical distribution of non-adjusted mortality and hospital admission showed an inner/coastal pattern but no North/South gradient was clear. Counties with higher economical development had significantly higher mortality and admission rates. However, healthcare resources distribution was not significantly associated with IHD hospital admission and mortality. When adjusted for age, gender, economic development and health resources distribution, there was still unexplained geographical variation both in hospital admissions and mortality rates.

Conclusion

A pattern in the geographic distribution of incidence and mortality of IHD was clear even after the adjustment for age and gender. Economical variables were the ones presenting the strongest association. These types of analysis may be very helpful for the definition of health policies, in particular to identify priority regions for disease prevention and guidelines for healthcare resources distribution.
Socio-demographic characteristics of women sustaining injuries during pregnancy: a study from the Danish National Birth Cohort
Authors: Virk Jasveer , Hsu Paul , Olsen Jørn
Published: July 2012

Objectives

To describe adverse birth outcomes associated with hospital-treated injuries that took place among women in the Danish National Birth Cohort.

Design

Longitudinal cohort study.

Setting

Denmark.

Participants

90 452 women and their offspring selected from the Danish National Birth Cohort.

Primary and secondary outcome measures

To determine if injured women were more likely to deliver an infant preterm, with low birth weight, stillborn or have a spontaneous abortion, the authors estimated HRs. ORs were generated to assess APGAR scores and infants born small for gestational age (SGA). Models were adjusted for maternal smoking and drinking during pregnancy, household socioeconomic status, eclampsia/pre-eclampsia or gestational diabetes status during pregnancy and maternal age at birth; estimates for preterm birth were also adjusted for prior history of preterm birth.

Results

In the cohort of 90 452 pregnant women, 3561 (3.9%) received medical treatment for an injury during pregnancy. Injured pregnant women were more likely to deliver infants that were stillborn or have pregnancies terminated by spontaneous abortion. The authors did not detect an adverse effect between injuries sustained during pregnancy and delivery of preterm, low birth weight or SGA infants, or infants with an APGAR score of <7.

Conclusions

The study shows that injuries occurring among women from an unselected population may not have an adverse effect on birth weight, gestational age, APGAR score or SGA status but may adversely affect the risk of stillbirth and spontaneous abortions in some situations.
Differential use of extended and immediate release quetiapine: a retrospective registry study of Finnish inpatients with schizophrenia spectrum and bipolar disorders
Authors: Hallinen Taru , Soini Erkki J , Granström Ola , Ovaskainen Yrjö , Leinonen Esa , Koponen Hannu J , Hänninen Kari
Published: July 2012

Objective

Extended release (XR) and immediate release (IR) quetiapine have differing dosing, titration and plasma concentration profiles. The authors assessed whether the use of quetiapine XR and IR in schizophrenia spectrum disorders (SCZ) and bipolar disorder (BD) differ.

Design

Retrospective non-interventional registry study.

Setting

Secondary healthcare.

Participants

All SCZ and BD (ICD-10 codes F20–F29, F30–F31) patients discharged between June 2008 and June 2010 from a Finnish psychiatric hospital with any use of quetiapine during their inpatient stay.

Primary and secondary outcome measures

Differences in patient characteristics between quetiapine XR and IR users were tested. To assess the profile of XR versus IR patients, logistic regressions were performed.

Results

43 patients used quetiapine XR, 58 used quetiapine IR and 55 used both formulations (n=156). 102 patients were diagnosed with SCZ and 54 with BD, with no significant differences between the quetiapine formulations. The mean daily dose of quetiapine XR was significantly higher than that of quetiapine IR (542 mg vs 328 mg; p<0.001). This was also true for the SCZ subgroup (XR: 593 mg vs IR: 338 mg; p<0.001) and the BD subgroup (XR: 466 mg vs IR: 308 mg; p=0.009). 48% of all quetiapine IR patients used a mean dose of ≤200 mg compared with 2% of XR patients. Injectable antipsychotics were combined with quetiapine IR but not with quetiapine XR (12% vs 0%; p=0.019). At discharge, quetiapine XR was used as monotherapy to a greater extent than IR (79% vs 44%; p=0.003). The odds for quetiapine XR use in hospital were lower with advancing age, substance abuse diagnosis and prior IR use.

Conclusions

Among SCZ and BD inpatients, quetiapine XR was more often used as monotherapy and in significantly higher doses than quetiapine IR. Differential use of the quetiapine formulations appears to depend, at least in part, on patient characteristics.
Multigene interactions and the prediction of depression in the Wisconsin Longitudinal Study
Authors: Roetker Nicholas S , Yonker James A , Lee Chee , Chang Vicky , Basson Jacob J , Roan Carol L , Hauser Taissa S , Hauser Robert M , Atwood Craig S
Published: July 2012

Objectives

Single genetic loci offer little predictive power for the identification of depression. This study examined whether an analysis of gene–gene (G × G) interactions of 78 single nucleotide polymorphisms (SNPs) in genes associated with depression and age-related diseases would identify significant interactions with increased predictive power for depression.

Design

A retrospective cohort study.

Setting

A survey of participants in the Wisconsin Longitudinal Study.

Participants

A total of 4811 persons (2464 women and 2347 men) who provided saliva for genotyping; the group comes from a randomly selected sample of Wisconsin high school graduates from the class of 1957 as well as a randomly selected sibling, almost all of whom are non-Hispanic white.

Primary outcome measure

Depression as determine by the Composite International Diagnostic Interview–Short-Form.

Results

Using a classification tree approach (recursive partitioning (RP)), the authors identified a number of candidate G × G interactions associated with depression. The primary SNP splits revealed by RP (ANKK1rs1800497 (also known asDRD2Taq1A) in men andDRD2rs224592 in women) were found to be significant as single factors by logistic regression (LR) after controlling for multiple testing (p=0.001 for both). Without considering interaction effects, only one of the five subsequent RP splits reached nominal significance in LR (FTOrs1421085 in women, p=0.008). However, after controlling for G × G interactions by running LR on RP-specific subsets, every split became significant and grew larger in magnitude (OR (before) → (after): men:GNRH1novel SNP: (1.43 → 1.57); women:APOC3rs2854116: (1.28 → 1.55),ACVR2Brs3749386: (1.11 → 2.17),FTOrs1421085: (1.32 → 1.65),IL6rs1800795: (1.12 → 1.85)).

Conclusions

The results suggest that examining G × G interactions improves the identification of genetic associations predictive of depression. 4 of the SNPs identified in these interactions were located in two pathways well known to impact depression: neurotransmitter (ANKK1andDRD2) and neuroendocrine (GNRH1andACVR2B) signalling. This study demonstrates the utility of RP analysis as an efficient and powerful exploratory analysis technique for uncovering genetic and molecular pathway interactions associated with disease aetiology.
Increase in self-reported migraine prevalence in the Danish adult population: a prospective longitudinal population-based study
Authors: Le Han , Tfelt-Hansen Peer , Skytthe Axel , Kyvik Kirsten Ohm , Olesen Jes
Published: July 2012

Objective

It is uncertain whether migraine prevalence has increased in modern society. The aim of this study was to assess any change in migraine prevalence over an 8-year period among the adult population in Denmark.

Design

Prospective longitudinal population-based study.

Setting

30 000 twin individuals were invited to participate in two cross-sectional questionnaire surveys containing validated questions to diagnose migraine in 1994 and 2002. The twins are representative of the Danish population with regard to migraine and other somatic diseases.

Participants

The 1994 cohort comprised 28 571 twin individuals aged 12–41 years and the 2002 cohort 31 865 twin individuals aged 20–71 years.

Outcome measures

Sex-, age- and subtype-specific incidence and lifetime prevalence as well as 1-year prevalence of migraine.

Results

1-year prevalence in 2002 was 12.3% for migraine, 4.1% for migraine with aura and 8.2% for migraine without aura. Lifetime prevalence of migraine was 16.1% in 1994 (aged 12–41 years) and 25.2% in 2002 (aged 20–71 years). Lifetime prevalence of migraine for age 20–41 was increased from 1994 to 2002 (18.5% vs 24.5%) by 32.2% (95% CI 27.0% to 37.3%; p<0.001). The difference was primarily seen in the population older than 32 years. The increase was especially evident in migraine with aura (5.6% vs 9.4%, p<0.001) but also a significant increase in migraine without aura was found (13.0% vs 15.1%, p<0.001). Eight-year period incidence rate of migraine was 0.141 corresponding to an average annual incidence rate of 17.6 per 1000 person-years.

Conclusions

Lifetime prevalence of migraine in Denmark increased substantially from 1994 to 2002. Part of the increase may be due to increased medical consultation resulting in increased rate of physician diagnosis or awareness due to previously participation in the 1994 survey. It is pertinent to study the environmental causes of the increase and to implement preventive measures.
Comparative assessment of CDS, CLSI disc diffusion and Etest techniques for antimicrobial susceptibility testing ofNeisseria gonorrhoeae: a 6-year study
Authors: Singh Vikram , Bala Manju , Kakran Monika , Ramesh V
Published: July 2012

Background

A variety of techniques are available for antimicrobial susceptibility testing ofNeisseria gonorrhoeae.

Objective

The aim of this study was to find a cost-effective, reliable and easily applicable microbiological method to detect antimicrobial susceptibilities ofN. gonorrhoeaein resource-poor countries.

Design

Prospective study.

Setting

Male and female STD clinic of Regional STD Teaching, Training and Research Centre, New Delhi, India.

Participants

N. gonorrhoeaeisolates from all male and female patients presenting with acute gonococcal urethritis and cervical discharge.

Material and methods

A total of 295 consecutiveN. gonorrhoeaeisolates during 2005–2010 was used to compare the Clinical and Laboratory Standards Institute (CLSI) and CDS disc diffusion technique with Etest by performing antimicrobial susceptibility testing in parallel for penicillin, tetracycline, ceftriaxone, ciprofloxacin and spectinomycin. WHO reference strains were used as controls.

Results

CDS disc diffusion zones of inhibition showed that complete percentage agreement for penicillin, ciprofloxacin and tetracycline was high with their analogous Etest minimal inhibitory concentrations in comparison to CLSI disc diffusion technique, that is, 91.5%, 92.9% and 99.3% versus 87.5%, 88.5% and 74.9%, respectively. CDS results had less number of major and minor category discrepancies in comparison to CLSI and CDS method showed excellent correlation coefficient (r=1) with Etest for all five antimicrobial agents tested in comparison to CLSI (r=0.92). It was very poor (r=0.61) by CLSI method for tetracycline. The correlation coefficients between the two methods and the Etest were identical if tetracycline was removed from the CLSI analysis.

Conclusions

The CDS technique is an attractive alternative forN. gonorrhoeaesusceptibility testing and is recommended for monitoring the antimicrobial susceptibility in less developed and resource-poor settings to facilitate enhanced antimicrobial resistance surveillance when the WHO Gonococcal Antimicrobial Surveillance Programme is undergoing expansion to meet the ongoing challenges of surveillance and control of gonococcal antimicrobial resistance.
Human semen quality in the new millennium: a prospective cross-sectional population-based study of 4867 men
Authors: Jørgensen Niels , Joensen Ulla Nordström , Jensen Tina Kold , Jensen Martin Blomberg , Almstrup Kristian , Olesen Inge Ahlmann , Juul Anders , Andersson Anna-Maria , Carlsen Elisabeth , Petersen Jørgen Holm , Toppari Jorma , Skakkebæk Niels E
Published: July 2012

Objectives

Considerable interest and controversy over a possible decline in semen quality during the 20th century raised concern that semen quality could have reached a critically low level where it might affect human reproduction. The authors therefore initiated a study to assess reproductive health in men from the general population and to monitor changes in semen quality over time.

Design

Cross-sectional study of men from the general Danish population. Inclusion criteria were place of residence in the Copenhagen area, and both the man and his mother being born and raised in Denmark. Men with severe or chronic diseases were not included.

Setting

Danish one-centre study.

Participants

4867 men, median age 19 years, included from 1996 to 2010.

Outcome measures

Semen volume, sperm concentration, total sperm count, sperm motility and sperm morphology.

Results

Only 23% of participants had optimal sperm concentration and sperm morphology. Comparing with historic data of men attending a Copenhagen infertility clinic in the 1940s and men who recently became fathers, these two groups had significantly better semen quality than our study group from the general population. Over the 15 years, median sperm concentration increased from 43 to 48 million/ml (p=0.02) and total sperm count from 132 to 151 million (p=0.001). The median percentage of motile spermatozoa and abnormal spermatozoa were 68% and 93%, and did not change during the study period.

Conclusions

This large prospective study of semen quality among young men of the general population showed an increasing trend in sperm concentration and total sperm count. However, only one in four men had optimal semen quality. In addition, one in four will most likely face a prolonged waiting time to pregnancy if they in the future want to father a child and another 15% are at risk of the need of fertility treatment. Thus, reduced semen quality seems so frequent that it may impair the fertility rates and further increase the demand for assisted reproduction.
An investigation of the effects of the common cold on simulated driving performance and detection of collisions: a laboratory study
Authors: Smith Andrew Paul , Jamson Samantha
Published: July 2012

Objective

The aim of the present research was to investigate whether individuals with a common cold showed impaired ability on a simulated driving task and the ability to detect potential collisions between moving objects.

Design

The study involved comparison of a healthy group with a group with colds. These scores were adjusted for individual differences by collecting further data when both groups were healthy and using these scores as covariates. On both occasions, volunteers rated their symptoms and carried out a simulated driving session. On the first occasion, volunteers also carried out a collision detection task.

Setting

University of Leeds Institute for Transport Studies.

Sample

Twenty-five students from the University of Leeds. Ten volunteers were healthy on both occasions and 15 had a cold on the first session and were healthy on the second.

Main outcome measures

In the collision detection task, the main outcomes were correct detections and response to a secondary identification task. In the simulated driving task, the outcomes were speed, lateral control, gap acceptance, overtaking behaviour, car following, vigilance and traffic light violations.

Results

Those with a cold detected fewer collisions and had a higher divided attention error than those who were healthy. Many basic driving skills were unimpaired by the illness. However, those with a cold were slower at responding to unexpected events and spent a greater percentage of time driving at a headway of <2 s.

Conclusions

The finding that having a common cold is associated with reduced ability to detect collisions and respond quickly to unexpected events is of practical importance. Further research is now required to examine the efficacy of information campaigns and countermeasures such as caffeine.
Views of policymakers, healthcare workers and NGOs on HIV pre-exposure prophylaxis (PrEP): a multinational qualitative study
Authors: Wheelock Ana , Eisingerich Andreas B , Gomez Gabriela B , Gray Emily , Dybul Mark R , Piot Peter
Published: July 2012

Objectives

To examine policymakers and providers' views on pre-exposure prophylaxis (PrEP) and their willingness to support its introduction, to inform policy and practice in this emerging field.

Design

Semistructured qualitative interview study.

Setting

Peru, Ukraine, India, Kenya, Uganda, Botswana and South Africa.

Participants

35 policymakers, 35 healthcare workers and 21 non-governmental organisation representatives involved in HIV prevention.

Results

Six themes emerged from the data: (1) perceived HIV prevention landscape: prevention initiatives needed to be improved and expanded; (2) PrEP awareness: 50 of 91 participants had heard of PrEP; (3) benefits of PrEP: one component of the combination prevention arsenal that could help prioritise HIV prevention, empower key populations and result in economic gains; (4) challenges of PrEP: regimen complexity, cost and cost-effectiveness, risk compensation, efficacy and effectiveness, stigmatisation and criminalisation, information and training and healthcare system capacity; (5) programmatic considerations: user eligibility, communication strategy, cost, distribution, medication and HIV testing compliance and (6) early versus late implementation: participants were divided as to whether they would support an early introduction of PrEP in their country or would prefer to wait until it has been successfully implemented in other countries, with around half of those we spoke to supporting each option. Very few said they would not support PrEP at all.

Conclusions

Despite the multiple challenges identified, there was general willingness to support the introduction of PrEP. Yet, strengthening existing HIV prevention efforts was also deemed necessary. Our results suggest that an effective PrEP programme would be delivered in healthcare facilities and involve non-governmental organisations and the community and consider the needs of mobile populations. Comprehensive information packages and training for users and providers would be critical. The cost of PrEP would be affordable and possibly segmented. Extensive counselling and innovative monitoring measures ought to be considered.
Touch screen computer health assessment in Australian general practice patients: a cross-sectional study protocol
Authors: Yoong Sze Lin , Carey Mariko Leanne , Sanson-Fisher Robert William , Russell Grant , Mazza Danielle , Makeham Meredith , Paul Christine Louise , Inder Kerry Jane , D'Este Catherine
Published: July 2012

Introduction

Cardiovascular disease (CVD) and cancer are leading causes of death globally. Early detection of cancer and risk factors for CVD may improve health outcomes and reduce mortality. General practitioners (GPs) are accessed by the majority of the population and play a key role in the prevention and early detection of chronic disease risk factors. This cross-sectional study aims to assess the acceptability of an electronic method of data collection in general practice patients. The study will describe the proportion screened in line with guidelines for CVD risk factors and cancer as well as report the prevalence of depression, lifestyle risk factors, level of provision of preconception care, cervical cancer vaccination and bone density testing. Lastly, the study will assess the level of agreement between GPs and patients perception regarding presence of risk factors and screening.

Methods and analysis

The study has been designed to maximise recruitment of GPs by including practitioners in the research team, minimising participation burden on GPs and offering remuneration for participation. Patient recruitment will be carried out by a research assistant located in general practice waiting rooms. Participants will be asked regarding the acceptability of the touch screen computer and to report on a range of health risk and preventive behaviours using the touch screen computer. GPs will complete a one-page survey indicating their perception of the presence of risk behaviours in their patients. Descriptive statistics will be generated to describe the acceptability of the touch screen and prevalence of health risk behaviours. Cohen's κ will be used to assess agreement between GP and patient perception of presence of health risk behaviours.

Ethics and dissemination

This study has been approved by the human research committees in participating universities. Findings will be disseminated via peer-reviewed publications, conference presentations as well as practice summaries provided to participating practices.