bluuebunny/crossref_metadata_2025 huggingface dataset:
The diagnostic accuracy of two methods for E6&7 mRNA detection in women with minor cytological abnormalities
Authors: KOLIOPOULOS GEORGE , CHRELIAS CHARALAMBOS , PAPPAS ASIMAKIS , MAKRIDIMA SOFIA , KOUNTOURIS EMMANUEL , ALEPAKI MARIA , SPATHIS ARIS , STATHOPOULOU VICKY , PANAYIOTIDES IOANNIS , PANAGOPOULOS PERICLES , KARAKITSOS PETROS , KASSANOS DIMITRIOS
Published: April 2012

Objective. To examine the diagnostic accuracy of nucleic acid sequence based amplification (NASBA) and flow cytometry for E6&7 human papillomavirus (HPV) mRNA detection in the triage of minor cytological abnormalities. Design. Prospective diagnostic accuracy study. Setting. Gynecology outpatient clinics of a university hospital. Population. 472 women with low‐grade squamous intraepithelial lesion (LSIL) or atypical squamous cells of undetermined significance (ASCUS). Methods. Residual material of the liquid‐based smears was tested by NASBA and by flow cytometry for E6&E7 mRNA expression. Histological diagnosis was used as reference standard. Main outcome measures. Accuracy indices of the two techniques and of type 16‐specific NASBA for the detection of cervical intraepithelial neoplasia (CIN) 2+ and CIN3+, accuracy indices at age >35 years, correlation between NASBA and flow, comparison between integrated and episomal high‐risk HPV infection for risk of CIN2+. Results. Both tests showed increased positivity rates with increasing severity of the lesion (p < 0.05, chi‐squared test for trend). There was a positive correlation between NASBA and flow results (phi coefficient = 0.325). NASBA‐positive cases were more likely to have CIN2+ than were NASBA‐negative/DNA‐positive for types 16, 18, 31, 33, 45 (25/73 vs. 4/52, p= 0.0004; Fisher‘s exact test). In the LSIL group the NASBA accuracy indices for CIN3+ were: sensitivity 75%, specificity 78.7% and positivity rate 20.8%, and for flow 77.8%, 64.5% and 35.9%, respectively. Conclusions. NASBA has favorable specificity and positivity rates for triaging LSIL prior to colposcopy. A relatively low sensitivity warrants cytological surveillance of the NASBA‐negative LSILs. Flow cytometry does not perform as well overall.
Attitudes and perceptions towards substances among people with mental disorders: a systematic review
Authors: Thornton L. K. , Baker A. L. , Johnson M. P. , Lewin T. J.
Published: April 2012

Thornton LK, Baker AL, Johnson MP, Lewin TJ. Attitudes and perceptions towards substances among people with mental disorders: A systematic review.

Objective:  To develop effective interventions for people with coexisting mental disorders (MD) and substance use, it may be beneficial to understand their attitudes and perceptions of substances.

Method:  A systematic literature search regarding attitudes and perceptions towards tobacco, alcohol or cannabis among people with MD was conducted. Studies’ methodological quality was assessed using the Newcastle–Ottawa Scale.

Results:  Twenty‐one papers were included in the review and found to have generally low methodological quality. Papers investigated reasons for substance use, substance use expectancies, substances’ perceived effects and reasons for quitting. People with psychotic disorders reported using substances primarily for relaxation and pleasure. Among people with mood disorders, alcohol was used primarily for social motives and tobacco for negative affect reduction.

Conclusion:  For substance use interventions among people with MD to be more effective, it may be necessary to tailor interventions specifically for this population and customize by substance type. Gaps in the literature regarding attitudes and perceptions towards substance use among people with MD were identified, which future research should aim to address. These include designing and conducting methodologically rigorous research, investigating perceived harmfulness and knowledge of substances, and broadening recruitment of participants to include people with MD other than psychosis.
The heritability of avoidant and dependent personality disorder assessed by personal interview and questionnaire
Authors: Gjerde L. C. , Czajkowski N. , Røysamb E. , Ørstavik R. E. , Knudsen G. P. , Østby K. , Torgersen S. , Myers J. , Kendler K. S. , Reichborn‐Kjennerud T.
Published: April 2012

Gjerde LC, Czajkowski N, Røysamb E, Ørstavik RE, Knudsen GP, Østby K, Torgersen S, Myers J, Kendler KS, Reichborn‐Kjennerud T. The heritability of avoidant and dependent personality disorder assessed by personal interview and questionnaire.

Objective:  Personality disorders (PDs) have been shown to be modestly heritable. Accurate heritability estimates are, however, dependent on reliable measurement methods, as measurement error deflates heritability. The aim of this study was to estimate the heritability of DSM‐IV avoidant and dependent personality disorder, by including two measures of the PDs at two time points.

Method:  Data were obtained from a population‐based cohort of young adult Norwegian twins, of whom 8045 had completed a self‐report questionnaire assessing PD traits. 2794 of these twins subsequently underwent a structured diagnostic interview for DSM‐IV PDs. Questionnaire items predicting interview results were selected by multiple regression, and measurement models of the PDs were fitted in Mx.

Results:  The heritabilities of the PD factors were 0.64 for avoidant PD and 0.66 for dependent PD. No evidence of common environment, that is, environmental factors that are shared between twins and make them similar, was found. Genetic and environmental contributions to avoidant and dependent PD seemed to be the same across sexes.

Conclusion:  The combination of both a questionnaire‐ and an interview assessment of avoidant and dependent PD results in substantially higher heritabilities than previously found using single‐occasion interviews only.
Chromium in leather footwear—risk assessment of chromium allergy and dermatitis
Authors: Thyssen Jacob P. , Strandesen Maria , Poulsen Pia B. , Menné Torkil , Johansen Jeanne D.
Published: April 2012

Background. Chromium‐tanned leather footwear, which releases >3 ppm hexavalent Cr(VI), may pose a risk of sensitizing and eliciting allergic dermatitis.

Objectives. To determine the content and potential release of chromium in leather footwear and to discuss the prevention of chromium contact allergy and dermatitis.

Methods. Sixty pairs of leather shoes, sandals and boots (20 children's, 20 men's, and 20 women's) were purchased in Copenhagen and examined with X‐ray fluorescence spectroscopy. Chromium was extracted according to the International Standard, ISO 17075. The detection level for Cr(VI) was 3 ppm.

Results. Chromium was identified in 95% of leather footwear products, the median content being 1.7% (range 0–3.3%). No association with store category or footwear category was found. A tendency for there to be a higher chromium content in footwear with high prices was shown (ptrend = 0.001). Cr(VI) was extracted from 44% of 18 footwear products, and, in three items, more than 10 ppm was extracted. One shoe had 62 ppm Cr(VI) extracted. Sandals seemed to be over‐represented among footwear with detectable Cr(VI). Cr(III) extraction reached a median value of 152 ppm.

Conclusions. Most leather footwear contained chromium. Cr(VI) was extracted from a high proportion of leather footwear; this poses a risk of sensitization.
Myo/Nog cells in normal, wounded and tumor‐bearing skin
Authors: Gerhart Jacquelyn , Hayes Candace , Scheinfeld Victoria , Chernick Michael , George‐Weinstein Mindy , Gilmour Susan
Published: April 2012

Murine and human skin were examined for the presence of Myo/Nog cells that were originally discovered in the chick embryo by their expression of MyoD mRNA, noggin and the G8 antigen. Myo/Nog cells are the primary source of noggin in telogen hair follicles. They are scarce within the interfollicular dermis and absent in the epidermis. Within 24 h following epidermal abrasion, Myo/Nog cells increase in number in the follicles and appear in the wound. Myo/Nog cells are also recruited to the stroma of tumors formed from v‐Ras‐transformed keratinocytes (Ker/Ras). Human squamous cell carcinomas and malignant melanomas contain significantly more Myo/Nog cells than basal cell carcinomas. Myo/Nog cells are distinct from macrophages, granulocytes and cells expressing alpha smooth muscle actin in the tumor stroma. Myo/Nog cells may be modulators of skin homoeostasis and wound healing, and potential diagnostic and therapeutic targets in skin cancer.
Absence of selenoprotein P but not selenocysteine lyase results in severe neurological dysfunction
Authors: Raman A. V. , Pitts M. W. , Seyedali A. , Hashimoto A. C. , Seale L. A. , Bellinger F. P. , Berry M. J.
Published: April 2012

Dietary selenium restriction in mammals causes bodily selenium to be preferentially retained in the brain relative to other organs. Almost all the known selenoproteins are found in brain, where expression is facilitated by selenocysteine (Sec)‐laden selenoprotein P. The brain also expresses selenocysteine lyase (Scly), an enzyme that putatively salvages Sec and recycles the selenium for selenoprotein translation. We compared mice with a genetic deletion of Scly to selenoprotein P (Sepp1) knockout mice for similarity of neurological impairments and whether dietary selenium modulates these parameters. We report that Scly knockout mice do not display neurological dysfunction comparable to Sepp1 knockout mice. Feeding a low‐selenium diet to Scly knockout mice revealed a mild spatial learning deficit without disrupting motor coordination. Additionally, we report that the neurological phenotype caused by the absence of Sepp1 is exacerbated in male vs. female mice. These findings indicate that Sec recycling via Scly becomes limiting under selenium deficiency and suggest the presence of a complementary mechanism for processing Sec. Our studies illuminate the interaction between Sepp1 and Scly in the distribution and turnover of body and brain selenium and emphasize the consideration of sex differences when studying selenium and selenoproteins in vertebrate biology.
A new model of Pde4d deficiency: genetic knock‐down of PDE4D enzyme in rats produces an antidepressant phenotype without spatial cognitive effects
Authors: Schaefer T. L. , Braun A. A. , Amos‐Kroohs R. M. , Williams M. T. , Ostertag E. , Vorhees C. V.
Published: April 2012

Phosphodiesterases (PDEs) are a superfamily of intracellular second messenger cyclic nucleotide hydrolyzing enzymes composed of 12 families. The Pde4 family has been implicated in depression and cognition, and PDE4 inhibitors have been evaluated as antidepressants and possible cognitive enhancers. Pde4d−/− mice show an antidepressant phenotype and learning enhancement on some tests, but not others as do mice treated with PDE4 inhibitors. Here, we report for the first time the behavioral phenotype of a new Pde4d knock‐down (KD) rat model of PDE4D deficiency. Consistent with other data on PDE4D deficiency, Pde4d KD rats showed depression resistance in the Porsolt forced swim test and hyperreactivity of the acoustic startle response with no differential response on prepulse inhibition, suggesting no sensorimotor gating defect. Pde4d KD rats also exhibited a small exploratory activity reduction but no difference following habituation, and no enhanced spatial learning or reference memory in the Morris water maze. A selective improvement in route‐based learning in the Cincinnati water maze was seen as well as enhanced contextual and cued fear conditioning and a more rapid rate of cued extinction from their higher freezing level that declined to wild‐type (WT) levels only after ∼20 extinction trials. The rat model confirms Pde4d's role in depression but not in spatial learning or memory enhancement and shows for the first time higher fear conditioning and altered extinction compared with controls. The new model provides a tool by which to better understand the role of PDE4D in neuropsychiatric disorders and for the development of alternate treatment approaches.
Functional analysis of the Notch ligand Jagged1 missense mutant proteins underlying Alagille syndrome
Authors: Tada Minoru , Itoh Satsuki , Ishii‐Watabe Akiko , Suzuki Takuo , Kawasaki Nana
Published: April 2012

Heterozygous mutations in the JAG1 gene, encoding Notch ligand Jagged1, cause Alagille syndrome (ALGS). As most of the mutations are nonsense or frameshift mutations producing inactive truncated proteins, haplo‐insufficiency is considered the major pathogenic mechanism of ALGS. However, the molecular mechanisms by which the missense mutations cause ALGS remain unclear. Here we analyzed the functional properties of four ALGS missense mutant proteins, P163L, R184H, G386R and C714Y, using transfected mammalian cells. P163L and R184H showed Notch‐binding activities similar to that of the wild‐type when assessed by immunoprecipitation. However, their trans‐activation and cis‐inhibition activities were almost completely impaired. These mutant proteins localized mainly to the endoplasmic reticulum (ER), suggesting that the mutations induced improper protein folding. Furthermore, the mutant proteins bound more strongly to the ER chaperone proteins calnexin and calreticulin than the wild‐type did. C714Y also localized to the ER, but possessed significant trans‐activation activity and lacked enhanced binding to the chaperones, indicating a less severe phenotype. The properties of G386R were the same as those of the wild‐type. Dominant‐negative effects were not detected for any mutant protein. These results indicate that accumulation in the ER and binding to the chaperones correlate with the impaired signal‐transduction activities of the missense mutant proteins, which may contribute to the pathogenic mechanism of ALGS. Our findings, which suggest the requirement for cell‐surface localization of Jagged1 for cis‐inhibition activities, also provide important information for understanding the molecular basis of Notch‐signaling pathways.

Structured digital abstract

 Jagged‐1physically interacts with Calreticulin and Calnexin by anti tag coimmunoprecipitation (View Interaction: 1, 2)

 Jagged‐1physically interacts with NOTCH3 by anti tag coimmunoprecipitation (View interaction)
Epidemiological aspects of pandemic influenza A(H1N1) virus from 2009 to 2011 in Iran
Authors: Yavarian Jila , Naseri Maryam , Shadab Azadeh , Shafiei Jandaghi Nazanin Z. , Mokhtari Azad Talat
Published: April 2012

Please cite this paper as: Yavarian et al. (2012). Epidemiological aspects of pandemic influenza A(H1N1) virus from 2009 to 2011 in Iran. Influenza and Other Respiratory Viruses 6(601), e74–e76.
Quantification of dust‐borne transmission of highly pathogenic avian influenza virus between chickens
Authors: Spekreijse Dieuwertje , Bouma Annemarie , Koch Guus , Stegeman Arjan
Published: April 2012

Please cite this paper as: Spekreijse et al. (2013) Quantification of dust‐borne transmission of highly pathogenic avian influenza virus between chickens. Influenza and Other Respiratory Viruses 7(2) 132–138.

Background  Understanding the transmission of highly pathogenic avian influenza virus (HPAIv) between poultry flocks is essential to prevent and control epidemics. Dust, produced in infected chicken flocks, has been hypothesized to be an important mechanical vector for between‐flock transmission of HPAIv.

Objectives  The aim of our study was to quantify the amount of virus shed by infected birds and its relation to deposition of virus in the environment and the rate of dust‐borne transmission between groups of chickens.

Methods  Four replicate experiments were performed, each replicate with two groups of 14 chickens housed in two separate rooms. In one group, chickens were inoculated with HPAIv. Ventilation forced the air from that room to the second (recipient) group through a tube. Deceased birds in the inoculated group were replaced with new susceptible birds up to day 10 p.i. Dust samples were collected daily. Trachea and cloaca swabs were collected daily to determine virus shedding and virus spread to the recipient group.

Results  The amount of virus detected in dust samples in the recipient room was, on average, 103·7 EID50/m3. Virus transmission from the inoculated to the recipient group occurred in two experiments. The transmission rate parameter for dust‐borne transmission was estimated at 0·08 new infections/infectious chicken/day.

Conclusions  The results of this study are a first step to elucidate the importance of dust‐borne transmission of HPAIv between flocks and help interpret environmental samples.
Serological evidence of swine influenza in Brazil
Authors: Rajão Daniela S. , Alves Fabiana , Del Puerto Helen L. , Braz Gissandra F. , Oliveira Fernanda G. , Ciacci‐Zanella Janice R. , Schaefer Rejane , dos Reis Jenner K. P. , Guedes Roberto M. C. , Lobato Zélia I. P. , Leite Rômulo C.
Published: April 2012

Please cite this paper as: Rajão et al. (2013). Serological evidence of swine influenza in Brazil. Influenza and Other Respiratory Viruses 7(2), 109–112.

The aim of this work was to detect serum antibodies specific to influenza viruses in swine in Brazil. Serum samples of 355 pigs from 17 herds in Minas Gerais state were tested by hemagglutination inhibition (HI) for antibodies against H1N1 swine (SIV) and human influenza viruses, and H3N2 SIV. HI revealed that 158 animals (44·5%) and 11 herds (64·7%) were positive for H1N1 SIV, 36 animals (10·1%) and four herds (23·5%) were positive for H3N2 SIV, and 136 animals (38·3%) and 10 herds (58·8%) were positive for H1N1 human. This study indicates that swine influenza is disseminated throughout Minas Gerais state, Brazil.
Estimating Potential E. coli Sources in a Watershed Using Spatially Explicit Modeling Techniques1
Authors: Riebschleager K.J. , Karthikeyan R. , Srinivasan R. , McKee K.
Published: April 2012

Riebschleager, K.J., R. Karthikeyan, R. Srinivasan, and K. McKee, 2012. Estimating Potential E. coli Sources in a Watershed Using Spatially Explicit Modeling Techniques. Journal of the American Water Resources Association (JAWRA) 48(4): 745‐761. DOI: 10.1111/j.1752‐1688.2012.00649.x

Abstract:  The Spatially Explicit Load Enrichment Calculation Tool (SELECT) was automated to characterize waste and the associated pathogens from various sources within a mixed land use watershed. Potential Escherichia coli loads in Lake Granbury watershed were estimated using spatially variable governing factors, such as land use, soil condition, and distance to streams. A new approach for characterizing E. coli loads resulting from malfunctioning on‐site wastewater treatment systems (OWTSs) was incorporated into SELECT along with the Pollutant Connectivity Factor (PCF) module. The PCF component was applied to identify areas contributing E. coli loads during runoff events by incorporating the influence of potential E. coli loading, runoff potential, and travel distance to waterbodies. Simulation results indicated livestock and wildlife are potential E. coli contributing sources in the watershed. The areas in which these sources are potentially contributing are not currently monitored for E. coli. The bacterial water quality violations seen around Lake Granbury are most likely the result of malfunctioning OWTSs and pet wastes. SELECT results demonstrate the need to evaluate each contributing source separately to effectively allocate site specific best management practices (BMPs) utilizing stakeholder inputs. It also serves as a powerful screening tool for determining areas where detailed investigation is merited.
Sustaining conservation values in selectively logged tropical forests: the attained and the attainable
Authors: Putz Francis E. , Zuidema Pieter A. , Synnott Timothy , Peña‐Claros Marielos , Pinard Michelle A. , Sheil Douglas , Vanclay Jerome K. , Sist Plinio , Gourlet‐Fleury Sylvie , Griscom Bronson , Palmer John , Zagt Roderick
Published: April 2012

Most tropical forests outside protected areas have been or will be selectively logged so it is essential to maximize the conservation values of partially harvested areas. Here we examine the extent to which these forests sustain timber production, retain species, and conserve carbon stocks. We then describe some improvements in tropical forestry and how their implementation can be promoted.

 A simple meta‐analysis based on >100 publications revealed substantial variability but that: timber yields decline by about 46% after the first harvest but are subsequently sustained at that level; 76% of carbon is retained in once‐logged forests; and, 85–100% of species of mammals, birds, invertebrates, and plants remain after logging. Timber stocks will not regain primary‐forest levels within current harvest cycles, but yields increase if collateral damage is reduced and silvicultural treatments are applied.

 Given that selectively logged forests retain substantial biodiversity, carbon, and timber stocks, this “middle way” between deforestation and total protection deserves more attention from researchers, conservation organizations, and policy‐makers. Improvements in forest management are now likely if synergies are enhanced among initiatives to retain forest carbon stocks (REDD+), assure the legality of forest products, certify responsible management, and devolve control over forests to empowered local communities.
A Stem Cell–Based Approach to Cartilage Repair
Authors: Johnson Kristen , Zhu Shoutian , Tremblay Matthew S. , Payette Joshua N. , Wang Jianing , Bouchez Laure C. , Meeusen Shelly , Althage Alana , Cho Charles Y. , Wu Xu , Schultz Peter G.
Published: April 2012

Osteoarthritis and Kartogenin

Osteoarthritis is characterized by progressive breakdown of articular cartilage and affects over 25 million people in the United States. Mesenchymal stem cells (MSCs), which reside in healthy and diseased joints, are multipotent adult stem cells that are able to differentiate into a variety of cell types.

Johnson
et al.

(p.
717
, published online 5 April) identified a small molecule, kartogenin, which was able to induce MSCs to differentiate into chondrocytes in vitro. When administered locally, kartogenin was efficacious in two animal models of osteoarthritis.
Magnetic Reconnection in the Near Venusian Magnetotail
Authors: Zhang T. L. , Lu Q. M. , Baumjohann W. , Russell C. T. , Fedorov A. , Barabash S. , Coates A. J. , Du A. M. , Cao J. B. , Nakamura R. , Teh W. L. , Wang R. S. , Dou X. K. , Wang S. , Glassmeier K. H. , Auster H. U. , Balikhin M.
Published: April 2012

Magnetic Reconnection

Magnetic reconnection (MR) has been observed in the magnetospheres of planets with an intrinsic magnetic field, such as Earth, Mercury, Jupiter, and Saturn. MR is a universal plasma process that occurs in regions of strong magnetic shear and converts magnetic energy into kinetic energy. On Earth, MR is responsible for magnetic storms and auroral events. Using data from the European Space Agency Venus Express spacecraft,

Zhang
et al.

(p.
567
, published online 5 April; see the Perspective by

Slavin

) present surprising evidence for MR in the magnetosphere of Venus, which is a nonmagnetized body.
Coupling Quantum Tunneling with Cavity Photons
Authors: Cristofolini Peter , Christmann Gabriel , Tsintzos Simeon I. , Deligeorgis George , Konstantinidis George , Hatzopoulos Zacharias , Savvidis Pavlos G. , Baumberg Jeremy J.
Published: April 2012

Tunneling Through with a Light Touch

Quantum tunneling underpins a host of essential techniques, such as scanning tunneling microscopy and quantum cascade lasers, as well as chemical reactions. The tunneling particles are normally electrons, and control of the tunneling process has generally been by electric fields. By coupling tunneling electrons with cavity photons trapped inside a semiconductor microcavity,

Cristofolini
et al.

(p.
704
, published online 5 April; see the Perspective by

Szymańska

) produced mixed states that then allowed direct optical control of the tunneling process. Such an optical-based approach to manipulating and controlling the tunneling process may find applications in quantum information science.
The Removal of RNA Primers from DNA Synthesized by the Reverse Transcriptase of the Retrotransposon Tf
1
Is Stimulated by Tf
1
Integrase
Authors: Herzig Eytan , Voronin Nickolay , Hizi Amnon
Published: April 2012

The Tf
1
retrotransposon represents a group of long terminal repeat retroelements that use an RNA self-primer for initiating reverse transcription while synthesizing the minus-sense DNA strand. Tf
1
reverse transcriptase (RT) was found earlier to generate the self-primer
in vitro
. Here, we show that this RT can remove from the synthesized cDNA the entire self-primer as well as the complete polypurine tract (PPT) sequence (serving as a second primer for cDNA synthesis). However, these primer removals, mediated by the RNase H activity of Tf
1
RT, are quite inefficient. Interestingly, the integrase of Tf
1
stimulated the specific Tf
1
RT-directed cleavage of both the self-primer and PPT, although there was no general enhancement of the RT's RNase H activity (and the integrase by itself is devoid of any primer cleavage). The RTs of two prototype retroviruses, murine leukemia virus and human immunodeficiency virus, showed only a partial and nonspecific cleavage of both Tf
1
-associated primers with no stimulation by Tf
1
integrase. Mutagenesis of Tf
1
integrase revealed that the complete Tf
1
integrase protein (excluding its chromodomain) is required for stimulating the Tf
1
RT primer removal activity. Nonetheless, a double mutant integrase that has lost its integration functions can still stimulate the RT's activity, though heat-inactivated integrase cannot enhance primer removals. These findings suggest that the enzymatic activity of Tf
1
integrase is not essential for stimulating the RT-mediated primer removal, while the proper folding of this protein is obligatory for this function. These results highlight possible new functions of Tf
1
integrase in the retrotransposon's reverse transcription process.
Functions of the Epstein-Barr Virus EBNA1 Protein in Viral Reactivation and Lytic Infection
Authors: Sivachandran Nirojini , Wang Xueqi , Frappier Lori
Published: April 2012

EBNA1 is the only nuclear Epstein-Barr virus (EBV) protein expressed in both latent and lytic modes of infection. While EBNA1 is known to play several important roles in latent infection, the reason for its continued expression in lytic infection is unknown. Here we identified two roles for EBNA1 in the reactivation of latent EBV to the lytic cycle in epithelial cells. First, EBNA1 depletion in latently infected cells was shown to positively contribute to spontaneous EBV reactivation, showing that EBNA1 has a role in suppressing reactivation. Second, when the lytic cycle was induced, EBNA1 depletion decreased lytic gene expression and DNA amplification, showing that it positively contributed to lytic infection. Since we have previously shown that EBNA1 disrupts promyelocytic leukemia (PML) nuclear bodies, we investigated whether this function could account for the effects of EBNA1 on lytic infection by repeating the experiments with cells lacking PML proteins. In the absence of PML, EBNA1 did not promote lytic infection, indicating that the EBNA1-mediated PML disruption is responsible for promoting lytic infection. In keeping with this conclusion, PML silencing was found to be sufficient to induce the EBV lytic cycle. Finally, by generating cells with single PML isoforms, we showed that individual PML isoforms were sufficient to suppress EBV lytic reactivation, although PML isoform IV (PML IV) was ineffective because it was most efficiently degraded by EBNA1. Our results provide the first function for EBNA1 in lytic infection and show that EBNA1 interactions with PML IV lead to a loss of PML nuclear bodies (NBs) that promotes lytic infection.
Residues within the C-Terminal Arm of the Herpes Simplex Virus 1 Glycoprotein B Ectodomain Contribute to Its Refolding during the Fusion Step of Virus Entry
Authors: Connolly Sarah A. , Longnecker Richard
Published: April 2012

Herpesvirus entry into cells requires coordinated interactions among several viral glycoproteins. The final membrane fusion step of entry is executed by glycoprotein B (gB), a class III viral fusion protein that is conserved across all herpesviruses. Fusion proteins are metastable proteins that mediate fusion by inserting into a target membrane and refolding from a prefusion to postfusion conformation to bring the viral and cell membranes together. Although the structure of gB has been solved in a conformation that likely represents its postfusion form, its prefusion structure and the details of how it refolds to execute fusion are unknown. The postfusion gB structure contains a trimeric coiled-coil at its core and a long C-terminal arm within the ectodomain packs against this coil in an antiparallel manner. This coil-arm complex is reminiscent of the six-helix bundle that provides the energy for fusion in class I fusogens. To determine the role of the coil-arm complex, we individually mutated residues in the herpes simplex virus 1 gB coil-arm complex to alanine and assessed the contribution of each residue to cell-cell and virus-cell fusion. Several coil mutations resulted in a loss of cell surface expression, indicating that the coil residues are important for proper processing of gB. Three mutations in the arm region (I671A, H681A, and F683A) reduced fusion without affecting expression. Combining these three arm mutations drastically reduced the ability of gB to execute fusion; however, fusion function could be restored by adding known hyperfusogenic mutations to the arm mutant. We propose that the formation of the coil-arm complex drives the gB transition to a postfusion conformation and the coil-arm complex performs a function similar to that of the six-helix bundle in class I fusion. Furthermore, we suggest that these specific mutations in the arm may energetically favor the prefusion state of gB.
Integrated Clinical, Pathologic, Virologic, and Transcriptomic Analysis of H5N1 Influenza Virus-Induced Viral Pneumonia in the Rhesus Macaque
Authors: Shinya Kyoko , Gao Yuwei , Cilloniz Cristian , Suzuki Yasuhiro , Fujie Masahiro , Deng Guohua , Zhu Qiyun , Fan Shufang , Makino Akiko , Muramoto Yukiko , Fukuyama Satoshi , Tamura Daisuke , Noda Takeshi , Eisfeld Amie J. , Katze Michael G. , Chen Hualan , Kawaoka Yoshihiro
Published: April 2012

Viral pneumonia has been frequently reported during early stages of influenza virus pandemics and in many human cases of highly pathogenic avian influenza (HPAI) H5N1 virus infection. To better understand the pathogenesis of this disease, we produced nonlethal viral pneumonia in rhesus macaques by using an HPAI H5N1 virus (A/Anhui/2/2005; referred to as Anhui/2). Infected macaques were monitored for 14 days, and tissue samples were collected at 6 time points for virologic, histopathologic, and transcriptomic analyses. Anhui/2 efficiently replicated in the lung from 12 h to 3 days postinfection (p.i.) and caused temporal but severe pneumonia that began to resolve by day 14. Lung transcriptional changes were first observed at 6 h, and increased expression of vascular permeability regulators and neutrophil chemoattractants correlated with increased serum leakage and neutrophil infiltrationin situ. Additional inflammatory, antiviral, and apoptotic genes were upregulated from 12 h, concurrent with viral antigen detection and increasing immune cell populations. A shift toward upregulation of acquired immunity was apparent after day 6. Expression levels of established immune cell molecular markers revealed remarkable similarity with pathological findings, indicating early and robust neutrophil infiltration, a slight delay in macrophage accumulation, and abundant late populations of T lymphocytes. We also characterized the putative mechanisms regulating a unique, pneumonia-associated biphasic fever pattern. Thus, this study is the first to use a comprehensive and integrative approach to delineate specific molecular mechanisms regulating influenza virus-induced pneumonia in nonhuman primates, an important first step toward better management of human influenza virus disease.
Characterization of the Viral
O
-Glycopeptidome: a Novel Tool of Relevance for Vaccine Design and Serodiagnosis
Authors: Cló Emiliano , Kračun Stjepan K. , Nudelman Aaron S. , Jensen Knud J. , Liljeqvist Jan-Åke , Olofsson Sigvard , Bergström Tomas , Blixt Ola
Published: April 2012

Viral envelope proteins mediate interactions with host cells, leading to internalization and intracellular propagation. Envelope proteins are glycosylated and are known to serve important functions in masking host immunity to viral glycoproteins. However, the viral infectious cycle in cells may also lead to aberrant glycosylation that may elicit immunity. Our knowledge of immunity to aberrant viral glycans and glycoproteins is limited, potentially due to technical limitations in identifying immunogenic glycans and glycopeptide epitopes. This work describes three different complementary methods for high-throughput screening and identification of potential immunodominant
O
-glycopeptide epitopes on viral envelope glycoproteins: (i) on-chip enzymatic glycosylation of scan peptides, (ii) chemical glycopeptide microarray synthesis, and (iii) a one-bead-one-compound random glycopeptide library. We used herpes simplex virus type 2 (HSV-2) as a model system and identified a simple
O
-glycopeptide pan-epitope,
501
PPA(GalNAc)TAPG
507
, on the mature gG-2 glycoprotein that was broadly recognized by IgG antibodies in HSV-2-infected individuals but not in HSV-1-infected or noninfected individuals. Serum reactivity to the extended sialyl-T glycoform was tolerated, suggesting that self glycans can participate in immune responses. The methods presented provide new insight into viral immunity and new targets for immunodiagnostic and therapeutic measures.
A 3-
O
-Sulfated Heparan Sulfate Binding Peptide Preferentially Targets Herpes Simplex Virus 2-Infected Cells
Authors: Ali Mohamed M. , Karasneh Ghadah A. , Jarding Min Jung , Tiwari Vaibhav , Shukla Deepak
Published: April 2012

Herpes simplex virus 2 (HSV-2) is the primary cause of genital herpes, which is one of the most common sexually transmitted viral infections worldwide and a major cofactor for human immunodeficiency virus infection. The lack of an effective vaccine or treatment and the emergence of drug-resistant strains highlight the need for developing new antivirals for HSV-2. Here, we demonstrate that a low-molecular-weight peptide isolated against 3-
O
-sulfated heparan sulfate (3-OS HS) can efficiently block HSV-2 infection. Treatment with the peptide inhibited viral entry and cell-to-cell spread both
in vitro
and
in vivo
using a mouse model of genital HSV-2 infection. Quite interestingly, the peptide showed a preferential binding to HSV-2-infected cells, with more than 200% increased binding compared to uninfected cells. Our additional results show that heparan sulfate expression is upregulated by 25% upon HSV-2 infection, which is a significant new finding that could be exploited for designing new diagnostic tests and treatment strategies against HSV-2-infected cells. In addition, our results also raise the possibility that 3-OS HS modifications within HS may be upregulated even more to accommodate for a significantly higher increase in the peptide binding to the infected cells.
A Pan-H1 Anti-Hemagglutinin Monoclonal Antibody with Potent Broad-Spectrum EfficacyIn Vivo
Authors: Tan Gene S. , Krammer Florian , Eggink Dirk , Kongchanagul Alita , Moran Thomas M. , Palese Peter
Published: April 2012

Seasonal epidemics caused by antigenic variations in influenza A virus remain a public health concern and an economic burden. The isolation and characterization of broadly neutralizing anti-hemagglutinin monoclonal antibodies (MAb) have highlighted the presence of highly conserved epitopes in divergent influenza A viruses. Here, we describe the generation and characterization of a mouse monoclonal antibody designed to target the conserved regions of the hemagglutinin of influenza A H1 viruses, a subtype that has caused pandemics in the human population in both the 20th and 21st centuries. By sequentially immunizing mice with plasmid DNA encoding the hemagglutinin of antigenically different H1 influenza A viruses (A/South Carolina/1/1918, A/USSR/92/1977, and A/California/4/2009), we isolated and identified MAb 6F12. Similar to other broadly neutralizing MAb previously described, MAb 6F12 has no hemagglutination inhibition activity against influenza A viruses and targets the stalk region of hemagglutinins. As designed, it has neutralizing activity against a divergent panel of H1 virusesin vitro, representing 79 years of antigenic drift. Most notably, MAb 6F12 prevented gross weight loss against divergent H1 viruses in passive transfer experiments in mice, both in pre- and postexposure prophylaxis regimens. The broad but specific activity of MAb 6F12 highlights the potent efficacy of monoclonal antibodies directed against a single subtype of influenza A virus.
Patterns of Acute Rhesus Cytomegalovirus (RhCMV) Infection Predict Long-Term RhCMV Infection
Authors: Assaf Basel T. , Mansfield Keith G. , Westmoreland Susan V. , Kaur Amitinder , Oxford Kristie L. , Diamond Don J. , Barry Peter A.
Published: April 2012

We previously reported that long-term rhesus cytomegalovirus (RhCMV) excretion in infected macaques was related to UL/b′ coding content. Acute biopsy specimens of the inoculation sites from the previous study have now been analyzed to determine whether there were acute phenotypic predictors of long-term RhCMV infection. Only in animals displaying acute endothelial tropism and neutrophilic inflammation was RhCMV excretion detected. The results imply that vaccinating against these early viral determinants would significantly impede long-term RhCMV infection.
Determinants in the Maturation of Rubella Virus P200 Replicase Polyprotein Precursor
Authors: Matthews Jason D. , Tzeng Wen-Pin , Frey Teryl K.
Published: April 2012

Rubella virus (RUBV), a positive-strand RNA virus, replicates its RNA within membrane-associated replication complexes (RCs) in the cytoplasm of infected cells. RNA synthesis is mediated by the nonstructural proteins (NSPs) P200 and its cleavage products, P150 and P90 (N and C terminal within P200, respectively), which are processed by a protease residing at the C terminus of P150. In this study of NSP maturation, we found that early NSP localization into foci appeared to target the membranes of the endoplasmic reticulum. During maturation, P150 and P90 likely interact within the context of P200 and remain in a complex after cleavage. We found that P150-P90 interactions were blocked by mutational disruption of an alpha helix at the N terminus (amino acids [aa] 36 to 49) of P200 and that these mutations also had an effect on NSP targeting, processing, and membrane association. While the P150-P90 interaction also required residues 1700 to 1900 within P90, focus formation required the entire RNA-dependent RNA polymerase (aa 1700 to 2116). Surprisingly, the RUBV capsid protein (CP) rescued RNA synthesis by several alanine-scanning mutations in the N-terminal alpha helix, and packaged replicon assays showed that rescue could be mediated by CP in the virus particle. We hypothesize that CP rescues these mutations as well as internal deletions of the Q domain within P150 and mutations in the 5′ and 3′
cis
-acting elements in the genomic RNA by chaperoning the maturation of P200. CP's ability to properly target the otherwise aggregated plasmid-expressed P200 provides support for this hypothesis.
Foamy Virus Pol Protein Expressed as a Gag-Pol Fusion Retains Enzymatic Activities, Allowing for Infectious Virus Production
Authors: Lee Eun-Gyung , Sinicrope Amber , Jackson Dana L. , Yu Shuyuarn F. , Linial Maxine L.
Published: April 2012

Foamy viruses (FV) synthesize Pol from a spliced
pol
mRNA independently of Gag, unlike orthoretroviruses, which synthesize Pol as a Gag-Pol protein that coassembles with Gag. We found that prototype FV (PFV) mutants expressing Gag and Pol only as a Gag-Pol protein without the spliced Pol contain protease activity equivalent to that of wild-type (WT) Pol. Regardless of the presence or absence of the spliced Pol, the PFV Gag-Pol proteins can assemble into virus-like particles (VLPs), in contrast to the orthoretroviral Gag-Pol proteins, which cannot form VLPs. However, the PFV Gag-Pol VLPs have aberrant morphologies and are not infectious. In the absence of the spliced Pol, coexpression of a PFV Gag-Pol protein with Gag can produce infectious virions. Our results suggest that enzymes encoded by PFV
pol
(protease, reverse transcriptase, and integrase) are enzymatically active if they are synthesized as part of a Gag-Pol protein.
Epitope Insertion at the N-Terminal Molecular Switch of the Rabbit Hemorrhagic Disease Virus T=3 Capsid Protein Leads to Larger T=4 Capsids
Authors: Luque Daniel , González José M. , Gómez-Blanco Josué , Marabini Roberto , Chichón Javier , Mena Ignacio , Angulo Iván , Carrascosa José L. , Verdaguer Nuria , Trus Benes L. , Bárcena Juan , Castón José R.
Published: April 2012

Viruses need only one or a few structural capsid proteins to build an infectious particle. This is possible through the extensive use of symmetry and the conformational polymorphism of the structural proteins. Using virus-like particles (VLP) from rabbit hemorrhagic disease virus (RHDV) as a model, we addressed the basis of calicivirus capsid assembly and their application in vaccine design. The RHDV capsid is based on a T=3 lattice containing 180 identical subunits (VP1). We determined the structure of RHDV VLP to 8.0-Å resolution by three-dimensional cryoelectron microscopy; in addition, we used San Miguel sea lion virus (SMSV) and feline calicivirus (FCV) capsid subunit structures to establish the backbone structure of VP1 by homology modeling and flexible docking analysis. Based on the three-domain VP1 model, several insertion mutants were designed to validate the VP1 pseudoatomic model, and foreign epitopes were placed at the N- or C-terminal end, as well as in an exposed loop on the capsid surface. We selected a set of T and B cell epitopes of various lengths derived from viral and eukaryotic origins. Structural analysis of these chimeric capsids further validates the VP1 model to design new chimeras. Whereas most insertions are well tolerated, VP1 with an FCV capsid protein-neutralizing epitope at the N terminus assembled into mixtures of T=3 and larger T=4 capsids. The calicivirus capsid protein, and perhaps that of many other viruses, thus can encode polymorphism modulators that are not anticipated from the plane sequence, with important implications for understanding virus assembly and evolution.
A meta-analysis to determine the effect on survival of platelet transfusions in patients with either spontaneous or traumatic antiplatelet medication-associated intracranial haemorrhage
Authors: Batchelor John S , Grayson Alan
Published: April 2012

Objectives

The aim of this study was to evaluate by meta-analysis the current level of evidence in order to establish the impact of a platelet transfusion on survival in patients on pre-injury antiplatelet agents who sustain an intracranial haemorrhage (either spontaneous or traumatic).

Design

This was a meta-analysis; the MEDLINE Database was searched using the PubMed interface and the Ovid interface. CINAHL and EMBASE Databases were also searched. The search was performed to identify randomised controlled trials (RCT)'s case-controlled studies or nested case-controlled studies. Comparing the outcome (death or survival) of patients with intracranial haemorrhage (ICH) and pre-injury antiplatelet agents who received a platelet transfusion against a similar cohort of patients who did not receive a platelet transfusion.

Results

499 citations were obtained from the PubMed search. 31 full articles were reviewed from 34 abstracts. 6 studies were found suitable for the meta-analysis. No randomised controlled studies were identified. 2 of the six studies were in patients with spontaneous ICH. The remaining four studies were in patients with traumatic intracranial haemorrhage. Significant heterogeneity was present between the studies, I2=58.276. The random effects model was therefore the preferred model, this produced a pooled OR for survival of 0.773 (95% CI 0.414 to 1.442).

Conclusions

The results of this meta-analysis has shown, based upon six small studies, that there was no clear benefit in terms of survival in the administration of a platelet transfusion to patients with antiplatelet-associated ICH. Further work is required in order to establish any potential benefit in the administration of a platelet transfusion in patients with spontaneous or traumatic intracranial haemorrhage who were on pre-injury antiplatelet agents.
Oral phenotype and scoring of vascular Ehlers–Danlos syndrome: a case–control study
Authors: Ferré François Côme , Frank Michael , Gogly Bruno , Golmard Lisa , Naveau Adrien , Chérifi Hafida , Emmerich Joseph , Gaultier Frédérick , Berdal Ariane , Jeunemaitre Xavier , Fournier Benjamin P J
Published: April 2012

Objective

Vascular Ehlers–Danlos syndrome (vEDS) is a rare genetic condition related to mutations in theCOL3A1gene, responsible of vascular, digestive and uterine accidents. Difficulty of clinical diagnosis has led to the design of diagnostic criteria, summarised in the Villefranche classification. The goal was to assess oral features of vEDS. Gingival recession is the only oral sign recognised as a minor diagnostic criterion. The authors aimed to check this assumption since bibliographical search related to gingival recession in vEDS proved scarce.

Design

Prospective case–control study.

Setting

Dental surgery department in a French tertiary hospital.

Participants

17 consecutive patients with genetically proven vEDS, aged 19–55 years, were compared with 46 age- and sex-matched controls.

Observations

Complete oral examination (clinical and radiological) with standardised assessment of periodontal structure, temporomandibular joint function and dental characteristics were performed.COL3A1mutations were identified by direct sequencing of genomic or complementary DNA.

Results

Prevalence of gingival recession was low among patients with vEDS, as for periodontitis. Conversely, patients showed marked gingival fragility, temporomandibular disorders, dentin formation defects, molar root fusion and increased root length. After logistic regression, three variables remained significantly associated to vEDS. These variables were integrated in a diagnostic oral score with 87.5% and 97% sensitivity and specificity, respectively.

Conclusions

Gingival recession is an inappropriate diagnostic criterion for vEDS. Several new specific oral signs of the disease were identified, whose combination may be of greater value in diagnosing vEDS.