bluuebunny/crossref_metadata_2025 huggingface dataset:
The corona of the daffodil Narcissus bulbocodium shares stamen‐like identity and is distinct from the orthodox floral whorls
Authors: Waters Mark T. , Tiley Anna M. M. , Kramer Elena M. , Meerow Alan W. , Langdale Jane A. , Scotland Robert W.
Published: February 2013

The structural homology of the daffodil corona has remained a source of debate throughout the history of botany. Over the years it has been separately referred to as a modified petal stipule, stamen and tepal. Here we provide insights from anatomy and molecular studies to clarify the early developmental stages and position of corona initiation in Narcissus bulbocodium. We demonstrate that the corona initiates as six separate anlagen from hypanthial tissue between the stamens and perianth. Scanning electron microscope images and serial sections demonstrate that corona initiation occurs late in development, after the other floral whorls are fully developed. To define more precisely the identity of the floral structures, daffodil orthologues of the ABC floral organ identity genes were isolated and expression patterns were examined in perianth, stamens, carpel, hypanthial tube and corona tissue. Coupled with in situ hybridisation experiments, these analyses showed that the expression pattern of the C‐class gene NbAGAMOUS in the corona is more similar to that of the stamens than that of the tepals. In combination, our results demonstrate that the corona of the daffodil N. bulbocodium exhibits stamen‐like identity, develops independently from the orthodox floral whorls and is best interpreted as a late elaboration of the region between the petals and stamens associated with epigyny and the hypanthium.
Tapetal dysplasia in a Swedish Vallhund dog
Authors: Scott Erin M. , Teixeira Leandro B.C. , Dubielzig Richard R. , Komáromy András M.
Published: February 2013

Objective

To describe the gross, histopathological, and ultrastructural findings in a dog with bilateral tapetal dysplasia.

Procedures

The globes of a 15‐year‐old neutered male Swedish Vallhund dog with a ventrally displaced tapetum in both eyes were fixed in 10% formalin and submitted to the Comparative Ocular Pathology Laboratory of Wisconsin for histological evaluation. Sections were stained with hematoxylin and eosin, Masson's trichrome, and Melan‐A immunohistochemistry (IHC), and tissues were subsequently processed for transmission electron microscopy.

Results

Bilateral fundic and gross examination revealed a tapetal fundus inferior to the optic nerve head (ONH) and a nontapetal fundus with mild scattering of tapetal tissue superior to the ONH. Histologically, there was decreased pigmentation of the retinal pigment epithelium with only a few melanin granules in the peripheral retina. The affected tapetum was relatively acellular and fibrous with occasional tapetal cells scattered throughout the inner choroid or displaced into the vascular outer choroid. Special stains revealed that the tapetum was mostly composed of collagen (Masson's trichrome) and failed to express Melan‐A (IHC) unlike a normal canine control tapetum. Ultrastructurally, the tapetum was markedly dysplastic both superior and inferior to the ONH with no uniformly arranged tapetal cells. The few cells identified within the tapetum contained irregularly arranged and disorganized electron‐dense structures within their cytoplasm, which were interpreted as dysplastic tapetal rodlets.

Conclusions

Based on microscopic and ultrastructural findings, this is the first report of tapetal dysplasia in a dog.
Evaluation of one‐ vs. two‐layered closure after wedge excision of 43 eyelid tumors in dogs
Authors: Romkes Gwendolyna , Klopfleisch Robert , Eule Johanna Corinna
Published: February 2013

Objective

The aim was to evaluate how two closure techniques after full‐thickness V‐shaped excision relate to wound recovery and how these techniques affect eyelid function and stability of the preocular tear film.

Animals studied

Thirty‐eight dogs with 43 eyelid tumors involving <25% of eyelid margin length were included in a prospective randomized trial.

Procedures

The method for wound closure was chosen randomly before the operation. In group A (n = 20), the wound was closed in one layer and in group B (n = 23) in two layers. The wound and eyelid structure were directly evaluated by slit‐lamp biomicroscopy several times postoperatively. Eyelid function was indirectly evaluated by interferometry, noninvasive tear film breakup time (nTFBUT), andSchirmer's tear test at the last examination, 5 weeks to 19 months postoperatively.

Results

No significant differences were found in group A vs. B although slightly more cases with perfect alignment of the eyelid were seen in group B. In both groups, neither slit‐lamp biomicroscopy, interferometry, nTFBUT, norSchirmer's tear test revealed negative influence on the eyelid function by the previous surgery. Histologically, 29 of 32 tumors were diagnosed as meibomian gland adenomas.

Conclusions

These results do not demonstrate a significant difference in wound recovery, eyelid structure, and function after wound closure in 1 vs. 2 layers.
Thromboelastographic evaluation of coagulative profiles in pig‐to‐monkey kidney xenotransplantation
Authors: Spiezia Luca , Boldrin Massimo , Radu Claudia , Bulato Cristiana , Bertini Diana , Bon Maria , Campello Elena , Vadori Marta , Galli Cesare , Gavasso Sabrina , Nottle Mark B. , Cowan Peter J. , Cozzi Emanuele , Simioni Paolo
Published: February 2013

Background

Activation of the clotting cascade is central in acute xenograft rejection (AHXR) that occurs when pig organs are transplanted into primates. The coagulopathy reported in this model is a very complex process that involves simultaneously coagulation factors, platelets and phospholipid‐bearing cells (i.e., leukocytes, red blood cells, and endothelial cells). Choosing whole blood for coagulation analysis theoretically appears more favorable compared with plasma. Whole blood rotation thromboelastometry (ROTEM®) is a point‐of‐care global coagulation analyzer able to evaluate the characteristics of clot formation and lysis by dynamic monitoring. The aim of this study was to record thromboelastographic profiles, performed by ROTEM®, in a series of immunosuppressed nephrectomized primates that received a life‐supporting kidney.

Methods

Of the eight primates, n = 4 received a pig kidney transgenic for human decay‐accelerating factor (hDAF/Gal+); n = 2, an α 1,3‐galactosyltransferase gene‐knockout (GT‐KO) pig kidney transgenic for human CD39, CD55, CD59 and fucosyltransferase (HTF); and n = 2, a GT‐KO pig kidney transgenic for hDAF. Blood samples were collected before and at least once per week after transplantation till euthanasia. Intrinsic (INTEM) and extrinsic (EXTEM) coagulation pathways and the function of fibrinogen (FIBTEM) were evaluated. Thromboelastographic parameters considered were clotting time (CT, seconds) and clot formation time (CFT, seconds) in INTEM and EXTEM and maximum clot firmness (MCF, mm) in FIBTEM. The correlations between CT in INTEM and activated partial thromboplastin time (aPTT), CT in EXTEM and PT, CFT in INTEM and EXTEM, and platelet counts and MCF in FIBTEM and fibrinogen plasma levels were also considered.

Results

In all animals, thromboelastographic profiles showed progressive prolongation of CT (activation of coagulative cascade) in INTEM. A close correspondence was observed between (i) the prolongation of the CFT values (propagation of clot formation), both in INTEM and EXTEM, and the decrease in platelet counts; (ii) the reduction in MCF values (clot firmness) ​​in FIBTEM and the decrease in fibrinogen plasma levels. No concordance between CT in INTEM and aPTT and between CT in EXTEM and PT was observed.

Conclusions

Our study demonstrated that ROTEM® analyzer could be a useful and complementary tool to study the consumptive coagulopathy, either “compensated” or “non‐compensated,” that takes place when transgenic pig kidneys are transplanted into primates. Larger and prospective studies are needed to confirm our results and to evaluate the role of ROTEM® to guide the management of consumptive coagulopathy in order to prolong the survival of the transplanted organ.
On admissible tensor products inp-adic Hodge theory
Authors: Di Matteo Giovanni
Published: February 2013

We prove that ifWandW′are non-zeroB-pairs whose tensor product is crystalline (or semi-stable or de Rham or Hodge–Tate), then there exists a characterμsuch thatW(μ−1) andW′(μ) are crystalline (or semi-stable or de Rham or Hodge–Tate). We also prove that ifWis aB-pair and ifFis a Schur functor (for example Symnor Λn) such thatF(W) is crystalline (or semi-stable or de Rham or Hodge–Tate) and if the rank ofWis sufficiently large, then there is a characterμsuch thatW(μ−1) is crystalline (or semi-stable or de Rham or Hodge–Tate). In particular, these results apply top-adic representations.
Heat Transfer in the Flow of a Cold, Axisymmetric Jet Over a Hot Sphere
Authors: Shu Jian-Jun , Wilks Graham
Published: February 2013

The heat-transfer characteristics of thin film flow over a hot sphere resulting from a cold vertical jet of liquid falling onto the surface have been investigated. The underlying physical features have been illustrated by numerical solutions of high accuracy based on the modified Keller box method. The solutions for film thickness distribution are good agreement with those obtained approximately by using the Pohlhausen integral momentum technique and observed experimentally by using water as working fluid, thus providing a basic confirmation of the validity of the results presented.
Comparison of the effects of downstream H2- and O2-based plasmas on the removal of photoresist, silicon, and silicon nitride
Authors: Thedjoisworo Bayu , Cheung David , Crist Vince
Published: February 2013

For the 45 nm technology node and beyond, there is a need to strip photoresist quickly while suppressing the loss of materials such as polycrystalline silicon (poly-Si) and silicon nitride (Si3N4). To achieve this goal, the authors characterized and compared the effects of downstream pure-H2, H2/N2, and O2/N2 plasmas on the etch behaviors of photoresist, poly-Si, and Si3N4. The addition of N2 to H2 plasma increases the photoresist ash rate to a maximum that is reached at ∼30–40% N2, and the ash rate drops with further addition of N2. At 30% N2 addition, the ash rate increases by a factor of ∼3 when compared to that obtained with pure-H2 plasma. For O2/N2 plasma, the photoresist ash rate also exhibits a maximum, which is attained with 5% N2 addition, and the ash rate drops drastically as more N2 is added. A small addition of N2 increases the H and O radical densities in the H2- and O2-based plasmas, respectively, resulting in the higher ash rates. The ash rate achieved by the O2/N2 chemistry is generally higher than that attained with the H2/N2 chemistry, and the difference becomes more significant at high temperatures. The activation energy for photoresist strip under O2/N2 plasma was measured to be ∼10 kcal/mol, which is higher when compared to the ∼5 kcal/mol measured for both the H2/N2 (30% N2) and the pure-H2 chemistries. At 300 °C, when compared to the O2-based chemistry, the H2-based chemistry was shown to remove Si3N4 with a much lower rate, ∼0.7 Å/min, highlighting the benefit of the latter in conserving material loss. The ability of the H2-based chemistry to suppress material loss and its nonoxidizing property could justify the trade off for its lower ash rates when compared to those obtained using the O2-based chemistry. For the H2-based chemistry, a small N2 addition to the H2 plasma was found to not only increase the ash rate but also suppress the Si etch rate by a factor of 8 to 22, depending on the temperature. Collectively, the H2/N2 chemistry shows a great promise for photoresist-strip applications in the advanced nodes, and it should be run at high temperatures (e.g., T ≥ 300 °C) to maximize the ash rate while still maintaining extremely low Si and Si3N4 losses.
Dilute Concentrations of a Psychiatric Drug Alter Behavior of Fish from Natural Populations
Authors: Brodin T. , Fick J. , Jonsson M. , Klaminder J.
Published: February 2013

Unintended Recipients of Antidepressants

Pharmaceuticals are used to treat a wide variety of ailments and conditions in humans. However, many animal species share physiologies, receptors, and pathways that may be acted upon by pharmaceutical compounds. Increasingly, pharmaceuticals are being found in natural aquatic systems. Such pharmaceutical pollution can cause mortality and alter development and reproduction of aquatic animals.

Brodin
et al.

(p.
814
) report that excreted drugs may also have far more subtle, yet eventually significant, impacts in natural systems. Benzodiazepines, which reduce anxiety in humans, alter social and foraging behavior in fish. European perch exposed to oxazepam were bolder, more active, less social and fed more rapidly.
Observation of Radiation Pressure Shot Noise on a Macroscopic Object
Authors: Purdy T. P. , Peterson R. W. , Regal C. A.
Published: February 2013

Macroscopic Uncertainty

According to the Heisenberg uncertainty principle, it is impossible to know both the position and the momentum of a microscopic particle with absolute certainty; pinpointing the location introduces an uncertainty in the velocity, which translates into position uncertainty at later times. Now,

Purdy
et al.

(p.
801
; see the Perspective by

Milburn

) have measured the position of a macroscopic object (a small, but visible-to-the-naked-eye membrane suspended in an optical cavity) at cryogenic temperatures and observed the uncertainty in its position caused by the recoiling photons used for the measurement.
Mr. Bayes Goes to Washington
The Signal and the Noise
Why So Many Predictions Fail—But Some Don't/The Art and Science of Prediction

by Nate Silver

Penguin, New York, 2012. 542 pp. $27.95, C$29.50. ISBN 9781594204111. Allen Lane, London. €25. ISBN 9781846147524.
Authors: Wang Sam , Campbell Benjamin C.
Published: February 2013

With a sense of fun, Silver explores good and bad forecasting in daily life, leisure activity, and science.
Beating Classical Computing Without a Quantum Computer
Authors: Franson James D.
Published: February 2013

Photons performing quantum random walks can be used to calculate matrix properties without actually implementing quantum logic gates.

[Also see Reports by

Childs
et al.

,

Broome
et al.

, and

Spring
et al.

]
European Budget Fight Moves to Parliament
Authors: Vogel Gretchen
Published: February 2013

European science won a welcome increase in funds, but it's still taking a hit.
Plagiarism Hunters Take Down Research Minister
Authors: Kupferschmidt Kai , Vogel Gretchen
Published: February 2013

Germany's minister for science and education is the latest in a string of officials here who have stepped down amid plagiarism allegations.
Cash-Strapped NASA Gets Many Ideas for 'Free' Telescopes
Authors: Bhattacharjee Yudhijit
Published: February 2013

The gift of two telescopes provides a chance to think big, but it will take money to turn those ideas into reality.
Faulty DNA Repair Linked to Ovarian Aging in Mice and Humans
Authors: Couzin-Frankel Jennifer
Published: February 2013

Several recent studies have pointed to DNA repair generally as something that falters with ovarian age, though details remain poorly understood.
Opsins: Not Just for Eyes
Authors: Pennisi Elizabeth
Published: February 2013

Studies in invertebrates are enriching our sense of how versatile and ancient these light-sensitive proteins are.
LIFE SCIENCE TECHNOLOGIES: Clinical Aspirations of Microarrays
Authors: May Mike
Published: February 2013

Although most microarray applications are currently research-use-only, this technology appears poised to move to the clinic for genomics-based applications. In fact, some products can already be used in medical diagnostics and many more are in development. For example, microarrays can be customized to detect small, specific genetic changes that indicate a particular disease. In the future, this technology will likely remain a useful tool for both research and clinical applications.
Immunity to Hepatitis B Virus (HBV) Infection Two Decades after Implementation of Universal Infant HBV Vaccination: Association of Detectable Residual Antibodies and Response to a Single HBV Challenge Dose
Authors: Spradling Philip R. , Xing Jian , Williams Roxanne , Masunu-Faleafaga Yolanda , Dulski Theresa , Mahamud Abdirahman , Drobeniuc Jan , Teshale Eyasu H.
Published: February 2013

Most persons who receive hepatitis B vaccine during infancy will have a level of antibody to hepatitis B surface antigen (anti-HBsAg) of <10 IU/liter 10 to 15 years later; however, most will demonstrate immune memory by an anamnestic response to a vaccine challenge dose. To determine whether there was a difference in anamnestic response among college students vaccinated during infancy, we compared anti-HBsAg levels after a 20-μg dose of Engerix-B in those with a residual anti-HBsAg level of 0 IU/liter and those with levels of 1 to 9 IU/liter. Anti-HBsAg was measured before (baseline) and 2 weeks after a challenge dose; a response was defined as a level of ≥10 IU/liter after the dose among those with <10 IU/liter at the baseline. Of the 153 students who completed the study, 130 (85%) had an anti-HBsAg level of <10 IU/liter at the baseline, 72 had a level of 0 IU/liter, and 58 had levels ranging from 1 to 9 IU/liter. Students with a levels of 1 to 9 IU/liter were more likely to respond to the challenge dose than those with a baseline anti-HBsAg level of 0 IU/liter (83% versus 50%;
P
< 0.001). The presence of any detectable anti-HBsAg among persons vaccinated in the remote past may indicate the persistence of immune memory.
Use of Nasopharyngeal Aspirate for Diagnosis of Pneumocystis Pneumonia
Authors: To Kelvin K. W. , Wong Sally C. Y. , Xu Ting , Poon Rosana W. S. , Mok Ka-Yi , Chan Jasper F. W. , Cheng Vincent C. C. , Chan Kwok-Hung , Hung Ivan F. N. , Yuen Kwok-Yung
Published: February 2013

Quantitative PCR on nasopharyngeal aspirate (NPA) can achieve high sensitivity and specificity in diagnosingPneumocystispneumonia (PCP) compared to microscopic examination of bronchoscopic specimens in a population with low HIV prevalence. Since NPA is a minimally invasive procedure, it is ideal as a screening test for PCP.
Conformational Flexibility of the Dengue Virus RNA-Dependent RNA Polymerase Revealed by a Complex with an Inhibitor
Authors: Noble Christian G. , Lim Siew Pheng , Chen Yen-Liang , Liew Chong Wai , Yap Lijian , Lescar Julien , Shi Pei-Yong
Published: February 2013

We report a highly reproducible method to crystallize the RNA-dependent RNA polymerase (RdRp) domain of dengue virus serotype 3 (DENV-3), allowing structure refinement to a 1.79-Å resolution and revealing amino acids not seen previously. We also present a DENV-3 polymerase/inhibitor cocrystal structure at a 2.1-Å resolution. The inhibitor binds to the RdRp as a dimer and causes conformational changes in the protein. The improved crystallization conditions and new structural information should accelerate structure-based drug discovery.
TRIM22 Inhibits Influenza A Virus Infection by Targeting the Viral Nucleoprotein for Degradation
Authors: Di Pietro Andrea , Kajaste-Rudnitski Anna , Oteiza Alexandra , Nicora Lucia , Towers Greg J. , Mechti Nadir , Vicenzi Elisa
Published: February 2013

Tripartite motif (TRIM) protein superfamily members are emerging as important effectors of the innate immune response against viral infections. In particular, TRIM22 was reported to exert antiviral activity against RNA viruses, such as hepatitis B virus (HBV), encephalomyocarditis virus (ECMV), and human immunodeficiency virus type 1 (HIV-1). We demonstrate here, for the first time, that TRIM22 is upregulated by influenza A virus (IAV) infection at both mRNA and protein levels in human alveolar epithelial A549 cells. Conversely, TRIM22 potently restricted IAV replication, in that prevention of TRIM22 expression by means of short hairpin RNA led to a 10-fold enhancement of IAV replication in these cells. Depletion of TRIM22 also reduced the anti-IAV activity of alpha interferon (IFN-α), suggesting that TRIM22 is an important IFN-stimulated gene that is required for maximal suppression of IAV by type I IFN. Furthermore, the IAV infectious titer decreased up to 100-fold in MDCK cells expressing exogenous human TRIM22. Restriction of IAV replication was accounted for by the interaction between TRIM22 and the viral nucleoprotein (NP), resulting in its polyubiquitination and degradation in a proteasome-dependent manner. Thus, TRIM22 represents a novel restriction factor upregulated upon IAV infection that curtails its replicative capacity in epithelial cells.
Worldwide Phylogenetic Relationship of Avian Poxviruses
Authors: Gyuranecz Miklós , Foster Jeffrey T. , Dán Ádám , Ip Hon S. , Egstad Kristina F. , Parker Patricia G. , Higashiguchi Jenni M. , Skinner Michael A. , Höfle Ursula , Kreizinger Zsuzsa , Dorrestein Gerry M. , Solt Szabolcs , Sós Endre , Kim Young Jun , Uhart Marcela , Pereda Ariel , González-Hein Gisela , Hidalgo Hector , Blanco Juan-Manuel , Erdélyi Károly
Published: February 2013

Poxvirus infections have been found in 230 species of wild and domestic birds worldwide in both terrestrial and marine environments. This ubiquity raises the question of how infection has been transmitted and globally dispersed. We present a comprehensive global phylogeny of 111 novel poxvirus isolates in addition to all available sequences from GenBank. Phylogenetic analysis of theAvipoxvirusgenus has traditionally relied on one gene region (4b core protein). In this study we expanded the analyses to include a second locus (DNA polymerase gene), allowing for a more robust phylogenetic framework, finer genetic resolution within specific groups, and the detection of potential recombination. Our phylogenetic results reveal several major features of avipoxvirus evolution and ecology and propose an updated avipoxvirus taxonomy, including three novel subclades. The characterization of poxviruses from 57 species of birds in this study extends the current knowledge of their host range and provides the first evidence of the phylogenetic effect of genetic recombination of avipoxviruses. The repeated occurrence of avian family or order-specific grouping within certain clades (e.g., starling poxvirus, falcon poxvirus, raptor poxvirus, etc.) indicates a marked role of host adaptation, while the sharing of poxvirus species within prey-predator systems emphasizes the capacity for cross-species infection and limited host adaptation. Our study provides a broad and comprehensive phylogenetic analysis of theAvipoxvirusgenus, an ecologically and environmentally important viral group, to formulate a genome sequencing strategy that will clarify avipoxvirus taxonomy.
Molecular Aspects and Comparative Genomics of Bacteriophage Endolysins
Authors: Oliveira Hugo , Melo Luís D. R. , Santos Sílvio B. , Nóbrega Franklin L. , Ferreira Eugénio C. , Cerca Nuno , Azeredo Joana , Kluskens Leon D.
Published: February 2013

Phages are recognized as the most abundant and diverse entities on the planet. Their diversity is determined predominantly by their dynamic adaptation capacities when confronted with different selective pressures in an endless cycle of coevolution with a widespread group of bacterial hosts. At the end of the infection cycle, progeny virions are confronted with a rigid cell wall that hinders their release into the environment and the opportunity to start a new infection cycle. Consequently, phages encode hydrolytic enzymes, called endolysins, to digest the peptidoglycan. In this work, we bring to light all phage endolysins found in completely sequenced double-stranded nucleic acid phage genomes and uncover clues that explain the phage-endolysin-host ecology that led phages to recruit unique and specialized endolysins.
Ubiquitin-Independent Proteasomal Degradation of Tumor Suppressors by Human Cytomegalovirus pp71 Requires the 19S Regulatory Particle
Authors: Winkler Laura L. , Hwang Jiwon , Kalejta Robert F.
Published: February 2013

Proteasomes generally degrade substrates tagged with polyubiquitin chains. In rare cases, however, proteasomes can degrade proteins without prior ubiquitination. For example, the human cytomegalovirus (HCMV) pp71 protein induces the proteasome-dependent, ubiquitin-independent degradation of the retinoblastoma (Rb) and Daxx proteins. These transcriptional corepressors and tumor suppressors inhibit the expression of cellular or viral genes that are required for efficient viral replication. Proteasomes are composed of a 20S catalytic core with or without one or two activator complexes, of which there are four different types. Here, we show that only one of these activators, the 19S regulatory particle that normally participates in ubiquitin-dependent protein degradation, is required for pp71-mediated degradation of Rb and Daxx. We report the unique use of a well-established route of substrate delivery to the proteasome by a viral protein to promote infection.
Venezuelan Equine Encephalitis Virus Replicon Particle Vaccine Protects Nonhuman Primates from Intramuscular and Aerosol Challenge with Ebolavirus
Authors: Herbert Andrew S. , Kuehne Ana I. , Barth James F. , Ortiz Ramon A. , Nichols Donald K. , Zak Samantha E. , Stonier Spencer W. , Muhammad Majidat A. , Bakken Russell R. , Prugar Laura I. , Olinger Gene G. , Groebner Jennifer L. , Lee John S. , Pratt William D. , Custer Max , Kamrud Kurt I. , Smith Jonathan F. , Hart Mary Kate , Dye John M.
Published: February 2013

There are no vaccines or therapeutics currently approved for the prevention or treatment of ebolavirus infection. Previously, a replicon vaccine based on Venezuelan equine encephalitis virus (VEEV) demonstrated protective efficacy against Marburg virus in nonhuman primates. Here, we report the protective efficacy of
Sudan virus
(SUDV)- and
Ebola virus
(EBOV)-specific VEEV replicon particle (VRP) vaccines in nonhuman primates. VRP vaccines were developed to express the glycoprotein (GP) of either SUDV or EBOV. A single intramuscular vaccination of cynomolgus macaques with VRP expressing SUDV GP provided complete protection against intramuscular challenge with SUDV. Vaccination against SUDV and subsequent survival of SUDV challenge did not fully protect cynomolgus macaques against intramuscular EBOV back-challenge. However, a single simultaneous intramuscular vaccination with VRP expressing SUDV GP combined with VRP expressing EBOV GP did provide complete protection against intramuscular challenge with either SUDV or EBOV in cynomolgus macaques. Finally, intramuscular vaccination with VRP expressing SUDV GP completely protected cynomolgus macaques when challenged with aerosolized SUDV, although complete protection against aerosol challenge required two vaccinations with this vaccine.
Comparative Characterization of Transfection- and Infection-Derived Simian Immunodeficiency Virus Challenge Stocks for
In Vivo
Nonhuman Primate Studies
Authors: Del Prete Gregory Q. , Scarlotta Matthew , Newman Laura , Reid Carolyn , Parodi Laura M. , Roser James D. , Oswald Kelli , Marx Preston A. , Miller Christopher J. , Desrosiers Ronald C. , Barouch Dan H. , Pal Ranajit , Piatak Michael , Chertova Elena , Giavedoni Luis D. , O'Connor David H. , Lifson Jeffrey D. , Keele Brandon F.
Published: February 2013

Simian immunodeficiency virus (SIV) stocks for
in vivo
nonhuman primate models of AIDS are typically generated by transfection of 293T cells with molecularly cloned viral genomes or by expansion in productively infected T cells. Although titers of stocks are determined for infectivity
in vitro
prior to
in vivo
inoculation, virus production methods may differentially affect stock features that are not routinely analyzed but may impact
in vivo
infectivity, mucosal transmissibility, and early infection events. We performed a detailed analysis of nine SIV stocks, comprising five infection-derived SIVmac251 viral swarm stocks and paired infection- and transfected-293T-cell-derived stocks of both SIVmac239 and SIVmac766. Representative stocks were evaluated for (i) virus content, (ii) infectious titer, (iii) sequence diversity and polymorphism frequency by single-genome amplification and 454 pyrosequencing, (iv) virion-associated Env content, and (v) cytokine and chemokine content by 36-plex Luminex analysis. Regardless of production method, all stocks had comparable particle/infectivity ratios, with the transfected-293T stocks possessing the highest overall virus content and infectivity titers despite containing markedly lower levels of virion-associated Env than infection-derived viruses. Transfected-293T stocks also contained fewer and lower levels of cytokines and chemokines than infection-derived stocks, which had elevated levels of multiple analytes, with substantial variability among stocks. Sequencing of the infection-derived SIVmac251 stocks revealed variable levels of viral diversity between stocks, with evidence of stock-specific selection and expansion of unique viral lineages. These analyses suggest that there may be underappreciated features of SIV
in vivo
challenge stocks with the potential to impact early infection events, which may merit consideration when selecting virus stocks for
in vivo
studies.
Norwalk Virus Minor Capsid Protein VP2 Associates within the VP1 Shell Domain
Authors: Vongpunsawad Sompong , Venkataram Prasad B. V. , Estes Mary K.
Published: February 2013

The major capsid protein of norovirus VP1 assembles to form an icosahedral viral particle. Despite evidence that the Norwalk virus (NV) minor structural protein VP2 is present in infectious virions, the available crystallographic and electron cryomicroscopy structures of NV have not revealed the location of VP2. In this study, we determined that VP1 associates with VP2 at the interior surface of the capsid, specifically with the shell (S) domain of VP1. We mapped the interaction site to amino acid 52 of VP1, an isoleucine located within a sequence motif IDPWI in the S domain that is highly conserved across norovirus genogroups. Mutation of this isoleucine abrogated VP2 incorporation into virus-like particles without affecting the ability for VP1 to dimerize and form particles. The highly basic nature of VP2 and its location interior to the viral particle are consistent with its potential role in assisting capsid assembly and genome encapsidation.
H3N2 Influenza Virus Infection Induces Broadly Reactive Hemagglutinin Stalk Antibodies in Humans and Mice
Authors: Margine Irina , Hai Rong , Albrecht Randy A. , Obermoser Gerlinde , Harrod A. Carson , Banchereau Jacques , Palucka Karolina , García-Sastre Adolfo , Palese Peter , Treanor John J. , Krammer Florian
Published: February 2013

Broadly neutralizing antibodies directed against the conserved stalk domain of the viral hemagglutinin have attracted increasing attention in recent years. However, only a limited number of stalk antibodies directed against group 2 influenza hemagglutinins have been isolated so far. Also, little is known about the general level of induction of these antibodies by influenza virus vaccination or infection. To characterize the anti-stalk humoral response in the mouse model as well as in humans, chimeric hemagglutinin constructs previously developed in our group were employed in serological assays. Whereas influenza virus infection induced high titers of stalk-reactive antibodies, immunization with inactivated influenza virus vaccines failed to do so in the mouse model. Analysis of serum samples collected from human individuals who were infected by influenza viruses also revealed the induction of stalk-reactive antibodies. Finally, we show that the hemagglutinin stalk-directed antibodies induced in mice and humans have broad reactivity and neutralizing activity
in vitro
and
in vivo
. The results of the study point toward the existence of highly conserved epitopes in the stalk domains of group 2 hemagglutinins, which can be targeted for the development of a universal influenza virus vaccine in humans.
A Single Amino Acid at the Hemagglutinin Cleavage Site Contributes to the Pathogenicity but Not the Transmission of Egyptian Highly Pathogenic H5N1 Influenza Virus in Chickens
Authors: Yoon Sun-Woo , Kayali Ghazi , Ali Mohamed A. , Webster Robert G. , Webby Richard J. , Ducatez Mariette F.
Published: February 2013

Highly pathogenic avian influenza (HPAI) virus H5N1 has been enzootic in Egypt since 2008. Virus-associated mortality (but not the number of cases) in humans and poultry seems to have decreased over time, but the reason for this remains unknown. We investigated the role of a single amino acid substitution in the hemagglutinin cleavage site on virus pathogenicity and transmission in chickens. The R325G substitution significantly reduced pathogenicity without altering the transmission efficiency of HPAI H5N1 virus.
Multiple Forms of Vector Manipulation by a Plant-Infecting Virus: Bemisia tabaci and Tomato Yellow Leaf Curl Virus
Authors: Liu Baiming , Preisser Evan L. , Chu Dong , Pan Huipeng , Xie Wen , Wang Shaoli , Wu Qingjun , Zhou Xuguo , Zhang Youjun
Published: February 2013

For many insect-vectored plant viruses, the relationship between feeding behavior and vector competence may prove integral to an understanding of the epidemiology of the resulting plant disease. While plant-infecting viruses are well known to change host plant physiology in a way that makes them more attractive to vectors, viral manipulation of the vectors themselves has only recently been reported. Previous research suggested that the rapid spread of
Tomato yellow leaf curl virus
(TYLCV) throughout China has been facilitated by its primary vector, the whitefly
Bemisia tabaci
. We conducted two experiments testing the impact of TYLCV infection of the host plant (tomato) and vector (
B. tabaci
biotypes B and Q) on whitefly feeding behavior. Whiteflies of biotypes B and Q both appeared to find TYLCV-infected plants more attractive, probing them more quickly and having a greater number of feeding bouts; this did not, however, alter the total time spent feeding. Viruliferous whiteflies fed more readily than uninfected whiteflies and spent more time salivating into sieve tube elements. Because vector salivation is essential for viral transmission, this virally mediated alteration of behavior should provide TYLCV a direct fitness benefit. This is the first report of such manipulation by a nonpropagative virus that belongs to an exclusively plant-infecting family of viruses (
Geminiviridae
). In the context of previous research showing that feeding on TYLCV-infected plants harms biotype B but helps biotype Q, the fact that both biotypes were equally affected by TYLCV also suggests that the virus may alter the biotype B-biotype Q competitive interaction in favor of biotype Q.
Tuberculosis of gallbladder neck: a cause of cholecystitis with cholelithiasis
Authors: Verma S K , Mishra Ashwini Kumar , Jaiswal Ashish Kumar
Published: February 2013

Tuberculosis of gallbladder neck is not a very common problem reported in the literature. Here, we report a case of gallbladder neck tuberculosis complicated with chronic cholecystitis with cholelithiasis in a 55-year-old woman. Diagnosis was made postoperatively on surgical biopsy.
A rare case of isolated complete diphallia and review of the literature
Authors: Kundal Vijay Kumar , Gajdhar Mufique , Shukla Arvind K , Kundal Raksha
Published: February 2013

A 3-year-old boy presented with complaints of abnormal genitalia. On examination, there was penile duplication with two completely separate phalluses, unequal in size, one with hypospadias and the other with meatus at the tip of the glans. Voiding cystourethrography revealed a normal sized single bladder but two urethras with no associated vesicoureteral reflux.
Severe rhabdomyolysis associated with a primary cytomegalovirus infection in an immunocompetent patient
Authors: Gindre Héloïse , Féasson Léonard , Auboyer Christian , Cathébras Pascal
Published: February 2013

Virus-induced rhabdomyolysis rarely induces respiratory failure. We discuss here a case of severe rhabdomyolysis with acute respiratory failure secondary to a cytomegalovirus (CMV) primary infection. We report a case of severe acute rhabdomyolysis, leading to respiratory failure and mechanical ventilation, associated with CMV primary infection in a young and otherwise healthy woman. We excluded other aetiologies such as metabolic myopathies, electrolyte disorders or Guillain-Barré syndrome with exhaustive researches. After 1 year, the patient recovered completely, apart from a slight muscle deconditioning. In this report, we compare our patient with five other similar cases found in the literature; our patient had the most severe presentation. The mechanism of acute viral-induced rhabdomyolysis remains elusive.
A qualitative study on clinical research in Finland: fragmented governance and volume in the 2000s
Authors: Hemminki Elina , Veerus Piret , Virtanen Jorma , Lehto Juhani
Published: February 2013

Objectives

Although concerns over clinical research have been expressed, the governance of clinical research has been little studied. The aim was to describe research policy, volume, funding and concerns over clinical research in Finland.

Design

A qualitative study and the data were collected from various sources, including documents, statistics and semistructured expert interviews.

Setting

Finland.

Results

We found no national policy for clinical research. Many actors were responsible for facilitating, directing, regulating and funding clinical research, but no actor had the main responsibility. Health professionals were the main drivers for clinical research. The role of the health ministry was small. The ministry distributed state money for clinical research in health services (EVO-money), but did not use it to direct research. Municipalities responsible for health services or national health insurance had little interest in clinical research. The Academy of Finland had had initiatives to promote clinical research, but they had not materialised in funding. Clinical research was common and internationally competitive, but its volume had declined relatively in the 2000s. Industry was an important private funder, mainly supporting drug trials made for licensing purposes. Drug trials without an outside sponsor (academic projects) declined between 2002 and 2010. The funding and its targeting and amount were no one's responsibility. Concerns over clinical research were similar as in other countries, but it had appeared late.

Conclusions

Our results suggest fragmented governance and funding in clinical research. The unsystematic research environment has not prevented clinical research from flourishing, but the public health relevance of the research carried out and its sustainability are unclear.
Strategy for treating selective serotonin reuptake inhibitor-resistant social anxiety disorder in the clinical setting: a randomised controlled trial protocol of cognitive behavioural therapy in combination with conventional treatment
Authors: Yoshinaga Naoki , Niitsu Tomihisa , Hanaoka Hideki , Sato Yasunori , Ohshima Fumiyo , Matsuki Satoshi , Kobori Osamu , Nakazato Michiko , Nakagawa Akiko , Iyo Masaomi , Shimizu Eiji
Published: February 2013

Introduction

Pharmacotherapy and cognitive behavioural therapy (CBT) are consistently effective as first-line treatments for social anxiety disorders (SADs). Nevertheless, pharmacotherapy is often the first choice in clinical practice. In many countries, the first line of pharmacotherapy involves the administration of a selective serotonin reuptake inhibitor (SSRI). Although a significant proportion of patients with SAD fail to respond to the initial SSRI administration, there is no standard approach to the management of SSRI-resistant SAD. This paper describes the study protocol for a randomised controlled trial to evaluate the clinical effectiveness of CBT as a next-step strategy, concomitant with conventional treatment, for patients with SSRI-resistant SAD.

Methods and analysis

This Prospective Randomized Open Blinded End-point study is designed with two parallel groups, with dynamic allocation at the individual level. The interventions for the two groups are conventional treatment, alone, and CBT combined with conventional treatment, for 16 weeks. The primary end-point of SAD severity will be assessed by an independent assessor using the Liebowitz Social Anxiety Scale, and secondary end-points include severity of other social anxieties, depressive severity and functional impairment. All measures will be assessed at weeks 0 (baseline), 8 (halfway point) and 16 (postintervention) and the outcomes will be analysed based on the intent-to-treat. Statistical analyses are planned for the study design stage so that field materials can be appropriately designed.

Ethics and dissemination

This study will be conducted at the academic outpatient clinic of Chiba University Hospital. Ethics approval was granted by the Institutional Review Board of Chiba University Hospital. All participants will be required to provide written informed consent. The trial will be implemented and reported in accordance with the recommendations of CONSORT.

Clinical Trial Registration Number

UMIN000007552.
The impact of smoke-free legislation on fetal, infant and child health: a systematic review and meta-analysis protocol
Authors: Been Jasper V , Nurmatov Ulugbek , van Schayck Constant P , Sheikh Aziz
Published: February 2013

Introduction

Second-hand smoke (SHS) exposure is estimated to kill 600 000 people worldwide annually. The WHO recommends that smoke-free indoor public environments are enforced through national legislation. Such regulations have been shown to reduce SHS exposure and, consequently, respiratory and cardiovascular morbidity. Evidence of particular health benefit in children is now emerging, including reductions in low birthweight deliveries, preterm birth and asthma exacerbations. We aim to comprehensively assess the impact of smoke-free legislation on fetal, infant and childhood outcomes. This can inform further development and implementation of global policy and strategies to reduce early life SHS exposure.

Methods

Two authors will search online databases (1975–present; no language restrictions) of published and unpublished/in-progress studies, and references and citations to articles of interest. We will consult experts in the field to identify additional studies. Studies should describe associations between comprehensive or partial smoking bans in public places and health outcomes among children (0–12 years): stillbirth, preterm birth, low birth weight, small for gestational age, perinatal mortality, congenital anomalies, bronchopulmonary dysplasia, upper and lower respiratory infections and wheezing disorders including asthma. The Cochrane Effectiveness Practice and Organisational Care (EPOC)-defined study designs are eligible. Study quality will be assessed using the Cochrane 7-domain-based evaluation for randomised and clinical trials, and EPOC criteria for quasiexperimental studies. Data will be extracted by two reviewers and presented in tabular and narrative form. Meta-analysis will be undertaken using random-effects models, and generic inverse variance analysis for adjusted effect estimates. We will report sensitivity analyses according to study quality and design characteristics, and subgroup analyses according to coverage of ban, age group and parental/maternal smoking status. Publication bias will be assessed.

Ethics and dissemination

Ethics assessment is not required.

Results

Will be presented in one manuscript. The protocol is registered with PROSPERO, registration number CRD42013003522.
Psychological distress mediates the association between daytime sleepiness and consumption of sweetened products: cross-sectional findings in a Catholic Middle-Eastern Canadian community
Authors: Moubarac Jean-Claude , Cargo Margaret , Receveur Olivier , Daniel Mark
Published: February 2013

Objective

To examine the associations between consumption of sweetened products, daytime sleepiness (DS) and psychological distress (PD) in a Catholic Middle-Eastern Canadian community, and to test the hypothesis that the association between DS and consumption of sweetened products is mediated by PD.

Design

A cross-sectional study.

Setting

A Catholic Middle-Eastern Canadian community.

Participants

186 men and women aged between 18 and 60 years.

Primary and secondary outcome measures

Sweetened product consumption was measured using a food frequency questionnaire (total sugars/day). DS and PD were measured using standardised questionnaires. The generalised linear model was used to estimate associations between sweetened product consumption, age, sex, self-reported body mass index, DS and PD. Baron and Kenny's four-step approach in addition to the Sobel test were used to establish mediation.

Results

Average DS score was 8.2 (SD=4.5) with 19.5% having excessive scores (>12). Mean PD score was 20.8 (SD=6.2) with 11.8% having high distress scores. Average consumption of sweetened products was 15.5 g/day (SD=13.9). Baron and Kenny's three steps to establish partial mediation were confirmed. First, DS was associated with consumption of sweetened products (p<0.03). Second, DS and PD were correlated (r=0.197; p<0.04). Third, PD was associated with consumption of sweetened products (p<0.01) when both PD and DS were entered as predictors in a multivariate regression. However, Baron and Kenny's fourth step to establish complete mediation was not met. The effect of DS on consumption of sweetened products controlling for PD was reduced, but it was not zero. Finally, the Sobel test was significant (2.14; p<0.03).

Conclusions

The association between DS and consumption of sweetened products in the Catholic Middle-Eastern Canadian community is partially mediated by psychological distress. Further work should test this mediation relationship in larger samples and verify the potential effects of other sleep variables in this relationship.