bluuebunny/crossref_metadata_2025 huggingface dataset:
Model‐based feasibility assessment and evaluation of prostate hyperthermia with a commercial MR‐guided endorectal HIFU ablation array
Authors: Salgaonkar Vasant A. , Prakash Punit , Rieke Viola , Ozhinsky Eugene , Plata Juan , Kurhanewicz John , Hsu I‐C. (Joe) , Diederich Chris J.
Published: February 2014

Purpose:

Feasibility of targeted and volumetric hyperthermia (40–45 °C) delivery to the prostate with a commercial MR‐guided endorectal ultrasound phased array system, designed specifically for thermal ablation and approved for ablation trials (ExAblate 2100, Insightec Ltd.), was assessed through computer simulations and tissue‐equivalent phantom experiments with the intention of fast clinical translation for targeted hyperthermia in conjunction with radiotherapy and chemotherapy.

Methods:

The simulations included a 3D finite element method based biothermal model, and acoustic field calculations for the ExAblate ERUS phased array (2.3 MHz, 2.3 × 4.0 cm2, ∼1000 channels) using the rectangular radiator method. Array beamforming strategies were investigated to deliver protracted, continuous‐wave hyperthermia to focal prostate cancer targets identified from representative patient cases. Constraints on power densities, sonication durations and switching speeds imposed by ExAblate hardware and software were incorporated in the models. Preliminary experiments included beamformed sonications in tissue mimicking phantoms under MR temperature monitoring at 3 T (GE Discovery MR750W).

Results:

Acoustic intensities considered during simulation were limited to ensure mild hyperthermia (Tmax < 45 °C) and fail‐safe operation of the ExAblate array (spatial and time averaged acoustic intensity ISATA < 3.4 W/cm2). Tissue volumes with therapeutic temperature levels (T > 41 °C) were estimated. Numerical simulations indicated that T > 41 °C was calculated in 13–23 cm3 volumes for sonications with planar or diverging beam patterns at 0.9–1.2 W/cm2, in 4.5–5.8 cm3 volumes for simultaneous multipoint focus beam patterns at ∼0.7 W/cm2, and in ∼6.0 cm3 for curvilinear (cylindrical) beam patterns at 0.75 W/cm2. Focused heating patterns may be practical for treating focal disease in a single posterior quadrant of the prostate and diffused heating patterns may be useful for heating quadrants, hemigland volumes or even bilateral targets. Treatable volumes may be limited by pubic bone heating. Therapeutic temperatures were estimated for a range of physiological parameters, sonication duty cycles and rectal cooling. Hyperthermia specific phasing patterns were implemented on the ExAblate prostate array and continuous‐wave sonications (∼0.88 W/cm2, 15 min) were performed in tissue‐mimicking material with real‐time MR‐based temperature imaging (PRFS imaging at 3.0 T). Shapes of heating patterns observed during experiments were consistent with simulations.

Conclusions:

The ExAblate 2100, designed specifically for thermal ablation, can be controlled for delivering continuous hyperthermia in prostate while working within operational constraints.
Refractive index measurement by prism autocollimation
Authors: Cheng Chao-Chia
Published: February 2014

An autocollimation-based method for measuring the refractive indices of solid or liquids using a Littrow prism is presented. Measurement accuracy is enhanced by use of a telescope. In solids, the refractive index is accurate to three decimal places. Similar accuracy is obtained in liquids by correcting for the wedge angle of the liquid container window. The proposed prismatic method confers high accuracy, compactness, and automation. It is suitable for index measurement applications in undergraduate laboratories.
Thermodynamics of bread baking: A two-state model
Authors: Zürcher Ulrich
Published: February 2014

Bread baking can be viewed as a complex physico-chemical process. It is governed by transport of heat and is accompanied by changes such as gelation of starch, the expansion of air cells within dough, and others. We focus on the thermodynamics of baking and investigate the heat flow through dough and find that the evaporation of excess water in dough is the rate-limiting step. We consider a simplified one-dimensional model of bread, treating the excess water content as a two-state variable that is zero for baked bread and a fixed constant for unbaked dough. We arrive at a system of coupled, nonlinear ordinary differential equations, which are solved using a standard Runge-Kutta integration method. The calculated baking times are consistent with common baking experience.
Resource Letter NTUC-1: Noether's Theorem in the Undergraduate Curriculum
Authors: Neuenschwander Dwight E.
Published: February 2014

This Resource Letter provides a guide to the literature, suitable for use in the undergraduate physics curriculum, on Noether's Theorem, which relates conservation laws to symmetries.
A Direct Quantitative Measure of Surface Mobility in a Glassy Polymer
Authors: Chai Y. , Salez T. , McGraw J. D. , Benzaquen M. , Dalnoki-Veress K. , Raphaël E. , Forrest J. A.
Published: February 2014

Polymer Film Behavior

An ongoing debate in the understanding of the behavior of thin-film glassy polymers is whether there is nanoconfinement of large molecules or enhanced mobility near a free surface.

Chai
et al.

(p.
994
; see the Perspective by

Chen
et al.

) prepared polymer films with a sharp step in the profile by depositing broken film fragments onto a uniform underlay. Atomic force microscopy revealed changes to the overall film profile with time at various temperatures. A transition was observed from localized motions to relaxation of the entire film at a temperature close to that of the bulk glass transition temperature.
A Better Route to Tech Standards
Authors: Lerner Josh , Tirole Jean
Published: February 2014

Prestandard price commitments, as well as government intervention, may help patent licensing and standard setting.
The Surface Mobility of Glasses
Authors: Chen Fei , Lam Chi-Hang , Tsui Ophelia K. C.
Published: February 2014

Surface diffusion on frozen polymer glasses is influenced by the surface dynamics of the glass itself.

[Also see Report by

Chai
et al.

]
Out of Beringia?
Authors: Hoffecker John F. , Elias Scott A. , O'Rourke Dennis H.
Published: February 2014

A shrub tundra refugium on the Bering land bridge may have played a pivotal role in the peopling of the Americas.
How Shape Affects Microtubule and Nanoparticle Assembly
Authors: Stevens Mark J.
Published: February 2014

Models of nanoparticle assembly can help explain aspects of the inverted structure that forms because of induced change in tubulin protein shape.
Welcome to Beringia
Authors: Pringle Heather
Published: February 2014

A flurry of studies suggests that instead of being simply a bridge from Asia to the Americas, Beringia may have beckoned the ancestors of the first Americans to linger.
Role of Antibodies in Protection Elicited by Active Vaccination with Genetically Inactivated Alpha Hemolysin in a Mouse Model of Staphylococcus aureus Skin and Soft Tissue Infections
Authors: Mocca Christopher P. , Brady Rebecca A. , Burns Drusilla L.
Published: February 2014

Due to the emergence of highly virulent community-associated methicillin-resistantStaphylococcus aureus(CA-MRSA) infections,S. aureushas become a major threat to public health. A majority of CA-MRSA skin and soft tissue infections in the United States are caused byS. aureusUSA300 strains that are known to produce high levels of alpha hemolysin (Hla). Therefore, vaccines that contain inactivated forms of this toxin are currently being developed. In this study, we sought to determine the immune mechanisms of protection for this antigen using a vaccine composed of a genetically inactivated form of Hla (HlaH35L). Using a murine model of skin and soft tissue infections (SSTI), we found that BALB/c mice were protected by vaccination with HlaH35L; however, Jh mice, which are deficient in mature B lymphocytes and lack IgM and IgG in their serum, were not protected. Passive immunization with anti-HlaH35L antibodies conferred protection against bacterial colonization. Moreover, we found a positive correlation between the total antibody concentration induced by active vaccination and reduced bacterial levels. Animals that developed detectable neutralizing antibody titers after active vaccination were significantly protected from infection. These data demonstrate that antibodies to Hla represent the major mechanism of protection afforded by active vaccination with inactivated Hla in this murine model of SSTI, and in this disease model, antibody levels correlate with protection. These results provide important information for the future development and evaluation ofS. aureusvaccines.
Development of Neutralizing Monoclonal Antibodies for Oncogenic Human Papillomavirus Types 31, 33, 45, 52, and 58
Authors: Brown Martha J. , Seitz Hanna , Towne Victoria , Müller Martin , Finnefrock Adam C.
Published: February 2014

Human papillomavirus (HPV) is the etiological agent for all cervical cancers, a significant number of other anogenital cancers, and a growing number of head and neck cancers. Two licensed vaccines offer protection against the most prevalent oncogenic types, 16 and 18, responsible for approximately 70% of cervical cancer cases worldwide and one of these also offers protection against types 6 and 11, responsible for 90% of genital warts. The vaccines are comprised of recombinantly expressed major capsid proteins that self-assemble into virus-like particles (VLPs) and prevent infection by eliciting neutralizing antibodies. Adding the other frequently identified oncogenic types 31, 33, 45, 52, and 58 to a vaccine would increase the coverage against HPV-induced cancers to approximately 90%. We describe the generation and characterization of panels of monoclonal antibodies to these five additional oncogenic HPV types, and the selection of antibody pairs that were high affinity and type specific and recognized conformation-dependent neutralizing epitopes. Such characteristics make these antibodies useful tools for monitoring the production and potency of a prototype vaccine as well as monitoring vaccine-induced immune responses in the clinic.
Genetic Diversity and Evolution of Salmonella enterica Serovar Enteritidis Strains with Different Phage Types
Authors: Zheng Jie , Pettengill James , Strain Errol , Allard Marc W. , Ahmed Rafiq , Zhao Shaohua , Brown Eric W.
Published: February 2014

Phage typing has been used for the epidemiological surveillance of
Salmonella enterica
serovar Enteritidis for over 2 decades. However, knowledge of the genetic and evolutionary relationships between phage types is very limited, making differences difficult to interpret. Here, single nucleotide polymorphisms (SNPs) identified from whole-genome comparisons were used to determine the relationships between some
S
. Enteritidis phage types (PTs) commonly associated with food-borne outbreaks in the United States. Emphasis was placed on the predominant phage types PT8, PT13a, and PT13 in North America. With >89,400 bp surveyed across 98
S
. Enteritidis isolates representing 14 distinct phage types, 55 informative SNPs were discovered within 23 chromosomally anchored loci. To maximize the discriminatory and evolutionary partitioning of these highly homogeneous strains, sequences comprising informative SNPs were concatenated into a single combined data matrix and subjected to phylogenetic analysis. The resultant phylogeny allocated most
S
. Enteritidis isolates into two distinct clades (clades I and II) and four subclades. Synapomorphic (shared and derived) sets of SNPs capable of distinguishing individual clades/subclades were identified. However, individual phage types appeared to be evolutionarily disjunct when mapped to this phylogeny, suggesting that phage typing may not be valid for making phylogenetic inferences. Furthermore, the set of SNPs identified here represents useful genetic markers for strain differentiation of more clonal
S
. Enteritidis strains and provides core genotypic markers for future development of a SNP typing scheme with
S
. Enteritidis.
Rolling Circle Amplification for Direct Detection of
rpoB
Gene Mutations in Mycobacterium tuberculosis Isolates from Clinical Specimens
Authors: Chen Xiaoyou , Wang Bin , Yang Wen , Kong Fanrong , Li Chuanyou , Sun Zhaogang , Jelfs Peter , Gilbert Gwendolyn L.
Published: February 2014

Rapid and accurate detection of multidrug resistance (MDR) in
Mycobacterium tuberculosis
is essential to improve treatment outcomes and reduce global transmission but remains a challenge. Rifampin (RIF) resistance is a reliable marker of MDR tuberculosis (TB) since by far the majority of RIF-resistant strains are also isoniazid (INH) resistant. We have developed a rapid, sensitive, and specific method for detecting the most common mutations associated with RIF resistance, in the RIF resistance determining region (RRDR) of
rpoB
, using a cocktail of six padlock probes and rolling circle amplification (RCA). We used this method to test 46 stored
M. tuberculosis
clinical isolates with known RIF susceptibility profiles (18 RIF resistant, 28 susceptible), a standard susceptible strain (H37Rv, ATCC 27294) and 78
M. tuberculosis
culture-positive clinical (sputum) samples, 59 of which grew RIF-resistant strains. All stored clinical isolates were correctly categorized, by the padlock probe/RCA method, as RIF susceptible or resistant; the sensitivity and specificity of the method, for direct detection of phenotypically RIF-resistant
M. tuberculosis
in clinical specimens, were 96.6 and 89.5%, respectively. This method is rapid, simple, and inexpensive and has the potential for high-throughput routine screening of clinical specimens for MDR
M. tuberculosis
, particularly in high prevalence settings with limited resources.
Performance of Panfungal- and Specific-PCR-Based Procedures for Etiological Diagnosis of Invasive Fungal Diseases on Tissue Biopsy Specimens with Proven Infection: a 7-Year Retrospective Analysis from a Reference Laboratory
Authors: Buitrago M. J. , Bernal-Martinez L. , Castelli M. V. , Rodriguez-Tudela J. L. , Cuenca-Estrella M.
Published: February 2014

A retrospective analysis of real-time PCR (RT-PCR) results for 151 biopsy samples obtained from 132 patients with proven invasive fungal diseases was performed. PCR-based techniques proved to be fast and sensitive and enabled definitive diagnosis in all cases studied, with detection of a total of 28 fungal species.
Modified CLSI Extended-Spectrum β-Lactamase (ESBL) Confirmatory Test for Phenotypic Detection of ESBLs among Enterobacteriaceae Producing Various β-Lactamases
Authors: Poulou Aggeliki , Grivakou Evgenia , Vrioni Georgia , Koumaki Vassiliki , Pittaras Theodoros , Pournaras Spyros , Tsakris Athanassios
Published: February 2014

The worldwide dissemination of
Enterobacteriaceae
producing AmpC β-lactamases and carbapenemases makes difficult the phenotypic detection of extended-spectrum β-lactamases (ESBLs), as they may be masked by these additional enzymes. A modification of the CLSI ESBL confirmatory test was developed and evaluated in a comparative study for its ability to successfully detect ESBLs among
Enterobacteriaceae
producing various carbapenemases (
Klebsiella pneumoniae
carbapenemase [KPC], VIM, NDM, and OXA-48) and plasmidic or derepressed AmpCs. The modified CLSI ESBL confirmatory test was performed with cefotaxime and ceftazidime disks with and without clavulanate, on which both boronic acid (BA) and EDTA were dispensed. A total of 162 genotypically confirmed ESBL-positive
Enterobacteriaceae
isolates (83 carbapenemase/ESBL producers, 25 AmpC/ESBL producers, and 54 ESBL-only producers) were examined. For comparison, 139 genotypically confirmed ESBL-negative
Enterobacteriaceae
isolates (94 of them possessed carbapenemases and 20 possessed AmpCs) were also tested. The standard CLSI ESBL confirmatory test was positive for 106 of the 162 ESBL producers (sensitivity, 65.4%) and showed false-positive results for 4 of the 139 non-ESBL producers (specificity, 97.1%). The modified CLSI ESBL confirmatory test detected 158 of 162 ESBL producers (sensitivity, 97.5%) and showed no false-positive results for non-ESBL producers (specificity, 100%). The findings of the study demonstrate that the modified CLSI ESBL confirmatory test using antibiotic disks containing both BA and EDTA accurately detects ESBLs in
Enterobacteriaceae
regardless of the coexistence of additional β-lactam resistance mechanisms.
Molecular Characterizations of PCR-Positive Mycoplasma pneumoniae Specimens Collected from Australia and China
Authors: Xue Guanhua , Wang Qinning , Yan Chao , Jeoffreys Neisha , Wang Liqiong , Li Shaoli , Gilbert Gwendolyn L. , Sun Hongmei
Published: February 2014

Mycoplasma pneumoniae
is an important cause of community-acquired pneumonia (CAP). In this study,
M. pneumoniae
strains in PCR-positive specimens collected from patients in Sydney, Australia (30 samples), and Beijing, China (83 samples), were characterized using multilocus variable-number tandem-repeat (VNTR) analysis (MLVA), P1-restriction fragment length polymorphism (RFLP) analysis, and sequencing of domain V of the 23S rRNA gene to compare genotype distribution and macrolide resistance rates between locations. Eighteen distinct MLVA types were identified in specimens from Sydney, of which 10 were known (types E, G, J, M, N, P, U, V, S, and X) and 8 previously unknown. Strains were equally distributed between P1-RFLP type 1 and type 2 variants. Among samples from Beijing, MLVA types E, G, J, P, U, X, and Z and four new types were identified. Most specimens belonged to P1-RFLP type 1. A nomenclature based on five VNTR loci is proposed to designate MLVA patterns. Macrolide resistance-associated mutations were identified in only 1 of 30 specimens (3.3%) from Sydney and 71 of 83 (85.5%) from Beijing (
P
< 0.05). This study demonstrated that although multiple individual
M. pneumoniae
strains were circulating in Beijing, the genotypes were less diverse than those in Sydney. However, the greatest regional difference was in the incidence of macrolide resistance, which may reflect differences in antibiotic use and/or measures in resistance control.
Development of a Two-Step Multiplex PCR Assay for Typing of Capsular Polysaccharide Synthesis Gene Clusters of Streptococcus suis
Authors: Okura Masatoshi , Lachance Claude , Osaki Makoto , Sekizaki Tsutomu , Maruyama Fumito , Nozawa Takashi , Nakagawa Ichiro , Hamada Shigeyuki , Rossignol Céline , Gottschalk Marcelo , Takamatsu Daisuke
Published: February 2014

We developed a practical and easy two-step multiplex PCR assay to aid in serotyping of
Streptococcus suis
. The assay accurately typed almost all of the serotype reference strains and field isolates of various serotypes and also identified the genotypes of capsular polysaccharide synthesis gene clusters of some serologically nontypeable strains.
Real-Time Whole-Genome Sequencing for Routine Typing, Surveillance, and Outbreak Detection of Verotoxigenic Escherichia coli
Authors: Joensen Katrine Grimstrup , Scheutz Flemming , Lund Ole , Hasman Henrik , Kaas Rolf S. , Nielsen Eva M. , Aarestrup Frank M.
Published: February 2014

Fast and accurate identification and typing of pathogens are essential for effective surveillance and outbreak detection. The current routine procedure is based on a variety of techniques, making the procedure laborious, time-consuming, and expensive. With whole-genome sequencing (WGS) becoming cheaper, it has huge potential in both diagnostics and routine surveillance. The aim of this study was to perform a real-time evaluation of WGS for routine typing and surveillance of verocytotoxin-producing
Escherichia coli
(VTEC). In Denmark, the Statens Serum Institut (SSI) routinely receives all suspected VTEC isolates. During a 7-week period in the fall of 2012, all incoming isolates were concurrently subjected to WGS using IonTorrent PGM. Real-time bioinformatics analysis was performed using web-tools (
www.genomicepidemiology.org
) for species determination, multilocus sequence type (MLST) typing, and determination of phylogenetic relationship, and a specific VirulenceFinder for detection of
E. coli
virulence genes was developed as part of this study. In total, 46 suspected VTEC isolates were characterized in parallel during the study. VirulenceFinder proved successful in detecting virulence genes included in routine typing, explicitly verocytotoxin 1 (
vtx1
), verocytotoxin 2 (
vtx2
), and intimin (
eae
), and also detected additional virulence genes. VirulenceFinder is also a robust method for assigning verocytotoxin (
vtx
) subtypes. A real-time clustering of isolates in agreement with the epidemiology was established from WGS, enabling discrimination between sporadic and outbreak isolates. Overall, WGS typing produced results faster and at a lower cost than the current routine. Therefore, WGS typing is a superior alternative to conventional typing strategies. This approach may also be applied to typing and surveillance of other pathogens.
Genome Scale Patterns of Recombination between Coinfecting Vaccinia Viruses
Authors: Qin Li , Evans David H.
Published: February 2014

Recombination plays a critical role in virus evolution. It helps avoid genetic decline and creates novel phenotypes. This promotes survival, and genome sequencing suggests that recombination has facilitated the evolution of human pathogens, including orthopoxviruses such as variola virus. Recombination can also be used to map genes, but although recombinant poxviruses are easily produced in culture, classical attempts to map the vaccinia virus (VACV) genome this way met with little success. We have sequenced recombinants formed when VACV strains TianTan and Dryvax are crossed under different conditions. These were a single round of growth in coinfected cells, five rounds of sequential passage, or recombinants obtained using leporipoxvirus-mediated DNA reactivation. Our studies showed that recombinants contain a patchwork of DNA, with the number of exchanges increasing with passage. Further passage also selected for TianTan DNA and correlated with increased plaque size. The recombinants produced through a single round of coinfection contain a disproportionate number of short conversion tracks (<1 kbp) and exhibited 1 exchange per 12 kbp, close to the ∼1 per 8 kbp in the literature. One by-product of this study was that rare mutations were also detected; VACV replication produces ∼1 × 10−8mutation per nucleotide copied per cycle of replication and ∼1 large (21 kbp) deletion per 70 rounds of passage. Viruses produced using DNA reactivation appeared no different from recombinants produced using ordinary methods. An attractive feature of this approach is that when it is combined with selection for a particular phenotype, it provides a way of mapping and dissecting more complex virus traits.

IMPORTANCEWhen two closely related viruses coinfect the same cell, they can swap genetic information through a process called recombination. Recombination produces new viruses bearing different combinations of genes, and it plays an important role in virus evolution. Poxviruses are a family of viruses that includes variola (or smallpox) virus, and although poxviruses are known to recombine, no one has previously mapped the patterns of DNAs exchanged between viruses. We coinfected cells with two different vaccinia poxviruses, isolated the progeny, and sequenced them. We show that poxvirus recombination is a very accurate process that assembles viruses containing DNA copied from both parents. In a single round of infection, DNA is swapped back and forth ∼18 times per genome to make recombinant viruses that are a mosaic of the two parental DNAs. This mixes many different genes in complex combinations and illustrates how recombination can produce viruses with greatly altered disease potential.
In VivoAblation of Type I Interferon Receptor from Cardiomyocytes Delays Coxsackieviral Clearance and Accelerates Myocardial Disease
Authors: Althof Nadine , Harkins Stephanie , Kemball Christopher C. , Flynn Claudia T. , Alirezaei Mehrdad , Whitton J. Lindsay
Published: February 2014

Acute coxsackievirus B3 (CVB3) infection is one of the most prevalent causes of acute myocarditis, a disease that frequently is identified only after the sudden death of apparently healthy individuals. CVB3 infects cardiomyocytes, but the infection is highly focal, even in the absence of a strong adaptive immune response, suggesting that virus spread within the heart may be tightly constrained by the innate immune system. Type I interferons (T1IFNs) are an obvious candidate, and T1IFN receptor (T1IFNR) knockout mice are highly susceptible to CVB3 infection, succumbing within a few days of challenge. Here, we investigated the role of T1IFNs in the heart using a mouse model in which the T1IFNR gene can be ablatedin vivo, specifically in cardiomyocytes. We found that T1IFN signaling into cardiomyocytes contributed substantially to the suppression of viral replication and infectious virus yield in the heart; in the absence of such signaling, virus titers were markedly elevated by day 3 postinfection (p.i.) and remained high at day 12 p.i., a time point at which virus was absent from genetically intact littermates, suggesting that the T1IFN-unresponsive cardiomyocytes may act as a safe haven for the virus. Nevertheless, in these mice the myocardial infection remained highly focal, despite the cardiomyocytes' inability to respond to T1IFN, indicating that other factors, as yet unidentified, are sufficient to prevent the more widespread dissemination of the infection throughout the heart. The absence of T1IFN signaling into cardiomyocytes also was accompanied by a profound acceleration and exacerbation of myocarditis and by a significant increase in mortality.

IMPORTANCEAcute coxsackievirus B3 (CVB3) infection is one of the most common causes of acute myocarditis, a serious and sometimes fatal disease. To optimize treatment, it is vital that we identify the immune factors that limit virus spread in the heart and other organs. Type I interferons play a key role in controlling many virus infections, but it has been suggested that they may not directly impact CVB3 infection within the heart. Here, using a novel line of transgenic mice, we show that these cytokines signal directly into cardiomyocytes, limiting viral replication, myocarditis, and death.
PrP Expression Level and Sensitivity to Prion Infection
Authors: Douet Jean-Yves , Lacroux Caroline , Corbière Fabien , Litaise Claire , Simmons Hugh , Lugan Séverine , Costes Pierrette , Cassard Hervé , Weisbecker Jean-Louis , Schelcher François , Andreoletti Olivier
Published: February 2014

Mice overexpressing the prion protein (PrP) sequence from various host species are widely used for measuring infectious titers in prion disease. However, the impact that the transgene expression level might have on the susceptibility to infection raises some concerns about the final biological relevance of these models. Here we report that endpoint titration of a sheep scrapie isolate in sheep and in mice overexpressing the ovine PrP results in similar estimates of the infectious titer.
Development of an Adenovirus-Based Respiratory Syncytial Virus Vaccine: Preclinical Evaluation of Efficacy, Immunogenicity, and Enhanced Disease in a Cotton Rat Model
Authors: Kim Eun , Okada Kaori , Beeler Judy A. , Crim Roberta L. , Piedra Pedro A. , Gilbert Brian E. , Gambotto Andrea
Published: February 2014

The lack of a vaccine against respiratory syncytial virus (RSV) is a challenging and serious gap in preventive medicine. Herein, we characterize the immunogenicity of an adenovirus serotype 5-based RSV vaccine encoding the fusion (F) protein (Ad5.RSV-F) and the protection provided following immunization with Ad5.RSV-F and assess its potential for producing enhanced disease in a cotton rat (CR) model. Animals were immunized intranasally (i.n.) and/or intramuscularly (i.m.) and subsequently challenged with RSV/A/Tracy (i.n.) to assess protection. Robust immune responses were seen in CRs vaccinated with Ad5.RSV-F given i.m. or i.n., and these responses correlated with reduced replication of the virus in noses and lungs after challenge. Neutralizing antibody responses following immunization with a single dose of Ad5.RSV-F at 1 × 10
11
viral particles (v.p.) elicited antibody titers 64- to 256-fold greater than those seen after natural infection. CRs boosted with Ad5.RSV-F i.n. 28 days after an i.m. dose also had significant increases in neutralizing antibody titers. Antibody affinity for different F-protein antigenic sites revealed substantial differences between antibodies elicited by Ad5.RSV-F and those seen after RSV infection; differences in antibody profiles were also seen between CRs given Ad5.RSV-F i.m. and CRs given Ad5.RSV-F i.n. Ad5.RSV-F priming did not result in enhanced disease following live-virus challenge, in contrast to the histopathology seen in CRs given the formalin-inactivated RSV/A/Burnett vaccine.

IMPORTANCE
Respiratory syncytial virus (RSV) is the most common cause of acute lower respiratory infection in infants and young children and a serious health threat in the immunocompromised and the elderly. Infection severity increased in children in an immunization trial, hampering the over 4-decade-long quest for a successful RSV vaccine. In this study, we show that a genetically engineered RSV-F-encoding adenoviral vector provides protective immunity against RSV challenge without enhanced lung disease in cotton rats (CRs). CRs were vaccinated under a number of different regimens, and the immunity induced by the recombinant adenoviral RSV vaccine administered by use of an intramuscular prime-intranasal boost regimen may provide the best protection for young infants and children at risk of RSV infection, since this population is naive to adenoviral preformed immunity. Overall, this report describes a potential RSV vaccine candidate that merits further evaluation in a phase I clinical study in humans.
Upolu virus and Aransas Bay virus, Two Presumptive Bunyaviruses, Are Novel Members of the Family Orthomyxoviridae
Authors: Briese Thomas , Chowdhary Rashmi , Travassos da Rosa Amelia , Hutchison Stephen K. , Popov Vsevolod , Street Craig , Tesh Robert B. , Lipkin W. Ian
Published: February 2014

Emerging and zoonotic pathogens pose continuing threats to human health and ongoing challenges to diagnostics. As nucleic acid tests are playing increasingly prominent roles in diagnostics, the genetic characterization of molecularly uncharacterized agents is expected to significantly enhance detection and surveillance capabilities. We report the identification of two previously unrecognized members of the family
Orthomyxoviridae
, which includes the influenza viruses and the tick-transmitted Thogoto and Dhori viruses. We provide morphological, serologic, and genetic evidence that
Upolu virus
(UPOV) from Australia and
Aransas Bay virus
(ABV) from North America, both previously considered potential bunyaviruses based on electron microscopy and physicochemical features, are orthomyxoviruses instead. Their genomes show up to 68% nucleotide sequence identity to Thogoto virus (segment 2; ∼74% at the amino acid level) and a more distant relationship to Dhori virus, the two prototype viruses of the recognized species of the genus
Thogotovirus
. Despite sequence similarity, the coding potentials of UPOV and ABV differed from that of Thogoto virus, instead being like that of Dhori virus. Our findings suggest that the tick-transmitted viruses UPOV and ABV represent geographically distinct viruses in the genus
Thogotovirus
of the family
Orthomyxoviridae
that do not fit in the two currently recognized species of this genus.

IMPORTANCE
Upolu virus
(UPOV) and
Aransas Bay virus
(ABV) are shown to be orthomyxoviruses instead of bunyaviruses, as previously thought. Genetic characterization and adequate classification of agents are paramount in this molecular age to devise appropriate surveillance and diagnostics. Although more closely related to Thogoto virus by sequence, UPOV and ABV differ in their coding potentials by lacking a proposed pathogenicity factor. In this respect, they are similar to Dhori virus, which, despite the lack of a pathogenicity factor, can cause disease. These findings enable further studies into the evolution and pathogenicity of orthomyxoviruses.
Anti-Lyssaviral Activity of Interferons κ and ω from the Serotine Bat, Eptesicus serotinus
Authors: He Xiaocui , Korytář Tomáš , Schatz Juliane , Freuling Conrad M. , Müller Thomas , Köllner Bernd
Published: February 2014

Interferons (IFNs) are cytokines produced by host cells in response to the infection with pathogens. By binding to the corresponding receptors, IFNs trigger different pathways to block intracellular replication and growth of pathogens and to impede the infection of surrounding cells. Due to their key role in host defense against viral infections, as well as for clinical therapies, the IFN responses and regulation mechanisms are well studied. However, studies of type I IFNs have mainly focused on alpha interferon (IFN-α) and IFN-β subtypes. Knowledge of IFN-κ and IFN-ω is limited. Moreover, most studies are performed in humans or mouse models but not in the original host of zoonotic pathogens. Bats are important reservoirs and transmitters of zoonotic viruses such as lyssaviruses. A few studies have shown an antiviral activity of IFNs in fruit bats. However, the function of type I IFNs against lyssaviruses in bats has not been studied yet. Here, IFN-κ and IFN-ω genes from the European serotine bat,
Eptesicus serotinus
, were cloned and functionally characterized.
E. serotinus
IFN-κ and IFN-ω genes are intronless and well conserved between microchiropteran species. The promoter regions of both genes contain essential regulatory elements for transcription factors.
In vitro
studies indicated a strong activation of IFN signaling by recombinant IFN-ω, whereas IFN-κ displayed weaker activation. Noticeably, both IFNs inhibit to different extents the replication of different lyssaviruses in susceptible bat cell lines. The present study provides functional data on the innate host defense against lyssaviruses in endangered European bats.

IMPORTANCE
We describe here for the first time the molecular and functional characterization of two type I interferons (IFN-κ and -ω) from European serotine bat (
Eptesicus serotinus
). The importance of this study is mainly based on the fact that very limited information about the early innate immune response against bat lyssaviruses in their natural host serotine bats is yet available. Generally, whereas the antiviral activity of other type I interferons is well studied, the functional involvement of IFN-κ and -ω has not yet been investigated.
Human Herpesvirus 6A Infection in CD46 Transgenic Mice: Viral Persistence in the Brain and Increased Production of Proinflammatory Chemokines via Toll-Like Receptor 9
Authors: Reynaud Joséphine M. , Jégou Jean-François , Welsch Jérémy C. , Horvat Branka
Published: February 2014

Human herpesvirus 6 (HHV-6) is widely spread in the human population and has been associated with several neuroinflammatory diseases, including multiple sclerosis. To develop a small-animal model of HHV-6 infection, we analyzed the susceptibility of several lines of transgenic mice expressing human CD46, identified as a receptor for HHV-6. We showed that HHV-6A (GS) infection results in the expression of viral transcripts in primary brain glial cultures from CD46-expressing mice, while HHV-6B (Z29) infection was inefficient. HHV-6A DNA persisted for up to 9 months in the brain of CD46-expressing mice but not in the nontransgenic littermates, whereas HHV-6B DNA levels decreased rapidly after infection in all mice. Persistence in the brain was observed with infectious but not heat-inactivated HHV-6A. Immunohistological studies revealed the presence of infiltrating lymphocytes in periventricular areas of the brain of HHV-6A-infected mice. Furthermore, HHV-6A stimulated the production of a panel of proinflammatory chemokines in primary brain glial cultures, including CCL2, CCL5, and CXCL10, and induced the expression of CCL5 in the brains of HHV-6A-infected mice. HHV-6A-induced production of chemokines in the primary glial cultures was dependent on the stimulation of toll-like receptor 9 (TLR9). Finally, HHV-6A induced signaling through human TLR9 as well, extending observations from the murine model to human infection. Altogether, this study presents a first murine model for HHV-6A-induced brain infection and suggests a role for TLR9 in the HHV-6A-initiated production of proinflammatory chemokines in the brain, opening novel perspectives for the study of virus-associated neuropathology.

IMPORTANCE
HHV-6 infection has been related to neuroinflammatory diseases; however, the lack of a suitable small-animal infection model has considerably hampered further studies of HHV-6-induced neuropathogenesis. In this study, we have characterized a new model for HHV-6 infection in mice expressing the human CD46 protein. Infection of CD46 transgenic mice with HHV-6A resulted in long-term persistence of viral DNA in the brains of infected animals and was followed by lymphocyte infiltration and upregulation of the CCL5 chemokine in the absence of clinical signs of disease. The secretion of a panel of chemokines was increased after infection in primary murine brain glial cultures, and the HHV-6-induced chemokine expression was inhibited when TLR9 signaling was blocked. These results describe the first murine model for HHV-6A-induced brain infection and suggest the importance of the TLR9 pathway in HHV-6A-initiated neuroinflammation.
Functional Analysis of Nuclear Localization Signals in VP1-2 Homologues from All Herpesvirus Subfamilies
Authors: Hennig T. , Abaitua F. , O'Hare P.
Published: February 2014

The herpes simplex virus (HSV) tegument protein VP1-2 contains an N-terminal nuclear localization signal (NLS) that is critical for capsid routing to the nuclear pore. Here we analyzed positionally conserved determinants in VP1-2 homologues from each of the alpha, beta, and gamma classes of human herpesviruses. The overall architectures of the VP1-2s were similar, with a conserved N-terminal ubiquitin-specific protease domain separated from an internal region by a linker that was quite poorly conserved in length and sequence. Within this linker region all herpesviruses contained a conserved, highly basic motif which nevertheless exhibited distinct class-specific features. The motif in HSV functioned as a monopartite NLS, while in varicella-zoster virus (VZV) activity required an adjacent basic section defining the motif as a bipartite NLS. Neither the beta- nor gammaherpesvirus VP1-2 motifs were identified by prediction algorithms, but they nevertheless functioned as efficient NLS motifs both in heterologous transfer assays and in HSV VP1-2. Furthermore, though with different efficiencies and with the exception of human herpesvirus 8 (HHV-8), these chimeric variants rescued the replication defect of an HSV mutant lacking its NLS motif. We demonstrate that the lysine at position 428 of HSV is critical for replication, with a single alanine substitution being sufficient to abrogate NLS function and virus growth. We conclude that the basic motifs of each of the VP1-2 proteins are likely to confer a similar function in capsid entry in the homologous setting and that while there is flexibility in the exact type of motif employed, specific individual residues are critical for function.

IMPORTANCE
To successfully infect cells, all herpesviruses, along with many other viruses, e.g., HIV, hepatitis B virus, and influenza virus, must navigate through the cytoplasmic environment and dock with nuclear pores for transport of their genomes into the nucleus. However, we still have a limited understanding of the detailed mechanisms involved. Insight into these events is needed and could offer opportunities for therapeutic intervention. This work investigated the role of a specific determinant in the structural protein VP1-2 in herpesvirus entry. We examined this determinant in representative VP1-2s from all herpesvirus subfamilies, demonstrated NLS function, dissected key residues, and showed functional relevance in rescuing replication of the mutant blocked in capsid navigation to the pore. The results are important and strongly support our conclusions of the generality that these motifs are crucial for entry of all herpesviruses. They also facilitate future analysis on selective host interactions and possible routes to disrupt function.
Polyangiitis overlap syndrome of granulomatosis with polyangiitis (Wegener's granulomatosis) and eosinophilic granulomatosis with polyangiitis (Churg-Strauss syndrome)
Authors: Uematsu Hironori , Takata Shohei , Sueishi Katsuo , Inoue Hiromasa
Published: February 2014

Polyangiitis overlap syndrome is defined as systemic vasculitis that cannot be classified into one of the well-defined vasculitic syndromes. In this report, a female patient who presented with vasculitis-like and asthmatic symptoms was diagnosed as having polyangiitis overlap syndrome of granulomatosis with polyangiitis (GPA; formerly known as Wegener's granulomatosis) and eosinophilic granulomatosis with polyangiitis (EGPA; formerly known as Churg-Strauss syndrome). The patient fulfilled the American College of Rheumatology diagnostic criteria for GPA and EGPA. She was successfully treated with immunosuppressants and steroids and has been in remission for 20 months. It is important to establish a proper diagnosis and introduce an appropriate treatment modality in patients with this rare and serious pathology to prevent irreversible organ damage.
Maxillary molar distalization with first class appliance
Authors: Ramesh Namitha , Palukunnu Biswas , Ravindran Nidhi , Nair Preeti P
Published: February 2014

Non-extraction treatment has gained popularity for corrections of mild-to-moderate class II malocclusion over the past few decades. The distalization of maxillary molars is of significant value for treatment of cases with minimal arch discrepancy and mild class II molar relation associated with a normal mandibular arch and acceptable profile. This paper describes our experience with a 16-year-old female patient who reported with irregularly placed upper front teeth and unpleasant smile. The patient was diagnosed to have angles class II malocclusion with moderate maxillary anterior crowding, deep bite of 4 mm on a skeletal class II base with an orthognathic maxilla and retrognathic mandible and normal growth pattern. She presented an ideal profile and so molar distalization was planned with the first-class appliance. Molars were distalised by 8 mm on the right and left quadrants and class I molar relation achieved within 4 months. The space gained was utilised effectively to align the arch and establish a class I molar and canine relation.
Huge trichobezoar causing obstructive jaundice
Authors: Verma Ankit , Sharma Sourabh , Tyagi Gaurav , Singh Sugandha
Published: February 2014

Trichobezoars are composed of hair or hair-like fibres that are commonly observed in the gastrointestinal tract of children and young women with psychiatric illnesses. Presentation of trichobezoars can be diverse. We report a case of a 29-year-old woman who presented with obstructive jaundice and a large upper abdominal lump. She was diagnosed with post-traumatic stress disorder on psychiatric evaluation. She underwent exploratory laparotomy and a large trichobezoar was removed. Her postoperative course was uneventful and she was referred to the psychiatric department for further management.
Insulinoma: rare yet important
Authors: Al-Saigh Taha Hasan
Published: February 2014

We report our experience with six cases of insulinoma that were managed at our hospital between 2009 and 2013. Four patients were women and two men, with mean age of 43 years and mean duration of symptoms before diagnosis of 5.7 years. Biochemical confirmation was made in all cases and preoperative localisation was successful in two cases only. In the other four, intraoperative localisation was made. Enucleation was performed in four patients and distal pancreatectomy in the other two. The tumour was proved benign in all patients by histopathology. Postoperatively, all patients were asymptomatic during a follow-up period ranging from 9 months to 4 years and both serum glucose and insulin were normal during that period. We think that careful preoperative workup and skilful surgical techniques can end the prolonged suffering of patients with insulinomas with excellent results.
Posterior reversible encephalopathy syndrome (PRES) in a patient with late postpartum eclampsia
Authors: Rijal Jharendra P , Giri Smith , Dawadi Suvash , Dahal Khagendra V
Published: February 2014

Posterior reversible encephalopathy syndrome (PRES) is a reversible neurological entity characterised by seizure, headaches, visual symptoms, impaired consciousness and other focal neurological findings. It is caused by a wide variety of causes ultimately leading to a vasogenic cerebral oedema of occipital and parietal lobes of the brain. We present here a young woman with headache, generalised tonic–clonic seizures and cortical blindness in a late postpartum stage. Reversibility of the symptoms and characteristic imaging findings led us to a diagnosis of PRES in our patient.
Cholelithiasis and the risk of liver cancer: results from cohort studies of 134 546 Chinese men and women
Authors: Vogtmann Emily , Shu Xiao-Ou , Li Hong-Lan , Chow Wong-Ho , Yang Gong , Ji Bu-Tian , Cai Hui , Yu Chang , Gao Yu-Tang , Zheng Wei , Xiang Yong-Bing
Published: February 2014

Background

Cholelithiasis and cholecystectomy have been proposed as risk factors for liver cancer, but findings have been inconsistent. We assessed this association using data from the Shanghai Women's and Men's Health Studies.

Methods

History of cholelithiasis and cholecystectomy were reported at baseline and follow-up interviews, and liver cancer diagnoses were ascertained from the Shanghai Cancer Registry and Vital Statistics Unit. Adjusted hazard ratios (aHRs) and 95% CIs were calculated after adjustment for potential confounders.

Results

A history of cholelithiasis and cholecystectomy was reported by 9.5% and 3.6% of participants at baseline, respectively. After a total of 859 882 person-years of follow-up for women and 391 093 for men, incident liver cancer was detected in 160 women and 252 men. A positive association was observed between a history of cholelithiasis or cholecystectomy and liver cancer in men (aHR 1.46; 95% CI 1.02 to 2.07) and women (aHR 1.55; 95% CI 1.06 to 2.26). Similar results were observed for cholelithiasis only, but cholecystectomy did not reach statistical significance. There was no strong evidence for detection bias of liver cancer due to cholelithiasis or cholecystectomy.

Conclusions

Our study suggests that cholelithiasis and possibly cholecystectomy may increase the risk of liver cancer.
Chronic pain after surgery: pathophysiology, risk factors and prevention
Authors: Reddi Danielle , Curran Natasha
Published: February 2014

Interest in chronic pain after surgery has grown since the finding that more than a fifth of patients attending chronic pain clinics cite surgery as the cause for their chronic pain. The problem is not limited to major surgery; even common minor procedures such as hernia repair have a significant risk of chronic pain. Surgical technique can influence the development of chronic postsurgical pain (CPSP) and techniques to minimise nerve injury should be used where possible. Central nervous system changes contribute to the development of persistent pain following surgical trauma and nerve injury. Pharmacological agents that interrupt the mechanisms contributing to central sensitisation may be helpful in reducing the incidence of CPSP. Psychosocial factors are also important in the development of chronic pain and should be addressed as part of a holistic approach to perioperative care.