bluuebunny/crossref_metadata_2025 huggingface dataset:
Cytomegalovirus antiviral resistance: characterization of results from clinical specimens
Authors: Kleiboeker S. , Nutt J. , Schindel B. , Dannehl J. , Hester J.
Published: June 2014

Background

Cytomegalovirus (CMV) infections are a major cause of disease among immunocompromised patients. Prolonged antiviral therapy is often necessary to prevent or treat CMV disease, and may lead to development of antiviral resistance (AVR). Timely identification of viral mutations conferring resistance is essential for effective patient management.

Methods

Amplification by polymerase chain reaction followed by bi‐directional nucleotide sequencing was performed for relevant regions of the CMV UL97 and UL54 genes. Results from 570 samples submitted to a commercial reference laboratory for testing were reviewed and characterized with respect to the frequency of mutations detected, association with viral load (VL), and consistency of results in a subset of patients with multiple samples. Only AVR mutations confirmed by marker transfer experiments were included in the analysis.

Results

AVR mutations were identified in 176 (30.9%) of the 570 samples evaluated. A total of 17 different UL97 mutations and 29 different UL54 mutations were detected. A single mutation per sample was most commonly observed, although 61 samples (10.7%) had >1 mutation, with 40 samples (7.0%) having mutations in both UL97 and UL54 genes. The VL of samples with AVR mutations ranged from 2.03–7.15 log10 copies/mL, and the VL did not differ significantly from samples without AVR mutations. A subset of patients (N = 85) had >1 sample tested, and 48.2% of these patients returned the same result for each sample analyzed, while the remainder had a different results.

Conclusions

Genomic mutations conferring resistance in CMV to antiviral drugs were commonly identified in samples submitted from clinical patients to a reference laboratory for AVR testing. Mutations were identified over the full range of VLs and no correlation was identified between VL and the presence of AVR mutations. In patients with multiple samples submitted for analysis, approximately half of the patients had samples with variable results when the initial result was compared to subsequent results.
Reproducibility of a quantitative cutaneous cytological technique
Authors: Udenberg Tyler J. , Griffin Craig E. , Rosenkrantz Wayne S. , Ghubash Rudayna M. , Angus John C. , Polissar Nayak L. , Neradilek Moni B.
Published: June 2014

Background

Cutaneous cytology is a valuable tool for diagnosis of canine superficial pyoderma. Current published reproducible techniques are semiquantitative.

Hypothesis/Objectives

The aim of this study was to evaluate the reproducibility of a quantitative method for skin surface cytology in dogs with superficial pyoderma.

Animals

Impression smears were collected from five normal dogs and 20 dogs with clinical and cytological evidence of superficial pyoderma.

Methods

Four investigators evaluated 10 oil immersion fields (OIF) on 25 slides, selecting fields with inflammatory cells, nuclear streaming and or keratinocytes under ×10 magnification. Investigators repeated blinded evaluations of all slides at least twice. For each OIF, polymorphonuclear leukocytes (PMNs), intracellular (IC) cocci, extracellular (EC) cocci, IC rods, EC rods and yeast were quantified. Nuclear streaming was scored as present or absent. For each parameter, within‐reader and between‐reader agreements were expressed by the intraclass correlation (ICC) value (≤0.20 poor, 0.21–0.40 fair, 0.41–0.60 moderate, 0.61–0.80 good and 0.81–1.00 excellent) or kappa statistic (κ).

Results

Reproducible parameters included: PMNs (ICC = 0.58), nuclear streaming (ICC = 0.68), EC cocci (ICC = 0.64) and IC cocci (ICC = 0.32). When qualified as present or absent, within‐reader κ for IC cocci was 0.71. The method demonstrated 93% sensitivity in identifying dogs with superficial pyoderma and 51% specificity in identifying normal dogs according to established criteria. However, if criteria for normal dogs were limited to the absence of PMNs and IC bacteria, sensitivity of 64% and specificity of 98% were demonstrated.

Conclusions and clinical importance

For several parameters, including PMNs, nuclear streaming, EC cocci and IC cocci, a reproducible, quantitative cytological technique was identified.
The establishment of Taraxacum officinale plants in grassland
Authors: Martinkova Z , Honek A
Published: June 2014

Factors involved in the establishment of Taraxacum officinale (dandelion) seedlings in grasslands and plant mortality in the first growing season were studied during 2008–2011. The study was conducted at 11 grassland sites that differed in the quantity of seed rain and the intensity of seedling predation by slugs. In April, of each study year, 10 plots open to the sedimentation of naturally dispersed seeds were established at each site, five plots of which were also sown with 250 T. officinale seeds each. The amount of natural seed rain, slug feeding activity and number of plants in each plot were then recorded at monthly intervals. Seed rain and seedling predation accounted for c. 50% of the variation in seedling establishment. Populations of established seedlings then declined through October, at which point they were at 25% of the original abundance on average. The percentage of plants that survived varied among sites and years. The temporal and spatial variation in T. officinale establishment and survival of young plants was enormous, even within a small area. Predation of seedlings by slugs can effectively prevent the establishment of T. officinale in grassland. This only occurred, however, at slug abundances not acceptable under conditions of public access.
Early postoperative treatment of thyroidectomy scars using botulinum toxin: A split‐scar, double‐blind randomized controlled trial
Authors: Kim Youn Sung , Lee Hyun Joo , Cho Sang Hyun , Lee Jeong Deuk , Kim Hei Sung
Published: June 2014

Operational scars, especially those located on the exposed parts of the body, can be distressing. Despite high demand for an early intervention to minimize surgical scars, there is yet no universal consensus on optimal treatment. A split‐scar, double‐blind randomized controlled trial was held to assess the safety and efficacy of early postoperative botulinum toxin type A (BTA) injection in surgical scars. A single session of treatment was performed where BTA was allocated to one half of the scar and 0.9% saline to the control half. Scars were assessed using the modified Stony Brook Scar Evaluation Scale (SBSES) with standardized photographs. Fifteen patients completed the study, and their data were analyzed. At 6 months' follow‐up, a significant improvement in SBSES score was noted for the BTA‐treated halves of the scars (p < 0.001), with minimal change on the saline‐treated side (p = 0.785). The mean calculated difference in SBSES scores (final/initial) between the BTA‐treated side and the saline‐treated side was also significant (p < 0.001). Early postoperative BTA injection was safe and effective in modulating thyroidectomy scars and may be a promising option for scar prevention.
Prediction Profiles for Nutritional Supplement Use Among Young German Elite Athletes
Authors: Dietz Pavel , Ulrich Rolf , Niess Andreas , Best Raymond , Simon Perikles , Striegel Heiko
Published: June 2014

Nutritional supplements (NS) are defined as concentrated sources of nutrients and other substances that have a nutritional or physiological effect and that are used in high frequency among athletes. The study aimed to create a prediction profile for young elite athletes to identify those athletes who have a higher relative risk for using NS. The second objective was to examine the hypothesis that the consumption of NS paves a gateway for the use of illicit drugs and doping substances. A self-designed anonymous paper-and-pencil questionnaire was used to examine the prevalence of NS consumption, doping, and illicit drug use in elite athletes with a mean age of 17 years (SD = 4 years). Logistic regression analysis was employed to assess whether NS consumption can be predicted by independent variables (e.g., biographical data, training characteristics, drug consumption behavior) to create the prediction profile for NS use. 55% and 5% of the athletes (n = 536) responded positively to having used NS and illicit drugs, respectively. Nutritional supplement consumption was positively correlated with age (OR: 1.92; CI: 1.21 to 3.05), the desire to enhance performance to become an Olympic or World Champion (OR: 3.72; CI: 2.33 to 6.01), and being educated about NS (OR: 2.76; CI: 1.73 to 4.45). It was negatively correlated with training frequency (OR: 0.55; CI: 0.35 to 0.86) and the use of nicotine (OR: 0.29; CI: 0.1 to 0.74) but did not correlate with illicit drug use and alcohol consumption. The present results show that NS are used on a large scale in elite sports. The prediction profile presented in this article may help to identify those athletes who have a high risk for using NS to plan potential education and prevention models more individually.
The Efficacy of a Self-Paced VO2max Test During Motorized Treadmill Exercise
Authors: Faulkner James , Mauger Alexis R. , Woolley Brandon , Lambrick Danielle
Published: June 2014

Purpose:

To assess the utility of a self-paced maximal oxygen uptake (VO2max) test (SPV) in eliciting an accurate measure of VO2max in comparison with a traditional graded exercise test (GXT) during motorized treadmill exercise.

Design:

This was a cross-sectional experimental study whereby recreationally trained men (n = 13, 25.5 ± 4.6 y) completed 2 maximal exercise tests (SPV, GXT) separated by a 72-h recovery period.

Methods:

The GXT was continuous and incremental, with prescribed 1-km/h increases every 2 min until the attainment of VO2max. The SPV consisted of 5 × 2-min stages of incremental exercise, which were self-selected and adjusted according to 5 prescribed RPE levels (RPE 11, 13, 15, 17, and 20).

Results:

Although no significant differences in VO2max were observed between the SPV and GXT (63.9 ± 3.3 cf 60.9 ± 4.6 mL · kg−1 · min−1, respectively, P > .05), the apparent 4.7% mean difference may be practically important. The 95% limits-of-agreement analysis was 3.03 ± 11.49 mL · kg−1 · min−1. Therefore, in the worst-case scenario, the GXT may underestimate measured VO2max as ascertained by the SPV by up to 19%. Conversely, the SPV could underestimate the GXT by 14%.

Conclusions:

The current study has shown that the SPV is an accurate measure of VO2max during exercise on a motorized treadmill and may provide a slightly higher VO2max value than that obtained from a traditional GXT. The higher VO2max during the SPV may be important when prescribing training or monitoring athlete progression.
Physical Activity for Campus Employees: A University Worksite Wellness Program
Authors: Butler Carling E. , Clark B. Ruth , Burlis Tamara L. , Castillo Jacqueline C. , Racette Susan B.
Published: June 2014

Background:

Workplaces provide ideal environments for wellness programming. The purpose of this study was to explore exercise self-efficacy among university employees and the effects of a worksite wellness program on physical activity, cardiorespiratory fitness, and cardiovascular disease (CVD) risk factors.

Methods:

Participants included 121 university employees (85% female). The worksite wellness program included cardiovascular health assessments, personal health reports, 8 weeks of pedometer-based walking and tracking activities, and weekly wellness sessions. Daily step count was assessed at baseline, Week 4, and Week 8. Exercise self-efficacy and CVD risk factors were evaluated at baseline and follow-up.

Results:

Daily step count increased from 6566 ± 258 (LSM ± SE) at baseline to 8605 ± 356 at Week 4 and 9107 ± 388 at Week 8 (P < .0001). Steps increased among normal weight, overweight, and obese subgroups. Exercise self-efficacy correlated with baseline steps (P < .05). Small improvements were observed in cardiorespiratory fitness, body mass index, blood pressure, blood glucose, total cholesterol, and triglycerides (all P < .01).

Conclusions:

A worksite wellness program was effective for improving physical activity, cardiorespiratory fitness, and CVD risk factors among university employees. Exercise barriers and outcome expectations were identified and have implications for future worksite wellness programming.
Identification of the giant impactor Theia in lunar rocks
Authors: Herwartz Daniel , Pack Andreas , Friedrichs Bjarne , Bischoff Addi
Published: June 2014

An analysis of motes of the Moon maker

How did the Moon form? According to the prevailing hypothesis, a Mars-sized body known as Theia smashed into Earth. Herwartz
et al.
analyzed fresh basalt samples from three Apollo landing sites and compared them with several samples of Earth's mantle. The oxygen isotope values measured in these lunar rocks differ significantly from the terrestrial material, supporting the giant-impact hypothesis.

Science
, this issue p.
1146
From aerosol-limited to invigoration of warm convective clouds
Authors: Koren Ilan , Dagan Guy , Altaratz Orit
Published: June 2014

Invigorating convection in warm clouds

Atmospheric aerosols—tiny airborne particles—affect the way clouds form and how they affect climate. Koren
et al.
investigated how the formation of warm clouds, such as those that form over the oceans, depends on pollution levels (see the Perspective by Remer). Aerosols affect cloud formation in cleaner air disproportionately more than in more polluted air. Before the widespread air pollution of the industrial era, it seems, warm convective clouds may have covered much less of the oceans than they do today.

Science
, this issue p.
1143
; see also p.
1089
Speed metal
Authors: Elliott Tim
Published: June 2014

Meteorite dating reveals that planetary core formation is a relatively fast process.

[Also see Report by

Kruijer
et al.

]
Just add aerosols
Authors: Remer Lorraine A.
Published: June 2014

Data from clean regions of the atmosphere show how little aerosol is needed to change clouds.

[Also see Report by

Koren
et al.

]
Science diplomacy with Cuba
Authors: Fink Gerald R. , Leshner Alan I. , Turekian Vaughan C.
Published: June 2014

Over a century ago, interactions between a Cuban scientist, Carlos Finlay, and a U.S. scientist, Jesse Lazear, led to an understanding of the role of the mosquito in transmitting yellow fever and to the development of effective countermeasures. Today, new infectious diseases confront both Cuba and the United States, but a longstanding diplomatic breach between the two nations now makes such valuable joint research more complicated, if not impossible. It is time for both governments to reconsider the rules that stand in the way of scientific collaboration, before a potentially deadly outbreak spreads through both countries and beyond. In particular, the U.S. government can make a relatively straightforward change to rules that would make collaboration much easier.
Lengthy RNAs earn respect as cellular players
Authors: Pennisi Elizabeth
Published: June 2014

A few of these molecules are clearly important, but just how many?
The killing ground
Authors: Balter Michael
Published: June 2014

Clues from a German coal mine show how early hunters lived, 300,000 years ago, and how their prey died.
Sensitivity of Real-Time PCR Performed on Dried Sera Spotted on Filter Paper for Diagnosis of Leptospirosis
Authors: Nhan T. X. , Teissier A. , Roche C. , Musso D.
Published: June 2014

In remote countries, leptospirosis confirmation is difficult because it requires the shipment of frozen samples to reference laboratories. The sensitivity of leptospirosis real-time PCR performed on filter paper-dried serum samples stored at ambient temperature was evaluated at 2 × 10
2
equivalent leptospires/ml (eqLeptospires/ml). This easy alternative procedure can be used to enhance the surveillance of leptospirosis.
Diagnostic Accuracy of a Molecular Drug Susceptibility Testing Method for the Antituberculosis Drug Ethambutol: a Systematic Review and Meta-Analysis
Authors: Cheng Song , Cui Zhenling , Li Yuanyuan , Hu Zhongyi
Published: June 2014

Ethambutol (EMB) is a first-line antituberculosis drug; however, drug resistance to EMB has been increasing. Molecular drug susceptibility testing (DST), based on theembBgene, has recently been used for rapid identification of EMB resistance. The aim of this meta-analysis was to establish the accuracy of molecular assay for detecting drug resistance to EMB. PubMed, Embase, and Web of Science were searched according to a written protocol and explicit study selection criteria. Measures of diagnostic accuracy were pooled using a random effects model. A total of 34 studies were included in the meta-analysis. The respective pooled sensitivities and specificities were 0.57 and 0.93 for PCR-DNA sequencing that targeted theembB306 codon, 0.76 and 0.89 for PCR-DNA sequencing that targeted theembB306, 406, and 497 codons, 0.64 and 0.70 for detectingMycobacterium tuberculosisisolates, 0.55 and 0.78 for detectingM. tuberculosissputum specimens using the GenoType MTBDRsltest, 0.57 and 0.87 for pyrosequencing, and 0.35 and 0.98 for PCR-restriction fragment length polymorphism. The respective pooled sensitivities and specificities were 0.55 and 0.92 when using a lower EMB concentration as the reference standard, 0.67 and 0.73 when using a higher EMB concentration as the reference standard, and 0.60 and 1.0 when using multiple reference standards. PCR-DNA sequencing using multiple sites of theembBgene as detection targets, includingembB306, 406, and 497, can be a rapid method for preliminarily screening for EMB resistance, but it does not fully replace phenotypic DST. Of the reference DST methods examined, the agreement rates were the best using MGIT 960 for molecular DST and using the proportion method on Middlebrook 7H10 media.
Cervical and Vaginal Flora Specimens Are Highly Concordant with Respect to Bacterial Vaginosis-Associated Organisms and Commensal Lactobacillus Species in Women of Reproductive Age
Authors: Smith William L. , Hedges Spencer R. , Mordechai Eli , Adelson Martin E. , Trama Jason P. , Gygax Scott E. , Kaunitz Andrew M. , Hilbert David W.
Published: June 2014

Matched vaginal and cervical specimens from 96 subjects were analyzed by quantitative PCR for the presence and concentration of bacterial vaginosis-associated microbes and commensal
Lactobacillus
spp. Detection of these microbes was 92% concordant, indicating that microbial floras at these body sites are generally similar.
Characteristics of Acute Nongonococcal Urethritis in Men Differ by Sexual Preference
Authors: Rane Vinita S. , Fairley Christopher K. , Weerakoon Ajith , Read Timothy H. , Fehler Glenda , Chen Marcus Y. , Bradshaw Catriona S.
Published: June 2014

Nongonococcal urethritis (NGU) is a common clinical syndrome, but no etiological agent is identified in a significant proportion of cases. Whether the spectrum of pathogens differs between heterosexual men (MSW) and men who have sex with men (MSM) is largely unstudied but of considerable clinical relevance. A retrospective review was done using the electronic medical record database of Melbourne Sexual Health Centre, Australia. Cases were first presentations of symptomatic acute NGU with ≥5 polymorphonuclear leukocytes (PMNL)/high-powered field (HPF) on urethral Gram stain between January 2006 and December 2011. First-stream urine was tested for
Chlamydia trachomatis
and
Mycoplasma genitalium
by PCR. Demographic, laboratory, and behavioral characteristics of cases were examined by univariate and multivariable analyses. Of 1,295 first presentations of acute NGU, 401 (32%; 95% confidence interval [CI] of 29 to 34%) had
C. trachomatis
and 134 (11%; 95% CI of 9 to 13%) had
M. genitalium
detected. MSM with acute NGU were less likely to have
C. trachomatis
(adjusted odds ratio [AOR] = 0.4; 95% CI of 0.3 to 0.6) or
M. genitalium
(AOR = 0.5; 95% CI of 0.3 to 0.8) and more likely to have idiopathic NGU (AOR = 2.4; 95% CI of 1.8 to 3.3), to report 100% condom use for anal/vaginal sex (AOR = 3.6; 95% CI of 2.7 to 5.0), or to have engaged in sexual activities other than anal/vaginal sex (AOR = 8.0; 95% CI of 3.6 to 17.8). Even when
C. trachomatis
or
M. genitalium
was detected, MSM were more likely than MSW to report consistent condom use (OR = 4.7; 95% CI of 2.6 to 8.3). MSM with acute NGU are less likely to have the established bacterial sexually transmitted infections (STIs) and more likely to report protected anal sex or sexual activity other than anal sex prior to symptom onset than MSW. These data suggest that the etiologic spectrum of pathogens differs between MSM and MSW in acute NGU and that relatively low-risk practices are capable of inducing acute NGU.
Competency Assessment of Microbiology Medical Laboratory Technologists in Ontario, Canada
Authors: Desjardins Marc , Fleming Christine Ann
Published: June 2014

Accreditation in Ontario, Canada, requires that licensed clinical laboratories participate in external quality assessment (also known as proficiency testing) and perform competency evaluation of their staff. To assess the extent of ongoing competency assessment practices, the Quality Management Program—Laboratory Services (QMP-LS) Microbiology Committee surveyed all 112 licensed Ontario microbiology laboratories. The questionnaire consisted of a total of 21 questions that included yes/no, multiple-choice, and short-answer formats. Participants were asked to provide information about existing programs, the frequency of testing, what areas are evaluated, and how results are communicated to the staff. Of the 111 responding laboratories, 6 indicated they did not have a formal evaluation program since they perform only limited bacteriology testing. Of the remaining 105 respondents, 87% perform evaluations at least annually or every 2 years, and 61% include any test or task performed, whereas 16% and 10% focus only on problem areas and high-volume complex tasks, respectively. The most common methods of evaluation were review of external quality assessment (EQA) challenges, direct observation, and worksheet review. With the exception of one participant, all communicate results to staff, and most take remedial action to correct the deficiencies. Although most accredited laboratories have a program to assess the ongoing competency of their staff, the methods used are not standardized or consistently applied, indicating that there is room for improvement. The survey successfully highlighted potential areas for improvement and allowed the QMP-LS Microbiology Committee to provide guidance to Ontario laboratories for establishing or improving existing microbiology-specific competency assessment programs.
Development of a Rapid Microarray-Based DNA Subtyping Assay for the Alleles of Shiga Toxins 1 and 2 of Escherichia coli
Authors: Geue Lutz , Stieber Bettina , Monecke Stefan , Engelmann Ines , Gunzer Florian , Slickers Peter , Braun Sascha D. , Ehricht Ralf
Published: June 2014

In this study, we developed a new rapid, economic, and automated microarray-based genotyping test for the standardized subtyping of Shiga toxins 1 and 2 of
Escherichia coli
. The microarrays from Alere Technologies can be used in two different formats, the ArrayTube and the ArrayStrip (which enables high-throughput testing in a 96-well format). One microarray chip harbors all the gene sequences necessary to distinguish between all Stx subtypes, facilitating the identification of single and multiple subtypes within a single isolate in one experiment. Specific software was developed to automatically analyze all data obtained from the microarray. The assay was validated with 21 Shiga toxin-producing
E. coli
(STEC) reference strains that were previously tested by the complete set of conventional subtyping PCRs. The microarray results showed 100% concordance with the PCR results. Essentially identical results were detected when the standard DNA extraction method was replaced by a time-saving heat lysis protocol. For further validation of the microarray, we identified the Stx subtypes or combinations of the subtypes in 446 STEC field isolates of human and animal origin. In summary, this oligonucleotide array represents an excellent diagnostic tool that provides some advantages over standard PCR-based subtyping. The number of the spotted probes on the microarrays can be increased by additional probes, such as for novel alleles, species markers, or resistance genes, should the need arise.
Probiotic-Associated Aspiration Pneumonia Due to Lactobacillus rhamnosus
Authors: Doern Christopher D. , Nguyen Sean T. , Afolabi Folashade , Burnham Carey-Ann D.
Published: June 2014

Lactobacilli are low-virulence, commensal organisms of the gastrointestinal and genitourinary tracts and are commonly used as “probiotic supplements.” Herein, we describe an episode of respiratory syncytial virus (RSV) bronchiolitis with bacterial superinfection secondary to administration ofLactobacillus rhamnosusin an 11-month-old female with trisomy 21.
Prevalence of Recovirus-Neutralizing Antibodies in Human Serum Samples
Authors: Farkas Tibor , Wong Ping Lun Cindy
Published: June 2014

To investigate recovirus infections and their association with zoonosis, the prevalence of the virus-neutralizing antibody against three recovirus serotypes was tested in the general population and in zookeepers. Neutralizing antibodies were detected in a significantly higher number of zookeepers than in the general population but with significantly lower titers than in macaques.
Time to Positivity and Detection of Growth in Anaerobic Blood Culture Vials Predict the Presence of Candida glabrata in Candidemia: a Two-Center European Cohort Study
Authors: Cobos-Trigueros Nazaret , Kaasch Achim J. , Soriano Alex , Torres Jorge-Luis , Vergara Andrea , Morata Laura , Zboromyrska Yuliya , De La Calle Cristina , Alejo Izaskun , Hernández Cristina , Cardozo Celia , Marco Franscesc , Del Río Ana , Almela Manel , Mensa Josep , Martínez José Antonio
Published: June 2014

This study shows the accuracy of exclusive or earlier growth in anaerobic vials to predict
Candida glabrata
in a large series of candidemic patients from two European hospitals using the Bactec 9240 system. Alternatively,
C. glabrata
can be predicted by a time to positivity cutoff value, which should be determined for each setting.
An Alphavirus-Based Adjuvant Enhances Serum and Mucosal Antibodies, T Cells, and Protective Immunity to Influenza Virus in Neonatal Mice
Authors: Khalil Syed Muaz , Tonkin Daniel R. , Snead Andrew T. , Parks Griffith D. , Johnston Robert E. , White Laura J.
Published: June 2014

Neonatal immune responses to infection and vaccination are biased toward TH2 at the cost of proinflammatory TH1 responses needed to combat intracellular pathogens. However, upon appropriate stimulation, the neonatal immune system can induce adult-like TH1 responses. Here we report that a new class of vaccine adjuvant is especially well suited to enhance early life immunity. The GVI3000 adjuvant is a safe, nonpropagating, truncated derivative of Venezuelan equine encephalitis virus that targets dendritic cells (DCs) in the draining lymph node (DLN) and produces intracellular viral RNA without propagating to other cells. RNA synthesis strongly activates the innate immune response so that in adult animals, codelivery of soluble protein antigens induces robust humoral, cellular, and mucosal responses. The adjuvant properties of GVI3000 were tested in a neonatal BALB/c mouse model using inactivated influenza virus (iFlu). After a single immunization, mice immunized with iFlu with the GVI3000 adjuvant (GVI3000-adjuvanted iFlu) had significantly higher and sustained influenza virus-specific IgG antibodies, mainly IgG2a (TH1), compared to the mice immunized with antigen only. GVI3000 significantly increased antigen-specific CD4+and CD8+T cells, primed mucosal immune responses, and enhanced protection from lethal challenge. As seen in adult mice, the GVI3000 adjuvant increased the DC population in the DLNs, caused activation and maturation of DCs, and induced proinflammatory cytokines and chemokines in the DLNs soon after immunization, including gamma interferon (IFN-γ), tumor necrosis factor alpha (TNF-α), granulocyte colony-stimulating factor (G-CSF), and interleukin 6 (IL-6). In summary, the GVI3000 adjuvant induced an adult-like adjuvant effect with an influenza vaccine and has the potential to improve the immunogenicity and protective efficacy of new and existing neonatal vaccines.

IMPORTANCEThe suboptimal immune responses in early life constitute a significant challenge for vaccine design. Here we report that a new class of adjuvant is safe and effective for early life immunization and demonstrate its ability to significantly improve the protective efficacy of an inactivated influenza virus vaccine in a neonatal mouse model. The GVI3000 adjuvant delivers a truncated, self-replicating viral RNA into dendritic cells in the draining lymph node. Intracellular RNA replication activates a strong innate immune response that significantly enhances adaptive antibody and cellular immune responses to codelivered antigens. A significant increase in protection results from a single immunization. Importantly, this adjuvant also primed a mucosal IgA response, which is likely to be critical for protection during many early life infections.
Transcriptional and Posttranscriptional Regulation of Cytokine Gene Expression in HIV-1 Antigen-Specific CD8+T Cells That Mediate Virus Inhibition
Authors: Payne Tamika L. , Blackinton Jeff , Frisbee Alyse , Pickeral Joy , Sawant Sheetal , Vandergrift Nathan A. , Freel Stephanie A. , Ferrari Guido , Keene Jack D. , Tomaras Georgia D.
Published: June 2014

The ability of CD8+T cells to effectively limit HIV-1 replication and block HIV-1 acquisition is determined by the capacity to rapidly respond to HIV-1 antigens. Understanding both the functional properties and regulation of an effective CD8+response would enable better evaluation of T cell-directed vaccine strategies and may inform the design of new therapies. We assessed the antigen specificity, cytokine signature, and mechanisms that regulate antiviral gene expression in CD8+T cells from a cohort of HIV-1-infected virus controllers (VCs) (<5,000 HIV-1 RNA copies/ml and CD4+lymphocyte counts of >400 cells/μl) capable of soluble inhibition of HIV-1. Gag p24 and Nef CD8+T cell-specific soluble virus inhibition was common among the VCs and correlated with substantial increases in the abundance of mRNAs encoding the antiviral cytokines macrophage inflammatory proteins MIP-1α, MIP-1αP (CCL3L1), and MIP-1β; granulocyte-macrophage colony-stimulating factor (GM-CSF); lymphotactin (XCL1); tumor necrosis factor receptor superfamily member 9 (TNFRSF9); and gamma interferon (IFN-γ). The induction of several of these mRNAs was driven through a coordinated response of both increased transcription and stabilization of mRNA, which together accounted for the observed increase in mRNA abundance. This coordinated response allows rapid and robust induction of mRNA messages that can enhance the CD8+T cells' ability to inhibit virus upon antigen encounter.

IMPORTANCEWe show that mRNA stability, in addition to transcription, is key in regulating the direct anti-HIV-1 function of antigen-specific memory CD8+T cells. Regulation at the level of RNA helps enable rapid recall of memory CD8+T cell effector functions for HIV-1 inhibition. By uncovering and understanding the mechanisms employed by CD8+T cell subsets with antigen-specific anti-HIV-1 activity, we can identify new strategies for comprehensive identification of other important antiviral genes. This will, in turn, enhance our ability to inhibit virus replication by informing both cure strategies and HIV-1 vaccine designs that aim to reduce transmission and can aid in blocking HIV-1 acquisition.
Binding of the Kaposi's Sarcoma-Associated Herpesvirus to the Ephrin Binding Surface of the EphA2 Receptor and Its Inhibition by a Small Molecule
Authors: Hahn Alexander S. , Desrosiers Ronald C.
Published: June 2014

The ephrin receptor tyrosine kinase A2 (EphA2) is an entry receptor for Kaposi's sarcoma-associated herpesvirus (KSHV) that is engaged by the virus through its gH/gL glycoprotein complex. We describe here that natural ephrin ligands inhibit the gH/gL-EphA2 interaction. The effects of point mutations within EphA2 demonstrated that KSHV gH/gL interacts with EphA2 through a restricted set of the same residues that mediate binding of A-type ephrins. Two previously described inhibitors of the EphA2 interaction with ephrin A5 also inhibited binding of KSHV gH/gL to EphA2. The more potent of the two compounds inhibited KSHV infection of blood vessel and lymphatic endothelial cells in the micromolar concentration range. Our results demonstrate that interaction of KSHV with EphA2 occurs in a fashion similar to that of the natural ephrin ligands. Our data further indicate a new avenue for drug development against KSHV.

IMPORTANCE
Our study reports two important findings. First, we show that KSHV engages its receptor, the receptor tyrosine kinase EphA2, at a site that overlaps the binding site of the natural ephrin ligands. Second, we demonstrate that KSHV infection of target cells can be blocked by a small-molecule inhibitor of the viral glycoprotein-EphA2 interaction. These findings represent a novel avenue for the development of strategies to treat KSHV-associated diseases.
Epithelial Immunization Induces Polyfunctional CD8+T Cells and Optimal Mousepox Protection
Authors: Hersperger Adam R. , Siciliano Nicholas A. , DeHaven Brian C. , Snook Adam E. , Eisenlohr Laurence C.
Published: June 2014

We assessed several routes of immunization with vaccinia virus (VACV) in protecting mice against ectromelia virus (ECTV). By a wide margin, skin scarification provided the greatest protection. Humoral immunity and resident-memory T cells notwithstanding, several approaches revealed that circulating, memory CD8+T cells primed via scarification were functionally superior and conferred enhanced virus control. Immunization via the epithelial route warrants further investigation, as it may also provide enhanced defense against other infectious agents.
Defining the Chemokine Basis for Leukocyte Recruitment during Viral Encephalitis
Authors: Michlmayr Daniela , McKimmie Clive S. , Pingen Marieke , Haxton Ben , Mansfield Karen , Johnson Nicholas , Fooks Anthony R. , Graham Gerard J.
Published: June 2014

The encephalitic response to viral infection requires local chemokine production and the ensuing recruitment of immune and inflammatory leukocytes. Accordingly, chemokine receptors present themselves as plausible therapeutic targets for drugs aimed at limiting encephalitic responses. However, it remains unclear which chemokines are central to this process and whether leukocyte recruitment is important for limiting viral proliferation and survival in the brain or whether it is predominantly a driver of coincident inflammatory pathogenesis. Here we examine chemokine expression and leukocyte recruitment in the context of avirulent and virulent Semliki Forest virus (SFV) as well as West Nile virus infection and demonstrate rapid and robust expression of a variety of inflammatory CC and CXC chemokines in all models. On this basis, we define a chemokine axis involved in leukocyte recruitment to the encephalitic brain during SFV infection. CXCR3 is the most active; CCR2 is also active but less so, and CCR5 plays only a modest role in leukocyte recruitment. Importantly, inhibition of each of these receptors individually and the resulting suppression of leukocyte recruitment to the infected brain have no effect on viral titer or survival following infection with a virulent SFV strain. In contrast, simultaneous blockade of CXCR3 and CCR2 results in significantly reduced mortality in response to virulent SFV infection. In summary, therefore, our data provide an unprecedented level of insight into chemokine orchestration of leukocyte recruitment in viral encephalitis. Our data also highlight CXCR3 and CCR2 as possible therapeutic targets for limiting inflammatory damage in response to viral infection of the brain.

IMPORTANCE
Brain inflammation (encephalitis) in response to viral infection can lead to severe illness and even death. This therefore represents an important clinical problem and one that requires the development of new therapeutic approaches. Central to the pathogenesis of encephalitis is the recruitment of inflammatory leukocytes to the infected brain, a process driven by members of the chemokine family. Here we provide an in-depth analysis of the chemokines involved in leukocyte recruitment to the virally infected brain and demonstrate that simultaneous blockade of two of these receptors, namely, CXCR3 and CCR2, does not alter viral titers within the brain but markedly reduces inflammatory leukocyte recruitment and enhances survival in a murine model of lethal viral encephalitis. Our results therefore highlight chemokine receptors as plausible therapeutic targets in treating viral encephalitis.
Post liver transplant presentation of needle-track metastasis of hepatocellular carcinoma following percutaneous liver biopsy
Authors: Joyce Daniel , Falk Gavin A , Gandhi Namita , Hashimoto Koji
Published: June 2014

Hepatocellular carcinoma (HCC) is one of the few malignant tumours often treated without prior histological confirmation (in the patient with cirrhosis). Contrast-enhanced cross-sectional imaging is frequently diagnostic of HCC with a high degree of accuracy. However, on occasion, a liver biopsy is required, a complication of which can be needle-track metastasis. We present the case of a 57-year-old man who had previously undergone a liver transplant; he was found to have abdominal wall metastasis at the site of a prior percutaneous biopsy. This is the second case until now date of needle-track metastasis that presented following liver transplantation.
Aggressive dentigerous cyst with ectopic central incisor
Authors: Jayam Cheranjeevi , Mitra Malay , Bandlapalli Anila , Jana Biswanath
Published: June 2014

Dentigerous cysts form from accumulation of fluid between reduced enamel epithelium and the crown of an unerupted tooth. They cause several difficulties such as swelling, non-eruption of the involved teeth, and displacement of adjacent teeth, and thus require early detection and prompt treatment. Treatment ranges from marsupialisation to enucleation. Enucleation is rarely used in children compared with marsupialisation. This paper discusses successful use of enucleation for treating a dentigerous cyst and explains the need for such a radical procedure.
Pyogenic hepatic abscess presenting years after a choledochojejunostomy: a rare clinical occurrence
Authors: Yu Megan , Mangaonkar Abhishek , Lovelace Candace , Ibe Michael
Published: June 2014

A 69-year-old Caucasian man presented with fever, chills/rigors and night sweats since 6 days. Blood cultures (4/4) initially reported Gram negative lactose-fermenting rods. Physical examination was fairly benign which included a normal abdominal examination. Laboratory tests were significant for an elevated white cell count, erythrocyte sedimentation rate and C reactive protein . Empirically, he was treated with piperacillin tazobactam. A chart review showed that he had undergone a choledochojejunostomy for a pancreatic head tumour 7 years before. We found a few reported cases of hepatic abscesses after choledochojejunostomy presenting years after the procedure. An abdominal CT scan confirmed our suspicion. Percutaneous drainage was performed and his antibiotics were switched to ciprofloxacin and metronidazole, based on the sensitivity report. The patient's clinical condition steadily improved.
Mediastinitis byActinomyces meyeriafter oesophageal stent placement
Authors: Branquinho Diogo Ferreira , Andrade Daniel Ramos , Almeida Nuno , Sofia Carlos
Published: June 2014

Actinomyces meyeriis a Gram-positive anaerobic forming bacterium of the genusActinomyces, part of the oral cavity's flora, and its classification remains an unresolved issue. It is an extremely rare cause of disease, occurring in middle-aged immunocompetent patients and frequently misdiagnosed as malignancy or lung abscess. A 56-year-old man diagnosed with oesophageal squamous cell carcinoma had an endoscopically placed stent to palliate his dysphagia. Two weeks later he presented with thoracalgia and fever, interpreted as a common lung infection. Owing to lack of improvement, additional examinations were undertaken revealing mediastinum involvement. Unlike the good prognosis usually associated with this infection, the patient eventually died, reflecting the aggressive nature of his underlying condition. To our knowledge, this is the first report of mediastinitis byA. meyeri, supporting the described propensity of this agent to disseminate, particularly to the thoracic cavity, although probably in this case with an iatrogenic contribution.
Posterior reversible encephalopathy syndrome resulting from repeat bortezomib usage
Authors: Nixon N A , Parhar K
Published: June 2014

Bortezomib is a chemotherapeutic agent that acts via proteasome inhibition resulting in cellular apoptosis and inhibition of angiogenesis. Although widely accepted as treatment of multiple myeloma and non-Hodgkin's lymphoma, it has also been shown to be efficacious in a variety of solid tumours such as pancreatic and colonic. Posterior reversible encephalopathy syndrome (PRES) is a neuroradiological syndrome characterised by vasogenic oedema involving the postero-occipital cortical and subcortical white matter resulting in visual disturbances, seizures and altered mental status. Although in most cases PRES is reversible with removal of the provoking condition or drug, if not appropriately recognised and treated it may lead to permanent and life-threatening sequelae such as intracerebral haemorrhage and ischaemic infarction. We report a case of PRES associated with bortezomib therapy and contrast it with four other previously reported cases. Recognition of this potentially severe neurological complication is important with the increasingly widespread use of bortezomib.
Horner's syndrome secondary to intervertebral disc herniation at the level of T1–2
Authors: Spacey Kate , Dannawi Zaher , Khazim R , Dannawi Z
Published: June 2014

A 54-year-old Caucasian woman presented with a 6 week history of periscapular pain and a T1 radiculopathy associated with Horner's syndrome. MRI of her cervicothoracic spine revealed an intervertebral disc herniation at the level of T1–2. During investigation she experienced some improvement in her symptoms and a conservative approach was pursued. At 6 months her pain and radiculopathy had resolved, and there was mild residual ptosis.
Environments, risk and health harms: a qualitative investigation into the illicit use of anabolic steroids among people using harm reduction services in the UK
Authors: Kimergård Andreas , McVeigh Jim
Published: June 2014

Objectives

The illicit use of anabolic steroids among the gym population continues to rise, along with the number of steroid using clients attending harm reduction services in the UK. This presents serious challenges to public health. Study objectives were to account for the experiences of anabolic steroid users and investigate how ‘risk environments’ produce harm.

Methods

Qualitative face-to-face interviews with 24 users of anabolic steroids engaged with harm reduction services in the UK.

Results

Body satisfaction was an important factor when deciding to start the use of anabolic steroids. Many users were unaware of the potential dangers of using drugs from the illicit market, whereas some had adopted a range of strategies to negotiate the hazards relating to the use of adulterated products, including self-experimentation to gauge the perceived efficacy and unwanted effects of these drugs. Viewpoints, first-hand anecdotes, norms and practices among groups of steroid users created boundaries of ‘sensible’ drug use, but also promoted practices that may increase the chance of harms occurring. Established users encouraged young users to go to harm reduction services but, at the same time, promoted risky injecting practices in the belief that this would enhance the efficacy of anabolic steroids.

Conclusions

Current steroid-related viewpoints and practices contribute to the risk environment surrounding the use of these drugs and may undermine the goal of current public health strategies including harm reduction interventions. The level of harms among anabolic steroid users are determined by multiple and intertwining factors, in addition to the harms caused by the pharmacological action or injury and illness associated with incorrect injecting techniques.
Do gastroenterologists monitor their patients taking 5-amino-salicylates following initiation of treatment
Authors: Siddique N , Farmer C , Muller A F
Published: June 2014

Background

5-Amino salicylate (5-ASA) medications may rarely be associated with a significant decline in renal function and interstitial nephritis. British Society of Gastroenterology guidelines advise regular renal function monitoring for patients taking these drugs.

Aim

To assess whether gastroenterologists in Kent were following best practice guidelines regarding the monitoring of their patients on 5-ASA therapy.

Methods

Using longitudinal community and regional pathology databases for the Kent population, our renal unit regularly screens a total population of 300 000 for evidence of renal disease. The data extracted are analysed using an automated computerised system to identify patients requiring intervention for kidney disease. All patients taking 5-ASA medication were identified from a population of 300 000. The pathology database was studied to identify the patients on 5-ASA treatment and whether they had had renal function tests.

Results

800 adult patients were identified taking 5-ASA therapy. 612 patients received 5-ASAs for 3 months or more, and these were included in the final analysis. 293 patients had no renal function checks while on treatment. 79 patients had renal function tests less than once every 4 years and 36 patients once every 2–4 years. 204 patients had renal function measurements in 50% or more of years of treatment, of whom 116 were checked every year. Some patients were started on treatment with abnormal results at baseline and some with identified kidney disease continued on their 5-ASAs.

Conclusions

The majority of patients receiving 5-ASA compounds do not have regular renal function monitoring. Clinicians are failing to follow best practice guidelines.