bluuebunny/crossref_metadata_2025_split huggingface dataset:
Influenza Virus Infectivity and Virulence following Ocular-Only Aerosol Inoculation of Ferrets
Authors: Belser Jessica A. , Gustin Kortney M. , Katz Jacqueline M. , Maines Taronna R. , Tumpey Terrence M. 
Published: June 2014

ABSTRACT
Respiratory pathogens have traditionally been studied by examining the exposure and infection of respiratory tract tissues. However, these studies typically overlook the role of ocular surfaces, which represent both a potential site of virus replication and a portal of entry for the establishment of a respiratory infection. To model transocular virus entry in a mammalian species, we established a novel inoculation method that delivers an aerosol inoculum exclusively to the ferret ocular surface. Using influenza virus as a representative respiratory pathogen, we found that both human and avian viruses mounted productive respiratory infections in ferrets following ocular-only aerosol inoculation, and we demonstrated that H5N1 virus can result in a fatal infection at doses below 10 PFU or with exposure times as short as 2 min. Ferrets inoculated by the ocular aerosol route with an avian (H7N7, H7N9) or human (H1N1, H3N2v) virus were capable of transmitting the virus to naïve animals in direct-contact or respiratory-droplet models, respectively. Our results reveal that ocular-only exposure to virus-containing aerosols constitutes a valid exposure route for a potentially fatal respiratory infection, even for viruses that do not demonstrate an ocular tropism, underscoring the public health implications of ocular exposure in clinical or occupational settings.

IMPORTANCE
In the absence of eye protection, the human ocular surface remains vulnerable to infection with aerosolized respiratory viruses. In this study, we present a way to inoculate laboratory mammals that excludes respiratory exposure, infecting ferrets only by ocular exposure to influenza virus-containing aerosols. This study demonstrates that the use of respiratory protection alone does not fully protect against influenza virus exposure, infection, and severe disease.
Intranasal P Particle Vaccine Provided Partial Cross-Variant Protection against Human GII.4 Norovirus Diarrhea in Gnotobiotic Pigs
Authors: Kocher Jacob , Bui Tammy , Giri-Rachman Ernawati , Wen Ke , Li Guohua , Yang Xingdong , Liu Fangning , Tan Ming , Xia Ming , Zhong Weiming , Jiang Xi , Yuan Lijuan 
Published: June 2014

ABSTRACT

Noroviruses (NoVs) are the leading cause of nonbacterial acute gastroenteritis worldwide in people of all ages. The P particle is a novel vaccine candidate derived from the protruding (P) domain of the NoV VP1 capsid protein. This study utilized the neonatal gnotobiotic pig model to evaluate the protective efficacies of primary infection, P particles, and virus-like particles (VLPs) against NoV infection and disease and the T cell responses to these treatments. Pigs either were vaccinated intranasally with GII.4/1997 NoV (VA387)-derived P particles or VLPs or were inoculated orally with a GII.4/2006b NoV variant. At postinoculation day (PID) 28, pigs either were euthanized or were challenged with the GII.4/2006b variant and monitored for diarrhea and virus shedding for 7 days. The T cell responses in intestinal and systemic lymphoid tissues were examined. Primary NoV infection provided 83% homologous protection against diarrhea and 49% homologous protection against virus shedding, while the P particle and VLP vaccines provided cross-variant protection (47% and 60%, respectively) against diarrhea. The protection rates against diarrhea are significantly inversely correlated with T cell expansion in the duodenum and are positively correlated with T cell expansion in the ileum and spleen. The P particle vaccine primed for stronger immune responses than VLPs, including significantly higher numbers of activated CD4
+
T cells in all tissues, gamma interferon-producing (IFN-γ
+
) CD8
+
T cells in the duodenum, regulatory T cells (Tregs) in the blood, and transforming growth factor β (TGF-β)-producing CD4
+
CD25
−
FoxP3
+
Tregs in the spleen postchallenge, indicating that P particles are more immunogenic than VLPs at the same dose. In conclusion, the P particle vaccine is a promising vaccine candidate worthy of further development.

IMPORTANCE
The norovirus (NoV) P particle is a vaccine candidate derived from the protruding (P) domain of the NoV VP1 capsid protein. P particles can be easily produced in
Escherichia coli
at high yields and thus may be more economically viable than the virus-like particle (VLP) vaccine. This study demonstrated, for the first time, the cross-variant protection (46.7%) of the intranasal P particle vaccine against human NoV diarrhea and revealed in detail the intestinal and systemic T cell responses by using the gnotobiotic pig model. The cross-variant protective efficacy of the P particle vaccine was comparable to that of the VLP vaccine in pigs (60%) and to the homologous protective efficacy of the VLP vaccine in humans (47%). NoV is now the leading cause of pediatric dehydrating diarrhea, responsible for approximately 1 million hospital visits for U.S. children and 218,000 deaths in developing countries. The P particle vaccine holds promise for reducing the disease burden and mortality.
The Herpes Simplex Virus 1 Virion Host Shutoff Protein Enhances Translation of Viral Late mRNAs by Preventing mRNA Overload
Authors: Dauber Bianca , Saffran Holly A. , Smiley James R. 
Published: June 2014

ABSTRACT

We recently demonstrated that the virion host shutoff (vhs) protein, an mRNA-specific endonuclease, is required for efficient herpes simplex virus 1 (HSV-1) replication and translation of viral true-late mRNAs, but not other viral and cellular mRNAs, in many cell types (B. Dauber, J. Pelletier, and J. R. Smiley, J. Virol. 85:5363–5373, 2011,
http://dx.doi.org/10.1128/JVI.00115-11
). Here, we evaluated whether the structure of true-late mRNAs or the timing of their transcription is responsible for the poor translation efficiency in the absence of vhs. To test whether the highly structured 5′ untranslated region (5′UTR) of the true-late gC mRNA is the primary obstacle for translation initiation, we replaced it with the less structured 5′UTR of the γ-actin mRNA. However, this mutation did not restore translation in the context of a vhs-deficient virus. We then examined whether the timing of transcription affects translation efficiency at late times. To this end, we engineered a vhs-deficient virus mutant that transcribes the true-late gene US11 with immediate-early kinetics (IEUS11-ΔSma). Interestingly, IEUS11-ΔSma showed increased translational activity on the US11 transcript at late times postinfection, and US11 protein levels were restored to wild-type levels. These results suggest that mRNAs can maintain translational activity throughout the late stage of infection if they are present before translation factors and/or ribosomes become limiting. Taken together, these results provide evidence that in the absence of the mRNA-destabilizing function of vhs, accumulation of viral mRNAs overwhelms the capacity of the host translational machinery, leading to functional exclusion of the last mRNAs that are made during infection.

IMPORTANCE
The process of mRNA translation accounts for a significant portion of a cell's energy consumption. To ensure efficient use of cellular resources, transcription, translation, and mRNA decay are tightly linked and highly regulated. However, during virus infection, the overall amount of mRNA may increase drastically, possibly overloading the capacity of the translation apparatus. Our results suggest that the HSV-1 vhs protein, an mRNA-specific endoribonuclease, prevents mRNA overload during infection, thereby allowing translation of late viral mRNAs. The requirement for vhs varies between cell types. Further studies of the basis for this difference likely will offer insights into how cells regulate overall mRNA levels and access to the translational apparatus.
STAT4 Deficiency Fails To Induce Lung Th2 or Th17 Immunity following Primary or Secondary Respiratory Syncytial Virus (RSV) Challenge but Enhances the Lung RSV-Specific CD8
+
T Cell Immune Response to Secondary Challenge
Authors: Dulek Daniel E. , Newcomb Dawn C. , Toki Shinji , Goliniewska Kasia , Cephus Jacqueline , Reiss Sara , Bates John T. , Crowe James E. , Boyd Kelli L. , Moore Martin L. , Zhou Weisong , Peebles R. Stokes 
Published: June 2014

ABSTRACT

Immune-mediated lung injury is a hallmark of lower respiratory tract illness caused by respiratory syncytial virus (RSV). STAT4 plays a critical role in CD4
+
Th1 lineage differentiation and gamma interferon (IFN-γ) protein expression by CD4
+
T cells. As CD4
+
Th1 differentiation is associated with negative regulation of CD4
+
Th2 and Th17 differentiation, we hypothesized that RSV infection of STAT4
−/−
mice would result in enhanced lung Th2 and Th17 inflammation and impaired lung Th1 inflammation compared to wild-type (WT) mice. We performed primary and secondary RSV challenges in WT and STAT4
−/−
mice and used STAT1
−/−
mice as a positive control for the development of RSV-specific lung Th2 and Th17 inflammation during primary challenge. Primary RSV challenge of STAT4
−/−
mice resulted in decreased T-bet and IFN-γ expression levels in CD4
+
T cells compared to those of WT mice. Lung Th2 and Th17 inflammation did not develop in primary RSV-challenged STAT4
−/−
mice. Decreased IFN-γ expression by NK cells, CD4
+
T cells, and CD8
+
T cells was associated with attenuated weight loss and enhanced viral clearance with primary challenge in STAT4
−/−
mice compared to WT mice. Following secondary challenge, WT and STAT4
−/−
mice also did not develop lung Th2 or Th17 inflammation. In contrast to primary challenge, secondary RSV challenge of STAT4
−/−
mice resulted in enhanced weight loss, an increased lung IFN-γ expression level, and an increased lung RSV-specific CD8
+
T cell response compared to those of WT mice. These data demonstrate that STAT4 regulates the RSV-specific CD8
+
T cell response to secondary infection but does not independently regulate lung Th2 or Th17 immune responses to RSV challenge.

IMPORTANCE
STAT4 is a protein critical for both innate and adaptive immune responses to viral infection. Our results show that STAT4 regulates the immune response to primary and secondary challenge with RSV but does not restrain RSV-induced lung Th2 or Th17 immune responses. These findings suggest that STAT4 expression may influence lung immunity and severity of illness following primary and secondary RSV infections.
A nationwide survey on the expectation of public healthcare providers on family medicine specialists in Malaysia—a qualitative analysis of 623 written comments
Authors: Chew Boon-How , Cheong Ai-Theng , Ismail Mastura , Hamzah Zuhra , A-Rashid Mohd-Radzniwan , Md-Yasin Mazapuspavina , Ali Norsiah 
Published: June 2014

ObjectiveTo examine the expectation of public healthcare providers/professionals (PHCPs) who are working closely with family medicine specialists (FMSs) at public health clinics.DesignCross-sectional study.SettingThis study is part of a larger national study on the perception of the Malaysian public healthcare professionals on FMSs.ParticipantsPHCPs from three categories of health facilities, namely hospitals, health clinics and health offices.Main outcome measuresQualitative analysis of written comments of respondents’ expectation of FMSs.ResultsThe participants’ response rate was 58% (780/1345) with an almost equal proportion from each public healthcare facility. We identified 21 subthemes for the 623 expectation comments. The six emerging themes are (1) need for more FMSs, (2) clinical roles and functions of FMSs, (3) administrative roles of FMSs, (4) contribution to community and public health, (5) attributes improvement and (6) research and audits. FMSs were expected to give attention to clinical duty. Delivering this responsibility with competence included having the latest medical knowledge in their own and others’ medical disciplines, practising evidence-based medicine in prehospital and posthospital care, better supervision of staff and doctors under their care, fostering effective teamwork, communicating more often with hospital specialists and making appropriate referral. Expectations ranged fromdefiniteandstrongfor more FMSs at the health clinics tolow expectationfor FMSs’ involvement in research; tomal-expectationon FMSs’ involvement in community and public health programmes.ConclusionsThere were some remarkable differences in expectations on FMSs from the three different PHCPs. These ranged from being clinically competent and administratively available for patients and staff at the health clinics, to mal-expectations on FMSs to engage in public health affairs. Relevant parties, including FMSs themselves, could take appropriate self-improvement initiatives to enhance public practice of family medicine and patient care.Trial registration numberNMRR ID: 08-12-1167.
Acute ischaemic stroke or transient ischaemic attack and the need for inpatient echocardiography
Authors: Menon Bijoy K , Coulter Jonathan I , Bal Simerpreet , Godzwon Catherine , Weeks Sarah , Hutchison Stuart , Hill Michael D , Coutts Shelagh B 
Published: June 2014

Abstract

Objectives
To determine the diagnostic yield of echocardiography and its utility in changing medical management; and to derive a risk score to guide its use in patients with in-hospital stroke or transient ischaemic attack (TIA).

Methods
We carried out a retrospective chart review from January 2009 to June 2010 of patients with acute ischaemic stroke or TIA who had undergone transthoracic echocardiography (TTE) or transoesophageal echocardiography (TOE). Clinical and imaging findings at baseline were noted and ‘potential clinically relevant findings’ identified on TTE and TOE. A multivariable logistic regression was used to identify predictors of potential clinically relevant findings on TTE or TOE and derive a risk score.

Results
Of 370 patients, 307 (83.0%) had TTE and 63 (17.0%) had additional TOE. Potential clinically relevant findings on echocardiography were noted in 28 (7.6%) patients. Change in medical management was noted in 19/307 (6.2%) patients on TTE and in 7/63 (11.1%) patients on TOE. Male sex (OR 3.05, 95% CI 1.19 to 7.84; p=0.021), abnormal admission ECG (OR 4.39, 95% CI 1.79 to 10.79; p=0.001), and embolic pattern imaging at baseline (OR 2.38, 95% CI 1.05 to 5.40; p=0.038) were independent predictors of findings on TTE or TOE. A risk score including these three variables had modest discrimination (c-statistic 0.69, 95% CI 0.59 to 0.80).

Conclusions
Echocardiography detected potential clinically relevant findings in a minority of patients (7.6%), but these findings changed medical management 90.5% of the time. A risk score using sex, ECG abnormality, and embolic pattern imaging at baseline could help predict which patients are more likely to have these echo findings.
Conjunctival bacterial and fungal flora in clinically normal sheep
Authors: Bonelli Francesca , Barsotti Giovanni , Attili Anna Rita , Mugnaini Linda , Cuteri Vincenzo , Preziuso Silvia , Corazza Michele , Preziuso Giovanna , Sgorbini Micaela 
Published: June 2014

ObjectivesThe aim was to identify conjunctival bacterial and fungal flora in clinically normal sheep.DesignProspective study.SettingTuscany.Participants100 eyes from 50 adult Massese female sheep were examined. The sheep included in the study were considered free of anterior ophthalmic abnormalities.Primary and secondary outcome measuresBacteria were identified by morphological assessment, Gram staining, biochemical tests. Identification of filamentous fungi was achieved at the genus level, andAspergillusspecies were identified based on keys provided by other authors. Yeast colonies were highlighted, but not identified.ResultsPositive cultures were obtained from 100/100 eyes for bacteria, and from 86/100 eyes for fungi. A total of 14 types of bacteria and 5 types of fungi were isolated. Yeasts were isolated from 13/100 eyes. The most frequent fungal isolates were saprophytic fungi.ConclusionsConjunctival bacterial and fungal flora of clinically normal eyes were reported in sheep. The positivity obtained for conjunctival bacteria was higher compared to findings in the literature by other authors in the same species (100 per cent v 40 per cent), while our results were in line with a recent work performed on mouflons (Ovis Musimon) with a 100 per cent positivity for bacterial conjunctival fornix. In our survey, Gram‐positive species were prevalent, as reported by other authors in different species. Few data are available in the literature regarding conjunctival fungal flora in healthy small ruminants. The prevalence of conjunctival fungal flora in this study was higher than findings reported in mouflons (86 per cent v 45 per cent). Differences in fungal prevalence may be due to different methods of managing herds, though further studies are required to verify this hypothesis. The similarities in bacterial and fungal isolates between sheep and mouflons suggest a genera pattern of conjunctival colonisation by bacteria and fungi.
Accurate ground-state variational wavefunction for the two-dimensional electron gas
Authors: Zhou Zhang-Wei , Sun Jiu-Xun , Khan Muhammad Ammar 
Published: June 2014

A variational wavefunction with two parameters is proposed for the two-dimensional electron gas (2DEG). The new wavefunction does not contain noninteger power of coordinate, and all integrals involving wavefunction can be analytically evaluated. This overcomes the shortcoming of the one proposed by Grinberg [ Phys. Rev. B 32, 4028 (1985)] which contains noninteger power of coordinate. The parameters have been determined by minimizing the average energy of an electron. The numerical results of energy and average separation of carriers for the ground state are in excellent agreement with the exact self-consistent (sc) results with average errors 0.21% and 0.57%. The accuracy is several times better than the wavefunction proposed of Grinberg with average errors 1.07% and 1.41%. The results also show that the triangular approximation for electric potential is correct as the density of 2DEG is low, but it deviates from the real situation as the density of 2DEG is high.
Correlation of Choroidal Thickness and Volume Measurements with Axial Length and Age Using Swept Source Optical Coherence Tomography and Optical Low-Coherence Reflectometry
Authors: Michalewski Janusz , Michalewska Zofia , Nawrocka Zofia , Bednarski Maciej , Nawrocki Jerzy 
Published: June 2014

Purpose. To report choroidal thickness and volume in healthy eyes using swept source optical coherence tomography (SS-OCT).Methods. A prospective observational study of 122 patients examined with swept source OCT (DRI-OCT, Topcon, Japan). In each eye, we performed 256 horizontal scans, 12 mm in length and centered on the fovea. We calculated choroidal thickness manually with a built-in caliper and automatically using DRI-OCT mapping software. Choroidal volume was also automatically calculated. We measured axial length with optical low-coherence reflectometry (Lenstar LS 900, Haag-Streit, Switzerland).Results. The choroid has focally increased thickness under the fovea. Choroid was thinnest in the outer nasal quadrant. In stepwise regression analysis, age was estimated as the most significant factor correlating with decreased choroidal thicknessF=23.146, P<0.001followed by axial lengthF=4.902, P=0.03. Refractive error was not statistically significantF=1.16, P=0.28.Conclusions. SS-OCT is the first commercially available system that can automatically create choroidal thickness and volume maps. Choroidal thickness is increased at the fovea and is thinnest nasally. Age and axial length are critical for the estimation of choroidal thickness and volume. Choroidal measurements derived from SS-OCT images have potential value for objectively documenting disease-related choroidal thickness abnormalities and monitoring progressive changes over time.
External Validation of a Multiplex Urinary Protein Panel for the Detection of Bladder Cancer in a Multicenter Cohort
Authors: Chen Li-Mei , Chang Myron , Dai Yunfeng , Chai Karl X. , Dyrskjøt Lars , Sanchez-Carbayo Marta , Szarvas Tibor , Zwarthoff Ellen C. , Lokeshwar Vinata , Jeronimo Carmen , Parker Alexander S. , Ross Shanti , Borre Michael , Ørntoft Torben F. , Jaeger Tobias , Beukers Willemien , Lopez Luis E. , Henrique Rui , Young Paul R. , Urquidi Virginia , Goodison Steve , Rosser Charles J. 
Published: June 2014

Abstract
Background: Because of the faltering sensitivity and/or specificity, urine-based assays currently have a limited role in the management of patients with bladder cancer. The aim of this study was to externally validate our previously reported protein biomarker panel from multiple sites in the United States and Europe.
Methods: This multicenter external validation study included a total of 320 subjects (bladder cancer = 183). The 10 biomarkers (IL8, MMP9, MMP10, SERPINA1, VEGFA, ANG, CA9, APOE, SDC1, and SERPINE1) were measured using commercial ELISA assays in an external laboratory. The diagnostic performance of the biomarker panel was assessed using receiver operator curves (ROC) and descriptive statistical values.
Results: Utilizing the combination of all 10 biomarkers, the area under the ROC for the diagnostic panel was noted to be 0.847 (95% confidence interval, 0.796–0.899), outperforming any single biomarker. The multiplex assay at optimal cutoff value achieved an overall sensitivity of 0.79, specificity of 0.79, positive prediction value of 0.73, and negative prediction value of 0.84 for bladder cancer classification. Sensitivity values of the diagnostic panel for high-grade bladder cancer, low-grade bladder cancer, muscle invasive bladder cancer, and non-muscle invasive bladder cancer were 0.81, 0.90, 0.95, and 0.77, respectively.
Conclusions: Urinary levels of the biomarker panel enabled discrimination of patients with bladder cancer and controls, and the levels of biomarker subsets were associated with advancing tumor grade and stage.
Impact: If proven to be reliable, urinary diagnostic biomarker assays can detect bladder cancer in a timely manner such that the patient can expect improvements in overall survival and quality of life. Cancer Epidemiol Biomarkers Prev; 23(9); 1804–12. ©2014 AACR.