bluuebunny/crossref_metadata_2025_split huggingface dataset:
Novel Less-Abundant Viral MicroRNAs Encoded by Herpes Simplex Virus 2 Latency-Associated Transcript and Their Roles in Regulating ICP34.5 and ICP0 mRNAs
Authors: Tang Shuang , Patel Amita , Krause Philip R. 
Published: November 2008

ABSTRACT
We recently identified an acutely and latently expressed viral microRNA (miRNA), miR-I, encoded by herpes simplex virus 2 (HSV-2) latency-associated transcript (LAT) through small RNA cloning and two miRNAs encoded by HSV-1 LAT through prediction. We now report the use of high-throughput sequencing technology to identify two additional relatively less-abundant viral miRNAs, miR-II and miR-III, encoded by HSV-2 LAT exon 2. miR-II includes two miRNAs, miR-II-5p and miR-II-3p, which are processed from the same miRNA precursor. miR-II and miR-III map antisense to the 5′ untranslated region of ICP34.5 and to the coding region of ICP0 exon 3, respectively. These novel miRNAs are conserved in different HSV-2 strains, and their presence in infected- and transfected-cell cultures was confirmed by Northern hybridization. All three HSV-2 LAT-encoded miRNAs map to genome locations similar to those of three out of four identified HSV-1 LAT-encoded miRNAs, but the sequences of these miRNAs are not conserved. The expression of LAT-encoded miRNAs is negatively regulated by ICP4, the major viral transactivator. We further show that, similar to miR-I, miR-II is able to efficiently silence the expression of ICP34.5, a key viral neurovirulence factor, and that miR-III is able to silence the expression of ICP0, a key viral transactivator. All these data suggest that LAT sequences likely contribute to HSV latency and reactivation through tight control of these LAT-encoded miRNAs and their viral targets.
Human Immunodeficiency Virus Type 2 (HIV-2)/HIV-1 Envelope Chimeras Detect High Titers of Broadly Reactive HIV-1 V3-Specific Antibodies in Human Plasma
Authors: Davis Katie L. , Bibollet-Ruche Frederic , Li Hui , Decker Julie M. , Kutsch Olaf , Morris Lynn , Salomon Aidy , Pinter Abraham , Hoxie James A. , Hahn Beatrice H. , Kwong Peter D. , Shaw George M. 
Published: November 2008

ABSTRACTDeciphering antibody specificities that constrain human immunodeficiency virus type 1 (HIV-1) envelope (Env) diversity, limit virus replication, and contribute to neutralization breadth and potency is an important goal of current HIV/AIDS vaccine research. Transplantation of discrete HIV-1 neutralizing epitopes into HIV-2 scaffolds may provide a sensitive, biologically functional context by which to quantify specific antibody reactivities even in complex sera. Here, we describe a novel HIV-2 proviral scaffold (pHIV-2KR.X7) into which we substituted the complete variable region 3 (V3) of theenvgene of HIV-1YU2or HIV-1Cconto yield the chimeric proviruses pHIV-2KR.X7YU2 V3 and pHIV-2KR.X7Ccon V3. These HIV-2/HIV-1 chimeras were replication competent and sensitive to selective pharmacological inhibitors of virus entry. V3 chimeric viruses were resistant to neutralization by HIV-1 monoclonal antibodies directed against the CD4 binding site, coreceptor binding site, and gp41 membrane proximal external region but exhibited striking sensitivity to HIV-1 V3-specific monoclonal antibodies, 447-52D and F425 B4e8 (50% inhibitory concentration of [IC50] <0.005 μg/ml for each). Plasma specimens from 11 HIV-1 clade B- and 10 HIV-1 clade C-infected subjects showed no neutralizing activity against HIV-2 but exhibited high-titer V3-specific neutralization against both HIV-2/HIV-1 V3 chimeras with IC50measurements ranging from 1:50 to greater than 1:40,000. Neutralization titers of B clade plasmas were as much as 1,000-fold lower when tested against the primary HIV-1YU2virus than with the HIV-2KR.X7YU2 V3 chimera, demonstrating highly effective shielding of V3 epitopes in the native Env trimer. This finding was replicated using a second primary HIV-1 strain (HIV-1BORI) and the corresponding HIV-2KR.X7BORI V3 chimera. We conclude that V3 is highly immunogenic in vivo, eliciting antibodies with substantial breadth of reactivity and neutralizing potential. These antibodies constrain HIV-1 Env to a structure(s) in which V3 epitopes are concealed prior to CD4 engagement but do not otherwise contribute to neutralization breadth and potency against most primary virus strains. Triggering of the viral spike to reveal V3 epitopes may be required if V3 immunogens are to be components of an effective HIV-1 vaccine.
Vaccinia Virus Extracellular Enveloped Virion Neutralization In Vitro and Protection In Vivo Depend on Complement
Authors: Benhnia Mohammed Rafii-El-Idrissi , McCausland Megan M. , Moyron Juan , Laudenslager John , Granger Steven , Rickert Sandra , Koriazova Lilia , Kubo Ralph , Kato Shinichiro , Crotty Shane 
Published: November 2008

ABSTRACTAntibody neutralization is an important component of protective immunity against vaccinia virus (VACV). Two distinct virion forms, mature virion and enveloped virion (MV and EV, respectively), possess separate functions and nonoverlapping immunological properties. In this study we examined the mechanics of EV neutralization, focusing on EV protein B5 (also called B5R). We show that neutralization of EV is predominantly complement dependent. From a panel of high-affinity anti-B5 monoclonal antibodies (MAbs), the only potent neutralizer in vitro (90% at 535 ng/ml) was an immunoglobulin G2a (IgG2a), and neutralization was complement mediated. This MAb was the most protective in vivo against lethal intranasal VACV challenge. Further studies demonstrated that in vivo depletion of complement caused a >50% loss of anti-B5 IgG2a protection, directly establishing the importance of complement for protection against the EV form. However, the mechanism of protection is not sterilizing immunity via elimination of the inoculum as the viral inoculum consisted of a purified MV form. The prevention of illness in vivo indicated rapid control of infection. We further demonstrate that antibody-mediated killing of VACV-infected cells expressing surface B5 is a second protective mechanism provided by complement-fixing anti-B5 IgG. Cell killing was very efficient, and this effector function was highly isotype specific. These results indicate that anti-B5 antibody-directed cell lysis via complement is a powerful mechanism for clearance of infected cells, keeping poxvirus-infected cells from being invisible to humoral immune responses. These findings highlight the importance of multiple mechanisms of antibody-mediated protection against VACV and point to key immunobiological differences between MVs and EVs that impact the outcome of infection.
Hepatitis C Virus Neuroinvasion: Identification of Infected Cells
Authors: Wilkinson Jeffrey , Radkowski Marek , Laskus Tomasz 
Published: November 2008

ABSTRACT
Hepatitis C virus (HCV) infection often is associated with cognitive dysfunction and depression. HCV sequences and replicative forms were detected in autopsy brain tissue and cerebrospinal fluid from infected patients, suggesting direct neuroinvasion. However, the phenotype of cells harboring HCV in brain remains unclear. We studied autopsy brain tissue from 12 HCV-infected patients, 6 of whom were coinfected with human immunodeficiency virus. Cryostat sections of frontal cortex and subcortical white matter were stained with monoclonal antibodies specific for microglia/macrophages (CD68), oligodendrocytes (2′,3′-cyclic nucleotide 3′-phosphodiesterase), astrocytes (glial fibrillary acidic protein [GFAP]), and neurons (neuronal-specific nuclear protein); separated by laser capture microscopy (LCM); and tested for the presence of positive- and negative-strand HCV RNA. Sections also were stained with antibodies to viral nonstructural protein 3 (NS3), separated by LCM, and phenotyped by real-time PCR. Finally, sections were double stained with antibodies specific for the cell phenotype and HCV NS3. HCV RNA was detected in CD68-positive cells in eight patients, and negative-strand HCV RNA, which is a viral replicative form, was found in three of these patients. HCV RNA also was found in astrocytes from three patients, but negative-strand RNA was not detected in these cells. In double immunostaining, 83 to 95% of cells positive for HCV NS3 also were CD68 positive, while 4 to 29% were GFAP positive. NS3-positive cells were negative for neuron and oligodendrocyte phenotypic markers. In conclusion, HCV infects brain microglia/macrophages and, to a lesser extent, astrocytes. Our findings could explain the biological basis of neurocognitive abnormalities in HCV infection.
Efficient Nuclear Export of Herpes Simplex Virus 1 Transcripts Requires both RNA Binding by ICP27 and ICP27 Interaction with TAP/NXF1
Authors: Johnson Lisa A. , Sandri-Goldin Rozanne M. 
Published: November 2008

ABSTRACT

Herpes simplex virus 1 (HSV-1) regulatory protein ICP27 has been reported to bind viral RNA and to interact with the nuclear export adaptor Aly/REF and the major cellular mRNA nuclear export receptor TAP/NXF1. Using in situ hybridization and in vitro export assays, we show here that poly(A)
+
RNA was retained in the nucleus of cells infected with viral ICP27 mutants that either cannot bind RNA or that do not interact with TAP/NXF1. Microarray analysis of nuclear and cytoplasmic RNA fractions demonstrated that efficient export of the majority of viral transcripts requires that ICP27 be able to bind RNA and to interact with TAP/NXF1. We conclude that ICP27 is the major export adaptor for HSV-1 mRNA and that it links bound transcripts to the TAP/NXF1 export receptor.
The C-Terminal Domain of ERp29 Mediates Polyomavirus Binding, Unfolding, and Infection
Authors: Rainey-Barger Emily K. , Mkrtchian Souren , Tsai Billy 
Published: November 2008

ABSTRACT
Penetration of the endoplasmic reticulum (ER) membrane by polyomavirus (PyV) is a decisive step in virus entry. We showed previously that the ER-resident factor ERp29 induces the local unfolding of PyV to initiate the ER membrane penetration process. ERp29 contains an N-terminal thioredoxin domain (NTD) that mediates its dimerization and a novel C-terminal all-helical domain (CTD) whose function is unclear. The NTD-mediated dimerization of ERp29 is critical for its unfolding activity; whether the CTD plays any role in PyV unfolding is unknown. We now show that three hydrophobic residues within the last helix of the ERp29 CTD that were individually mutated to either lysine or alanine abolished ERp29's ability to stimulate PyV unfolding and infection. This effect was not due to global misfolding of the mutant proteins, as they dimerize and do not form aggregates or display increased protease sensitivity. Moreover, the mutant proteins stimulated secretion of the secretory protein thyroglobulin with an efficiency similar to that of wild-type ERp29. Using a cross-linking coimmunoprecipitation assay, we found that the physical interaction of the ERp29 CTD mutants with PyV is inefficient. Our data thus demonstrate that the ERp29 CTD plays a crucial role in PyV unfolding and infection, likely by serving as part of a substrate-binding domain.
In Vivo Fitness Cost of the M184V Mutation in Multidrug-Resistant Human Immunodeficiency Virus Type 1 in the Absence of Lamivudine
Authors: Paredes Roger , Sagar Manish , Marconi Vincent C. , Hoh Rebecca , Martin Jeffrey N. , Parkin Neil T. , Petropoulos Christos J. , Deeks Steven G. , Kuritzkes Daniel R. 
Published: November 2008

ABSTRACT

Lamivudine therapy selects for the M184V mutation. Although this mutation reduces the replicative capacity of human immunodeficiency virus in vitro, its impact on viral fitness in vivo has not been well defined. We used quantitative allele-specific PCR to precisely calculate the fitness differences between the mutated M184V virus and one that had reverted to the wild type in a cohort of patients by selectively interrupting reverse transcriptase inhibitor therapy, and we found that the M184V variants were consistently 4 to 8% less fit than the wild type in the absence of drug. After a lag phase of variable duration, wild-type variants emerged due to continued evolution of
pol
and back mutation rather than through emergence of an archived wild-type variant.
Inhibitory effect of rapamycin and dexamethasone on production of IL-17 and IFN-γ in Vogt–Koyanagi–Harada patients
Authors: Yang K , Wen J , Liu X , Kijlstra A , Chen L , Chi W , Zhou H , Huang X , Yang P 
Published: November 2008

Aims:To evaluate the effect of rapamycin (RAPA) and dexamethasone (DEX) on the production of IL-17 and IFN-γ by peripheral blood mononuclear cells (PBMCs) from Vogt–Koyanagi–Harada (VKH) patients and healthy individuals.Methods:Blood samples were drawn from 10 active VKH patients and 10 healthy individuals. PBMCs were cultured with or without anti-CD3 and anti-CD28 antibodies in the presence or absence of different concentrations of RAPA or DEX for 72 h. IL-17 and IFN-γ concentrations in the supernatants were measured by enzyme-linked immunosorbent assay (ELISA).Results:The expression of IL-17 and IFN-γ was significantly increased in active VKH patients compared with that in healthy controls. Both RAPA and DEX were able to significantly inhibit the production of IL-17 and IFN-γ by PBMCs from patients and healthy controls. RAPA was able to completely inhibit IL-17 production at a dosage of 10 ng/ml but only partially suppressed IFN-γ production even at a much higher concentration (1000 ng/ml). DEX inhibited the production of both IL-17 and IFN-γ by approximately 70%.Conclusions:This study indicates that both RAPA and DEX inhibit the production of IL-17 and IFN-γ by PBMCs. RAPA is much stronger in inhibiting the production of IL-17 than DEX.
Predictive index to differentiate invasive squamous cell carcinoma from preinvasive ocular surface lesions by impression cytology
Authors: Barros J N , Lowen M S , Ballalai P L , Mascaro V L D M , Gomes J Á P , Martins M C 
Published: November 2008

Background/aims:In the literature, no cytological features have been identified that reliably differentiate invasive squamous cell carcinoma (SCC) from preinvasive lesions in impression cytology (IC) samples. The aim was to identify cytological features related to malignancy and apply them in a quantitative model to determine an index score with the best predictive power to differentiate SCC from preinvasive ocular surface lesions by IC.Methods:39 patients with ocular surface epithelial lesions were enrolled. IC was obtained from all lesions before surgical excision. Specimens with atypical cells were evaluated regarding 11 cytological parameters based on the 2001 Bethesda system.Results:Histopathological diagnosis was pterygium in one case, actinic keratosis in nine cases, intraepithelial neoplasia in nine cases and SCC in 20 cases. Analysis of the receiver operating characteristic curve revealed that a predictive index score (cut-off point) ⩾4.25 presented the best relationship between sensitivity and specificity in identifying SCC (sensitivity of 95%, specificity of 93%, positive predictive value of 95% and negative predictive value of 93%).Conclusion:The scoring system model presented is suitable for clinical practice in differentiating SCC from preinvasive ocular surface lesions by IC and can be better evaluated with prospective use.
A prospective study of punch biomechanics and cognitive function for amateur boxers
Authors: Stojsih S , Boitano M , Wilhelm M , Bir C 
Published: November 2008

ObjectiveTo evaluate several biomechanical factors of the head during a sparring session and their link to cognitive function.DesignInstrumented Boxing Headgear (IBH) was used for data collection during four 2 min sparring sessions. Neurocognitive assessment was measured using the ImPACT© Concussion management software. A baseline neurocognitive test was obtained from each athlete prior to sparring; two additional tests were obtained and compared with the baseline.SettingMale and female amateur boxers.ParticipantsData were collected from 30 male and 30 female amateur boxers.Main outcome measurementsHead accelerations (translational and rotational), injury severity indexes (Head Injury Criteria (HIC) and Gadd Severity Index (GSI)) and cognitive function scores.ResultsPeak translational and rotational acceleration values were 191gand 17 156 rad/s2, respectively, for males and 184gand 13 113 rad/s2, respectively, for females. The peak HIC and GSI values for males were 1652 and 2292, respectively, and for females 1079 and 1487, respectively. There was no significant difference in the neurocognitive scores between genders. A decrease was exhibited in the delayed memory postbout scores. All other scores either increased or did not significantly decrease when compared with the baseline.ConclusionsThe majority of impacts experienced by both genders were under the threshold for mild head injury. There was a statistically significant difference between peak translational and rotational acceleration, HIC and GSI when comparing genders. When analysing cognitive functions, there was no statistical difference between genders.
The effects of acute dynamic exercise on haemostasis in first class Scottish football referees
Authors: Peat E E , Dawson M , McKenzie A , Hillis W S 
Published: November 2008

PurposePhysical fitness may confer protection from thrombosis; however, exercise-induced platelet activation may be involved in the triggering of acute vascular events. This study aimed to assess haemostatic responses to acute exercise in trained and sedentary middle-aged subjects.Methods21 first class Scottish football referees and 15 sedentary controls performed a treadmill exercise test. Blood sampling was performed before, immediately after and 30 minutes post-exercise. Samples were analysed for platelet count, prothrombin time, activated partial thromboplastin time (APTT) and serum fibrinogen. Platelet activation was assessed using flow cytometry with CD62 (P-selectin) and antifibrinogen antibodies at rest and in response to ADP and epinephrine.ResultsMean maximal oxygen consumption (Vo2) (ml/kg per minute) achieved was 47.23 (5.02) in the referees and 30.1 (5.2) in sedentary controls. Total platelet count (×10−9/l) increased immediately post-exercise (228.2 (40.5), 278.6 (48.9) p=0.001) remaining elevated at 30 minutes in both groups. APTT (s) was reduced immediately post-exercise (32.15 (3.1), 29.7 (3.94) p=0.001) with a further reduction seen at 30 minutes (32.15 (3.1), 28.4 (3.31) p=0.001). In the referees, percentage CD62 expression increased immediately post-exercise (0.688 (0.52), 1.42 (1.3) p=0.008). Percentage antifibrinogen expression increased post-exercise (5.19 (4.31), 13.01 (14.24) p=0.017), with a further increase at 30 minutes (5.19 (4.31), 20.47 (26.8) p=0.02). Similar trends were seen in sedentary controls.ConclusionThis study suggests that in an older athletic population, physical fitness does not protect against the prothrombotic effects of exercise. These data suggest that during a football match when referees achieve approximately 80% of peak Vo2(23) they may be at risk of significant platelet activation. Prophylactic platelet inhibition should be considered in this group after appropriate screening and risk stratification.
Promoting physical activity in children: the stepwise development of the primary school-based JUMP-in intervention applying the RE-AIM evaluation framework
Authors: De Meij J S B , Chinapaw M J M , Kremers S P J , Van der wal M F , Jurg M E , Van Mechelen W 
Published: November 2008

BackgroundThere is a lack of effective intervention strategies that promote physical activity (PA) in school children. Furthermore, there is a gap between PA intervention research and the delivery of programmes in practice. Evaluation studies seldom lead to adaptations in interventions that are subsequently evaluated by implementation on a wider scale. The stepwise development and study of JUMP-in aims to add knowledge to better understand how, when and for whom intervention effects (or lack of effects) occur.MethodsThis paper describes the stepwise development of JUMP-in, a Dutch school-based multi-level intervention programme, aimed at the promotion of PA behaviour in 6–12-year-old children. JUMP-in incorporates education, sports, care and policy components. JUMP-in consists of six programme components:Pupil Follow-up Monitoring System;School sports clubs;In-class exercises with “The Class Moves!”;Personal workbook “This is the way you move!”;Parental Information Services;Extra lessons in physical education, Motor RemedialTeaching and extra care.The process and effect outcomes of a pilot study were translated into an improved programme and intervention organisation, using the RE-AIM framework (Reach, Efficacy, Adoption, Implementation and Maintenance).This paper presents the process and results of the application of this framework, which resulted in a widescale implementation of JUMP-in.ResultsThe application of the RE-AIM framework resulted in challenges and remedies for an improved JUMP-in intervention. The remedies required changes at three different levels: (1) the content of the programme components; (2) the organisation and programme management; and (3) the evaluation design.ConclusionsConsidering factors that determine the impact of PA interventions in ‘real life’ is of great importance. The RE-AIM framework appeared to be a useful guide by which process and effect outcomes could be translated into an improved programme content and organisation.
Downscaling Climate over Complex Terrain: High Finescale (&lt;1000 m) Spatial Variation of Near-Ground Temperatures in a Montane Forested Landscape (Great Smoky Mountains)*
Authors: Fridley Jason D. 
Published: November 2008

AbstractLandscape-driven microclimates in mountainous terrain pose significant obstacles to predicting the response of organisms to atmospheric warming, but few if any studies have documented the extent of such finescale variation over large regions. This paper demonstrates that ground-level temperature regimes in Great Smoky Mountains National Park (Tennessee and North Carolina) vary considerably over fine spatial scales and are only partially linked to synoptic weather patterns and environmental lapse rates. A 120-sensor network deployed across two watersheds in 2005–06 exhibited finescale (&lt;1000-m extent) temperature differences of over 2°C for daily minima and over 4°C for daily maxima. Landscape controls over minimum temperatures were associated with finescale patterns of soil moisture content, and maximum temperatures were associated with finescale insolation differences caused by topographic exposure and vegetation cover. By linking the sensor array data to 10 regional weather stations and topographic variables describing site radiation load and moisture content, multilevel spatial models of 30-m resolution were constructed to map daily temperatures across the 2090-km2 park, validated with an independent 50-sensor network. Maps reveal that different landscape positions do not maintain relative differences in temperature regimes across seasons. Near-stream locations are warmer in the winter and cooler in the summer, and sites of low elevation more closely track synoptic weather patterns than do wetter high-elevation sites. This study suggests a strong interplay between near-ground heat and water balances and indicates that the influence of past and future shifts in regional temperatures on the park’s biota may be buffered by soil moisture surfeits from high regional rainfall.