bluuebunny/crossref_metadata_2025 huggingface dataset:
Public Trusts
Authors: Barratt John
Published: June 2006

Public, as distinct from charitable, trusts were authoritatively recognised in 1827, when the House of Lords determined that the permanent basis of equitable jurisdiction protected ‘public money’ from unlawful application, on the same basis as charitable funds were protected. These trusts made the relevant public functionaries subject to equitable remedies, but the remedies were displaced from equitable practice by jurisdictional fusion and by administrative audit surcharges. In the twentieth century they survived unrecognised in the ambiguously phrased ‘local authority fiduciary duty to ratepayers’. Recently, public trusts have again been implicitly acknowledged, and the distinctive remedy of personal liability to make good loss arising from misapplication has been applied. However the right to litigate is confined to current fund‐holders in fulfilment of their protective trust duty and to the Attorney General in the general public interest in countering unlawful appropriations of public money.
Mortgagee Powers Rhetoric
Authors: Conaglen Matthew
Published: June 2006

This article addresses the rhetoric of land law's interpretive community at the time of, and following, the House of Lords' decision in Boland. It is argued, contrary to the views of other commentators, that the decision was not strongly motivated by a concern to see social justice done between commercial lenders and wives but, rather, was a sensible and socially realistic application of clear statutory provisions understood within the context of land law orthodoxy. Furthermore, decisions following Boland which have been said to have sapped its strength also cohere comfortably within that community's stable rhetoric. Consequently, it cannot be argued that proportionality in human rights law requires a return to the supposed ‘high point’ of Boland– when considered carefully, the evidence, both from Boland itself and from land law's wider interpretive community, shows that such a high point did not exist.
Feminist praxis in a trade union gender project
Authors: Ledwith Sue
Published: June 2006

Research as political feminist engagement is explored through analysis of a collaborative west–east European gender equality project with trade union women. Similarities with and differences between forms of labourist and feminist research praxis are discussed. These are brought together through the Freirian concept of conscientizacao or conscientisation, a dialogic pedagogy which has at its core the dialectic of reflection and action. It is proposed that such a framework can be usefully deployed for gendering analysis of ‘engaged’ industrial relations.
Functional map and age‐related differences in the human face: nonimmunologic contact urticaria induced by hexyl nicotinate
Authors: Marrakchi Slaheddine , Maibach Howard I.
Published: June 2006

Variation in human skin reactivity to various irritants in association with age and body region has been reported. Hexyl nicotinate (HN), a lipophilic nicotinate ester, was used to induce nonimmunologic contact urticaria in human volunteers of 2 age groups: 10 young subjects [24–34 years, mean ± standard deviation (SD) 29.8 ± 3.9 years] and 10 older volunteers (66–83 years, mean ± SD 73.6 ± 17.4 years); and to define skin function and potential age‐related differences in various facial areas. About 5 mM of HN in ethanol was applied to 8 locations on the face, neck, and volar forearm. A laser Doppler flowmeter was used to determine baseline blood flow and to monitor the skin blood flow changes after HN application. In the contralateral areas, stratum corneum turnover was determined using 5% dansyl chloride in petrolatum. In the young group, the perioral area exhibited the strongest reaction to HN. In the older group, the chin was the most sensitive site. In both the groups, the forearm was the least responsive. The older group demonstrated a stronger reaction than the younger group in 3 sites (forehead, cheek, and nasolabial area). Stratum corneum turnover was slower in the nasolabial area and in the forearm in both age groups, whereas the fastest was in the perioral area and the chin in the younger group and in the chin and the forehead in the older group. Compared to the older group, the younger group showed a slower stratum corneum turnover in the nose and the neck. This study demonstrates the regional and the age‐related variability of the stratum corneum turnover and the skin reactions to HN. These observations may help explain some aspects of the cutaneous intolerance in skin care of the face.
Effectiveness of Chinese Teams: The Role of Conflict Types and Conflict Management Approaches
Authors: Tjosvold Dean , Law Kenneth S. , Sun Haifa
Published: June 2006

Findings based on 186 teams involving 689 employees, working in twelve Chinese state-owned factories in three cities, indicated that a cooperative in contrast to a competitive approach was related to perceived team effectiveness, as measured by both team managers and team members. The role of conflict types for team effectiveness, on the other hand, is ambiguous. Furthermore, conflict management approaches affect team perceptions of relational and task conflict. Results suggest that a cooperative conflict management approach may be equally useful for Chinese work teams, as it is for teams in the Western context.
Cultural Boundary of Expectancy Theory-Based Performance Management: A Commentary on DeNisi and Pritchard's Performance Improvement Model
Authors: Zhang Kai , Song Lynda Jiwen , Hackett Rick D. , Bycio Peter
Published: June 2006

We consider Chinese cultural, psychological and social factors that are likely to place boundaries on the applicability of DeNisi and Pritchard's motivational framework and discuss how these cultural barriers play out at each of the links in their model. We conclude with a summary and a discussion of some of the broader implications of applying models of work motivation to a Chinese economy in transition.
Comparative pharmacokinetics of Panadol Extend and immediate‐release paracetamol in a simulated overdose model
Authors: Tan Clifford , Graudins Andis
Published: June 2006

Background:  Panadol Extend is a modified‐release paracetamol formulation in which each 665 mg tablet contains 69% slow‐release and 31% immediate‐release paracetamol. There are no data on Panadol Extend pharmacokinetics in overdose. It is unknown whether the paracetamol treatment nomogram can be used to make decisions regarding the toxicity of this product in overdose.

Objective:  To compare the pharmacokinetics of Panadol Extend and immediate‐release paracetamol (APAP‐IR) in simulated overdose model in healthy volunteers.

Methods:  Cross‐over study using a dose of 90 mg/kg ideal body weight of Panadol Extend or APAP‐IR in seven healthy volunteers. Serum paracetamol concentrations were measured over 12 h. Maximal paracetamol concentration (Cmax), time to Cmax (Tmax), area under the curve (AUC) and elimination half‐life (t1/2) were compared.

Results:  Mean paracetamol dose was 73 mg/kg actual body weight. Panadol Extend produced lower Cmax (0.208 mmol/L ± 0.02 vs 0.48 mmol/L ± 0.02, P = 0.0001) and AUC0−12 h when compared with APAP‐IR. Tmax was delayed with Panadol Extend (2.83 h ± 0.26 vs 0.94 h ± 0.17, P = 0.0001). Absorption was complete in all subjects by 4 h post ingestion in both study arms. Elimination t1/2 was 1.69 ± 0.09 h for APAP‐IR and 1.65 ± 0.13 h for Panadol Extend (not significant).

Conclusions:  Reductions in Panadol Extend Cmax and AUC0−12 h might be related to elimination occurring during the absorption phase. In this model of Panadol Extend moderate overdose, Tmax was significantly delayed. In larger overdoses, time to peak paracetamol levels might be further delayed, because of continuing absorption from the formulation. Therefore, the paracetamol treatment nomogram might not reliably predict hepatotoxicity from Panadol Extend if paracetamol levels are measured too early.
Airway: Australian Resuscitation Council Guideline 2006
Authors:   Australian Resuscitation Council
Published: June 2006

This guideline is applicable to adults, children and infants.

4 Airway
Breathing: Australian Resuscitation Council Guideline 2006
Authors:   Australian Resuscitation Council
Published: June 2006

This guideline is applicable to adults, children and infants.

5 Breathing
Rural‐Urban Disparities in Health‐Related Quality of Life Within Disease Categories of Veterans
Authors: Weeks William B. , Wallace Amy E. , Wang Stanley , Lee Austin , Kazis Lewis E.
Published: June 2006

ABSTRACT: 
Context: Compared to their urban counterparts, rural veterans have been found to have lower health‐related quality of life. Purpose: To determine whether these disparities persist when examining disease categories of rural and urban veterans. Methods: We obtained survey data on 748,216 veterans who were current or anticipated Veterans Health Administration patients. Using International Classification of Diseases (ICD)‐9CM codes, we determined whether these veterans had diagnoses that fell into any of 30 physical health disease categories, and we used ZIP codes to determine whether veterans lived in rural or urban settings. We compared rural to urban prevalence of disease categories as well as urban to rural health‐related quality‐of‐life physical health component summary scores (PCS) and mental health component summary scores (MCS) for each disease category. Findings: Physical diagnoses were significantly more prevalent in the rural veteran population for most disease categories examined. For every disease category examined, PCS were significantly lower for veterans who lived in rural, compared to urban, settings (P < .001 for all); rural veterans also experienced lower MCS for all disease categories although differences were modest. Differences persisted after controlling for sociodemographic factors. Conclusions: Compared to the urban veteran population, within disease categories, the rural veteran population experiences higher disease prevalence and lower physical and mental quality‐of‐life scores. Policymakers should anticipate greater health care demands from the rural veteran population and work to meet that demand.
The Role of Rural Health Clinics in Hospitalization due to Ambulatory Care Sensitive Conditions: A Study in Nebraska
Authors: Zhang Wanqing , Mueller Keith J. , Chen Li‐Wu , Conway Kevin
Published: June 2006

ABSTRACT: 
Context: Hospitalization due to ambulatory care sensitive conditions (ACSCs) is often used as an indicator for measuring access to primary care. Rural health clinics (RHCs) provide basic primary care services for rural residents in health professional shortage areas (HPSAs). The relationship between RHCs and ACSCs is unclear. Purpose: The purpose of this study was to examine the relationship between the presence of RHCs in rural HPSAs and the likelihood of having an acute or chronic ACSC as the reason for hospitalization. Methods: Nebraska hospital discharge data (1999‐2001) and the 2003 Area Resource File were used in this analysis. A multilevel logistic regression analysis was used to examine the relationship between the presence of RHCs in rural HPSAs and the likelihood of having an ACSC as the reason for hospitalization, after controlling for individual characteristics and county‐level contextual factors stratified by 3 age groups. The eligibility for logistic regression was limited to patients from 28 rural Nebraska counties designated as HPSAs in 2001. Patients with commercial payers were excluded from the study. Findings: Elderly patients residing in rural Nebraska HPSAs with at least one RHC were significantly less likely to have a hospitalization due to chronic ACSCs. Conclusions: The presence of RHC is a significant factor associated with fewer hospitalizations for chronic ACSCs among the rural elderly residing in HPSAs.
Financial Indicators for Critical Access Hospitals
Authors: Pink George H. , Holmes G. Mark , D’Alpe Cameron , Strunk Lindsay A. , McGee Patrick , Slifkin Rebecca T.
Published: June 2006

ABSTRACT: 
Context: There is a growing recognition of the need to measure and report hospital financial performance. However, there exists little comparative financial indicator data specifically for critical access hospitals (CAHs). CAHs differ from other hospitals on a number of dimensions that might affect appropriate indicators of performance, including differences in Medicare reimbursement, limits on bed size and average length of stay, and relaxed staffing rules. Purpose: To develop comparative financial indicators specifically designed for CAHs using Medicare cost report data. Methods: A technical advisory group of individuals with extensive experience in rural hospital finance and operations provided advice to a research team from the University of North Carolina at Chapel Hill. Twenty indicators deemed appropriate for assessment of CAH financial condition were chosen and formulas determined. Issues 1 and 2 of the CAH Financial Indicators Report were mailed to the chief executive officers of 853 CAHs in the summer of 2004 and 1,092 CAHs in the summer of 2005, respectively. Each report included indicator values specifically for their CAH, indicator medians for peer groups, and an evaluation form. Findings: Chief executive officers found the indicators to be useful and the underlying formulas to be appropriate. The multiple years of data provide snapshots of the industry as a whole, rather than trend data for a constant set of hospitals. Conclusions: The CAH Financial Indicators Report is a useful first step toward comparative financial indicators for CAHs.
Urban‐Rural Differences in the Availability of Hospital Information Technology Applications: A Survey of Georgia Hospitals
Authors: Culler Steven D , Atherly Adam , Walczak Sandra , Davis Anne , Hawley Jonathan N. , Rask Kimberly J. , Naylor Vi , Thorpe Kenneth E.
Published: June 2006

ABSTRACT: 
Context: Information technology (IT) has been identified as a potential tool for improving the safety of health care delivery. Purpose: To determine if there are significant differences between urban and rural community hospitals in the availability of selected IT functional applications and technological devices. Methods: A mailed survey of community hospitals in Georgia assessing the current availability of IT applications (54.6% response rate). Findings: Georgia hospitals reported having 63.6% of 56 possible functional applications computerized but only 52.9% of 41 technological devices. Compared to rural hospitals, urban hospitals had significantly more functional applications computerized (38.0 vs 31.8, P = .031) and technological devices available (23.9 vs 18.2, P = .016). Urban hospitals had significantly (P < .05) more IT applications available in 4 areas: emergency room services (7 of 10), surgical/operating room (8 of 12), laboratory (7 of 12), and radiology (5 of 11). Overall, the availability of IT applications was bimodal in rural hospitals: over 40% of rural hospitals had adopted over 70% of all applications, while approximately 26% of rural hospitals had adopted less than 30%. Conclusions: Some of the observed urban/rural differences in availability of IT applications may be due to differences in the scope of services provided by rural hospitals, in particular laboratories, radiology departments, emergency rooms, and surgery/operating rooms. Nevertheless, the bimodal distribution of IT applications adopted in rural hospitals raises concerns about the ability of selected rural hospitals to take advantage of regional data‐sharing initiatives and maintain quality of patient care in the future.
Characteristics of Rural and Urban Cadaveric Organ Transplant Donors and Recipients
Authors: Weeks William B. , Lushkov Gili , Nelson William A. , Wallace Amy E.
Published: June 2006

ABSTRACT: 
Context: Health disparities have been found when comparing rural and urban populations. Purpose: To compare characteristics of rural and urban cadaveric transplant donors and recipients. Methods: We used deidentified individual‐level data on 55,929 cadaveric transplant donor‐recipient exchanges between 2000 and 2003 and examined the relative rates of donating and receiving cadaveric transplants for rural compared to urban residents, as defined by ZIP Codes. Findings: When compared to their urban counterparts, rural organ donors were more likely to have died from head trauma, drowning, motor vehicle accidents, or suicide and less likely to have died from cerebrovascular events, cardiac events, or homicide (P < .001 for all). Rural transplant recipients had lower levels of educational attainment and were less likely to have had the transplant financed by private insurance (P < .001 for all). While we found no statistical difference in days wait to organ transplantation, rural residents were more likely to donate than to receive cadaveric organs (P < .001). Conclusions: The differences in organ donation that we found warrant further exploration.
Dental Services for Migrant and Seasonal Farmworkers in US Community/Migrant Health Centers
Authors: Lukes Sherri M. , Simon Bret
Published: June 2006

ABSTRACT: 
Context: Migrant and seasonal farmworkers are recognized as a medically underserved population, yet little information on need, access, and services is available—particularly with regard to oral health care. Purpose: This study describes the facilities, services, staffing, and patient characteristics of US dental clinics serving migrant and seasonal farmworkers, and identifies trends and issues that may impede or improve dental care access and service. Methods: National databases were used to identify community and migrant health centers providing oral health care to migrant and seasonal farmworkers. Mailed surveys collected information on clinic history, operational details, services provided, patient demographics, employment and resource needs, and perceived barriers to care. Findings: Among the 81 respondents (response rate 41%), hours of operation varied from 1 evening a week to more than 40 hours a week; 52% had no evening hours. Almost all the clinics offered preventive, diagnostic, and basic restorative dental services, and roughly two thirds also offered complex restorative services. Patients most frequently sought emergency dental care (44%) followed by basic restorative services (32%) and preventive services (26%). The dentist position was the most difficult to fill, and new funding sources were cited as the most important resource need. Respondents perceived cost of services, lack of transportation, and limited clinic hours as primary barriers to care. Conclusions: While some barriers to care have been almost universally addressed (eg, language), there is evidence that some impediments remain and may present significant obstacles to a broad improvement in oral health care for migrant and seasonal farmworkers.
Hepatitis C Virus Entry Depends on Clathrin-Mediated Endocytosis
Authors: Blanchard Emmanuelle , Belouzard Sandrine , Goueslain Lucie , Wakita Takaji , Dubuisson Jean , Wychowski Czeslaw , Rouillé Yves
Published: June 2006

Due to difficulties in cell culture propagation, the mechanisms of hepatitis C virus (HCV) entry are poorly understood. Here, postbinding cellular mechanisms of HCV entry were studied using both retroviral particles pseudotyped with HCV envelope glycoproteins (HCVpp) and the HCV clone JFH-1 propagated in cell culture (HCVcc). HCVpp entry was measured by quantitative real-time PCR after 3 h of contact with target cells, and HCVcc infection was quantified by immunoblot analysis and immunofluorescence detection of HCV proteins expressed in infected cells. The functional role of clathrin-mediated endocytosis in HCV entry was assessed by small interfering RNA-mediated clathrin heavy chain depletion and with chlorpromazine, an inhibitor of clathrin-coated pit formation at the plasma membrane. In both conditions, HCVpp entry and HCVcc infection were inhibited. HCVcc infection was also inhibited by pretreating target cells with bafilomycin A1 or chloroquine, two drugs known to interfere with endosome acidification. These data indicate that HCV enters target cells by clathrin-mediated endocytosis, followed by a fusion step from within an acidic endosomal compartment.
Heparan Sulfate Proteoglycan Binding Properties of Adeno-Associated Virus Retargeting Mutants and Consequences for Their In Vivo Tropism
Authors: Perabo Luca , Goldnau Daniela , White Kathryn , Endell Jan , Boucas Jorge , Humme Sibille , Work Lorraine M. , Janicki Hanna , Hallek Michael , Baker Andrew H. , Büning Hildegard
Published: June 2006

Adeno-associated virus type 2 (AAV-2) targeting vectors have been generated by insertion of ligand peptides into the viral capsid at amino acid position 587. This procedure ablates binding of heparan sulfate proteoglycan (HSPG), AAV-2's primary receptor, in some but not all mutants. Using an AAV-2 display library, we investigated molecular mechanisms responsible for this phenotype, demonstrating that peptides containing a net negative charge are prone to confer an HSPG nonbinding phenotype. Interestingly, in vivo studies correlated the inability to bind to HSPG with liver and spleen detargeting in mice after systemic application, suggesting several strategies to improve efficiency of AAV-2 retargeting to alternative tissues.
Sequestration and Protection of Double-Stranded RNA by the Betanodavirus B2 Protein
Authors: Fenner Beau J. , Goh Winnie , Kwang Jimmy
Published: June 2006

Betanodavirus B2 belongs to a group of functionally related proteins from the sense-strand RNA virus family
Nodaviridae
that suppress cellular RNA interference. The B2 proteins of insect alphanodaviruses block RNA interference by binding to double-stranded RNA (dsRNA), thus preventing Dicer-mediated cleavage and the subsequent generation of short interfering RNAs. We show here that the fish betanodavirus B2 protein also binds dsRNA. Binding is sequence independent, and maximal binding occurs with dsRNA substrates greater than 20 bp in length. The binding of B2 to long dsRNA is sufficient to completely block Dicer cleavage of dsRNA in vitro. Protein-protein interaction studies indicated that B2 interacts with itself and with other dsRNA binding proteins, the interaction occurring through binding to shared dsRNA substrates. Induction of the dsRNA-dependent interferon response was not antagonized by B2, as the interferon-responsive
Mx
gene of permissive fish cells was induced by wild-type viral RNA1 but not by a B2 mutant. The induction of
Mx
instead relied solely on viral RNA1 accumulation, which is impaired in the B2 mutant. Hyperediting of virus dsRNA and site-specific editing of 5-HT2C mRNA were both antagonized by B2. RNA editing was not, however, observed in transfected wild-type or B2 mutant RNA1, suggesting that this pathway does not contribute to the RNA1 accumulation defect of the B2 mutant. We thus conclude that betanodavirus B2 is a dsRNA binding protein that sequesters and protects both long and short dsRNAs to protect betanodavirus from cellular RNA interference.
TORC1 and TORC2 Coactivators Are Required for Tax Activation of the Human T-Cell Leukemia Virus Type 1 Long Terminal Repeats
Authors: Siu Yeung-Tung , Chin King-Tung , Siu Kam-Leung , Yee Wai Choy Elizabeth , Jeang Kuan-Teh , Jin Dong-Yan
Published: June 2006

Human T-cell leukemia virus type 1 (HTLV-1) Tax protein activates viral transcription from the long terminal repeats (LTR). Mechanisms through which Tax activates LTR have been established, but coactivators of this process remain to be identified and characterized. Here we show that all three members of the TORC family of transcriptional regulators are coactivators of Tax for LTR-driven expression. TORC coactivation requires CREB, but not ATF4 or other bZIP factors. Tax physically interacts with TORC1, TORC2, and TORC3 (TORC1/2/3), and the depletion of TORC1/2/3 inhibited Tax activity. TORC coactivation can be further enhanced by transcriptional coactivator p300. In addition, coactivators in the p300 family are required for full activity of Tax independently of TORC1/2/3. Thus, both TORC and p300 families of coactivators are essential for optimal activation of HTLV-1 transcription by Tax.
Sequence Analysis and Organization of theNeodiprion abietisNucleopolyhedrovirus Genome
Authors: Duffy Simon P. , Young Aaron M. , Morin Benoit , Lucarotti Christopher J. , Koop Ben F. , Levin David B.
Published: June 2006

Of 30 baculovirus genomes that have been sequenced to date, the only nonlepidopteran baculoviruses include the dipteranCulex nigripalpusnucleopolyhedrovirus and two hymenopteran nucleopolyhedroviruses that infect the sawfliesNeodiprion lecontei(NeleNPV) andNeodiprion sertifer(NeseNPV). This study provides a complete sequence and genome analysis of the nucleopolyhedrovirus that infects the balsam fir sawflyNeodiprion abietis(Hymenoptera, Symphyta, Diprionidae). TheN. abietisnucleopolyhedrovirus (NeabNPV) is 84,264 bp in size, with a G+C content of 33.5%, and contains 93 predicted open reading frames (ORFs). Eleven predicted ORFs are unique to this baculovirus, 10 ORFs have a putative sequence homologue in the NeleNPV genome but not the NeseNPV genome, and 1 ORF (neab53) has a putative sequence homologue in the NeseNPV genome but not the NeleNPV genome. Specific repeat sequences are coincident with major genome rearrangements that distinguish NeabNPV and NeleNPV. Genes associated with these repeat regions encode a common amino acid motif, suggesting that they are a family of repeated contiguous gene clusters. Lepidopteran baculoviruses, similarly, have a family of repeated genes called thebrogene family. However, there is no significant sequence similarity between the NeabNPV andbrogenes. Homologues of early-expressed genes such asie-1andlef-3were absent in NeabNPV, as they are in the previously sequenced hymenopteran baculoviruses. Analyses of ORF upstream sequences identified potential temporally distinct genes on the basis of putative promoter elements.
The Double-Stranded RNA Binding Protein 76:NF45 Heterodimer Inhibits Translation Initiation at the Rhinovirus Type 2 Internal Ribosome Entry Site
Authors: Merrill Melinda K. , Gromeier Matthias
Published: June 2006

Poliovirus (PV) plus-strand RNA genomes initiate translation in a cap-independent manner via an internal ribosome entry site (IRES) in their 5′ untranslated region. Viral translation is codetermined by cellular IRES
trans
-acting factors, which can influence viral propagation in a cell-type-specific manner. Engineering of a poliovirus recombinant devoid of neuropathogenic properties but highly lytic in malignant glioma cells was accomplished by exchange of the cognate poliovirus IRES with its counterpart from human rhinovirus type 2 (HRV2), generating PV-RIPO. Neuroblast:glioma heterokaryon analyses revealed that loss of neurovirulence is due to
trans
-dominant repression of PV-RIPO propagation in neuronal cells. The double-stranded RNA binding protein 76 (DRBP76) was previously identified to bind to the HRV2 IRES in neuronal cells and to inhibit PV-RIPO translation and propagation (M. Merrill, E. Dobrikova, and M. Gromeier, J. Virol. 80:3347-3356, 2006). The results of size exclusion chromatography indicate that DRBP76 heterodimerizes with nuclear factor of activated T cells, 45 kDa (NF45), in neuronal but not in glioma cells. The DRBP76:NF45 heterodimer binds to the HRV2 IRES in neuronal but not in glioma cells. Ribosomal profile analyses show that the heterodimer preferentially associates with the translation apparatus in neuronal cells and arrests translation at the HRV2 IRES, preventing PV-RIPO RNA assembly into polysomes. Results of this study suggest that the DRBP76:NF45 heterodimer selectively blocks HRV2 IRES-driven translation initiation in neuron-derived cells.
Removal of Arginine 332 Allows Human TRIM5α To Bind Human Immunodeficiency Virus Capsids and To Restrict Infection
Authors: Li Yuan , Li Xing , Stremlau Matthew , Lee Mark , Sodroski Joseph
Published: June 2006

Human TRIM5α (TRIM5α
hu
) only modestly inhibits human immunodeficiency virus type 1 (HIV-1) and does not inhibit simian immunodeficiency virus (SIV
mac
). Alteration of arginine 332 in the TRIM5α
hu
B30.2 domain to proline, the residue found in rhesus monkey TRIM5α, has been shown to create a potent restricting factor for both HIV-1 and SIV
mac.
Here we demonstrate that the potentiation of HIV-1 inhibition results from the removal of a positively charged residue at position 332 of TRIM5α
hu.
The increase in restricting activity correlated with an increase in the ability of TRIM5α
hu
mutants lacking arginine 332 to bind HIV-1 capsid complexes. A change in the cyclophilin A-binding loop of the HIV-1 capsid decreased TRIM5α
hu
R332P binding and allowed escape from restriction. The ability of TRIM5α
hu
to restrict SIV
mac
could be disrupted by the presence of any charged residue at position 332. Thus, charged residues in the v1 region of the TRIM5α
hu
B30.2 domain can modulate capsid binding and restriction potency. Therapeutic strategies designed to neutralize arginine 332 of TRIM5α
hu
might potentiate the innate resistance of human cells to HIV-1 infection.
Increased Immunogenicity of Human Immunodeficiency Virus gp120 Engineered To Express Galα1-3Galβ1-4GlcNAc-R Epitopes
Authors: Abdel-Motal Ussama , Wang Shixia , Lu Shan , Wigglesworth Kim , Galili Uri
Published: June 2006

The glycan shield comprised of multiple carbohydrate chains on the human immunodeficiency virus (HIV) envelope glycoprotein gp120 helps the virus to evade neutralizing antibodies. The present study describes a novel method for increasing immunogenicity of gp120 vaccine by enzymatic replacement of sialic acid on these carbohydrate chains with Galα1-3Galβ1-4GlcNAc-R (α-gal) epitopes. These epitopes are ligands for the natural anti-Gal antibody constituting ∼1% of immunoglobulin G in humans. We hypothesize that vaccination with gp120 expressing α-gal epitopes (gp120αgal) results in in vivo formation of immune complexes with anti-Gal, which targets vaccines for effective uptake by antigen-presenting cells (APC), due to interaction between the Fc portion of the antibody and Fcγ receptors on APC. This in turn results in effective transport of the vaccine to lymph nodes and effective processing and presentation of gp120 immunogenic peptides by APC for eliciting a strong anti-gp120 immune response. This hypothesis was tested in α-1,3-galactosyltransferase knockout mice, which produce anti-Gal. Mice immunized with gp120αgalproduced anti-gp120 antibodies in titers that were >100-fold higher than those measured in mice immunized with comparable amounts of gp120 and effectively neutralized HIV. T-cell response, measured by ELISPOT, was much higher in mice immunized with gp120αgalthan in mice immunized with gp120. It is suggested that gp120αgalcan serve as a platform for anti-Gal-mediated targeting of additional vaccinating HIV proteins fused to gp120αgal, thereby creating effective prophylactic vaccines.
Transmission of Simian Immunodeficiency Virus SIVcpz and the Evolution of Infection in the Presence and Absence of Concurrent Human Immunodeficiency Virus Type 1 Infection in Chimpanzees
Authors: Heeney Jonathan L. , Rutjens Erik , Verschoor Ernst J. , Niphuis Henk , ten Haaft Peter , Rouse Scott , McClure Hazel , Balla-Jhagjhoorsingh Sunita , Bogers Willy , Salas Mary , Cobb Kathy , Kestens Luc , Davis David , van der Groen Guido , Courgnaud Valerie , Peeters Martine , Murthy Krishna K.
Published: June 2006

Current data suggest that the human immunodeficiency virus type 1 (HIV-1) epidemic arose by transmission of simian immunodeficiency virus (SIV) SIVcpz from a subspecies of common chimpanzees (Pan troglodytes troglodytes) to humans. SIVcpz of chimpanzees is itself a molecular chimera of SIVs from two or more different monkey species, suggesting that recombination was made possible by coinfection of one individual animal with different lentiviruses. However, very little is known about SIVcpz transmission and the susceptibility to lentivirus coinfection of its natural host, the chimpanzee. Here, it is revealed that either infected plasma or peripheral blood mononuclear cells readily confer infection when exposure occurs by the intravenous or mucosal route. Importantly, the presence of preexisting HIV-1 infection did not modify the kinetics of SIVcpz infection once it was established by different routes. Although humoral responses appeared as early as 4 weeks postinfection, neutralization to SIVcpz-ANT varied markedly between animals. Analysis of the SIVcpzenvsequence over time revealed the emergence of genetic viral variants and persistent SIVcpz RNA levels of between 104and 105copies/ml plasma regardless of the presence or absence of concurrent HIV-1 infection. These unique data provide important insight into possible routes of transmission, the kinetics of acute SIVcpz infection, and how readily coinfection with SIVcpz and other lentiviruses may be established as necessary preconditions for potential recombination.
Regulation of Adeno-Associated Virus DNA Replication by the Cellular TAF-I/Set Complex
Authors: Pegoraro Gianluca , Marcello Alessandro , Myers Michael P. , Giacca Mauro
Published: June 2006

The Rep proteins of the adeno-associated virus (AAV) are required for viral replication in the presence of adenovirus helper functions and as yet poorly characterized cellular factors. In an attempt to identify such factors, we purified Flag-Rep68-interacting proteins from human cell lysates. Several polypeptides were identified by mass spectrometry, among which was ANP32B, a member of the acidic nuclear protein 32 family which takes part in the formation of the template-activating factor I/Set oncoprotein (TAF-I/Set) complex. The N terminus of Rep was found to specifically bind the acidic domain of ANP32B; through this interaction, Rep was also able to recruit other members of the TAF-I/Set complex, including the ANP32A protein and the histone chaperone TAF-I/Set. Further experiments revealed that silencing of ANP32A and ANP32B inhibited AAV replication, while overexpression of all of the components of the TAF-I/Set complex increased de novo AAV DNA synthesis in permissive cells. Besides being the first indication that the TAF-I/Set complex participates in wild-type AAV replication, these findings have important implications for the generation of recombinant AAV vectors since overexpression of the TAF-I/Set components was found to markedly increase viral vector production.
Rapid Pathogenesis Induced by a Vesicular Stomatitis Virus Matrix Protein Mutant: Viral Pathogenesis Is Linked to Induction of Tumor Necrosis Factor Alpha
Authors: Publicover Jean , Ramsburg Elizabeth , Robek Michael , Rose John K.
Published: June 2006

Vesicular stomatitis virus (VSV) matrix (M) protein blocks host mRNA export from the nucleus and thereby inhibits interferon induction in infected cells. M mutants with mutations of methionine 51 (M51) lack this shutoff function. We examined pathogenesis of a VSV M mutant with a deletion of M51 (VSVΔM51) after intranasal infection of BALB/c mice and found an unexpected phenotype. Mice that received VSVΔM51 experienced a more rapid but overall less severe weight loss than mice that received the recombinant wild-type VSV (rwtVSV). Rapid weight loss was not explained by faster initial replication because VSVΔM51 replication was controlled faster than rwtVSV replication in the lungs and did not spread systemically like rwtVSV. This faster control of VSVΔM51 correlated with a more rapid induction of interferon in the lung. Because tumor necrosis factor alpha (TNF-α) is associated with weight loss, we examined TNF-α induction in mice infected with rwtVSV or VSVΔM51. We found more-rapid induction of TNF-α by the mutant at early times after infection, while rwtVSV induced more TNF-α later in infection. This result suggested that TNF-α induction might explain both the rapid weight loss caused by the mutant and the overall greater weight loss caused by the rwtVSV. Using TNF-α knockout mice (C57BL/6 background), we showed that weight loss following rwtVSV infection was greatly reduced in the absence of TNF-α. Although the rapid weight loss caused by VSVΔM51 was less pronounced in C57BL/6 mice, it was eliminated in the absence of TNF-α. These results indicate a role for TNF-α in the pathogenesis of VSV.
PKR and RNase L Contribute to Protection against Lethal West Nile Virus Infection by Controlling Early Viral Spread in the Periphery and Replication in Neurons
Authors: Samuel Melanie A. , Whitby Kevin , Keller Brian C. , Marri Anantha , Barchet Winfried , Williams Bryan R. G. , Silverman Robert H. , Gale Michael , Diamond Michael S.
Published: June 2006

West Nile virus (WNV) is a neurotropic, mosquito-borne flavivirus that can cause lethal meningoencephalitis. Type I interferon (IFN) plays a critical role in controlling WNV replication, spread, and tropism. In this study, we begin to examine the effector mechanisms by which type I IFN inhibits WNV infection. Mice lacking both the interferon-induced, double-stranded-RNA-activated protein kinase (PKR) and the endoribonuclease of the 2′,5′-oligoadenylate synthetase-RNase L system (PKR−/−× RL−/−) were highly susceptible to subcutaneous WNV infection, with a 90% mortality rate compared to the 30% mortality rate observed in congenic wild-type mice. PKR−/−× RL−/−mice had increased viral loads in their draining lymph nodes, sera, and spleens, which led to early viral entry into the central nervous system (CNS) and higher viral burden in neuronal tissues. Although mice lacking RNase L showed a higher CNS viral burden and an increased mortality, they were less susceptible than the PKR−/−× RL−/−mice; thus, we also infer an antiviral role for PKR in the control of WNV infection. Notably, a deficiency in both PKR and RNase L resulted in a decreased ability of type I IFN to inhibit WNV in primary macrophages and cortical neurons. In contrast, the peripheral neurons of the superior cervical ganglia of PKR−/−× RL−/−mice showed no deficiency in the IFN-mediated inhibition of WNV. Our data suggest that PKR and RNase L contribute to IFN-mediated protection in a cell-restricted manner and control WNV infection in peripheral tissues and some neuronal subtypes.
Processing of Human Cytomegalovirus UL37 Mutant Glycoproteins in the Endoplasmic Reticulum Lumen prior to Mitochondrial Importation
Authors: Mavinakere Manohara S. , Williamson Chad D. , Goldmacher Victor S. , Colberg-Poley Anamaris M.
Published: June 2006

The human cytomegalovirus (HCMV) UL37 glycoprotein (gpUL37) is internally cleaved and its products divergently traffic to mitochondria or are retained in the secretory pathway. To define the requirements for gpUL37 cleavage, residues −1 and −3 of the consensus endoplasmic reticulum (ER) signal peptidase I site within exon 3 (UL37x3) were replaced by bulky tyrosines (gpUL37 cleavage site mutant I). Internal cleavage of this UL37x3 mutant was inhibited, verifying usage of the consensus site at amino acids (aa) 193/194. The full-length mitochondrial species of gpUL37 cleavage site mutant I was N glycosylated and endoglycosidase H sensitive, indicating that ER translocation and processing took place prior to its mitochondrial importation. Moreover, these results suggest that internal cleavage of gpUL37 is not necessary for its N glycosylation. Partial deletion or disruption of the UL37 hydrophobic core immediately upstream of the cleavage site resulted in decreased protein abundance, suggesting that the UL37x3 hydrophobic α-helix contributes to either correct folding or stability of gpUL37. Insertion of the UL37x3 hydrophobic core and cleavage site into pUL37M, a splice variant of gpUL37 which lacks these sequences and is neither proteolytically cleaved nor N glycosylated, resulted in its internal cleavage and N glycosylation. Its NH2-terminal fragment, pUL37M-NH2, was detected more abundantly in mitochondria, while its N-glycosylated C-terminal fragment, gpUL37M-COOH, was detected predominantly in the ER in a manner analogous to that of gpUL37 cleavage products. These results indicate that UL37x3 aa 178 to 205 are prerequisite for gpUL37 internal cleavage and alter UL37 protein topology allowing N glycosylation of its C-terminal sequences. In contrast, the NH2-terminal UL37x1 hydrophobic leader, present in pUL37x1, pUL37M, and gpUL37, is not cleaved from mature UL37 protein, retaining a membrane anchor for UL37 isoforms during trafficking. Taken together, these results suggest that HCMV gpUL37 undergoes sequential trafficking, during which it is ER translocated, processed, and then mitochondrially imported.
Epstein-Barr Virus Shed in Saliva Is High in B-Cell-Tropic Glycoprotein gp42
Authors: Jiang R. , Scott R. S. , Hutt-Fletcher L. M.
Published: June 2006

Epstein-Barr virus is an orally transmitted human herpesvirus that infects epithelial cells and establishes latency in memory B lymphocytes. Movement of virus between the two cell types is facilitated by changes in amounts of an envelope glycoprotein, gp42, which are effected by interaction of gp42 with HLA class II in a B cell. Here we used the differential ability of virus to bind to CD21-positive B cells and CD21-negative epithelial cells, which is also influenced by levels of gp42, to determine that the majority of virus shed in saliva is derived from an HLA class II-negative cell.
Comparative Analysis of 22 Coronavirus HKU1 Genomes Reveals a Novel Genotype and Evidence of Natural Recombination in Coronavirus HKU1
Authors: Woo Patrick C. Y. , Lau Susanna K. P. , Yip Cyril C. Y. , Huang Yi , Tsoi Hoi-Wah , Chan Kwok-Hung , Yuen Kwok-Yung
Published: June 2006

We sequenced and compared the complete genomes of 22 strains of coronavirus HKU1 (CoV HKU1) obtained from nasopharyngeal aspirates of patients with respiratory tract infections over a 2-year period. Phylogenetic analysis of 24 putative proteins and polypeptides showed that the 22 CoV HKU1 strains fell into three clusters (genotype A, 13 strains; genotype B, 3 strains and genotype C, 6 strains). However, different phylogenetic relationships among the three clusters were observed in different regions of their genomes. From nsp4 to nsp6, the genotype A strains were clustered with the genotype B strains. For nsp7 and nsp8 and from nsp10 to nsp16, the genotype A strains were clustered with the genotype C strains. From hemagglutinin esterase (HE) to nucleocapsid (N), the genotype B strains were clustered closely with the genotype C strains. Bootscan analysis showed possible recombination between genotypes B and C from nucleotide positions 11500 to 13000, corresponding to the nsp6-nsp7 junction, giving rise to genotype A, and between genotypes A and B from nucleotide positions 21500 to 22500, corresponding to the nsp16-HE junction, giving rise to genotype C. Multiple alignments further narrowed the sites of crossover to a 143-bp region between nucleotide positions 11750 and 11892 and a 29-bp region between nucleotide positions 21502 and 21530. Genome analysis also revealed various numbers of tandem copies of a perfect 30-base acidic tandem repeat (ATR) which encodes NDDEDVVTGD and various numbers and sequences of imperfect repeats in the N terminus of nsp3 inside the acidic domain upstream of papain-like protease 1 among the 22 genomes. All 10 CoV HKU1 strains with incomplete imperfect repeats (1.4 and 4.4) belonged to genotype A. The present study represents the first evidence for natural recombination in coronavirus associated with human infection. Analysis of a single gene is not sufficient for the genotyping of CoV HKU1 strains but requires amplification and sequencing of at least two gene loci, one from nsp10 to nsp16 (e.g.,
pol
or helicase) and another from HE to N (e.g., spike or N). Further studies will delineate whether the ATR is useful for the molecular typing of CoV HKU1.
Activation of Triggering Receptor Expressed on Myeloid Cells-1 on Human Neutrophils by Marburg and Ebola Viruses
Authors: Mohamadzadeh Mansour , Coberley Sadie S. , Olinger Gene G. , Kalina Warren V. , Ruthel Gordon , Fuller Claudette L. , Swenson Dana L. , Pratt William D. , Kuhns Douglas B. , Schmaljohn Alan L.
Published: June 2006

Marburg virus (MARV) and Ebola virus (EBOV), members of the viral familyFiloviridae, cause fatal hemorrhagic fevers in humans and nonhuman primates. High viral burden is coincident with inadequate adaptive immune responses and robust inflammatory responses, and virus-mediated dysregulation of early host defenses has been proposed. Recently, a novel class of innate receptors called the triggering receptors expressed in myeloid cells (TREM) has been discovered and shown to play an important role in innate inflammatory responses and sepsis. Here, we report that MARV and EBOV activate TREM-1 on human neutrophils, resulting in DAP12 phosphorylation, TREM-1 shedding, mobilization of intracellular calcium, secretion of proinflammatory cytokines, and phenotypic changes. A peptide specific to TREM-1 diminished the release of tumor necrosis factor alpha by filovirus-activated human neutrophils in vitro, and a soluble recombinant TREM-1 competitively inhibited the loss of cell surface TREM-1 that otherwise occurred on neutrophils exposed to filoviruses. These data imply direct activation of TREM-1 by filoviruses and also indicate that neutrophils may play a prominent role in the immune and inflammatory responses to filovirus infections.
Analyses of Phosphorylation Events in the Rubella Virus Capsid Protein: Role in Early Replication Events
Authors: Law LokMan J. , Ilkow Carolina S. , Tzeng Wen-Pin , Rawluk Matthew , Stuart David T. , Frey Teryl K. , Hobman Tom C.
Published: June 2006

The Rubella virus capsid protein is phosphorylated prior to virus assembly. Our previous data are consistent with a model in which dynamic phosphorylation of the capsid regulates its RNA binding activity and, in turn, nucleocapsid assembly. In the present study, the process of capsid phosphorylation was examined in further detail. We show that phosphorylation of serine 46 in the RNA binding region of the capsid is required to trigger phosphorylation of additional amino acid residues that include threonine 47. This residue likely plays a direct role in regulating the binding of genomic RNA to the capsid. We also provide evidence which suggests that the capsid is dephosphorylated prior to or during virus budding. Finally, whereas the phosphorylation state of the capsid does not directly influence the rate of synthesis of viral RNA and proteins or the assembly and secretion of virions, the presence of phosphate on the capsid is critical for early events in virus replication, most likely the uncoating of virions and/or disassembly of nucleocapsids.
Optimal Long-Term Humoral Responses to Replication-Defective Herpes Simplex Virus Require CD21/CD35 Complement Receptor Expression on Stromal Cells
Authors: Brockman Mark A. , Verschoor Admar , Zhu Jia , Carroll Michael C. , Knipe David M.
Published: June 2006

Replication-defective herpes simplex virus (HSV) strains elicit durable immune responses and protect against virulent HSV challenge in mice, despite being unable to establish latent infection in neuronal cells. Mechanisms for generating long-lived immunity in the absence of viral persistence remain uncertain. In animals immunized with replication-defective HSV, durable serum immunoglobulin G (IgG) responses were elicited. Surprisingly, Western blot analyses revealed that the specificities of antiviral IgG changed over time, and antibody reactivity to some viral proteins was detected only very late. Thus, some of the durable IgG activity appeared to be contributed by either new or significantly enhanced antibody responses at late times. Following immunization, radiation bone marrow-chimeric mice lacking complement receptors CD21 and CD35 on stromal cells elicited only short-lived serum IgG and failed to mount recall responses to subsequent HSV exposure. Our results suggest that complement-mediated retention of viral antigens by stromal cells, such as follicular dendritic cells, is critical for optimal maintenance of antibody responses and B-cell memory following vaccination with replication-defective HSV.