bluuebunny/crossref_metadata_2025 huggingface dataset:
Application of harmonic radar technology to monitor tree snail dispersal
Authors: Hall Kevin T. , Hadfield Michael G.
Published: March 2009

Abstract.Planned conservation efforts for tree snails of the endangered genusAchatinella, endemic to the island of O'ahu, Hawai'i, will include translocations among the remaining wild and captive‐bred populations. In order to establish optimal levels of artificial migration among neighboring groups of snails within fragmented populations, efforts to determine natural dispersal rates through direct observation were initiated. Capture–mark–recapture (CMR) efforts have proved inadequate for obtaining the requisite dispersal estimates, due to low recapture probabilities. In addition, snail dispersal beyond the boundaries of a finite CMR study site was indistinguishable from mortality. In the preliminary study reported here, both the low recapture probability and dispersal detection problems of past CMR efforts were addressed by using harmonic radar tracking. This approach yielded rough dispersal estimates that were unattainable using CMR alone by providing 100% recapture rates even beyond the normal survey area boundaries. Extensive snail movements within clusters of connected trees were frequently observed after tracking for merely a few hours, although movements between unconnected trees were rare and recorded only after monthly survey intervals. Just 11 out of 40 tracked snails made between‐tree movements (average distance of 4.94±1.52 m) during the entire 7‐month study, and provided the only data utilizable for inferring gene flow in and out of subpopulations. Meteorological data loggers were deployed when tracking began to look for an association between such snail movement and weather fluctuations. The resultant data indicate that increases in both wind gusts and humidity facilitate dispersal (R2=0.77, p‐value <0.001), and that passive wind dispersal alone may be responsible for many snail movements (R2=0.59, p‐value=0.0014). Despite having provided coarse estimates of short‐term dispersal and corresponding wind influences, the limitations of the radar method can be substantial.
Thermal emission from the stellar-mass black hole binary XTE J1118+480 in the low/hard state
Authors: Reis R. C. , Miller J. M. , Fabian A. C.
Published: March 2009

We report on the detection of a thermal disc component from the stellar-mass black hole binary XTE J1118+480 in the canonical low/hard state. The presence of a thermal component with a temperature of approximately 0.21 keV in the Chandra spectra of XTE J1118+480 is found at more than the 14σ confidence level. Based on this evidence, we argue that the accretion disc in XTE J1118+480 is not truncated far from the central black hole in contrast with previous claims.
Retinoblastoma: An overview for advanced practice nurses
Authors: Canty Caroline Anne
Published: March 2009

Purpose: To provide an overview of the current literature surrounding the detection and management of retinoblastoma as well as care for the long‐term survivor to serve as a guide for advanced practice nurses.

Data sources: Comprehensive review of the literature on retinoblastoma including case studies, journal reviews, retrospective studies, and briefs.

Conclusions: Retinoblastoma is a pediatric intraocular malignancy usually detected in the primary care setting. Typically asymptomatic, primary care providers will often identify the presence of retinoblastoma upon routine eye exam. Prompt diagnosis and commencement of treatment are paramount as delay in diagnosis leads to higher morbidity and mortality. Treatment options vary depending on the extent of disease.

Implications for practice: Although current survival rates for retinoblastoma exceed 90%, individual visual outcome and survival are dependent upon early detection and prompt referral. In addition to research and education of survivors and families, advanced practice nurses play a key role in early detection and maintaining the current survival rate.
Nurse practitioner‐led multidisciplinary teams to improve chronic illness care: The unique strengths of nurse practitioners applied to shared medical appointments/group visits
Authors: Watts Sharon A. , Gee Julie , O’Day Mary Ellen , Schaub Kimberley , Lawrence Renee , Aron David , Kirsh Susan
Published: March 2009

Purpose: To describe the roles of nurse practitioners (NPs) in a novel model of healthcare delivery for patients with chronic disease: shared medical appointments (SMAs)/group visits based on the chronic care model (CCM). To map the specific skills of NPs to the six elements of the CCM: self‐management, decision support, delivery system design, clinical information systems, community resources, and organizational support.

Data sources: Case studies of three disease‐specific multidisciplinary SMAs (diabetes, heart failure, and hypertension) in which NPs played a leadership role.

Conclusions: NPs have multiple roles in development, implementation, and sustainability of SMAs as quality improvement interventions. Although the specific skills of NPs map out all six elements of the CCM, in our context, they had the greatest role in self‐management, decision support, and delivery system design.

Implications for practice: With the increasing numbers of patients with chronic illnesses, healthcare systems are increasingly challenged to provide necessary care and empower patients to participate in that care. NPs can play a key role in helping to meet these challenges.
Parental disconnect between perceived and actual weight status of children: A metasynthesis of the current research
Authors: Doolen Jessica , Alpert Patricia T. , Miller Sally K.
Published: March 2009

Purpose: Obesity is one of the most common chronic diseases in childhood. Many studies offer a variety of explanations for the alarming increase in childhood obesity; however, none discuss why an apparent disconnect exists in parental perceptions of their child’s weight status. The purpose of this article was to review the current research literature on parental perceptions about their children’s weight.

Data source: The articles included in this review were retrieved through a literature search using PubMed. Key words used to obtain relevant articles include childhood obesity, childhood overweight, and parental perception.

Conclusions: Several studies looked at parental perceptions of childhood obesity generated from the United Kingdom, Australia, Italy, and the United States. Universally, parents were more likely to misperceive their child’s weight. This was especially true for parents who were themselves overweight.

Implications for practice: If parents do not recognize their child as at risk for overweight or overweight, they cannot intervene to diminish the risk factors for pediatric obesity and its related complications. More research is needed to identify why this phenomenon occurs. Only then can effective interventions be initiated.
Family nurse practitioner clinical requirements: Is the best recommendation 500 hours?
Authors: Bray Christell O. , Olson Karen Koozer
Published: March 2009

Purpose: The purpose of the article was to stimulate further dialog around the recommended number of clinical hours required for students in family nurse practitioner (FNP) programs.

Data sources: Nursing literature was reviewed for research supporting the current clinical hour requirement. Both authors are faculty in FNP programs that provided experiential data for the article.

Conclusions and implications for practice: There are no scientific data upon which to base the number of required clinical hour. Researchers need to identify what type and how many clinical hours are needed to prepare competent new FNP graduates.
Latent autoimmune diabetes of adulthood (LADA): An often misdiagnosed type of diabetes mellitus
Authors: Appel Susan J , Wadas Theresa M , Rosenthal Richard S , Ovalle Fernando
Published: March 2009

Purpose: The purpose of this article is to raise awareness about a frequently misdiagnosed form of diabetes, latent autoimmune diabetes of adulthood (LADA), to describe its clinical and epidemiological characteristics, and to compare them to those of the more common and widely known types of diabetes, type 1 diabetes mellitus (DM) and type 2 DM.

Data sources: A review of the pertinent literature describing the features of LADA from 2000–2007 is summarized.

Conclusions: LADA is a rather common and often underrecognized form of diabetes whose clinical presentation falls somewhere between that of type 1 DM and type 2 DM. From a pathophysiological perspective, it is more closely related to type 1 DM, and some have even used the term type 1.5 diabetes to refer to it; however, it is most often misdiagnosed and treated as type 2 DM.

Implications for practice: Nurse practitioners (NPs) should always consider alternate diagnoses when patients with newly or previously identified adult‐onset diabetes mellitus do not fit the traditional stereotype of type 2 DM (i.e., overweight with signs of insulin resistance and a significant family history of diabetes). Statistically, strong consideration must be given to the diagnosis of LADA, especially in those who are of normal weight, show little evidence of insulin resistance, and have hardly any family history of diabetes. Knowing the patient’s exact diabetes type can give the NP a much greater understanding of the natural history of the patient’s disease, the changes that may occur as the patient ages, and how to optimally manage their diabetes to minimize complications. Likewise, when a patient is correctly diagnosed, they can be empowered to manage their diabetes with the appropriate therapies.
Building Mind, Brain, and Education Connections: The View From the Upper Valley
Authors: Coch Donna , Michlovitz Stephen A. , Ansari Daniel , Baird Abigail
Published: March 2009

ABSTRACT— This article describes the efforts of a small group of educators and researchers to build a model for making connections across mind, brain, and education. With a common goal of sharing, strengthening, and building useable knowledge about child and adolescent learning and development, we focused on questions of mutual interest to educators and researchers. We describe our efforts to develop a common vocabulary and language and to create opportunities for dialogue and discussion, including classes and talks for in‐service and preservice teachers, research laboratories open to in‐service and preservice teachers, local conferences that provided a context for educator and researcher interactions, and researcher outreach in the local education community at the administrative, classroom, and student levels. These activities represent concrete mechanisms by which links might be forged between educators and researchers within the context of Mind, Brain, and Education.
Motivational Orientation, Error Monitoring, and Academic Performance in Middle Childhood: A Behavioral and Electrophysiological Investigation
Authors: Fisher Kelly R. , Marshall Peter J. , Nanayakkara Ajantha R.
Published: March 2009

ABSTRACT— Previous research suggests that academic motivation orientation relates to students’ causal interpretations about academic outcomes and their emotional reactions to those outcomes. The current study examines how student motivation may relate to certain neurophysiological systems that are thought to underlie the processing of successes and failures. In the cognitive neuroscience literature, the error‐related negativity (ERN) in the event‐related potential has been associated with error processing and the degree of an individual’s emotional investment in his or her performance. The current study examined the relation between academic motivational characteristics and ERN amplitude during a speeded reaction time task in 3rd‐ to 5th‐grade students (n = 17). Intrinsically oriented students displayed larger amplitude ERN responses and made more internally directed attributions about their task performance. The findings suggest that students with high intrinsic orientation attribute performance to personal control and that their error‐monitoring system is more strongly engaged by performance errors.
Lack of association between the MTHFR (C677T) polymorphism and atopic disease
Authors: Thuesen Betina Heinsbaek , Husemoen Lise Lotte Nystrup , Fenger Mogens , Linneberg Allan
Published: March 2009

Background:  Impaired folate metabolism has been suggested as a potential risk factor for the development of asthma and atopic disease. However, there have been conflicting reports on the potential association between atopic disease and a common polymorphism of the methylene‐tetrahydrofolate reductase (MTHFR)‐gene, a well‐known marker of impaired folate metabolism.

Objectives:  The aim of this study was to investigate the association between the MTHFR (C677T) polymorphism and different outcome variables of asthma and atopic disease.

Methods:  This study was a population‐based study of 1189 participants aged 15–77 years living in Copenhagen, the Capital of Denmark. Examinations included measurements of specific IgE and skin prick tests against inhalant allergens, metacholine bronchial hyper‐reactivity, and serum eosinophilic cationic protein, and a self‐administered questionnaire about diagnoses and symptoms of allergy and asthma. In addition, participants were genotyped for the MTHFR (C677T) polymorphism.

Results:  None of the examined outcomes were significantly associated with the MTHFR (C677T) polymorphism.

Conclusions:  The results of this study using detailed objective markers of atopic disease do not support the hypothesis that impaired folate metabolism as reflected by the MTHFR genotype is involved in the development of atopic disease.

Please cite this paper as: Thuesen BH, Husemoen LLN, Fenger M and Linneberg A. Lack of association between the MTHFR (C677T) polymorphism and atopic disease. The Clinical Respiratory Journal 2009; 3: 102–108.
Internet Alternatives to Traditional Proctored Testing: Where Are We Now?
Authors: Tippins Nancy T.
Published: March 2009

Early in 2006, my colleagues and I published an article on unproctored Internet testing (UIT) in employment settings (Tippins et al., 2006). The purpose of that article was to identify the issues surrounding UIT and the ways in which those issues might be resolved. The panel of experts addressed a number of important questions about (a) the uses and applications of UIT and (b) the major issues and known problems associated with UIT, including test security, examinee identification, cheating, ethical use of tests, subgroup and cultural issues, standardization, and context effects. In addition, the panelists attempted to predict the future of UIT, highlight the research needed to facilitate UIT and provide advice to practitioners contemplating UIT. In the present article, “UIT” is used to refer to Internet-based testing completed by a candidate without a traditional human proctor. Thus, nontraditional forms of or alternatives to proctoring may be in place, such as quantitative analyses of response patterns, the use of video cameras, or follow-up testing with traditional proctoring.
Development and characterization of 15 polymorphic microsatellite loci isolated from Rafinesque's big‐eared bat, Corynorhinus rafinesquii
Authors: PIAGGIO ANTOINETTE J. , FIGUEROA JULIA A. , PERKINS SUSAN L.
Published: March 2009

We developed and characterized 15 microsatellite markers for Rafinesque's big‐eared bat, Corynorhinus rafinesquii. In a population from Tennessee, the number of alleles per locus ranged from three to 13 and observed heterozygosities were 0.35 to 0.97 per locus. These loci will provide appropriate variability for estimation of population connectivity, demographic parameters, and genetic diversity for this species of concern.
Understanding the contribution of Guyton's large circulatory model to long‐term control of arterial pressure
Authors: Montani Jean‐Pierre , Van Vliet Bruce N.
Published: March 2009

With the publication in 1972 of a large computer model of circulatory control, Guyton and colleagues challenged the then prevailing views on how blood pressure and cardiac output were controlled. At that time, it was widely accepted that the heart controlled cardiac output and that peripheral resistance controlled arterial blood pressure. By incorporating the empirically demonstrated concepts of blood flow autoregulation and the pressure–natriuresis relationship into their mathematical model, Guyton and colleagues were able to develop a number of revolutionary concepts. Guyton's circulatory model was particularly instrumental in exploring the linkage between blood pressure and sodium balance and in demonstrating an overriding importance of renal salt and water balance in setting the long‐term blood pressure level. In both the model and experimental data, any long‐lasting imbalance between salt intake and salt excretion leads to a progressive alteration of the degree of filling of the vascular system and thus to parallel changes in blood pressure. In turn, changes in blood pressure alter sodium excretion, opposing the initial salt imbalance. Although Guyton's model does not include the most recent cardiovascular discoveries, the concepts underlying the basic functioning of the cardiovascular system can serve as a well‐built basis for the development of new, large and integrative cardiovascular models.
Aerodynamic Analysis of a Vehicle Tanker
Authors: Buil Ramon Miralbes , Herrer Luis Castejon
Published: March 2009

The aim of this article is the presentation of a series of aerodynamic improvements for semitrailer tankers, which reduce the aerodynamic resistance of these vehicles, and, consequently, result in a positive impact on fuel consumption, which is substantially reduced (up to 11%). To make the analysis the computational fluid dynamics (CFD) methodology, using FLUENT, has been used since it allows simulating some geometries and modifications of the geometry without making physical prototypes that considerably increase the time and the economical resources needed. Three improvements are studied: the aerodynamic front, the undercarriage skirt, and the final box adaptor. First they are studied in isolation, so that the independent contribution of each improvement can be appreciated, while helping in the selection of the most convenient one. With the aerodynamic front the drag coefficient has a reduction of 6.13%, with the underskirt 9.6%, and with the boat tail 7.72%. Finally, all the improvements are jointly examined, resulting in a decrease of up to 23% in aerodynamic drag coefficient.
Considerations on a Mass-Based System Representation of a Pneumatic Cylinder
Authors: Thomas M. Brian , Maul Gary P.
Published: March 2009

Pneumatic actuators can be advantageous over electromagnetic and hydraulic actuators in many servo motion applications. The difficulty in their practical use comes from the highly nonlinear dynamics of the actuator and control valve. Previous works have used the cylinder’s position, velocity, and internal pressure as state variables in system models. This paper replaces pressure in the state model with the mass of gas in each chamber of the cylinder, giving a better representation of the system dynamics. Under certain circumstances, the total mass of gas in the cylinder may be assumed to be constant. This allows development of a reduced-order system model.
Evaluation of the Dynamic Transfer Matrix of Cavitating Inducers by Means of a Simplified “Lumped-Parameter” Model
Authors: Cervone Angelo , Tsujimoto Yoshinobu , Kawata Yutaka
Published: March 2009

The paper will present an analytical model for the evaluation of the pressure and flow rate oscillations in a given axial inducer test facility. The proposed reduced order model is based on several simplifying assumptions and takes into account the facility design and the dynamic properties of the tested inducer. The model has been used for evaluating the dynamic performance of a prototype of the LE-7 engine liquid oxygen (LOX) inducer, in tests carried out under given external flow rate excitations. The main results of these calculations will be shown, including the expected oscillations under a wide range of operational conditions and the influence of facility design. Calculations showed that the only way to obtain the two linearly independent test conditions, necessary for evaluating the inducer transfer matrix, is by changing the facility suction line: Any other changes in the facility design would result ineffective. Some other important design indications provided by the analytical model will be presented in the paper.
Pressurized Flow in a Mesostructured Silica Modified by Silane Groups
Authors: Punyamurtula Venkata K. , Han Aijie , Qiao Yu
Published: March 2009

By applying a quasihydrostatic pressure, water or electrolyte solution can be compressed into a surface treated MSU-H mesoporous silica. Based on the pressure-volume curves, thermodynamic and kinetic characteristics of the pressurized flow are analyzed. For pure water based system, continuum theory explains the testing data quite well but fails to capture the rate effect. For electrolyte solution based system, the classic interface theory breaks down, probably due to the unique ion behaviors in the nanoenvironment.
Numerical and Experimental Investigations of Steam Condensation in LP Part of a Large Power Turbine
Authors: Wróblewski Włodzimierz , Dykas Sławomir , Gardzilewicz Andrzej , Kolovratnik Michal
Published: March 2009

This paper presents the experimental investigations of steam flow with condensation in the blading system of the low-pressure (LP) part of a 360 MW turbine. To this end, special probes were used, which provided flow visualization opportunities including localization of the front of condensation, determining distributions of pressure, temperature, velocity, and flow angle in the inter-row gaps, measurements of water droplet concentration and sizes. The measurements have proved that the condensation process in the LP turbine might be of heterogeneous nature, depending on the concentration of chemical impurities in steam. The measurement results constituted the basis for computational fluid dynamics (CFD) flow calculations, which were performed using the time-dependent 3D Reynolds averaged Navier–Stokes equations coupled with two-equation turbulence model (k-ω SST) and additional conservation equations for the liquid phase. The set of governing equations has been closed by a “local” real gas equation of state. The condensation phenomena were modeled on the basis of the classical nucleation theory. The heterogeneous condensation model on the insoluble and soluble impurities was implemented into presented CFD code. The system of governing equations was solved by means of a finite volume method on a multiblock structured grid. The obtained numerical results and experimental data were compared and discussed.
Structure of the Anaphase-Promoting Complex/Cyclosome Interacting with a Mitotic Checkpoint Complex
Authors: Herzog Franz , Primorac Ivana , Dube Prakash , Lenart Peter , Sander Björn , Mechtler Karl , Stark Holger , Peters Jan-Michael
Published: March 2009

Once all chromosomes are connected to the mitotic spindle (bioriented), anaphase is initiated by the protein ubiquitylation activity of the anaphase-promoting complex/cyclosome (APC/C) and its coactivator Cdc20 (APC/C
Cdc20
). Before chromosome biorientation, anaphase is delayed by a mitotic checkpoint complex (MCC) that inhibits APC/C
Cdc20
. We used single-particle electron microscopy to obtain three-dimensional models of human APC/C in various functional states: bound to MCC, to Cdc20, or to neither (apo-APC/C). These experiments revealed that MCC associates with the Cdc20 binding site on APC/C, locks the otherwise flexible APC/C in a “closed” state, and prevents binding and ubiquitylation of a wide range of different APC/C substrates. These observations clarify the structural basis for the inhibition of APC/C by spindle checkpoint proteins.
Human Substantia Nigra Neurons Encode Unexpected Financial Rewards
Authors: Zaghloul Kareem A. , Blanco Justin A. , Weidemann Christoph T. , McGill Kathryn , Jaggi Jurg L. , Baltuch Gordon H. , Kahana Michael J.
Published: March 2009

The brain's sensitivity to unexpected outcomes plays a fundamental role in an organism's ability to adapt and learn new behaviors. Emerging research suggests that midbrain dopaminergic neurons encode these unexpected outcomes. We used microelectrode recordings during deep brain stimulation surgery to study neuronal activity in the human substantia nigra (SN) while patients with Parkinson's disease engaged in a probabilistic learning task motivated by virtual financial rewards. Based on a model of the participants' expected reward, we divided trial outcomes into expected and unexpected gains and losses. SN neurons exhibited significantly higher firing rates after unexpected gains than unexpected losses. No such differences were observed after expected gains and losses. This result provides critical support for the hypothesized role of the SN in human reinforcement learning.
A Recessive Mutation in the APP Gene with Dominant-Negative Effect on Amyloidogenesis
Authors: Di Fede Giuseppe , Catania Marcella , Morbin Michela , Rossi Giacomina , Suardi Silvia , Mazzoleni Giulia , Merlin Marco , Giovagnoli Anna Rita , Prioni Sara , Erbetta Alessandra , Falcone Chiara , Gobbi Marco , Colombo Laura , Bastone Antonio , Beeg Marten , Manzoni Claudia , Francescucci Bruna , Spagnoli Alberto , Cantù Laura , Del Favero Elena , Levy Efrat , Salmona Mario , Tagliavini Fabrizio
Published: March 2009

β-Amyloid precursor protein (APP) mutations cause familial Alzheimer's disease with nearly complete penetrance. We found an APP mutation [alanine-673→valine-673 (A673V)] that causes disease only in the homozygous state, whereas heterozygous carriers were unaffected, consistent with a recessive Mendelian trait of inheritance. The A673V mutation affected APP processing, resulting in enhanced β-amyloid (Aβ) production and formation of amyloid fibrils in vitro. Co-incubation of mutated and wild-type peptides conferred instability on Aβ aggregates and inhibited amyloidogenesis and neurotoxicity. The highly amyloidogenic effect of the A673V mutation in the homozygous state and its anti-amyloidogenic effect in the heterozygous state account for the autosomal recessive pattern of inheritance and have implications for genetic screening and the potential treatment of Alzheimer's disease.
RSY-1 Is a Local Inhibitor of Presynaptic Assembly in
C. elegans
Authors: Patel Maulik R. , Shen Kang
Published: March 2009

As fundamental units of neuronal communication, chemical synapses are composed of presynaptic and postsynaptic specializations that form at specific locations with defined shape and size. Synaptic assembly must be tightly regulated to prevent overgrowth of the synapse size and number, but the molecular mechanisms that inhibit synapse assembly are poorly understood. We identified regulator of synaptogenesis–1 (RSY-1) as an evolutionarily conserved molecule that locally antagonized presynaptic assembly. The loss of RSY-1 in
Caenorhabditis elegans
led to formation of extra synapses and recruitment of excessive synaptic material to presynaptic sites. RSY-1 directly interacted with and negatively regulated SYD-2/liprin-alpha, a master assembly molecule that recruits numerous synaptic components to presynaptic sites. RSY-1 also bound and regulated SYD-1, a synaptic protein required for proper functioning of SYD-2. Thus, local inhibitory mechanisms govern synapse formation.
The Initial Stages of Template-Controlled CaCO
3
Formation Revealed by Cryo-TEM
Authors: Pouget Emilie M. , Bomans Paul H. H. , Goos Jeroen A. C. M. , Frederik Peter M. , de With Gijsbertus , Sommerdijk Nico A. J. M.
Published: March 2009

Biogenic calcium carbonate forms the inorganic component of seashells, otoliths, and many marine skeletons, and its formation is directed by an ordered template of macromolecules. Classical nucleation theory considers crystal formation to occur from a critical nucleus formed by the assembly of ions from solution. Using cryotransmission electron microscopy, we found that template-directed calcium carbonate formation starts with the formation of prenucleation clusters. Their aggregation leads to the nucleation of amorphous nanoparticles in solution. These nanoparticles assemble at the template and, after reaching a critical size, develop dynamic crystalline domains, one of which is selectively stabilized by the template. Our findings have implications for template-directed mineral formation in biological as well as in synthetic systems.
Evidence of a Functional B-Cell Immunodeficiency in Adults Who Experience Serogroup C Meningococcal Disease
Authors: Foster Rachel A. , Carlring Jennifer , McKendrick Michael W. , Lees Andrew , Borrow Ray , Read Robert C. , Heath Andrew W.
Published: March 2009

After adolescence, the incidence of meningococcal disease decreases with age as a result of the cumulative immunizing effect of repeated nasopharyngeal colonization. Nevertheless, some adults succumb to meningococcal disease, so we hypothesized that this is due to a subtle functional immunological defect. Peripheral blood lymphocytes derived from survivors of serogroup C meningococcal disease and from age- and sex-matched controls were incubated with a polyclonal B-cell activator containing anti-immunoglobulin D (α-δ-dex) employed to mimic antigen-specific stimuli encountered during immune responses to bacterial polysaccharides, with and without T-cell activation (using anti-CD3/anti-CD28). Subsequent proliferation and activation of T and B lymphocytes were measured. In patients, T-cell responses to polyclonal stimuli and the delivery of T-cell help to B cells were unimpaired. Levels of B-cell proliferation in response to α-δ-dex stimulation alone were low in all samples but were significantly lower in patients than in controls, and these differences were more pronounced with the addition of T-cell help. The data are consistent with the presence of a subtle immunodeficiency in adults who have exhibited susceptibility to meningococcal disease. This defect is manifested as an impaired B-cell response to T-cell-independent type 2 antigens analogous to bacterial capsular polysaccharide.
An Improved Whole-Blood Gamma Interferon Assay Based on the CFP21-MPT64 Fusion Protein
Authors: Fu Ruiling , Wang Chun , Shi Chunwei , Lu Mengji , Fang Zhengming , Lu Jia , Wang Fang , Fan Xionglin
Published: March 2009

Differentiation of latent tuberculosis infection (LTBI) from a healthy, unexposed population plays a vital role in the strategy of controlling and eliminating tuberculosis (TB). Both CFP21 and MPT64, antigens encoded by the RD2 region which are restricted in the
Mycobacterium tuberculosis
complex, are TB-specific diagnostic candidate antigens. In this study, we designed a fusion protein by linking both CFP21 and MPT64 with a 15-amino-acid peptide, (G
4
S
1
)
3
, and overexpressed the fusion protein in
Escherichia coli
. A new whole-blood gamma interferon assay based on the recombinant fusion protein, CFP21-MPT64 (rCM-WBIA), was developed and compared with the tuberculin skin test (TST) for screening of LTBI in household contacts of patients with sputum-positive TB. rCM-WBIA had a slightly higher sensitivity (66.7%; 24/36 contacts) than that of the TST (61.1%; 22/36 contacts) for household contacts. We found that rCM-WBIA had a very high sensitivity (90.9%) and specificity (71.4%) for LTBI detection compared with TST. The overall agreement between rCM-WBIA and TST was 83.3% (
k
= 0.64); rCM-WBIA positivity was associated with a larger TST induration. These results suggest that rCM-WBIA, based on the recombinant fusion protein CFP21-MPT64, is a promising alternative diagnostic tool for detection of LTBI.
Universal Detection and Identification of Avian Influenza Virus by Use of Resequencing Microarrays
Authors: Lin Baochuan , Malanoski Anthony P. , Wang Zheng , Blaney Kate M. , Long Nina C. , Meador Carolyn E. , Metzgar David , Myers Christopher A. , Yingst Samuel L. , Monteville Marshall R. , Saad Magdi D. , Schnur Joel M. , Tibbetts Clark , Stenger David A.
Published: March 2009

Zoonotic microbes have historically been, and continue to emerge as, threats to human health. The recent outbreaks of highly pathogenic avian influenza virus in bird populations and the appearance of some human infections have increased the concern of a possible new influenza pandemic, which highlights the need for broad-spectrum detection methods for rapidly identifying the spread or outbreak of all variants of avian influenza virus. In this study, we demonstrate that high-density resequencing pathogen microarrays (RPM) can be such a tool. The results from 37 influenza virus isolates show that the RPM platform is an effective means for detecting and subtyping influenza virus, while simultaneously providing sequence information for strain resolution, pathogenicity, and drug resistance without additional analysis. This study establishes that the RPM platform is a broad-spectrum pathogen detection and surveillance tool for monitoring the circulation of prevalent influenza viruses in the poultry industry and in wild birds or incidental exposures and infections in humans.
PCR Diagnosis of
Opisthorchis viverrini
and
Haplorchis taichui
Infections in a Lao Community in an Area of Endemicity and Comparison of Diagnostic Methods for Parasitological Field Surveys
Authors: Lovis Leonore , Mak Tippi K. , Phongluxa Khampheng , Soukhathammavong Phonepasong , Sayasone Somphou , Akkhavong Kongsap , Odermatt Peter , Keiser Jennifer , Felger Ingrid
Published: March 2009

Opisthorchiasis is a major public health problem in Southeast Asia. Affected individuals often have mixed infections with the liver fluke
Opisthorchis viverrini
and minute intestinal flukes such as
Haplorchis taichui
. The usual methods of diagnosing these infections involve the demonstration of fluke eggs in stool samples under light microscopy, but sensitivity and specificity are low. We developed two PCR tests that detect and discriminate between
O. viverrini
and
H. taichui
infections. PCR tests were validated by stool samples from purged individuals. We then applied the PCR tests to estimate the prevalence of
O. viverrini
and
H. taichui
infections from a random sample of individuals selected from a community in an area of endemicity in Khong District, Laos. PCR results were compared with those from the Kato-Katz (KK) method and the formalin-ether concentration technique (FECT). When validated with purge results, PCR tests of
O. viverrini
and
H. taichui
had sensitivities of 93.7% (95% confidence interval [CI], 85.8 to 97.9%) and 73.3% (95% CI, 60.3 to 83.9%) and could detect as little as 0.75 pg DNA and 1.32 ng DNA, respectively. The PCR-determined community prevalences of
O. viverrini
and
H. taichui
infections were 63.9% (95% CI, 54.1 to 72.9%) and 30.6% (95% CI, 22.1 to 40.2%), respectively. Using PCR as the gold standard to detect
O. viverrini
, three KK thick smears performed comparably well, whereas one KK smear and FECT were poorer (sensitivities of 91.4% [95% CI, 81.0 to 97.1%,], 62.3% [95% CI, 49.8 to 73.7%], and 49.3% [95% CI, 37.0 to 61.6%], respectively). PCR may be a valuable and sensitive diagnostic tool, particularly for low-intensity
O. viverrini
and
H. taichui
infections.
Rapid and Accurate Diagnosis of Human Intestinal Spirochetosis by Fluorescence In Situ Hybridization
Authors: Schmiedel Dinah , Epple Hans-Jörg , Loddenkemper Christoph , Ignatius Ralf , Wagner Jutta , Hammer Bettina , Petrich Annett , Stein Harald , Göbel Ulf B. , Schneider Thomas , Moter Annette
Published: March 2009

Human intestinal spirochetosis (HIS) is associated with overgrowth of the large intestine by spirochetes of the genus
Brachyspira
. The microbiological diagnosis of HIS is hampered by the fastidious nature and slow growth of
Brachyspira
spp. In clinical practice, HIS is diagnosed histopathologically, and a significant portion of cases may be missed. Fluorescence in situ hybridization (FISH) is a molecular method that allows the visualization and identification of single bacteria within tissue sections. In this study, we analyzed intestinal biopsy samples from five patients with possible HIS. All specimens yielded positive results by histopathological techniques. PCR amplification and sequencing of the 16S rRNA gene were performed. Sequences of two isolates clustered in the group of
Brachyspira aalborgi
, whereas in three cases, the sequences were highly similar to that of
Brachyspira pilosicoli
. Three phylotypes showed mismatches at distinct nucleotide positions with
Brachyspira
sp. sequences published previously. In addition, culture for
Brachyspira
was successful in three cases. On the basis of these data, we designed and evaluated a
Brachyspira
genus-specific 16S rRNA-directed FISH probe that detects all of the
Brachyspira
spp. published to date. FISH of biopsy samples resulted in strong, unequivocal signals of brush-like formations at the crypt surfaces. This technique allowed simultaneous visualization of single spirochetes and their identification as
Brachyspira
spp. In conclusion, FISH provides a fast and accurate technique for the visualization and identification of intestinal spirochetes in tissue sections. It therefore represents a valuable tool for routine diagnosis of HIS.
Epstein-Barr Virus-Related Diarrhea or Exacerbation of Inflammatory Bowel Disease: Diagnostic Dilemma
Authors: Weinberg Ido , Neuman Tzahi , Margalit Maya , Ayman Fadeela , Wolf Dana G. , Ben-Yehuda Arie
Published: March 2009

While the presence of Epstein-Barr virus (EBV) in colonic specimens from patients with inflammatory bowel disease (IBD) has been documented, diarrhea secondary to gastrointestinal involvement by EBV in the context of primary EBV infection in patients with IBD has not been reported. We describe a patient with IBD who presented with diarrhea and primary EBV infection and propose a role for colonic involvement by EBV in the pathogenesis of his symptoms.
Comparison of Routine Bench and Molecular Diagnostic Methods in Identification of
Burkholderia pseudomallei
Authors: Hodgson Kelly , Engler Cathy , Govan Brenda , Ketheesan Natkunam , Norton Robert
Published: March 2009

This study compared the identification of
Burkholderia pseudomallei
with that of related organisms. Bench tests and latex agglutination were compared with molecular identification. Using bench tests and latex agglutination alone, 100% (30/30) of
B. pseudomallei
isolates were correctly identified. Amoxicillin-clavulanate susceptibility testing was also a good and simple discriminatory test.
Regulation of Episomal Gene Expression by KRAB/KAP1-Mediated Histone Modifications
Authors: Barde Isabelle , Laurenti Elisa , Verp Sonia , Groner Anna Claire , Towne Christopher , Padrun Viviane , Aebischer Patrick , Trumpp Andreas , Trono Didier
Published: March 2009

KAP1 is an essential cofactor of KRAB zinc finger proteins, a family of vertebrate-specific epigenetic repressors of largely unknown functions encoded in the hundreds by the mouse and human genomes. So far, KRAB/KAP1-mediated gene regulation has been studied within the environment of chromosomal DNA. Here we demonstrate that KRAB/KAP1 regulation is fully functional within the context of episomal DNA, such as adeno-associated viral and nonintegrated lentiviral vectors, and is correlated with histone modifications typically associated with this epigenetic regulator.
The Marmoset Model of GB Virus B Infections: Adaptation to Host Phenotypic Variation
Authors: Weatherford Trudie , Chavez Deborah , Brasky Kathleen M. , Lanford Robert E.
Published: March 2009

Worldwide, approximately 170 million people are chronically infected with hepatitis C virus (HCV), and chronic infection frequently progresses to serious liver disease, including cirrhosis and hepatocellular carcinoma. GB virus B (GBV-B), the virus phylogenetically most closely related to HCV, causes hepatitis in tamarins. We have demonstrated the suitability of the tamarin as a host for GBV-B and as a surrogate nonhuman primate model for HCV infection, and we have initiated studies of GBV-B infection in a closely related species, the common marmoset (
Callithrix jacchus
). Here, we demonstrate that marmosets exhibit two phenotypes upon infection with GBV-B: the susceptible phenotype and the partially resistant phenotype. In addition, we identify changes that may correlate with adaptation of the virus to the partially resistant host. GBV-B was serially passaged five times through 14 marmosets as one lineage and two times through 6 marmosets as a second lineage. Virus adapted to the marmosets and eventually exhibited robust infections in two separate lineages, lineages 1 and 2. A third lineage was initiated with a molecular clone, and again, susceptible and partially resistant phenotypes were observed. Three isolates were fully sequenced (from lineage 1), and 21 nucleotide changes were observed, with six amino acid changes. We speculate that the marmoset partially resistant phenotype may be due to a polymorphism in the marmoset population that affects critical virus-host interactions and that wild-type GBV-B is capable of rapidly adapting to this altered host.
Selection of a Simian-Human Immunodeficiency Virus Strain Resistant to a Vaginal Microbicide in Macaques
Authors: Dudley Dawn M. , Wentzel Jennifer L. , Lalonde Matthew S. , Veazey Ronald S. , Arts Eric J.
Published: March 2009

PSC-RANTES binds to CCR5, inhibits human immunodeficiency virus type 1 (HIV-1) entry, and has been shown as a vaginal microbicide to protect rhesus macaques from a simian-human immunodeficiency virus chimera (SHIV
SF162-p3
) infection in a dose-dependent manner. In this study,
env
gene sequences from SHIV
SF162-p3
-infected rhesus macaques treated with PSC-RANTES were analyzed for possible drug escape variants. Two specific mutations located in the V3 region of gp120 (K315R) and C-helical domain of gp41 (N640D) were identified in a macaque (m584) pretreated with a 100 μM dose of PSC-RANTES. These two
env
mutations were found throughout infection (through week 77) but were found at only low frequencies in the inoculating SHIV
SF162-p3
stock and in the other SHIV
SF162-p3
-infected macaques. HIV-1
env
genes from macaque m584 (
env
m584
) and from inoculating SHIV
SF162-p3
(
env
p3
) were cloned into an HIV-1 backbone. Increases in 50% inhibitory concentrations to PSC-RANTES with
env
m584
were modest (sevenfold) and most pronounced in cells expressing rhesus macaque CCR5 as compared to human CCR5. Nonetheless, virus harboring
env
m584
, unlike inoculating virus
env
p3
, could replicate even at the highest tissue culture PSC-RANTES concentrations (100 nM). Dual-virus competitions revealed a dramatic increase in fitness of chimeric virus containing
env
m584
(K315R/N640D) over that containing
env
p3
, but again, only in rhesus CCR5-expressing cells. This study is the first to describe the immediate selection and infection of a drug-resistant SHIV variant in the face of a protective vaginal microbicide, PSC-RANTES. This rhesus CCR5-specific/PSC- RANTES resistance selection is particularly alarming given the relative homogeneity of the SHIV
SF162-p3
stock compared to the potential exposure to a heterogeneous HIV-1 population in human transmission.
Virion Incorporation of the Herpes Simplex Virus Type 1 Tegument Protein VP22 Occurs via Glycoprotein E-Specific Recruitment to the Late Secretory Pathway
Authors: Stylianou Julianna , Maringer Kevin , Cook Rachelle , Bernard Emmanuelle , Elliott Gillian
Published: March 2009

The mechanism by which herpesviruses acquire their tegument is not yet clear. One model is that outer tegument proteins are recruited by the cytoplasmic tails of viral glycoproteins. In the case of herpes simplex virus tegument protein VP22, interactions with the glycoproteins gE and gD have been shown. We have previously shown that the C-terminal half of VP22 contains the necessary signal for assembly into the virus. Here, we show that during infection VP22 interacts with gE and gM, as well as its tegument partner VP16. However, by using a range of techniques we were unable to demonstrate VP22 binding to gD. By using pulldown assays, we show that while the cytoplasmic tails of both gE and gM interact with VP22, only gE interacts efficiently with the C-terminal packaging domain of VP22. Furthermore, gE but not gM can recruit VP22 to the Golgi/
trans
-Golgi network region of the cell in the absence of other virus proteins. To examine the role of the gE-VP22 interaction in infection, we constructed a recombinant virus expressing a mutant VP22 protein with a 14-residue deletion that is unable to bind gE (ΔgEbind). Coimmunoprecipitation assays confirmed that this variant of VP22 was unable to complex with gE. Moreover, VP22 was no longer recruited to its characteristic cytoplasmic trafficking complexes but exhibited a diffuse localization. Importantly, packaging of this variant into virions was abrogated. The mutant virus exhibited poor growth in epithelial cells, similar to the defect we have observed for a VP22 knockout virus. These results suggest that deletion of just 14 residues from the VP22 protein is sufficient to inhibit binding to gE and hence recruitment to the viral envelope and assembly into the virus, resulting in a growth phenotype equivalent to that produced by deleting the entire reading frame.
Phosphorylation of the U
L
31 Protein of Herpes Simplex Virus 1 by the U
S
3-Encoded Kinase Regulates Localization of the Nuclear Envelopment Complex and Egress of Nucleocapsids
Authors: Mou Fan , Wills Elizabeth , Baines Joel D.
Published: March 2009

Herpes simplex virus 1 nucleocapsids bud through the inner nuclear membrane (INM) into the perinuclear space to obtain a primary viral envelope. This process requires a protein complex at the INM composed of the U
L
31 and U
L
34 gene products. While it is clear that the viral kinase encoded by the U
S
3 gene regulates the localization of pU
L
31/pU
L
34 within the INM, the molecular mechanism by which this is accomplished remains enigmatic. Here, we have determined the following. (i) The N terminus of pU
L
31 is indispensable for the protein's normal function and contains up to six serines that are phosphorylated by the U
S
3 kinase during infection. (ii) Phosphorylation at these six serines was not essential for a productive infection but was required for optimal viral growth kinetics. (iii) In the presence of active U
S
3 kinase, changing the serines to alanine caused the pU
L
31/pU
L
34 complex to aggregate at the nuclear rim and caused some virions to accumulate aberrantly in herniations of the nuclear membrane, much as in cells infected with a U
S
3 kinase-dead mutant. (iv) The replacement of the six serines of pU
L
31 with glutamic acid largely restored the smooth distribution of pU
L
34/pU
L
31 at the nuclear membrane and precluded the accumulation of virions in herniations whether or not U
S
3 kinase was active but also precluded the optimal primary envelopment of nucleocapsids. These observations indicate that the phosphorylation of pU
L
31 by pU
S
3 represents an important regulatory event in the virion egress pathway that can account for much of pU
S
3's role in nuclear egress. The data also suggest that the dynamics of pU
L
31 phosphorylation modulate both the primary envelopment and the subsequent fusion of the nascent virion envelope with the outer nuclear membrane.
Region Required for Protein Expression from the Stop-Start Pentanucleotide in the M Gene of Influenza B Virus
Authors: Hatta Masato , Kohlmeier Candice K. , Hatta Yasuko , Ozawa Makoto , Kawaoka Yoshihiro
Published: March 2009

Segment 7 of influenza B virus encodes two proteins, M1 and BM2. BM2 is expressed from a stop-start pentanucleotide, in which the BM2 initiation codon overlaps with the M1 stop codon. Here, we demonstrate that 45 nucleotides of the 3′ end of the M1 coding region, but not the 5′ end of the BM2 coding region, are sufficient for the efficient expression of the downstream protein. Placing these 45 nucleotides and the stop-start pentanucleotide in between the coding sequences induced the expression of at least three noninfluenza proteins, suggesting the utility of this system for expressing multiple proteins from one mRNA.