bluuebunny/crossref_metadata_2025 huggingface dataset:
SECONDARY AND TERTIARY HYPERPARATHYROIDISM: ROLE OF PREOPERATIVE LOCALIZATION
Authors: Lai Eric C. H. , Ching Alex S. C. , Leong Heng Tat
Published: September 2007

Background:  The usefulness of both technetium Tc‐99m sestamibi (MIBI) scintigraphy and ultrasonography (USG) scan for the detection of enlarged parathyroid glands secondary to renal hyperparathyroidism is rarely addressed.

Methods:  A retrospective study from July 1999 to June 2005 was carried out on patients with secondary and tertiary hyperparathyroidism to determine the role of preoperative localization.

Results:  In the 5 years, 73 patients with renal hyperparathyroidism underwent initial bilateral neck exploration with total parathyroidectomy. Four patients underwent neck exploration with parathyroidectomy for persistent hyperparathyroidism. Two patients underwent neck exploration with parathyroidectomy for recurrent hyperparathyroidism. For patients with initial secondary/tertiary hyperparathyroidism, MIBI scintigraphy correctly showed 101 of 276 (36.6%) surgically confirmed enlarged parathyroids, whereas USG scan showed 99 of 276 (35.9%) surgically confirmed enlarged parathyroids. For persistent or recurrent secondary/tertiary hyperparathyroidism, MIBI scintigraphy and USG scan had sensitivity of 100 and 50%, respectively.

Conclusions:  In conclusion, preoperative localization studies have a limited value when used before first neck exploration in secondary/tertiary hyperparathyroidism because of the poor results in identifying all parathyroid glands. In persistent/recurrent hyperparathyroidism, it may play a useful role in localization of the missed or ectopic parathyroid gland.
Mobilizing Market Forces to Combat Global Environmental Change: Lessons from UN–Private Sector Partnerships in China
Authors: Khoday Kishan
Published: September 2007

While globalization has brought far‐reaching benefits to communities around the world in the form of increasing foreign investment and trade, and reduced levels of poverty, the externalities of the global market have also taken on greater prominence. In particular, issues of global environmental change now stand as a central concern for governments around the world, with increasing threats to the sustainability of hard‐won development gains. While international frameworks such as the United Nations conventions on climate change and biodiversity have been enacted to take joint action on issues of common concern, a major challenge has been to enact effective implementation regimes to achieve results on the ground. One hope lays in the forces of the market itself, engaging global market forces and the role of the private sector to facilitate a global shift to sustainable growth and business practices. This article analyses this challenge and emerging opportunities for market‐based approaches to implement international environmental law through a case study of China and the innovative partnerships being forged there between the UN, governmental and private institutions.
Constructing British Criminology
Authors: SOOTHILL KEITH , PEELO MOIRA
Published: September 2007

Abstract: How has knowledge been constructed in British criminology since the 1960s? While histories of theory are plentiful and, due to such activities as the Research Assessment Exercise, awareness of citation counts has grown, we have become interested in a less formal – harder to assess – area of knowledge construction. Our questions have formed around the ways in which current, practising criminologists perceive the development of their discipline (if it is sufficiently unitary to be called such), and what has influenced them more directly. In so doing, we are attempting to tap into the creative impact on criminology and criminologists of the range of studies that do not necessarily figure as largely in international citation studies. In collecting from fellow‐criminologists a sense of which studies and writers have both shaped criminology and influenced their own thinking, we have arrived at a paradoxical picture of British criminology: one in which there is tension between how current practitioners present a highly‐fragmented, wide‐ranging set of influences, yet do so within a discipline in which there appears to be constant repetition of similar questions over time.
Go for it – life is full of opportunities. Promising results of the YARi working model with young people
Authors: Saarnio Tuula , Nissinen Pasi
Published: September 2007

This article describes a framework of the YARi model, developed as part of the European Union's (EU) Leonardo da Vinci project, called Youth at Risk, international (2003–2006). In the article we present the results of the implementation of the YARi working model. The research was conducted as a comparative case study and applied a qualitative inductive analysis of the content. The YARi working model was implemented simultaneously in four EU countries – Finland (Espoo and Vihti), Italy (Palermo and Naples), Holland (Nijmegen) and the UK (Bradford) – between November 2004 and April 2005.
Post‐IPO Operating Performance, Venture Capital and the Bubble Years
Authors: Coakley Jerry , Hadass Leon , Wood Andrew
Published: September 2007

Abstract:  We analyse the post‐issue operating performance of 316 venture‐backed and 274 non‐venture UK IPOs 1985–2003. The finding of a statistically significant five‐year, operational decline exhibited over the full sample period is not robust. Rather it is driven by the dramatic underperformance during the 1998–2000 bubble years while IPOs perform normally in the remaining years. Cross‐section regression results indicate support for venture capital certification in the non‐bubble years but a significantly negative relationship between operating performance and venture capitalist board representation during the bubble years. The bubble year underperformance is explained by market timing and by low quality companies taking advantage of investor sentiment.
The timescale and mechanisms of fault sealing and water‐rock interaction after an earthquake
Authors: CLAESSON L. , SKELTON A. , GRAHAM C. , MÖRTH C.‐M.
Published: September 2007

Hydrogeochemical monitoring of a basalt‐hosted aquifer, which contains Ice Age meteoric water and is situated at 1220 m below sea level in the Tjörnes Fracture Zone, northern Iceland, has been ongoing since July 2002. Based on hydrogeochemical changes following an earthquake of magnitude (Mw) 5.8 on 16 September 2002, we constrained the timescales of post‐seismic fault sealing and water–rock interaction. We interpret that the earthquake ruptured a hydrological barrier, permitting a rapid influx of chemically and isotopically distinct Ice Age meteoric water from a second aquifer. During the two subsequent years, we monitored a chemical and isotopic recovery towards pre‐earthquake aquifer compositions, which we interpret to have been mainly facilitated by fault‐sealing processes. This recovery was interrupted in November 2004 by a second rupturing event, which was probably induced by two minor earthquakes and which reopened the pathway to the second aquifer. We conclude that the timescale of fault sealing was approximately 2 years and that the approach to isotopic equilibrium (from global meteoric water line) was approximately 18% after >104 years.
Treatment of Hypertension and Dyslipidemia or Their Combination Among US Managed‐Care Patients
Authors: Dutro Michael P. , Gerthoffer Tracey D. , Peterson Eric D. , Tang Simon S. K. , Goldberg George A.
Published: September 2007

The authors examined treatment rates in managed‐care patients with hypertension (HTN) only or dyslipidemia (DYS) only compared with patients who had both (HTN+DYS). A retrospective, cross‐sectional claims analysis was performed in a 2002 US national managed‐care database of 1.23 million continuously eligible members aged 18 years or older. Median age was 44.0 years, 8.8% were aged 65 years or older, and 53.2% were women. Study criteria identified 354,324 patients, 32.9% with HTN only, 34.7% with DYS only, and 32.4% with HTN+DYS. Overall, 49.7% of HTN patients had DYS and 48.3% of DYS patients had HTN. Patients with HTN+DYS were significantly older, more likely to have cardiovascular comorbidities, and more likely to use medications and hospital facilities than were patients with HTN only or DYS only (P<.01). About two‐thirds of patients with HTN only received 1 or more prescription for an antihypertensive medication, compared with three‐quarters of those with HTN+DYS. Fewer than half of patients with DYS only and approximately two‐thirds with HTN+DYS received a cholesterol‐lowering agent.
Is Echo‐Determined Left Ventricular Geometry Associated with Ventricular Filling and Midwall Shortening in Hypertensive Ventricular Hypertrophy?
Authors: Palmiero Pasquale , Maiello Maria , Nanda Navin C
Published: September 2007

Background: The correlation between left ventricular (LV) geometry, mass, diastolic function, and midwall fractional shortening (MFS) in hypertensive patients with left ventricular hypertrophy (LVH) is not well established owing to limited diffusion of MFS evaluation. The aim of the study was to evaluate this correlation in 1887 consecutive hypertensive patients, all affected by LVH (mean age 66 years, 924 males), with LV ejection fraction (LVEF) >45% for early detection of ventricular dysfunction rather than LVEF and diastolic function impairment. Methods and Results: All patients underwent M‐B mode echocardiography and PW‐Doppler evaluation. LV geometry and mass were compared with Doppler‐determined mitral flow and tissue velocities. LV geometry was eccentric (E) for 1018 subjects (53.9%) and concentric (C) for 869 (46.1%). There was no difference concerning LV diastolic dysfunction (P: n.s.) between 576 (30.6%) of the ELVH and 368 (19.4%) of the CLVH patients. The following parameters showed significant statistical differences: LV MFS impairment (P < 0.01) between 86 (4.6%) of the ELVH and 177 (9.4%) of the CLVH patients. LV MFS impairment rate was higher in 171 patients without LV diastolic dysfunction (9.1%), than in 92 patients affected (4.9%, P < 0.02). In CLVH patients, a higher prevalence of LV MFS impairment was observed in 143 without LV diastolic dysfunction (7.6%), than in 34 patients affected (1.8%, P < 0.01). In ELVH patients, a lower prevalence of LV MFS impairment was observed in 28 without diastolic dysfunction (1.5%), than in 58 patients affected (3.1%, P < 0,03). Conclusion: Midwall LV impairment, an independent predictor of cardiac death and morbidity in hypertensive patients, can allow early identification of patients with LV dysfunction even when LVEF or assessment of diastolic function are normal. LV MFS impairment rate is higher in CLVH patients, and even higher when considering only those CLVH patients with no diastolic dysfunction. These results suggest that the ventricular dysfunction with normal LVEF is not always due to diastolic dysfunction, but often to systolic dysfunction as assessed by MFS impairment, an important early sign of ventricular dysfunction in hypertensive patients, even when diastolic function is normal.
Immunosuppression and Squamous Cell Carcinoma: A Focus on Solid Organ Transplant Recipients
Authors: Herman Sara , Rogers Heather D. , Ratner Désirée
Published: September 2007

As transplant medicine advances, new immunosuppressive regimens are increasing the long‐term survival of solid organ transplant recipients (SOTRs). This growing population is at significantly increased risk for developing cutaneous malignancies, particularly squamous cell carcinoma (SCC), as a result of chronic immunosuppression. Conventional risk factors for the development of skin cancer, including fair skin type, advanced age, sun exposure, and genetic predisposition, also play a crucial role in the initiation and progression of SCC in SOTRs. Immunosuppressed patients develop more aggressive and more numerous SCCs than immunocompetent individuals, however. It is important to understand the mechanisms underlying immunosuppression‐mediated SCC development to identify prognostic markers and to develop effective prevention and treatment strategies. This article addresses the fundamental differences between SCC in SOTRs and those in the general population, focusing on the role that immunosuppression plays in the pathogenesis of this malignancy.
Rheumatoid Arthritis Associated With Pemphigus Foliaceus in a Patient Not Taking Penicillamine
Authors: Sáez‐de‐Ocariz Marimar , Granados Julio , Yamamoto‐Furusho Jesús K. , López‐Martínez Alondra , Vega‐Memije María Elisa
Published: September 2007

A 50‐year‐old‐woman presented with a history of crusted plaques on her face and trunk. On physical examination, she had a disseminated dermatosis involving the face, chest, trunk, abdomen, and both ankles. She had erythematous and scaly plaques on both cheeks (Figure 1). In both ears and on the rest of the body, she presented with scarce intact vesicles and blisters, a few eroded and crusted vesicles, and residual hyperpigmented macules. The onset of the dermatosis occurred 6 months before diagnosis, affecting the chest, followed by further spreading of vesicles on the body. She denied ever having developed ulcerative or erosive lesions on mucosal surfaces. The presence of blisters were followed by the development of crusts and hyperpigmented macules. One month before visiting our service, the patient had been receiving treatment with dapsone, with slight improvement of the cutaneous lesions. Biopsy results of one of the scaly plaques on the cheeks demonstrated a subcorneal blister with a few acantholytic cells and a scarce inflammatory infiltrate in the papillary dermis (Figure 2). Because of the clinical picture, especially the erythematous lesions on both cheeks, tests for antinuclear antibodies were performed to rule out pemphigus erythematosus, and the results were negative. Besides the cutaneous manifestations, the patient had had signs and symptoms of arthritis for at least 11 years; she had received sporadic relief with the nonsteroidal anti‐inflammatory drug ketoprofen. She reported generalized morning rigidity lasting for 2 hours and arthralgias in the temporomandibular joints, elbows, wrists, metacarpophalangeal joints, knees, and ankles. On examination, she had phlogosis and synovial thickening in the elbows, wrists, metacarpophalangeal joints, knees, and ankles. She also had flexion contractures on both elbows as well as ulnar deviation of the left hand and swan‐neck deformity in several fingers. Radiographic examination of the hands demonstrated generalized osteopenia, decreased articular spaces in metacarpophalangeal and proximal interphalangeal joints, and slight erosion in the carpal bones (Figure 3). Results of rheumatoid factor testing were positive (521 IU/mL). The diagnoses of both pemphigus foliaceus and rheumatoid arthritis were made. We performed high‐resolution DNA typing of HLA‐DRB1 polymorphism using sequence‐specific oligonucleotides,1 and found that the patient carried HLA DRB1*0404 and *1406.

The patient had erythematous and scaly plaques on both cheeks.

image

Biopsy revealed a subcorneal blister with a few acantholytic cells and a scarce inflammatory infiltrate in the papillary dermis.

image

Radiography of the hands, revealed generalized osteopenia, decreased articular spaces in metacarpophalangeal and proximal interphalangeal joints, and slight erosion in the carpal bones.

image
Updates on bisphosphonates and potential pathobiology of bisphosphonate‐induced jaw osteonecrosis
Authors: Sarin J , DeRossi SS , Akintoye SO
Published: September 2007

Osteonecrosis of the jaws is a major complication associated with long‐term use of bisphosphonates. While osteonecrosis can arise from other precipitating conditions, bisphosphonate‐induced jaw osteonecrosis (BJON) is highly associated with long‐term administration of pamidronate (Aredia®) and zoledronic acid (Zometa®), which are two intravenous bisphosphonate formulations. The underlying pathogenesis of BJON and its site‐specific presentation still remain to be fully elucidated. This review will discuss clinically available bisphosphonates, current opinions, pathogenesis, and management guidelines for bisphosphonate‐induced jaw osteonecrosis.
GRB 060206 and the quandary of achromatic breaks in afterglow light curves
Authors: Curran P. A. , van der Horst A. J. , Wijers R. A. M. J. , Starling R. L. C. , Castro-Tirado A. J. , Fynbo J. P. U. , Gorosabel J. , Järvinen A. S. , Malesani D. , Rol E. , Tanvir N. R. , Wiersema K. , Burleigh M. R. , Casewell S. L. , Dobbie P. D. , Guziy S. , Jakobsson P. , Jelínek M. , Laursen P. , Levan A. J. , Mundell C. G. , Näränen J. , Piranomonte S.
Published: September 2007

Gamma-ray burst afterglow observations in the Swift era have a perceived lack of achromatic jet breaks compared with the BeppoSAX era. We present our multi-wavelength analysis of GRB060206 as an illustrative example of how inferences of jet breaks from optical and X-ray data might differ. The results of temporal and spectral analyses are compared, and attempts are made to fit the data within the context of the standard blast wave model. We find that while the break appears more pronounced in the optical and evidence for it from the X-ray alone is weak, the data are actually consistent with an achromatic break at about 16 h. This break and the light curves fit standard blast wave models, either as a jet break or as an injection break. As the pre-Swift sample of afterglows are dominated by optical observations, and in the Swift era most well-sampled light curves are in the X-ray, caution is needed when making a direct comparison between the two samples, and when making definite statements on the absence of achromatic breaks.
A Template‐assembled Model of the N‐peptide Helix Bundle from HIV‐1 Gp‐41 with High Affinity for C‐peptide
Authors: Xu Weiming , Taylor John W.
Published: September 2007

The infection of target cells by HIV‐1 is initiated by fusion of the viral and cell membranes, which is mediated by the viral glycoproteins, gp120 and gp41. After initial cell binding by gp120, the folding of gp41 to form a stable six‐helix bundle structure is directly associated with membrane fusion. This helix bundle is composed of an α‐helical trimer of gp41 N‐peptide, with three copies of the α‐helical gp41 C‐peptide folded onto it in an antiparallel orientation. This report describes the synthesis and study of an N‐peptide three‐helix bundle structure, KTA‐3N29b, that is assembled on a threefold symmetric template derived from Kemp’s triacid. At neutral pH and in the presence of physiologic salt, KTA‐3N29b, exists as a monomer with high helix content. Binding isotherms measured by circular dichroism spectropolarimetry indicate that KTA‐3N29b binds three copies of the C‐peptide native sequence, with a KD of about 260 nm. These features of KTA‐3N29b demonstrate that this templated three‐helix bundle serves as a functional model for the native N‐peptide structure that will allow detailed studies of the folding and thermodynamic stability of the gp41 six‐helix bundle, and may aid the future development of potent HIV‐1 fusion inhibitors and immunogens.
Pre‐harvest Interventions to Reduce the Shedding ofE. coliO157 in the Faeces of Weaned Domestic Ruminants: A Systematic Review
Authors: Sargeant J. M. , Amezcua M. R. , Rajic A. , Waddell L.
Published: September 2007

Our objective was to use formal systematic review methods to evaluate the efficacy of interventions to reduce faecal shedding ofEscherichia coliO157 in post‐weaned ruminants by increasing animal resistance. The methodology consisted of an extensive search to identify all potentially relevant research, screening of titles and abstracts for relevance to the research question, quality assessment of relevant research, extraction of data from research of sufficient quality, and qualitative summarization of results. The interventions evaluated included probiotics, vaccination, antimicrobials, sodium chlorate, bacteriophages and other feed additives. There was evidence of efficacy for the probiotic combinationLactobacillus acidophilusNP51 (NPC 747) andPropionibacterium freudenreichiiand for sodium chlorate in feed or water. The effectiveness of vaccination varied among studies and among vaccine protocols and there was no consistent evidence to suggest that antibiotic use was associated with a decrease in faecal shedding ofE. coliO157, or that current industry uses of antimicrobials were associated with increased faecal shedding. There were an insufficient number of studies available to address the effectiveness of bacteriophages and several other feed additives. In general, few of the primary studies evaluated the interventions under commercial housing conditions with a natural disease challenge, there were inconsistencies in the results among study designs and in some cases among studies within study designs, and a relatively large proportion of publications were excluded based on quality assessment criteria. Few studies reported on associations between the proposed intervention and production parameters, such as average daily gain and feed: gain ratio. While the results suggest that some interventions may be efficacious, there are knowledge gaps in our understanding of the efficacy of pre‐harvest interventions to increase animal resistance toE. coliO157 that require further targeted research.
Penetration of Amphotericin B Lipid Formulations into Pleural Effusion
Authors: Weiler Stefan , Bellmann-Weiler Rosa , Joannidis Michael , Bellmann Romuald
Published: September 2007

The penetration of the amphotericin B (AMB) lipid formulations (liposomal AMB, AMB colloidal dispersion, and AMB lipid complex formulations) into pleural effusions in seven critically ill patients was assessed. AMB was detected in all pleural effusion samples at concentrations ranging from 0.02 to 0.43 μg/ml. The penetration ratio was 3 to 44%.
Toll-Like Receptor 2 Controls the Gamma Interferon Response toFrancisella tularensisby Mouse Liver Lymphocytes
Authors: Hong Kee-Jong , Wickstrum Jason R. , Yeh Hung-Wen , Parmely Michael J.
Published: September 2007

The production of gamma interferon (IFN-γ) is a key step in the protective innate immune response toFrancisella tularensis. Natural killer cells and T cells in the liver are important sources of this cytokine during primaryF. tularensisinfections, and interleukin-12 (IL-12) appears to be an essential coactivating cytokine for hepatic IFN-γ expression. The present study was undertaken to determine whether or not macrophages (Mφ) or dendritic cells (DC) provide coactivating signals for the liver IFN-γ response in vitro, whether IL-12 mediates these effects, and whether Toll-like receptor (TLR) signaling is essential to induce this costimulatory activity. Both bone marrow-derived Mφ and DC significantly augmented the IFN-γ response ofF. tularensis-challenged liver lymphocytes in vitro. While both cell types produced IL-12p40 in response toF. tularensischallenge, only DC secreted large quantities of IL-12p70. DC from both IL-12p35-deficient and TLR2-deficient mice failed to produce IL-12p70 and did not costimulate liver lymphocytes for IFN-γ production in response to viableF. tularensisorganisms. Conversely, liver lymphocytes from TLR2-deficient mice cocultured with wild-type accessory cells produced IFN-γ at levels comparable to those for wild-type hepatic lymphocytes. These findings indicate that TLR2 controls hepatic lymphocyte IFN-γ responses toF. tularensisby regulating DC IL-12 production. While Mφ also coinduced hepatic IFN-γ production in response toF. tularensis, they did so in a fashion less dependent on TLR2.
The
Mycobacterium bovis
BCG Cyclic AMP Receptor-Like Protein Is a Functional DNA Binding Protein In Vitro and In Vivo, but Its Activity Differs from That of Its
M. tuberculosis
Ortholog, Rv3676
Authors: Bai Guangchun , Gazdik Michaela A. , Schaak Damen D. , McDonough Kathleen A.
Published: September 2007

Mycobacterium tuberculosis
Rv3676 encodes a cyclic AMP (cAMP) receptor-like protein (CRP
Mt
) that has been implicated in global gene regulation and may play an important role during tuberculosis infection. The CRP
Mt
ortholog in
Mycobacterium bovis
BCG, CRP
BCG
, is dysfunctional in an
Escherichia coli
CRP competition assay and has been proposed as a potential source of
M. bovis
BCG's attenuation. We compared CRP
BCG
and CRP
Mt
in vitro and in vivo, in
M. bovis
BCG and
M. tuberculosis
, to evaluate CRP
BCG
's potential function in a mycobacterial system. Both proteins formed dimers in mycobacterial lysates, bound to the same target DNA sequences, and were similarly affected by the presence of cAMP in DNA binding assays. However, CRP
Mt
and CRP
BCG
differed in their relative affinities for specific DNA target sequences and in their susceptibilities to protease digestion. Surprisingly, CRP
BCG
DNA binding activity was stronger than that of CRP
Mt
both in vitro and in vivo, as measured by electrophoretic mobility shift and chromatin immunoprecipitation assays. Nutrient starvation-associated regulation of several CRP
Mt
regulon members also differed between
M. bovis
BCG and
M. tuberculosis
. We conclude that CRP
BCG
is a functional cAMP-responsive DNA binding protein with an in vivo DNA binding profile in
M. bovis
BCG similar to that of CRP
Mt
in
M. tuberculosis
. However, biologically significant functional differences may exist between CRP
BCG
and CRP
Mt
with respect to gene regulation, and this issue warrants further study.
Galectin-3 Modulates Immune and Inflammatory Responses during Helminthic Infection: Impact of Galectin-3 Deficiency on the Functions of Dendritic Cells
Authors: Breuilh Laetitia , Vanhoutte François , Fontaine Josette , van Stijn Caroline M. W. , Tillie-Leblond Isabelle , Capron Monique , Faveeuw Christelle , Jouault Thierry , van Die Irma , Gosset Philippe , Trottein François
Published: September 2007

Galectin-3 (Gal-3) is a multifunctional β-galactoside-binding lectin that senses self-derived and microbial glycoconjugates. Although Gal-3 is important in immune reactions and host defense in some experimental models, the function of Gal-3 during helminthic diseases (e.g., schistosomiasis) is still elusive. We show that, compared to wild-typeSchistosoma mansoni-infected mice, infected Gal-3−/−mice have a reduced number of T and B lymphocytes in the spleen, develop reduced liver granulomas at 7 weeks (acute phase) and 14 weeks (chronic phase) postinfection, and mount a biased cellular and humoral Th1 response. In an attempt to understand this latter phenomenon, we studied the role of endogenous Gal-3 in dendritic cells (DCs), the most potent antigen-presenting cells, both in vitro and in vivo. Although Gal-3 deficiency in DCs does not impact their differentiation and maturation processes, it greatly influences the strength (but not the nature) of the adaptive immune response that they trigger, suggesting that Gal-3 deficiency in some other cell types may be important during murine schistosomiasis. As a whole, this study implies that Gal-3 is a modulator of the immune/inflammatory responses during helminthic infection and reveals for the first time that Gal-3 expression in DCs is pivotal to control the magnitude of T-lymphocyte priming.
Feasibility of a Molecular Screening Method for Detection ofSalmonella entericaandCampylobacter jejuniin a Routine Community-Based Clinical Microbiology Laboratory
Authors: Schuurman T. , de Boer R. F. , van Zanten E. , van Slochteren K. R. , Scheper H. R. , Dijk-Alberts B. G. , Möller A. V. M. , Kooistra-Smid A. M. D.
Published: September 2007

Conventional diagnostic methods for the detection ofSalmonella entericaandCampylobacter jejuniare laborious and time-consuming procedures, resulting in final results, for the majority of specimens, only after 3 to 4 days. Molecular detection can improve the time to reporting of the final results from several days to the next day. However, molecular assays for the detection of gastrointestinal pathogens directly from stool specimens have not made it into the routine clinical microbiology laboratory. In this study we have assessed the feasibility of a real-time PCR-based molecular screening method (MSM), aimed atS. entericaandC. jejuni, in the daily practice of a routine clinical microbiology laboratory. We have prospectively analyzed 2,067 stool specimens submitted for routine detection of gastrointestinal bacterial pathogens over a 7-month period. The MSM showed 98 to 100% sensitivity but routine culture showed only 77.8 to 86.8% sensitivity when an extended “gold standard” that included all culture-positive and all MSM-positive specimens, as confirmed by an independent secondary PCR of a different target gene, was used. An overall improvement in the rate of detection of both pathogens of 15 to 18% was observed. Both approaches performed nearly identically with regard to the specificity, positive predictive value, and negative predictive value, with the values for MSM being 99.7%, 93.1 to 96.6%, and 99.8 to 100%, respectively, and those for routine culture being 100%, 100%, and 97.6 to 99.5%, respectively. Finally, the final results were reported between 3 and 4 days earlier for negative specimens compared to the time of reporting of the results of routine culture. Positive specimens, on the other hand, required an additional 2 days to obtain a final result compared to the time required for routine culture, although preliminary MSM PCR-positive results were reported, on average, 2.9 to 3.8 days before the final routine culture results were reported. In conclusion, MSM can be incorporated into the daily practice of a routine clinical microbiology laboratory with ease. Furthermore, it provides an improvement in the screening forS. entericaandC. jejuniand substantially improves the time to the reporting of negative results.
Accuracy of
Burkholderia pseudomallei
Identification Using the API 20NE System and a Latex Agglutination Test
Authors: Amornchai Premjit , Chierakul Wirongrong , Wuthiekanun Vanaporn , Mahakhunkijcharoen Yuvadee , Phetsouvanh Rattanaphone , Currie Bart J. , Newton Paul N. , van Vinh Chau Nguyen , Wongratanacheewin Surasakdi , Day Nicholas P. J. , Peacock Sharon J.
Published: September 2007

In an evaluation of the API 20NE for the identification of
Burkholderia
spp., 792/800 (99%)
Burkholderia pseudomallei
and 17/19 (89%)
B. cepacia
isolates were correctly identified but 10
B. mallei
and 98
B. thailandensis
isolates were not correctly identified. A latex agglutination test was positive for 796/800 (99.5%)
B. pseudomallei
isolates and negative for 120 other oxidase-positive gram-negative bacilli.
Genetic Diversity of Paired Middle-Ear and Pharyngeal Nontypeable
Haemophilus influenzae
Isolates from Children with Acute Otitis Media
Authors: Berrens Zachary J. , Marrs Carl F. , Pettigrew Melinda M. , Sandstedt Sara A. , Patel Mayuri , Gilsdorf Janet R.
Published: September 2007

Pulsed-field gel electrophoresis was used to determine genetic diversities of multiple nontypeable
Haemophilus influenzae
isolates from throat and ear specimens of eight children with otitis media. From five children, all ear and throat isolates were identical. The bacterial populations in these specimens showed less diversity than populations in throat isolates of healthy children.
Isolation and Genetic Characterization of a Coinfection of Non-O157 Shiga Toxin-Producing
Escherichia coli
Authors: Gilmour Matthew W. , Tabor Helen , Wang Gehua , Clark Clifford G. , Tracz Dobryan M. , Olson Adam B. , Mascarenhas Mariola , Karmali Mohamed A. , Mailman Tim , Ng Lai-King
Published: September 2007

A coinfection of O177:NM and O55:H7 Shiga toxin-producing
Escherichia coli
(STEC) was identified for a child with acute bloody diarrhea and hemolytic uremic syndrome by using culture and serotype-specific molecular reagents. The profile of O157-related genetic islands revealed that the O55:H7 isolate was highly similar to O157 STEC whereas the O177:NM isolate lacked several fimbrial O islands and non-locus-of-enterocyte-effacement effector determinants. However, both STEC serotypes are known to cause serious disease, and the significant repertoire of virulence determinants in both strains made it impossible to determine their individual contributions to the clinical symptoms.
Use of Peptide Nucleic Acid-Fluorescence In Situ Hybridization for Definitive, Rapid Identification of Five Common
Candida
Species
Authors: Reller Megan E. , Mallonee Amanda B. , Kwiatkowski Nicole P. , Merz William G.
Published: September 2007

We investigated a 2.5-h peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) assay with five
Candida
species-specific probes to identify
Candida
colonies and compared it to standard 2-h to 5-day phenotypic identification methods. Suspensions were made and slides were prepared and read for fluorescence per the manufacturer's instructions. Sensitivity was 99% (109/110), and specificity was 99% (129/130). PNA-FISH can rapidly identify those
Candida
species isolated most frequently.
Yield of Stool Culture with Isolate Toxin Testing versus a Two-Step Algorithm Including Stool Toxin Testing for Detection of Toxigenic
Clostridium difficile
Authors: Reller Megan E. , Lema Clara A. , Perl Trish M. , Cai Mian , Ross Tracy L. , Speck Kathleen A. , Carroll Karen C.
Published: September 2007

We examined the incremental yield of stool culture (with toxin testing on isolates) versus our two-step algorithm for optimal detection of toxigenic
Clostridium difficile
. Per the two-step algorithm, stools were screened for
C. difficile
-associated glutamate dehydrogenase (GDH) antigen and, if positive, tested for toxin by a direct (stool) cell culture cytotoxicity neutralization assay (CCNA). In parallel, stools were cultured for
C. difficile
and tested for toxin by both indirect (isolate) CCNA and conventional PCR if the direct CCNA was negative. The “gold standard” for toxigenic
C. difficile
was detection of
C. difficile
by the GDH screen or by culture and toxin production by direct or indirect CCNA. We tested 439 specimens from 439 patients. GDH screening detected all culture-positive specimens. The sensitivity of the two-step algorithm was 77% (95% confidence interval [CI], 70 to 84%), and that of culture was 87% (95% CI, 80 to 92%). PCR results correlated completely with those of CCNA testing on isolates (29/29 positive and 32/32 negative, respectively). We conclude that GDH is an excellent screening test and that culture with isolate CCNA testing detects an additional 23% of toxigenic
C. difficile
missed by direct CCNA. Since culture is tedious and also detects nontoxigenic
C. difficile
, we conclude that culture is most useful (i) when the direct CCNA is negative but a high clinical suspicion of toxigenic
C. difficile
remains, (ii) in the evaluation of new diagnostic tests for toxigenic
C. difficile
(where the best reference standard is essential), and (iii) in epidemiologic studies (where the availability of an isolate allows for strain typing and antimicrobial susceptibility testing).
Modification of Cellular Autophagy Protein LC3 by Poliovirus
Authors: Taylor Matthew P. , Kirkegaard Karla
Published: September 2007

Poliovirus infection remodels intracellular membranes, creating a large number of membranous vesicles on which viral RNA replication occurs. Poliovirus-induced vesicles display hallmarks of cellular autophagosomes, including delimiting double membranes surrounding the cytosolic lumen, acquisition of the endosomal marker LAMP-1, and recruitment of the 18-kDa host protein LC3. Autophagy results in the covalent lipidation of LC3, conferring the property of membrane association to this previously microtubule-associated protein and providing a biochemical marker for the induction of autophagy. Here, we report that a similar modification of LC3 occurs both during poliovirus infection and following expression of a single viral protein, a stable precursor termed 2BC. Therefore, one of the early steps in cellular autophagy, LC3 modification, can be genetically separated from the induction of double-membraned vesicles that contain the modified LC3, which requires both viral proteins 2BC and 3A. The existence of viral inducers that promote a distinct aspect of the formation of autophagosome-like membranes both facilitates the dissection of this cellular process and supports the hypothesis that this branch of the innate immune response is directly subverted by poliovirus.
Interaction of Decay-Accelerating Factor with Coxsackievirus B3
Authors: Hafenstein Susan , Bowman Valorie D. , Chipman Paul R. , Kelly Carol M. Bator , Lin Feng , Medof M. Edward , Rossmann Michael G.
Published: September 2007

Many entero-, parecho-, and rhinoviruses use immunoglobulin (Ig)-like receptors that bind into the viral canyon and are required to initiate viral uncoating during infection. However, some of these viruses use an alternative or additional receptor that binds outside the canyon. Both the coxsackievirus-adenovirus receptor (CAR), an Ig-like molecule that binds into the viral canyon, and decay-accelerating factor (DAF) have been identified as cellular receptors for coxsackievirus B3 (CVB3). A cryoelectron microscopy reconstruction of a variant of CVB3 complexed with DAF shows full occupancy of the DAF receptor in each of 60 binding sites. The DAF molecule bridges the canyon, blocking the CAR binding site and causing the two receptors to compete with one another. The binding site of DAF on CVB3 differs from the binding site of DAF on the surface of echoviruses, suggesting independent evolutionary processes.
Characterization of the Intracellular Deproteinized Relaxed Circular DNA of Hepatitis B Virus: an Intermediate of Covalently Closed Circular DNA Formation
Authors: Guo Haitao , Jiang Dong , Zhou Tianlun , Cuconati Andrea , Block Timothy M. , Guo Ju-Tao
Published: September 2007

Covalently closed circular DNA (cccDNA) of hepatitis B virus (HBV) is formed by conversion of capsid-associated relaxed circular DNA (rcDNA) via unknown mechanisms and exists in the nucleus of the infected hepatocyte as a minichromosome that serves as the transcription template for viral RNAs. To study the molecular pathway of cccDNA formation and its regulation by viral and cellular factors, we have established a cell line that supports the replication of an envelope protein-deficient HBV genome in a tetracycline-inducible manner. Following induction of HBV replication, the cells accumulate higher levels of cccDNA as well as larger amounts of deproteinized rcDNA (DP-rcDNA) than cells that replicate wild-type HBV genomes. These results indicate that HBV envelope proteins negatively regulate cccDNA formation, and conversion of DP-rcDNA into cccDNA is a rate-limiting step of cccDNA formation in HepG2 cells. Detailed analyses reveal the following: (i) DP-rcDNA exists in both cytoplasm and nucleus; (ii) while nuclear DP-rcDNA is sensitive to DNase I digestion, a small fraction of cytoplasmic DP-rcDNA is DNase I resistant; (iii) both DNase I-sensitive and -resistant cytoplasmic DP-rcDNAs cosediment with capsids and can be immunoprecipitated with HBV core antibody; and (iv) a primer extension assay maps the 5′ end of the minus strand of DP-rcDNA at the authentic end of virion rcDNA. Hence, our results favor a hypothesis that the removal of viral polymerase protein covalently linked to the 5′ end of the minus-strand DNA occurs inside the capsid in the cytoplasm and most possibly via a reaction that cleaves the phosphodiester bond between the tyrosine of the polymerase and the 5′ phosphoryl group of minus-strand DNA.
Fas Ligand Interactions Contribute to CD8+T-Cell-Mediated Control of West Nile Virus Infection in the Central Nervous System
Authors: Shrestha Bimmi , Diamond Michael S.
Published: September 2007

West Nile virus (WNV) is a neurotropic flavivirus that causes encephalitis, most frequently in elderly and immunocompromised humans. Previous studies demonstrated that CD8+T cells utilize perforin-dependent cytolytic mechanisms to limit WNV infection. Nonetheless, the phenotype of perforin-deficient CD8+T cells was not as severe as that of an absence of CD8+T cells, suggesting additional effector control mechanisms. In this study, we evaluated the contribution of Fas-Fas ligand (FasL) interactions to CD8+T-cell-mediated control of WNV infection. Notably, the cell death receptor Fas was strongly upregulated on neurons in culture and in vivo after WNV infection.gldmice that were functionally deficient in FasL expression showed increased susceptibility to lethal WNV infection. Although antigen-specific priming of CD8+T cells in peripheral lymphoid tissues was normal ingldmice, increased central nervous system (CNS) viral burdens and delayed clearance were observed. Moreover, the adoptive transfer of WNV-primed wild-type but notgldCD8+T cells to recipient CD8−/−orgldmice efficiently limited infection in the CNS and enhanced survival rates. Overall, our data suggest that CD8+T cells also utilize FasL effector mechanisms to contain WNV infection in Fas-expressing neurons in the CNS.
B7 Costimulation Molecules Expressed from the Herpes Simplex Virus 2 Genome Rescue Immune Induction in B7-Deficient Mice
Authors: Thebeau Lydia G. , Vagvala Sri P. , Wong Yee M. , Morrison Lynda A.
Published: September 2007

The interaction between B7 costimulation molecules on antigen-presenting cells and CD28 on antigen-responsive T cells is essential for T-cell activation and maturation of immune responses to herpes simplex virus (HSV) infection. Vaccine-induced immune responses also depend upon adequate upregulation of B7 costimulation molecules, but this signal may be limiting for replication-defective virus vaccines. We investigated whether expression of B7 costimulation molecules by a prototypical replication-defective antiviral vaccine could enhance immune responses to the vaccine and whether B7-1 and B7-2 would be similarly effective. We altered an ICP8−replication-defective strain of HSV type 2 (HSV-2), 5BlacZ, to encode either murine B7-1 or B7-2. B7 molecule expression was detected on the surface of cells infected in vitro and at the RNA level in tissue of immunized mice. Immunization of B7-1/B7-2 knockout mice with B7-encoding virus modestly expanded the number of gamma interferon-producing T cells and significantly augmented class-switched HSV-specific antibody responses compared with the parental virus. Mice immunized with either B7-expressing virus showed less replication of challenge virus in the genital mucosa than mice immunized with 5BlacZ, markedly fewer signs of genital and neurological disease, and little weight loss. Virtually all mice immunized with B7-encoding virus survived challenge with a large dose of HSV-2, whereas most 5BlacZ-immunized mice succumbed to infection. These results indicate that protective immune responses can be enhanced by the inclusion of host B7 costimulation molecules in a prototypical replication-defective HSV vaccine against HSV-2 genital infection and that B7-1 and B7-2 induce immune responses with similar capacities to fight HSV-2 infection.
High Fidelity of Murine Hepatitis Virus Replication Is Decreased in nsp14 Exoribonuclease Mutants
Authors: Eckerle Lance D. , Lu Xiaotao , Sperry Steven M. , Choi Leena , Denison Mark R.
Published: September 2007

Replication fidelity of RNA virus genomes is constrained by the opposing necessities of generating sufficient diversity for adaptation and maintaining genetic stability, but it is unclear how the largest viral RNA genomes have evolved and are maintained under these constraints. A coronavirus (CoV) nonstructural protein, nsp14, contains conserved active-site motifs of cellular exonucleases, including DNA proofreading enzymes, and the severe acute respiratory syndrome CoV (SARS-CoV) nsp14 has 3′-to-5′ exoribonuclease (ExoN) activity in vitro. Here, we show that nsp14 ExoN remarkably increases replication fidelity of the CoV murine hepatitis virus (MHV). Replacement of conserved MHV ExoN active-site residues with alanines resulted in viable mutant viruses with growth and RNA synthesis defects that during passage accumulated 15-fold more mutations than wild-type virus without changes in growth fitness. The estimated mutation rate for ExoN mutants was similar to that reported for other RNA viruses, whereas that of wild-type MHV was less than the established rates for RNA viruses in general, suggesting that CoVs with intact ExoN replicate with unusually high fidelity. Our results indicate that nsp14 ExoN plays a critical role in prevention or repair of nucleotide incorporation errors during genome replication. The established mutants are unique tools to test the hypothesis that high replication fidelity is required for the evolution and stability of large RNA genomes.
Affinity Thresholds for Membrane Fusion Triggering by Viral Glycoproteins
Authors: Hasegawa Kosei , Hu Chunling , Nakamura Takafumi , Marks James D. , Russell Stephen J. , Peng Kah-Whye
Published: September 2007

Enveloped viruses trigger membrane fusion to gain entry into cells. The receptor affinities of their attachment proteins vary greatly, from 10
−4
M to 10
−9
M, but the significance of this is unknown. Using six retargeted measles viruses that bind to Her-2/neu with a 5-log range in affinity, we show that receptor affinity has little impact on viral attachment but is nevertheless a key determinant of infectivity and intercellular fusion. For a given cell surface receptor density, there is an affinity threshold above which cell-cell fusion proceeds efficiently. Suprathreshold affinities do not further enhance the efficiency of membrane fusion.
Random T-Cell Receptor Recruitment in Human Immunodeficiency Virus Type 1 (HIV-1)-Specific CD8
+
T Cells from Genetically Identical Twins Infected with the Same HIV-1 Strain
Authors: Yu Xu G. , Lichterfeld Mathias , Williams Katie L. , Martinez-Picado Javier , Walker Bruce D.
Published: September 2007

Human immunodeficiency virus type 1 (HIV-1) cytotoxic T-lymphocyte escape mutations represent both a major reason for loss of HIV immune control and a considerable challenge for HIV-1 vaccine design. Previous data suggest that initial HIV-1-specific CD8
+
T-cell responses are determined largely by viral and host genetics, but the mechanisms influencing the subsequent viral evolution are unclear. Here, we show a random recruitment of T-cell receptor (TCR) alpha and beta clonotypes of the initial HIV-1-specific CD8
+
T cells during primary infection in two genetically identical twins infected simultaneously with the same virus, suggesting that stochastic TCR recruitment of HIV-1-specific CD8
+
T cells contributes to the diverse and unpredictable HIV-1 sequence evolution.
Novel TRIM5 Isoforms Expressed byMacaca nemestrina
Authors: Brennan Greg , Kozyrev Yury , Kodama Toshiaki , Hu Shiu-Lok
Published: September 2007

The TRIM5 family of proteins contains a RING domain, one or two B boxes, and a coiled-coil domain. TheTRIM5α isoform also encodes a C-terminal B30.2(SPRY) domain, differences within which define the breadth and potency of TRIM5α-mediated retroviral restriction. BecauseMacaca nemestrinaanimals are susceptible to some human immunodeficiency virus (HIV) isolates, we sought to determine if differences exist in theTRIM5gene and transcripts of these animals. We identified a two-nucleotide deletion (Δ2) in the transcript at the 5′ terminus of exon 7 in allM. nemestrinaTRIM5 cDNA clones examined. This frameshift results in a truncated protein of 300 amino acids lacking the B30.2(SPRY) domain, which we have named TRIM5θ. This deletion is likely due to a single nucleotide polymorphism that alters the 3′ splice site between intron 6 and exon 7. In some clones, a deletion of the entire 27-nucleotide exon 7 (Δexon7) resulted in the restoration of the TRIM5 open reading frame and the generation of another novel isoform,TRIM5η. There are 18 amino acid differences betweenM. nemestrinaTRIM5η andMacaca mulattaTRIM5α, some of which are at or near locations previously shown to affect the breadth and potency of TRIM5α-mediated restriction. Infectivity assays performed on permissive CrFK cells stably transduced with TRIM5η or TRIM5θ show that these isoforms are incapable of restricting either HIV type 1 (HIV-1) or simian immunodeficiency virus infection. The expression of TRIM5 alleles incapable of restricting HIV-1 infection may contribute to the previously reported increased susceptibility ofM. nemestrinato HIV-1 infection in vivo.