bluuebunny/crossref_metadata_2025 huggingface dataset:
Putative Role of Carbon Monoxide Signaling Pathway in Penile Erectile Function
Authors: Abdel Aziz Mohamed T. , Mostafa Taymour , Atta Hazem , Wassef Mohamed A. , Fouad Hanan H. , Rashed Laila A. , Sabry Dina
Published: January 2009

Introduction

Erectile response depends on nitric oxide (NO) generated by NO synthase (NOS) enzyme of the nerves and vascular endothelium in the cavernous tissue. NO activates soluble guanylate cyclase (sGC), leading to the production of cyclic guanosine monophosphate (cGMP). cGMP activates cGMP-dependent protein kinase that activates Ca2+/ATPase pump that activates Ca2+/K efflux pump extruding Ca2+ across the plasma membrane with consequent smooth muscle cell relaxation. A role similar to that of NOS/NO signaling has been postulated for carbon monoxide (CO) produced in mammals from heme catabolism by heme oxygenase (HO) enzyme.

Aim

To assess CO signaling pathway for erectile function by reviewing published studies.

Methods

A systematic review of published studies on this affair based on Pubmed and Medical Subject Heading databases, with search for all concerned articles.

Main Outcome Measures

Documentation of positive as well as negative criteria of CO/HO signaling focused on penile tissue.

Results

The concept that HO-derived CO could play a role in mediating erectile function acting in synergism with, or as a potentiator for, NOS/NO signaling pathway is gaining momentum. CO/HO signaling pathway has been shown to partially mediate the actions of oral phosphodiesterase type 5 inhibitors. In addition, it was shown that the use of CO releasing molecules potentiated cavernous cGMP levels. However, increased CO production or release was reported to be associated, in some studies, with vasoconstriction.

Conclusion

This review sheds a light on the significance of cavernous tissue CO signaling pathway that may pave the way for creation of therapeutic modalities based on this pathway.
Disappearance of Female Genital Mutilation from the Bedouin Population of Southern Israel
Authors: Halila Suhil , Belmaker R.H. , Rabia Yunis Abu , Froimovici Miron , Applebaum Julia
Published: January 2009

Introduction

Recently, clinicians in Southern Israel perceived that the practice of female genital mutilation had disappeared entirely in the Bedouin population. We previously studied the prevalence of this practice in 1995.

Aim

We decided to survey again the Bedouin population focusing on those tribes previously reported to perform this practice.

Methods

Eighty percent of the interviews were done by an Arabic-speaking psychiatrist and 20% were done by an Arabic speaking nurse in the gynecologic clinic of a large Bedouin township or the gynecologic clinic of a smaller Bedouin township. Women were asked if they would be willing to answer a few questions about their past and if they were willing to have the gynecologist, with no additional procedure, note whether any operation had been performed on their genitalia.

Main Outcome Measures

Physical examination by gynecologist and an oral questionnaire.

Results

One hundred and thirty two women were examined. No cases of any scarring of the kind reported in the previous study were found on physical examination.

Conclusions

FGM has apparently disappeared over 15 years in a population in which it was once prevalent.
Psychobiological Correlates of Extramarital Affairs and Differences Between Stable and Occasional Infidelity Among Men with Sexual Dysfunctions
Authors: Fisher Alessandra D. , Corona Giovanni , Bandini Elisa , Mannucci Edoardo , Lotti Francesco , Boddi Valentina , Forti Gianni , Maggi Mario
Published: January 2009

Introduction

The relationship between extramarital affairs and male sexual dysfunctions (SDs) has not been completely clarified.

Aim

The aim of the present study is to describe how the reported presence of extramarital affairs vs. no report differentiates men attending medical care for SDs.

Methods

A nonselected series of 2,592 heterosexual (mean age 55 ± 12.5 years) male patients with SD was studied with Structured Interview on Erectile Dysfunction (SIEDY). Furthermore, a complete physical examination and a series of biochemical, hormonal, and penile vascular evaluations were performed.

Main Outcome Measures

Several hormonal, biochemical, and instrumental parameters were investigated along with items derived from SIEDY.

Results

Infidelity is associated with relational problems within the primary couple, particularly in those with a stable secondary relationship. Furthermore, a higher androgenization is present in men with infidelity. Patients with SD reporting extramarital affairs had higher stress at work, a longer primary relationship span, and higher risk of conflicts within the primal couple and within the family. In addition, infidel men showed a higher risk of partner’s illness and partner’s hypoactive sexual desire. Among organic factors, a lower prevalence of hypogonadism was observed in infidel men. These subjects showed a better sexual function than the rest of the sample. In addition, they reported a higher intercourse frequency in the previous 3 months (considering both the primary and secondary relationship), a lower prevalence of hypoactive sexual desire (HSD), and a lower feeling of discomfort or guilt about autoerotism. Finally, when SIEDY scales were considered, SIEDY Scale 2 (relational domain) and Scale 3 (intrapsychic domain) scores were significantly higher in infidel subjects when compared with the rest of the sample.

Conclusions

Infidelity is associated with relational problems within the primary couple, particularly in those with a stable secondary relationship. Furthermore, a higher androgenization is present in men who commit infidelity.
Treatment of Lifelong Vaginismus in Traditional Islamic Couples: A Prospective Study
Authors: Yasan Aziz , Akdeniz Nurten
Published: January 2009

Introduction

As sexual values and the meaning of sexuality are culturally dependent, the efficacy of sex therapy models may not necessarily be similar across cultures.

Aim

The aim was to address two questions; the first was to identify whether a group of patients with lifelong vaginismus living in traditional Islamic culture benefited from the sex therapy. The second question addressed was how Muslim culture affects sex therapy in the treatment of vaginismus.

Methods

Data were obtained from all patients with lifelong vaginismus who attended an outpatient clinic in the course of 1 year. Forty-four couples were investigated in the initial session. Thirty-six couples who completed the treatment were assessed after 3 months.

Main Outcome Measures

In the final evaluation, pleasurable full vaginal penetration after active penile insertion was accepted as a successful outcome treatment for the present study.

Results

In the study, 36 patients who had completed treatment were determined, 29 of whom had successful outcome of treatment, while eight dropped out. The model building process variable resulted in “married by matchmaker without consent (OR = 0.060, CI = 0.046–0.771, P = 0.031)” and “not allowing pelvic examination (OR = 0.124, CI = 0.016–0.941, P = 0.044)” as negative predictors for successful outcome of treatment.

Conclusion

Sex therapy is a feasible method of treatment for vaginismus within the cultural environment investigated, although some modifications may be needed in some setting such as for those married by matchmaker without their consent.
Resolution of Recurrent Ischemic Priapism After Celiac Plexus Block: Case Report Evidence for its Neurogenic Origin?
Authors: Burnett Arthur L. , Santiago Carlos S.
Published: January 2009

Introduction

Recurrent ischemic priapism likely has a neurogenic basis for some clinical presentations.

Aim

To describe a phenomenon of resolved recurrent ischemic priapism in a patient following celiac plexus block for recurrent upper abdominal pain.

Methods

The case report profiled a 44-year-old Caucasian man who developed recurrent ischemic priapism after retroperitoneal surgery. He had failed multiple conventional interventions for his priapism. After he underwent a celiac plexus block for recurrent upper abdominal pain, his recurrent priapism episodes immediately stopped.

Main Outcome Measures

Clinical history regarding priapism occurrences.

Results

The patient obtained a resolution of his priapism after celiac plexus block, without recurrences at a 6-month follow-up assessment.

Conclusions

Celiac plexus block successfully led to resolution of recurrent ischemic priapism of probable neurogenic origin associated with spinal nerve outflow injury.
The Effect of Dual-Rate Suspension Damping on Vehicle Response to Transient Road Inputs
Authors: Waters T. P. , Hyun Y. , Brennan M. J.
Published: January 2009

The acceleration response of road vehicles to shock inputs from road irregularities such as bumps and hollows is an important consideration in the design of vehicle suspensions and damping characteristics, in particular. In this paper, the influence of the damper on the shock response of a simple vehicle model is considered. An analysis is presented of a single degree-of-freedom model subjected to a transient displacement input. Simple approximate expressions are given for the peak acceleration during an impulse of both short and long durations compared to the natural period, from which the role of the damper is clearly apparent. For impulses of short duration the peak acceleration occurs during the impulse and is shown to be approximately proportional to the damping ratio. Corollary to this, the peak acceleration can be reduced by switching the damper to a lower value during the impulse. The potential benefits of doing so are illustrated through numerical simulation, and a simple formula is given for the maximum possible reduction in peak acceleration. The results are also contrasted with those of a conventional dual-rate automotive damper model. The switchable damper is found to offer sufficient benefit to warrant further investigation.
Acoustic Radiation From an Infinite Laminated Composite Cylindrical Shell With Doubly Periodic Rings
Authors: Yin X. W. , Liu L. J. , Hua H. X. , Shen R. Y.
Published: January 2009

Acoustic radiation from a point-driven, infinite fluid-loaded, laminated composite shell, which is reinforced by doubly periodic rings, is investigated theoretically. The theory is based on the classical laminated composite shell theory, the Helmholtz equation, and the boundary conditions at the shell-fluid interface as well as at the junctions between the shell and the rings. The rings interact with the shell only through normal forces. The solution for the radial displacement in wave number domain is developed by using Mace’s method (1980, “Sound Radiation Form a Plate Reinforced by Two Sets of Parallel Stiffeners,” J. Sound Vib., 71(3), pp. 435–441) for an infinite flat plate. The stationary phase approximate is then employed to find the expression for the far-field pressure. Numerical results are presented for discussion of the effects of lamination schemes, Poisson’s ratios, ply angles, and damping on the far-field acoustic radiation, which may lend themselves to better understanding the characteristics of acoustic radiation from the laminated composite shells. In addition, the helical wave spectra of the stiffened cylinders are presented, in which the effects of wave number conversion due to the periodic rings are obviously identified as additional bright patterns.
Effect of Nonhomogeneity on Vibration of Orthotropic Rectangular Plates of Varying Thickness Resting on Pasternak Foundation
Authors: Lal Roshan , Dhanpati
Published: January 2009

Free transverse vibrations of nonhomogeneous orthotropic rectangular plates of varying thickness with two opposite simply supported edges (y=0 and y=b) and resting on two-parameter foundation (Pasternak-type) have been studied on the basis of classical plate theory. The other two edges (x=0 and x=a) may be any combination of clamped and simply supported edge conditions. The nonhomogeneity of the plate material is assumed to arise due to the exponential variations in Young’s moduli and density along one direction. By expressing the displacement mode as a sine function of the variable between simply supported edges, the fourth order partial differential equation governing the motion of such plates of exponentially varying thickness in another direction gets reduced to an ordinary differential equation with variable coefficients. The resulting equation is then solved numerically by using the Chebyshev collocation technique for two different combinations of clamped and simply supported conditions at the other two edges. The lowest three frequencies have been computed to study the behavior of foundation parameters together with other plate parameters such as nonhomogeneity, density, and thickness variation on the frequencies of the plate with different aspect ratios. Normalized displacements are presented for a specified plate. A comparison of results with those obtained by other methods shows the computational efficiency of the present approach.
Experimental Identification of Generalized Proportional Viscous Damping Matrix
Authors: Adhikari S. , Phani A. Srikantha
Published: January 2009

A simple and easy-to-implement algorithm to identify a generalized proportional viscous damping matrix is developed in this work. The chief advantage of the proposed technique is that only a single drive-point frequency response function (FRF) measurement is needed. Such FRFs are routinely measured using the standard techniques of an experimental modal analysis, such as impulse test. The practical utility of the proposed identification scheme is illustrated on three representative structures: (1) a free-free beam in flexural vibration, (2) a quasiperiodic three-cantilever structure made by inserting slots in a plate in out-of-plane flexural vibration, and (3) a point-coupled-beam system. The finite element method is used to obtain the mass and stiffness matrices for each system, and the damping matrix is fitted to a measured variation of the damping (modal damping factors) with the natural frequency of vibration. The fitted viscous damping matrix does accommodate for any smooth variation of damping with frequency, as opposed to the conventional proportional damping matrix. It is concluded that a more generalized viscous damping matrix, allowing for a smooth variation of damping as a function of frequency, can be accommodated within the framework of standard finite element modeling and vibration analysis of linear systems.
An Improvement of a Nonclassical Numerical Method for the Computation of Fractional Derivatives
Authors: Diethelm Kai
Published: January 2009

Standard methods for the numerical calculation of fractional derivatives can be slow and memory consuming due to the nonlocality of the differential operators. Yuan and Agrawal (2002, “A Numerical Scheme for Dynamic Systems Containing Fractional Derivatives,” ASME J. Vibr. Acoust., 124, pp. 321–324) have proposed a more efficient approach for operators whose order is between 0 and 1 that differs substantially from the traditional concepts. It seems, however, that the accuracy of the results can be poor. We modify the approach, adapting it better to the properties of the problem, and show that this leads to a significantly improved quality. Our idea also works for operators of order greater than 1.
The Aryl Hydrocarbon Nuclear Translocator Alters CD30-Mediated NF-κB–Dependent Transcription
Authors: Wright Casey W. , Duckett Colin S.
Published: January 2009

Expression and signaling of CD30, a tumor necrosis factor receptor family member, is up-regulated in numerous lymphoid-derived neoplasias, most notably anaplastic large-cell lymphoma (ALCL) and Hodgkin's lymphoma. To gain insight into the mechanism of CD30 signaling, we used an affinity purification strategy that led to the identification of the aryl hydrocarbon receptor nuclear translocator (ARNT) as a CD30-interacting protein that modulated the activity of the RelB subunit of the transcription factor nuclear factor κB (NF-κB). ALCL cells that were deficient in ARNT exhibited defects in RelB recruitment to NF-κB–responsive promoters, whereas RelA recruitment to the same sites was potentiated, resulting in the augmented expression of these NF-κB–responsive genes. These findings indicate that ARNT functions in concert with RelB in a CD30-induced negative feedback mechanism.
Historical Warnings of Future Food Insecurity with Unprecedented Seasonal Heat
Authors: Battisti David. S. , Naylor Rosamond L.
Published: January 2009

Higher growing season temperatures can have dramatic impacts on agricultural productivity, farm incomes, and food security. We used observational data and output from 23 global climate models to show a high probability (>90%) that growing season temperatures in the tropics and subtropics by the end of the 21st century will exceed the most extreme seasonal temperatures recorded from 1900 to 2006. In temperate regions, the hottest seasons on record will represent the future norm in many locations. We used historical examples to illustrate the magnitude of damage to food systems caused by extreme seasonal heat and show that these short-run events could become long-term trends without sufficient investments in adaptation.
tasselseed1
Is a Lipoxygenase Affecting Jasmonic Acid Signaling in Sex Determination of Maize
Authors: Acosta Iván F. , Laparra Hélène , Romero Sandra P. , Schmelz Eric , Hamberg Mats , Mottinger John P. , Moreno Maria A. , Dellaporta Stephen L.
Published: January 2009

Sex determination in maize is controlled by a developmental cascade leading to the formation of unisexual florets derived from an initially bisexual floral meristem. Abortion of pistil primordia in staminate florets is controlled by a
tasselseed
-mediated cell death process. We positionally cloned and characterized the function of the sex determination gene
tasselseed1
(
ts1
). The TS1 protein encodes a plastid-targeted lipoxygenase with predicted 13-lipoxygenase specificity, which suggests that TS1 may be involved in the biosynthesis of the plant hormone jasmonic acid. In the absence of a functional
ts1
gene, lipoxygenase activity was missing and endogenous jasmonic acid concentrations were reduced in developing inflorescences. Application of jasmonic acid to developing inflorescences rescued stamen development in mutant
ts1
and
ts2
inflorescences, revealing a role for jasmonic acid in male flower development in maize.
Mispredicting Affective and Behavioral Responses to Racism
Authors: Kawakami Kerry , Dunn Elizabeth , Karmali Francine , Dovidio John F.
Published: January 2009

Contemporary race relations are marked by an apparent paradox: Overt prejudice is strongly condemned, yet acts of blatant racism still frequently occur. We propose that one reason for this inconsistency is that people misunderstand how they would feel and behave after witnessing racism. The present research demonstrates that although people predicted that they would be very upset by a racist act, when people actually experienced this event they showed relatively little emotional distress. Furthermore, people overestimated the degree to which a racist comment would provoke social rejection of the racist. These findings suggest that racism may persevere in part because people who anticipate feeling upset and believe that they will take action may actually respond with indifference when faced with an act of racism.
Structure of a Type IV Secretion System Core Complex
Authors: Fronzes Rémi , Schäfer Eva , Wang Luchun , Saibil Helen R. , Orlova Elena V. , Waksman Gabriel
Published: January 2009

Type IV secretion systems (T4SSs) are important virulence factors used by Gram-negative bacterial pathogens to inject effectors into host cells or to spread plasmids harboring antibiotic resistance genes. We report the 15 angstrom resolution cryo–electron microscopy structure of the core complex of a T4SS. The core complex is composed of three proteins, each present in 14 copies and forming a ∼1.1-megadalton two-chambered, double membrane–spanning channel. The structure is double-walled, with each component apparently spanning a large part of the channel. The complex is open on the cytoplasmic side and constricted on the extracellular side. Overall, the T4SS core complex structure is different in both architecture and composition from the other known double membrane–spanning secretion system that has been structurally characterized.
Simpson's Paradox in a Synthetic Microbial System
Authors: Chuang John S. , Rivoire Olivier , Leibler Stanislas
Published: January 2009

The maintenance of “public” or “common good” producers is a major question in the evolution of cooperation. Because nonproducers benefit from the shared resource without bearing its cost of production, they may proliferate faster than producers. We established a synthetic microbial system consisting of two
Escherichia coli
strains of common-good producers and nonproducers. Depending on the population structure, which was varied by forming groups with different initial compositions, an apparently paradoxical situation could be attained in which nonproducers grew faster within each group, yet producers increased overall. We show that a simple way to generate the variance required for this effect is through stochastic fluctuations via population bottlenecks. The synthetic approach described here thus provides a way to study generic mechanisms of natural selection.
Grappling with the Gulf
Authors: Greenbaum Dov , Gerstein Mark
Published: January 2009

Science for Lawyers
.
Eric York Drogin, Ed.
. Section of Science and Technology Law, American Bar Association, Chicago, 2008. 364 pp. Paper, $129.95. ISBN 9781590319260.

The contributors offer an introductory survey of 13 applied scientific disciplines from the perspective of their uses in legal matters.
Bacterial Particle Endocytosis by Epithelial Cells Is Selective and Enhanced by Tumor Necrosis Factor Receptor Ligands
Authors: Clark Robin T. , Hope Andrew , Lopez-Fraga Marta , Schiller Neal , Lo David D.
Published: January 2009

Bacterial pathogens use virulence strategies to invade epithelial barriers, but active processes of epithelial cells may also contribute to the endocytosis of microbial particles. To focus on the latter, we studied the uptake of fixed and fluorescently labeled bacterial particles in intestinal and bronchoepithelial cell cultures and found it to be enhanced in Caco-2BBe and NCI-H292 cells after treatment with tumor necrosis factor alpha and an agonist antibody against the lymphotoxin beta receptor. Confocal fluorescence microscopy, flow cytometry, and transmission electron microscopy revealed that
Staphylococcus aureus
and
Yersinia enterocolitica
were readily endocytosed, although there was scant uptake of
Shigella sonnei
,
Salmonella enterica
serovar Typhimurium, and
Klebsiella pneumoniae
particles. Endocytosed
Staphylococcus
was often associated with cytoplasmic claudin-4 vesicles; this was not found for
Yersinia
, suggesting that cytokine treatment upregulated two distinct endocytosis pathways. Interestingly, when
Staphylococcus
and
Yersinia
were coincubated with epithelial monolayers, the cells were unlikely to take up
Yersinia
unless they had also endocytosed large numbers of
Staphylococcus
particles, although the two bacteria were apparently processed in distinct compartments. Cytokine treatment induced an upregulation and redistribution of β1 integrin to the apical surface of NCI-H292 cells; consistent with this effect, treatment with anti-β1 integrin antibody blocked uptake of both
Yersinia
and
Staphylococcus
in NCI-H292 and Caco-2BBe cells. Our results suggest that capture of bacterial particles by mucosal epithelial cells is selective and that different endocytic mechanisms are enhanced by proinflammatory cytokines.
Relationship between Human Immunodeficiency Virus Type 1 Coinfection, Anemia, and Levels and Function of Antibodies to Variant Surface Antigens in Pregnancy-Associated Malaria
Authors: Jaworowski Anthony , Fernandes Liselle A. , Yosaatmadja Francisca , Feng Gaoqian , Mwapasa Victor , Molyneux Malcolm E. , Meshnick Steven R. , Lewis Jenny , Rogerson Stephen J.
Published: January 2009

Human immunodeficiency virus type 1 (HIV-1) coinfection decreases antibodies to variant surface antigens implicated in pregnancy-associated malaria (VSA-PAM) caused by
Plasmodium falciparum
. The effect of HIV-1 on antibody functions that may protect mothers from pregnancy-associated malaria is unknown. Sera from multigravid pregnant women with malaria and HIV-1 coinfection (
n
= 58) or malaria alone (
n
= 29) and from HIV-1-infected (
n
= 102) or -uninfected (
n
= 54) multigravidae without malaria were analyzed for anti-VSA-PAM antibodies by flow cytometry, the ability to inhibit adhesion to chondroitin sulfate A, or to opsonize CS2-infected erythrocytes for phagocytosis by THP-1 cells. In women with malaria, anti-VSA-PAM levels correlated better with opsonic activity (
r
= 0.60) than with adhesion-blocking activity (
r
= 0.33). In univariate analysis, HIV-1 coinfection was associated with lower opsonic activity but not adhesion-blocking activity or anti-VSA-PAM levels. Malaria-infected women with anemia (hemoglobin levels of <11.0 g/dl) had lower opsonic activity than nonanemic women (
P
= 0.007) independent of HIV-1 status. By multivariate analysis, in malaria-infected women, anemia (but not HIV status) was associated with opsonic activity. In women without malaria, opsonic activity was not associated with either anemia or HIV-1 status. In multigravid pregnant women with malaria, impaired serum opsonic activity may contribute to anemia and possibly to the decreased immunity to pregnancy-associated malaria associated with HIV-1.
Humoral Immune Responses of White-Tailed Deer (
Odocoileus virginianus
) to
Mycobacterium bovis
BCG Vaccination and Experimental Challenge with
M. bovis
Authors: Nol P. , Lyashchenko K. P. , Greenwald R. , Esfandiari J. , Waters W. R. , Palmer M. V. , Nonnecke B. J. , Keefe T. J. , Thacker T. C. , Rhyan J. C. , Aldwell F. E. , Salman M. D.
Published: January 2009

Monitoring of the kinetics of production of serum antibodies to multiple mycobacterial antigens can be useful as a diagnostic tool for the detection of
Mycobacterium bovis
infection as well as for the characterization of disease progression and the efficacy of intervention strategies in several species. The humoral immune responses to multiple
M. bovis
antigens by white-tailed deer vaccinated with BCG orally via a lipid-formulated bait (
n
= 5), orally in liquid form (
n
= 5), and subcutaneously (
n
= 6) were evaluated over time after vaccination and after experimental challenge with virulent
M. bovis
and were compared to the responses by unvaccinated deer (
n
= 6). Antibody responses were evaluated by using a rapid test (RT), a multiantigen print immunoassay (MAPIA), a lipoarabinomannan enzyme-linked immunosorbent assay (LAM-ELISA), and immunoblotting to whole-cell sonicate and recombinant antigen MPB83. MAPIA and RT detected minimal to no antibody responses over those at the baseline to multiple
M. bovis
antigens in vaccinated white-tailed deer after challenge. This was in contrast to the presence of more readily detectable antibody responses in nonvaccinated deer with more advanced disease. The LAM-ELISA results indicated an overall decrease in the level of production of detectable antibodies against lipoarabinomannan-enriched mycobacterial antigen in vaccinated animals compared to that in nonvaccinated animals after challenge. Immunoblot data were inconsistent but did suggest the occurrence of unique antibody responses by certain vaccinated groups to Ag85 and HSP70. These findings support further research toward the improvement and potential use of antibody-based assays, such as MAPIA, RT, and LAM-ELISA, as tools for the antemortem assessment of disease progression in white-tailed deer in both experimental and field vaccine trials.
Detection of Anthrax Toxin by an Ultrasensitive Immunoassay Using Europium Nanoparticles
Authors: Tang Shixing , Moayeri Mahtab , Chen Zhaochun , Harma Harri , Zhao Jiangqin , Hu Haijing , Purcell Robert H. , Leppla Stephen H. , Hewlett Indira K.
Published: January 2009

We developed a europium nanoparticle-based immunoassay (ENIA) for the sensitive detection of anthrax protective antigen (PA). The ENIA exhibited a linear dose-dependent pattern within the detection range of 0.01 to 100 ng/ml and was approximately 100-fold more sensitive than enzyme-linked immunosorbent assay (ELISA). False-positive results were not observed with serum samples from healthy adults, mouse plasma without PA, or plasma samples collected from mice injected with anthrax lethal factor or edema factor alone. For the detection of plasma samples spiked with PA, the detection sensitivities for ENIA and ELISA were 100% (11/11 samples) and 36.4% (4/11 samples), respectively. The assay exhibited a linear but qualitative correlation between the PA injected and the PA detected in murine blood (
r
= 0.97731;
P
< 0.0001). Anthrax PA was also detected in the circulation of mice infected with spores from a toxigenic Sterne-like strain of
Bacillus anthracis
, but only in the later stages of infection. These results indicate that the universal labeling technology based on europium nanoparticles and its application may provide a rapid and sensitive testing platform for clinical diagnosis and laboratory research.
Development of Fully Automated Determination of Marker-Specific Immunoglobulin G (IgG) Avidity Based on the Avidity Competition Assay Format: Application for Abbott Architect Cytomegalovirus and Toxo IgG Avidity Assays
Authors: Curdt Ingo , Praast Gerald , Sickinger Eva , Schultess Jan , Herold Iris , Braun Hans Bertram , Bernhardt Stephanie , Maine Gregory T. , Smith Darwin D. , Hsu Stephen , Christ Heike M. , Pucci Dominick , Hausmann Michael , Herzogenrath Jörg
Published: January 2009

Determination of the avidity of immunoglobulin G (IgG) directed against a specific marker has become an established diagnostic tool for identifying or excluding acute infections with pathogens. A novel assay format termed AVIcomp (avidity competition based on mass action) circumventing the conventional chaotropic format has been developed for determination of the avidity of marker-specific IgG in patient specimens. Its applications for cytomegalovirus (CMV) and
Toxoplasma gondii
are presented. Specific high-avidity IgG from the patient specimen is selectively blocked using a soluble antigen in a sample pretreatment reagent, and the amount of remaining specific low-avidity IgG is determined relative to that in an untreated control. The comparison of the conventional chaotropic format, represented by the Radim CMV IgG Avidity assay, and the newly developed AVIcomp method, as exemplified by the Architect CMV IgG Avidity assay, on blood drawn within 4 months after seroconversion revealed a sensitivity of 100% (97.3% by an alternative calculation) for the AVIcomp format versus 87.5% (75.7% by an alternative calculation) for the chaotropic avidity assay. The specificity on 312 CMV IgG reactive and CMV IgM nonreactive specimens from pregnant women was 100% for the AVIcomp assay and 99.7% for the conventional avidity assay. The Architect Toxo IgG Avidity assay showed an agreement of 97.2% with the bioMérieux Vidas Toxo IgG Avidity Assay employing chaotropic reagents. These performance data suggest that the AVIcomp format shows superior sensitivity and equivalent specificity for the determination of IgG avidity to assays based on the chaotropic method and that the AVIcomp format may also be applicable to other disease states.
Comparison of Viral Isolation and Multiplex Real-Time Reverse Transcription-PCR for Confirmation of Respiratory Syncytial Virus and Influenza Virus Detection by Antigen Immunoassays
Authors: Liao R. S. , Tomalty L. L. , Majury A. , Zoutman D. E.
Published: January 2009

We evaluated the Prodesse ProFlu-1 real-time reverse transcription-PCR multiplex assay with the SmartCycler instrument for the detection of human respiratory syncytial virus (RSV) and influenza A and B viruses in comparison to conventional cell culture and antigen immunoassays with the BD Directigen A+B and Binax NOW RSV assays over two successive respiratory virus seasons. Ninety-two percent of the 361 specimens tested were nasopharyngeal aspirates obtained from individual patients, of which 119 were positive for RSV and 59 were positive for influenza virus. The median age of the patients whose specimens were positive for RSV and influenza virus were 6.3 months and 42.4 years, respectively. The specificity of all of the methods tested was ≥99%, and the individual sensitivities of NOW RSV, RSV culture, Directigen A+B, influenza virus culture, and the Proflu-1 PCR for influenza/RSV were 82% (95% confidence interval [CI], 73 to 88), 57% (95% CI, 44 to 69), 59% (95% CI, 44 to 72), 54% (95% CI, 38 to 69), and 98% (95% CI, 93 to 100)/95% (95% CI, 85 to 99), respectively. In a clinical setting where viral isolation is performed to confirm rapid antigen immunoassay results for these common respiratory viruses, one-step real-time reverse transcriptase PCR testing can be a more sensitive and timely confirmatory method.
Unreliability of Results of PCR Detection of
Helicobacter pylori
in Clinical or Environmental Samples
Authors: Sugimoto Mitsushige , Wu Jeng-Yih , Abudayyeh Suhaib , Hoffman Jill , Brahem Hajer , Al-Khatib Khaldun , Yamaoka Yoshio , Graham David Y.
Published: January 2009

The aim of this study was to compare published
Helicobacter pylori
primer pairs for their ability to reliably detect
H. pylori
in gastric biopsy specimens and salivary samples. Detection limits of the 26 PCR primer pairs previously described for detection of
H. pylori
DNA in clinical samples were determined. Sensitivity and specificity were determined using primers with detection limits of <100 CFU/ml using 50
H. pylori
-positive and -negative (by concordance by culture and histology) coded gastric biopsy specimens. These results were then confirmed with gastric biopsy specimens and saliva from patients with confirmed
H. pylori
status. Five of the twenty-six previously reported primer pairs (HP64-f/HP64-r, HP1/HP2, EHC-U/EHC-L, VAG-F/VAG-R, and ICT37/ICT38) had detection limits of <100 CFU/ml in the presence of gastric tissue. None had 100% specificity or sensitivity; all produced false-positive results. The HP64-f/HP64-r for
ureA
and HP1/HP2 for 16S rRNA individually had sensitivities and specificities of >90% with gastric biopsy specimens. No combinations of primer pairs improved the results. Using these five primer pairs, 54% of the positive saliva samples were determined to be false positive; both the HP64-f/HP64-r and the HP1/HP2 sets produced false positives with saliva. We conclude that clinicians should not rely on results using current PCR primers alone to decide the
H. pylori
status of an individual patient or as a basis for treatment decisions. The results of studies based on PCR identification of
H. pylori
in environmental samples should be viewed with caution. Possibly, specific primers sets can be identified based on the presence of multiple putative virulence factor genes.
Needle-to-Incubator Transport Time: Logistic Factors Influencing Transport Time for Blood Culture Specimens
Authors: Kerremans J. J. , van der Bij A. K. , Goessens W. , Verbrugh H. A. , Vos M. C.
Published: January 2009

The maximum recommended transport time for blood cultures is 4 h [L. S. Garcia (ed.),
2007 Update: Clinical Microbiology Procedures Handbook
, 2nd ed., 2007]. In a previous study, we found that the average transport time was 10 h. In this cohort study, we measured transport times for blood cultures in a larger sample and identified predictors for transport times. A total of 4,322 blood cultures from 1,313 patients were included. The median transport time was 3.5 h, with 47% of cultures exceeding the recommended 4 h. Off-site location and type of clinical specialty were the most important predictors of long transport times. Cultures collected during weekend days or on wards at the largest distances from the laboratory were also associated with long transport times.
A Systems Biology Approach To Identify the Combination Effects of Human Herpesvirus 8 Genes on NF-κB Activation
Authors: Konrad Andreas , Wies Effi , Thurau Mathias , Marquardt Gaby , Naschberger Elisabeth , Hentschel Sonja , Jochmann Ramona , Schulz Thomas F. , Erfle Holger , Brors Benedikt , Lausen Berthold , Neipel Frank , Stürzl Michael
Published: January 2009

Human herpesvirus 8 (HHV-8) is the etiologic agent of Kaposi's sarcoma and primary effusion lymphoma. Activation of the cellular transcription factor nuclear factor-kappa B (NF-κB) is essential for latent persistence of HHV-8, survival of HHV-8-infected cells, and disease progression. We used reverse-transfected cell microarrays (RTCM) as an unbiased systems biology approach to systematically analyze the effects of HHV-8 genes on the NF-κB signaling pathway. All HHV-8 genes individually (
n
= 86) and, additionally, all K and latent genes in pairwise combinations (
n
= 231) were investigated. Statistical analyses of more than 14,000 transfections identified ORF75 as a novel and confirmed K13 as a known HHV-8 activator of NF-κB. K13 and ORF75 showed cooperative NF-κB activation. Small interfering RNA-mediated knockdown of ORF75 expression demonstrated that this gene contributes significantly to NF-κB activation in HHV-8-infected cells. Furthermore, our approach confirmed K10.5 as an NF-κB inhibitor and newly identified K1 as an inhibitor of both K13- and ORF75-mediated NF-κB activation. All results obtained with RTCM were confirmed with classical transfection experiments. Our work describes the first successful application of RTCM for the systematic analysis of pathofunctions of genes of an infectious agent. With this approach, ORF75 and K1 were identified as novel HHV-8 regulatory molecules on the NF-κB signal transduction pathway. The genes identified may be involved in fine-tuning of the balance between latency and lytic replication, since this depends critically on the state of NF-κB activity.
Sindbis Virus-Based Measles DNA Vaccines Protect Cotton Rats against Respiratory Measles: Relevance of Antibodies, Mucosal and Systemic Antibody-Secreting Cells, Memory B Cells, and Th1-Type Cytokines as Correlates of Immunity
Authors: Pasetti Marcela F. , Ramirez Karina , Resendiz-Albor Aldo , Ulmer Jeffrey , Barry Eileen M. , Levine Myron M.
Published: January 2009

Measles remains an important cause of pediatric morbidity and mortality in developing countries, especially among infants who are too young to receive the current licensed live attenuated measles vaccine. We developed two Sindbis virus DNA vaccines encoding the measles virus hemagglutinin (pMSIN-H) and fusion proteins (pMSINH-FdU) and examined their immunogenicities and protective efficacies when administered alone or followed by the live measles virus vaccine in cotton rats. Neutralizing antibodies, mucosal and systemic antibody-secreting cells, memory B cells, and gamma interferon-secreting T cells developed after priming and increased after boosting. pMSIN-H priming conferred 100% protection against pulmonary measles, whereas pMSINH-FdU protected only in conjunction with the live measles virus vaccine boost.
Borna Disease Virus Requires Cholesterol in both Cellular Membrane and Viral Envelope for Efficient Cell Entry
Authors: Clemente Roberto , de Parseval Aymeric , Perez Mar , de la Torre Juan C.
Published: January 2009

Borna disease virus (BDV), the prototypic member of the familyBornaviridaewithin the orderMononegavirales, provides an important model for the investigation of viral persistence within the central nervous system (CNS) and of associated brain disorders. BDV is highly neurotropic and enters its target cell via receptor-mediated endocytosis, a process mediated by the virus surface glycoprotein (G), but the cellular factors and pathways determining BDV cell tropism within the CNS remain mostly unknown. Cholesterol has been shown to influence viral infections via its effects on different viral processes, including replication, budding, and cell entry. In this work, we show that cell entry, but not replication and gene expression, of BDV was drastically inhibited by depletion of cellular cholesterol levels. BDV G-mediated attachment to BDV-susceptible cells was cholesterol independent, but G localized to lipid rafts (LR) at the plasma membrane. LR structure and function critically depend on cholesterol, and hence, compromised structural integrity and function of LR caused by cholesterol depletion likely inhibited the initial stages of BDV cell internalization. Furthermore, we also show that viral-envelope cholesterol is required for BDV infectivity.
A Basic Patch on α-Adaptin Is Required for Binding of Human Immunodeficiency Virus Type 1 Nef and Cooperative Assembly of a CD4-Nef-AP-2 Complex
Authors: Chaudhuri Rittik , Mattera Rafael , Lindwasser O. Wolf , Robinson Margaret S. , Bonifacino Juan S.
Published: January 2009

A critical function of the human immunodeficiency virus type 1 Nef protein is the downregulation of CD4 from the surfaces of infected cells. Nef is believed to act by linking the cytosolic tail of CD4 to the endocytic machinery, thereby increasing the rate of CD4 internalization. In support of this model, weak binary interactions between CD4, Nef, and the endocytic adaptor complex, AP-2, have been reported. In particular, dileucine and diacidic motifs in the C-terminal flexible loop of Nef have been shown to mediate binding to a combination of the α and σ2 subunits of AP-2. Here, we report the identification of a potential binding site for the Nef diacidic motif on α-adaptin. This site comprises two basic residues, lysine-297 and arginine-340, on the α-adaptin trunk domain. The mutation of these residues specifically inhibits the ability of Nef to bind AP-2 and downregulate CD4. We also present evidence that the diacidic motif on Nef and the basic patch on α-adaptin are both required for the cooperative assembly of a CD4-Nef-AP-2 complex. This cooperativity explains how Nef is able to efficiently downregulate CD4 despite weak binary interactions between components of the tripartite complex.
Partial Protection of Simian Immunodeficiency Virus (SIV)-Infected Rhesus Monkeys against Superinfection with a Heterologous SIV Isolate
Authors: Yeh Wendy W. , Jaru-ampornpan Pimkwan , Nevidomskyte Daiva , Asmal Mohammed , Rao Srinivas S. , Buzby Adam P. , Montefiori David C. , Korber Bette T. , Letvin Norman L.
Published: January 2009

Although there is increasing evidence that individuals already infected with human immunodeficiency virus type 1 (HIV-1) can be infected with a heterologous strain of the virus, the extent of protection against superinfection conferred by the first infection and the biologic consequences of superinfection are not well understood. We explored these questions in the simian immunodeficiency virus (SIV)/rhesus monkey model of HIV-1/AIDS. We infected cohorts of rhesus monkeys with either SIVmac251 or SIVsmE660 and then exposed animals to the reciprocal virus through intrarectal inoculations. Employing a quantitative real-time PCR assay, we determined the replication kinetics of the two strains of virus for 20 weeks. We found that primary infection with a replication-competent virus did not protect against acquisition of infection by a heterologous virus but did confer relative control of the superinfecting virus. In animals that became superinfected, there was a reduction in peak replication and rapid control of the second virus. The relative susceptibility to superinfection was not correlated with CD4
+
T-cell count, CD4
+
memory T-cell subsets, cytokine production by virus-specific CD8
+
or CD4
+
cells, or neutralizing antibodies at the time of exposure to the second virus. Although there were transient increases in viral loads of the primary virus and a modest decline in CD4
+
T-cell counts after superinfection, there was no evidence of disease acceleration. These findings indicate that an immunodeficiency virus infection confers partial protection against a second immunodeficiency virus infection, but this protection may be mediated by mechanisms other than classical adaptive immune responses.
Papillomavirus E2 Proteins and the Host Brd4 Protein Associate with Transcriptionally Active Cellular Chromatin
Authors: Jang Moon Kyoo , Kwon Deukwoo , McBride Alison A.
Published: January 2009

The interaction of papillomavirus E2 proteins with cellular Brd4 protein is important for transcriptional regulation of viral genes and partitioning of viral genomes. Bovine papillomavirus type 1 (BPV-1) E2 binds cellular chromatin in complex with Brd4 in both mitotic and interphase cells. To identify specific sites of E2 interaction on cellular chromatin, a genome-wide chromatin immunoprecipitation-on-chip analysis was carried out using human promoter sequences. Both E2 and Brd4 were found bound to most transcriptionally active promoters in C33A cells. These promoters were also bound by RNA polymerase II and were modified by histone H3 acetylation and K4 trimethylation, all indicators of active transcription. E2 binding strongly correlated with Brd4 and RNA polymerase II occupancy and H3K4me3 modification at all human promoters, indicating that E2 bound to active promoters. E2 binding did not correlate with the presence of consensus E2 binding sites in the promoters. Furthermore, the mRNA levels of E2-bound cellular genes were not significantly changed by E2 expression. Thus, the papillomavirus E2 proteins bind to transcriptionally active cellular genes but do not change their activity. We propose that this may be a way for the virus to ensure that the viral genome is retained in transcriptionally active regions of the nucleus to escape silencing. Therefore, E2-mediated tethering of viral genomes to host chromatin has multiple roles: to partition the viral genome to daughter cells, to ensure that the genomes are retained in the nucleus, and to make certain that the genomes are retained in functionally active nuclear domains.
Mitogen-Activated Protein Kinase-Activated Kinase RSK2 Plays a Role in Innate Immune Responses to Influenza Virus Infection
Authors: Kakugawa Satoshi , Shimojima Masayuki , Goto Hideo , Horimoto Taisuke , Oshimori Naoki , Neumann Gabriele , Yamamoto Tadashi , Kawaoka Yoshihiro
Published: January 2009

Viral infections induce signaling pathways in mammalian cells that stimulate innate immune responses and affect cellular processes, such as apoptosis, mitosis, and differentiation. Here, we report that the ribosomal protein S6 kinase alpha 3 (RSK2), which is activated through the “classical” mitogen-activated protein kinase pathway, plays a role in innate immune responses to influenza virus infection. RSK2 functions in the regulation of cell growth and differentiation but was not known to play a role in the cellular antiviral response. We have found that knockdown of RSK2 enhanced viral polymerase activity and growth of influenza viruses. Influenza virus infection stimulates NK-κB- and beta interferon-dependent promoters. This stimulation was reduced in RSK2 knockdown cells, suggesting that RSK2 executes its effect through innate immune response pathways. Furthermore, RSK2 knockdown suppressed influenza virus-induced phosphorylation of the double-stranded RNA-activated protein kinase PKR, a known antiviral protein. These findings establish a role for RSK2 in the cellular antiviral response.