bluuebunny/crossref_metadata_2025 huggingface dataset:
German Castles, Customs, and Culture: Introducing a New Approach to the Undergraduate Culture Course
Authors: Lalande John F.
Published: August 2010

Courses on the culture of the German‐speaking world (GSW) have long dotted the landscape of undergraduate course offerings at North American colleges and universities. The primary purpose of this article is to share information about a new kind of undergraduate culture course that uses castles as a vehicle for introducing students to past and present aspects of GSW culture.
The Contributions of Talker Familiarity and Individual Talker Characteristics
Authors: Schierloh Maren , Hayes‐Harb Rachel
Published: August 2010

The article details a two‐phase study investigating the relative contributions of talker‐familiarity and talker‐intelligibility to L2 listening comprehension by learners in a classroom setting. Students from beginning to low‐intermediate German FL classes participated in German language listening tasks, being exposed to speech by their German teacher (familiar talker) and other, unfamiliar talkers. Findings were consistent in both phases of the study: the relative intelligibility of particular talkers was more predictive to L2 listening accuracy than familiarity with the talker. Follow‐up investigations revealed a relationship between speech rate and listening comprehension accuracy.
German Television in the United States and Abroad1
Authors: Ecke Peter
Published: August 2010

In this article, I review the options that teachers and students have to obtain German TV in the U.S.: through satellite and cable networks and by streaming, downloading, or recording TV programs through the internet. I also address how one can access TV programs over the air in the German‐speaking countries by using a notebook PC and TV card.
Learners' Descriptions of German Pronunciation, Vocabulary, and Grammar: A Folk Linguistic Account
Authors: Chavez Monika
Published: August 2010

Following a folk linguistic approach, this investigation of first‐, second‐ and fourth‐year learners' accounts of German found that (1) few had held pre‐conceived notions about German prior to language study; (2) most pre‐conceived notions concerned German pronunciation; (3) pre‐conceived notions about vocabulary were most likely to influence the decision to study German; and (4) among current perceptions of German, learners (a) believe German to be more “systematic” than English; (b) are virtually exclusively concerned with rules of “accuracy” (not appropriateness); (c) tend to judge the “merits” of German rules in comparison with English but also the Romance languages; (d) consider grammar more different between English and German than vocabulary and pronunciation; and (e) register no observable differences across different years of study or between German and non‐German majors.
The Role of Explicit Information in Instructed SLA: An On‐Line Study with Processing Instruction and German Accusative Case Inflections
Authors: Culman Hillah , Henry Nicholas , VanPatten Bill
Published: August 2010

The present study reports the findings of an experiment on the effects of explicit information on the learning of German case markings. Fifty‐nine learners of first‐ and second‐year German received computer‐based processing instruction on German accusative case marking and word order. These learners were divided into two groups: one received explicit information on the nature and form of case marking in German prior to the treatment, and one group did not. We measured the effects of explicit information by tracking correct responses on the computer as participants made their way through the activities. Analyses revealed that explicit information had an effect: those who received explicit information began to correctly respond to stimulus sentences (i.e., began to correctly indicate who did what to whom) sooner than those who did not. These results contradict previous research and suggest a hidden role for explicit information within processing instruction.
Service Learning in Germany: A Four‐Week Summer Teaching Program in Saxony‐Anhalt
Authors: Ducate Lara
Published: August 2010

This article describes a four‐week service learning abroad program. Students from a U.S. university assist and teach in elementary school English classes for one month in Saxony‐Anhalt, Germany while living with their German partner teachers. The program description and students' feedback illustrate that this program provides students with the opportunity to improve their cultural and language competence through immersion in the German culture away from their American peers. Students report feeling engaged in the lives of their host families and schools and gain new understandings of the German culture through the perspectives they encounter during their month of volunteering.
The New Cosmopolitan Monolingualism: On Linguistic Citizenship in Twenty‐First Century Germany
Authors: Gramling David
Published: August 2010

In the early years of the twenty‐first century, being German has become a matter of linguistic competence and performance. An acute shift in citizenship statutes at the end of the 1990s brought about a peripatetic departure from Germany's “right of blood” (ius sanguinis) toward a French‐inspired “right of territory” (ius soli). Yet in the nine years since the policy's implementation, a paradigm quite removed from territorial citizenship has taken hold—one that I will outline as a ius linguarum, or “right of languages.” This article analyzes the civic discourse on German language use as it has evolved from the late 1990s in immigration statutes, press discourse, school reform initiatives, and national service awards. Together, these developments serve as interlocking case studies in the emergence of a new cosmopolitan monolingualism—amid the fluctuating conditions of European integration and economic globalization. The article concludes with some speculations on the impact of a ius linguarum for teachers of literature and language in the German Studies context.
“Bridging the Gap” within a Class: Managing a Range of Proficiency Levels in a Third‐Year German Course
Authors: O'Brien Traci S.
Published: August 2010

While much attention has been paid to the gap between beginning language instruction and literature courses, there is another gap which presents significant challenges to college instructors, namely the gap in proficiency levels between students in the same third‐year course. This article argues that bridging this particular gap can productively challenge students as well as increase retention rates by fostering individualized learning goals. In addition, it makes practical suggestions for creating and selecting exercises from a commonly‐used advanced college textbook in order to accommodate students of various ability levels in the same class.
Bridging the Gap: A Literacy‐Oriented Approach to Teaching the Graphic Novel Der erste Frühling
Authors: Bridges Elizabeth
Published: August 2010

This article examines the phenomenon of graphic novels generally, and in particular, their utility in second‐language contexts via the example of the graphic adaptation of Klaus Kordoris Der erste Frühling. This text lends itself especially to bridging thematic gaps in 20th‐century German literature courses, as well as the “language/literature divide,” which is a current topic in language pedagogy. The article provides specific classroom techniques, in‐class tasks, and writing assignments that will accommodate the varying student proficiency levels that often challenge instructors in second‐ and third‐year college German courses.
Two Thematic Units for the Middle School Curriculum: An Initiative by the Kinder lernen Deutsch Steering Committee's Writing Team1
Authors: Busch Iris , Freimann‐Cavanaugh Corinna , Eichler Ester
Published: August 2010

The Kinder lernen Deutsch Committee (KLD) is a standing committee of the AATG that has existed since 1987 and that was originally charged to support the advocacy of German in grades K‐8. With generous funding by the Ständige Arbeitsgruppe Deutsch als Fremdsprache (StADaF) from the German government and the Goethe‐Institut, the Kinder lernen Deutsch (KLD) Steering Committee has made great progress in many directions. The contribution here represents an initiative by a writing team of the committee to provide two curricular proposals to address the glaring omission of usable and useful classroom materials for middle school learners of German. The writing team applied the concepts reflected by Wiggins and McTighe in Understanding by Design as well as the thematic planning framework of Helena Curtain and Carol Ann Dahlberg. The thematic units entitled “Die Umwelt und ich” and “Oma Annas Schatzkoffer: ein neues Leben in Amerika” provide a design template that can be used for developing additional thematic units for middle school learners.
Machine Translation in the German Classroom: Detection, Reaction, Prevention
Authors: Steding Sören
Published: August 2010

There are many websites today that offer free machine translations and although beginning students of German are not always proficient enough to judge the quality of these translations or to fully understand certain translation results, they use these services nonetheless for their assignments. The problem for the educator is to distinguish between machine translation and genuine student work. Depending on the assignment and the language level, this task can be very easy or quite difficult. This article describes approaches for dealing with texts that appear to be machine translated rather than “hand‐crafted.”
Does Vagueness Exclude Knowledge?
Authors: BARNETT DAVID
Published: August 2010

On two standard views of vagueness, vagueness as to whether Harry is bald entails that nobody knows whether Harry is bald—either because vagueness is a type of missing truth, and so there is nothing to know, or because vagueness is a type of ignorance, and so even though there is a truth of the matter, nobody can know what that truth is. Vagueness as to whether Harry is bald does entail that nobodyclearlyknows that Harry is bald and that nobodyclearlyknows that Harry is not bald. But it does not entail that nobodyknowsthat Harry is bald or that nobodyknowsthat Harry is not bald. Hence, the two standard views of vagueness are mistaken.
Faultless Disagreement and Aesthetic Realism
Authors: SCHAFER KARL
Published: August 2010

It has recently been argued that certain areas of discourse, such as discourse about matters of taste, involve a phenomenon of ‘‘faultless disagreement’’ that rules out giving a standard realist or contextualist semantics for them. Thus, it is argued, we are left with no choice but to consider more adventurous semantic alternatives for these areas, such as a semantic account that involves relativizing truth to perspectives or contexts of assessment. I argue that the sort of faultless disagreement present in these cases is in fact compatible with a realist treatment of their semantics. Then I briefly consider other considerations that might be thought to speak against realism about these areas of discourse. I conclude with the tentative suggestion that realism about matters of taste is far more plausible (at least in some cases) than most philosophers believe today.
Creep Damage and the Remaining Life Concept
Authors: Woodford D. A.
Published: August 2010

Creep damage is described in terms of both density changes, which measure the nucleation and growth of grain boundary voids, and also changes in mechanical properties. In both cases the damage is related to the creep test variables. A loading sequence effect is demonstrated, implying that the linear life fraction rule is not applicable to creep rupture under changing stress conditions. Based on these observations, a new approach to life prediction for nonsteady temperature and stress is proposed. The principles of the new model, which is based on remaining life rather than used life, are described. It is shown how this approach requires no prior commitment to a particular damage law and that the behavior of low alloy steels, in laboratory tests and also after service operation, is consistent with the remaining life concept.
Remaining Creep or Stress-Rupture Life Under Nonsteady Temperature and Stress
Authors: Roberts B. W. , Ellis F. V. , Bynum J. E.
Published: August 2010

This paper presents a historical review of the assessment of remaining creep life for components operating under stress at elevated temperatures. Topics considered in this review include the effects of multiaxial stress state on creep deformation and rupture, empirical methods of summing creep damage, empirical methods of evaluating elevated temperature strength, testing of service exposed material, and microstructural observations of creep damage. While considerable understanding of the damage process has been gained in laboratory testing, only limited success has been attained in quantifying damage of service exposed materials designed to operate in the long time and low stress regime permitted by industrial codes.
Creep of Metals and Plastics Under Combined Stresses, A Review
Authors: Findley W. N. , Cho U. W. , Ding J. L.
Published: August 2010

Prior research on creep of metals and plastics under combined stresses is reviewed. Recent and current research in this area are described. Emphasis of the review is on experimental investigations of creep under multiaxial stresses since 1950.
Effects of Nonlinear Stress-Strain Rate Relation on Deformation and Fracture of Materials in Creep Range
Authors: Ohtani Ryuichi , Taira Shuji
Published: August 2010

Fundamental and practical investigations on high-temperature strength of materials have made significant contributions to industrial development for the last twenty-five years. However, an additional effort is required to make clear the effects of dominant factors on the deformation and fracture of materials, since the knowledge obtained to date are not enough to explain the overall phenomena of high temperature strength. In this paper the importance of the effects of nonlinearlity and time-dependence of stress-strain relation on the well-known creep behaviors are reviewed and reexamined on the basis of our laboratory study conducted over the last twenty years.
Viscoplasticity Based on Total Strain. The Modelling of Creep With Special Considerations of Initial Strain and Aging
Authors: Krempl Erhard
Published: August 2010

A previously proposed theory is specialized for the uniaxial state of stress and its prediction for creep and relaxation is analyzed in detail. Constant true stress and constant load creep tests are simulated in the presence and absence of thermal aging together with the constant strain relaxation test. The signs of the creep rate and its time derivative as well as the relaxation rate and its time derivative are introduced as criteria. The constant load creep test can reproduce the normal creep curves and nonclassical creep curves (ε˙ > 0; ε¨ > 0 for all ε and for σ0 > 0). The capabilities of the constant true stress test are limited to primary creep if the work-hardening slope is positive. When aging is introduced almost any creep curve can be reproduced in both tests. The importance of initial strain is discussed and demonstrated by room temperature creep tests on Type 304 Stainless Steel. It is suggested that the initial strains together with the creep curves be reported in the future. Poisson’s ratio in creep needs to be measured and tests are proposed which will enable a quantitative assessment of aging.
Creating Polytope Representations of Design Spaces for Visual Exploration Using Consistency Techniques
Authors: Devanathan Srikanth , Ramani Karthik
Published: August 2010

Understanding the limits of a design is an important aspect of the design process. When mathematical models are constructed to describe a design concept, the limits are typically expressed as constraints involving the variables of that concept. The set of values for the design variables that do not violate constraints constitute the design space of that concept. In this work, we transform a parametric design problem into a geometry problem thereby enabling computational geometry algorithms to support design exploration. A polytope-based representation is presented to geometrically approximate the design space. The design space is represented as a finite set of (at most) three-dimensional (possibly nonconvex) polytopes, i.e., points, intervals, polygons, and polyhedra. The algorithm for constructing the design space is developed by interpreting constraint-consistency algorithms as computational-geometric operations and consequently extending (3,2)-consistency algorithm for polytope representations. A simple example of a fingernail clipper design is used to illustrate the approach.
Integration of Sustainability Into Early Design Through the Function Impact Matrix
Authors: Devanathan Srikanth , Ramanujan Devarajan , Bernstein William Z. , Zhao Fu , Ramani Karthik
Published: August 2010

The issue of environmental sustainability, which is unprecedented in both magnitude and complexity, presents one of the biggest challenges faced by modern society. Design engineers can make significant contributions by incorporating environmental awareness into product and process development. It is critical that engineers make a paradigm shift in product design from centering on cost and performance to balancing economic, environmental, and societal considerations. Although there have been quite a few designs for environment (or ecodesign) tools developed, so far, these tools have only achieved limited industrial penetration. The present-day methods are either too qualitative to offer concrete solutions and not effective for designers with limited experience or too quantitative, costly, and time consuming. Thus, current ecodesign tools cannot be implemented during the early design phases. This paper develops a novel, semiquantitative ecodesign methodology that is targeted specifically toward the early stages of the design process. The new methodology is a combination of environmental life cycle assessment and visual tools such as quality function deployment, functional-component matrix, and Pugh chart. Since the early design process is function-oriented, a new visual tool called the function impact matrix has been developed to correlate environmental impacts with product function. Redesign of office staplers for reduced carbon footprint has been selected as a case study to demonstrate the use of the proposed approach. Life cycle assessment results confirm that the new stapler design generated using this methodology promotes improved environmental performance.
Progressive Design Processes and Bounded Rational Designers
Authors: Herrmann Jeffrey W.
Published: August 2010

This paper presents a method for assessing the quality of a progressive design process by measuring the profitability of the product that the process generates. The proposed approach uses separations, a type of problem decomposition, to model progressive design processes. The subproblems in the separations correspond roughly to phases in the progressive design processes. The proposed method simulates the choices of a bounded rational designer for each subproblem using different search algorithms. This paper presents a simple two-variable problem to help describe the approach and then applies the approach to assess motor design processes. Different types and versions of these search processes are considered to determine if the results are robust to the decision-making model. The results indicate that well-designed progressive design processes are the best way to generate profitable product designs. Methods for assessing the quality of engineering design processes can be used to guide improvements to engineering design processes and generate more valuable products.
A Generalized Constraint Model for Two-Dimensional Beam Flexures: Nonlinear Load-Displacement Formulation
Authors: Awtar Shorya , Sen Shiladitya
Published: August 2010

To utilize beam flexures in constraint-based flexure mechanism design, it is important to develop qualitative and quantitative understanding of their constraint characteristics in terms of stiffness and error motions. This paper provides a highly generalized yet accurate closed-form parametric load-displacement model for two-dimensional beam flexures, taking into account the nonlinearities arising from load equilibrium applied in the deformed configuration. In particular, stiffness and error motions are parametrically quantified in terms of elastic, load-stiffening, kinematic, and elastokinematic effects. The proposed beam constraint model incorporates a wide range of loading conditions, boundary conditions, initial curvature, and beam shape. The accuracy and effectiveness of the proposed beam constraint model is verified by nonlinear finite elements analysis.
A Generalized Constraint Model for Two-Dimensional Beam Flexures: Nonlinear Strain Energy Formulation
Authors: Awtar Shorya , Sen Shiladitya
Published: August 2010

The beam constraint model (BCM), presented previously, captures pertinent nonlinearities to predict the constraint characteristics of a generalized beam flexure in terms of its stiffness and error motions. In this paper, a nonlinear strain energy formulation for the beam flexure, consistent with the transverse-direction load-displacement and axial-direction geometric constraint relations in the BCM, is presented. An explicit strain energy expression, in terms of beam end displacements, that accommodates generalized loading conditions, boundary conditions, initial curvature, and beam shape, is derived. Using energy-based arguments, new insight into the BCM is elucidated by fundamental relations among its stiffness, constraint, and energy coefficients. The presence of axial load in the geometric constraint and strain energy expressions—a unique attribute of distributed compliance flexures that leads to the elastokinematic effect—is highlighted. Using the principle of virtual work, this strain energy expression for a generalized beam is employed in determining the load-displacement relations, and therefore constraint characteristics, of a flexure mechanism comprising multiple beams. The benefit of this approach is evident in its mathematical efficiency and succinctness, which is to be expected with the use of energy methods. All analytical results are validated to a high degree of accuracy via nonlinear finite element analysis.
Aggregation and Multilevel Design for Systems: Finding Guidelines
Authors: Saari Donald G.
Published: August 2010

All areas of engineering have a need to find appropriate aggregated outcomes for systems. Issues range from decision problems, “divide-and-conquer” approaches that include aspects of multidisciplinary design optimization and the effects of a division of labor for, perhaps, a design project, the inefficiencies that can accompany multidisciplinary projects involving, say, design, manufacturing, and sales, to the complexities of multiscale design, analysis, and even nanotechnology. But as shown, if the adopted approach (e.g., management choices, divide-and-conquer methodology, modeling of the biology/physics, decision rule, etc.) satisfies particular accepted practices, then certain complexities and inefficiencies must be anticipated. A disturbing corollary is that even should “success” appear to have been achieved with an approach that satisfies these conditions, it need not be as firm as expected. Ways to improve methodologies must avoid the specified conditions.
Response to Comment on “The Silicate-Mediated Formose Reaction: Bottom-Up Synthesis of Sugar Silicates”
Authors: Lambert Joseph B. , Gurusamy-Thangavelu Senthil Andavan , Ma Kuangbiao
Published: August 2010

We reported that silicate mediates the formation of four-carbon and six-carbon sugars from simple two- and three-carbon molecules. Kim and Benner’s contention that silicate has little impact on these reactions is based on experiments using harsher conditions of temperature, pH, silicate concentration, and reaction time than in our study, thus confounding any comparisons between our results.
Linear Arrays of Nuclear Envelope Proteins Harness Retrograde Actin Flow for Nuclear Movement
Authors: Luxton G. W. Gant , Gomes Edgar R. , Folker Eric S. , Vintinner Erin , Gundersen Gregg G.
Published: August 2010

Line Up for Movement

The nuclei of animal cells can move to specific locations and help to polarize migrating and differentiating cells.

Luxton
et al.

(p.
956
; see the Perspective by

Starr

) found that linear arrays of nuclear membrane proteins assembled on, and moved with, actin cables toward the rear of the cell during nuclear movement in polarizing fibroblasts. Interfering with the components of these linear arrays prevented nuclear movement and centrosome reorientation. Thus, nuclear membrane proteins assemble into actin-dependent arrays during force transduction.
mTOR-Dependent Synapse Formation Underlies the Rapid Antidepressant Effects of NMDA Antagonists
Authors: Li Nanxin , Lee Boyoung , Liu Rong-Jian , Banasr Mounira , Dwyer Jason M. , Iwata Masaaki , Li Xiao-Yuan , Aghajanian George , Duman Ronald S.
Published: August 2010

Antidepressant Action of Ketamine

In contrast to the weeks or months of treatment required for standard antidepressant medication, ketamine administration produces an antidepressant response within 4 to 6 hours in depressed patients. What lies behind the rapid actions of ketamine?

Li
et al.

(p.
959
; see the Perspective by

Cryan and O'Leary

) found that ketamine administration resulted in fast activation of mammalian target of rapamycin (mTOR) signaling and increased levels of synaptic proteins in the rat prefrontal cortex. Ketamine rapidly increased the density and function of the dendritic spines of layer V pyramidal neurons in the prefrontal cortex. Thus, the behavioral actions of ketamine in models of depression and antidepressant response are dependent on mTOR signaling.
Conserved Fungal LysM Effector Ecp6 Prevents Chitin-Triggered Immunity in Plants
Authors: de Jonge Ronnie , Peter van Esse H. , Kombrink Anja , Shinya Tomonori , Desaki Yoshitake , Bours Ralph , van der Krol Sander , Shibuya Naoto , Joosten Matthieu H. A. J. , Thomma Bart P. H. J.
Published: August 2010

Fungal Defenses

One of the major driving forces of evolution is the constant arms race between plants and animals and the microbial pathogens that infect them. The fungus
Cladosporium fulvum
causes leaf mold on tomato plants. One of the ways tomato plants sense infections by
C. fulvum
is by detecting chitin, a component of fungal cell walls. In response, the fungus has evolved strategies to evade detection.

De Jonge
et al.

(p.
953
) have now identified one such mechanism in
C. fulvum
, mediated by the effector protein Ecp6. Secreted Ecp6 is able to bind to chitin oligosaccharides that are released upon degradation of the fungal cell wall and sequester them so that they are not detected by tomato chitin receptors. Proteins with domain structure similar to Ecp6 are conserved throughout the fungal kingdom, which suggests that chitin sequestration may represent a general mechanism used by fungi to evade immune detection.
Unidirectional Emission of a Quantum Dot Coupled to a Nanoantenna
Authors: Curto Alberto G. , Volpe Giorgio , Taminiau Tim H. , Kreuzer Mark P. , Quidant Romain , van Hulst Niek F.
Published: August 2010

Directed Emission

The atomlike features of quantum dots—the discrete energy levels and subsequent emission of light at discrete wavelengths—make them key building blocks in optoelectronic circuits and optical communication. However, the emitted light tends to be omnidirectional, which limits applications that require accurate transmission between sender and receiver.

Curto
et al.

(p.
930
; see the Perspective by

Giessen and Lippitz

) have designed an optical antenna, a shrunk-down version of the Yagi-Uda design used in microwave and radio communication, and show that coupling the quantum dot to the antenna provides control over the direction of the emitted light.
Drought-Induced Reduction in Global Terrestrial Net Primary Production from 2000 Through 2009
Authors: Zhao Maosheng , Running Steven W.
Published: August 2010

Reversing the Trend

Terrestrial net primary productivity (NPP, the amount of atmospheric carbon fixed by plants and accumulated as biomass) increased from 1982 through 1999, which has been attributed to factors such as nitrogen deposition, CO
2
fertilization, forest regrowth, and climatic changes.
Zhao and Running
(p.
940
) used satellite data to estimate global terrestrial NPP over the past decade and found that the earlier trend has been reversed and that NPP has been decreasing. Combining this result with climate change data suggests that large-scale droughts are responsible for the decline. Future widespread droughts caused by global warming may thus further weaken the terrestrial carbon sink.
Sustainability and Sources of Wealth
The Plundered Planet
Why We Must—and How We Can—Manage Nature for Global Prosperity / How to Reconcile Prosperity with Nature

Paul Collier

Oxford University Press, New York, 2010. 287 pp. $24.95. ISBN 9780195395259. Allen Lane, London. £20. ISBN 9781846142239.
Authors: Low Bobbi S.
Published: August 2010

Collier addresses social, economic, and environmental problems resulting from the current mismanagement and improper exploitation of resources.
A Tale of Two Jets
Authors: Yunes Nicolás
Published: August 2010

Numerical models suggest that black hole mergers may produce a strong, potentially detectable electromagnetic counterpart to their emitted gravitational waves.
The Case of the Missing $470 Million in BP's Promised Research Fund
Authors: Schenkman Lauren
Published: August 2010

On 24 May, BP pledged $500 million over the next 10 years to support independent research to answer questions about what the oil might do to deep-sea corals, wetland birds, and young fish. But just 3 weeks later, a presidential directive put the money on hold. Now scientists are worried that what many saw as their best hope for a long-term, comprehensive research plan will disappear along with the oil slick.
Senator Builds His Legacy With University of Alabama Earmarks
Authors: Mervis Jeffrey
Published: August 2010

Senator Richard Shelby (R–AL) has single-handedly implemented an ersatz federal academic infrastructure program for the University of Alabama, Tuscaloosa, campus over the past decade. Although most science policymakers believe earmarks weaken U.S. science by placing the preferences of individual lawmakers above those of the peer-review process, Shelby says the earmarks he has secured for several of the state's universities have been good for Alabama's economy and its scientists.
Serological Protection Induced by
Haemophilus influenzae
Type b Conjugate Vaccine in Mexican Children: Is a Booster Dose of the Vaccine Needed?
Authors: Rodriguez Romeo S. , Mascarenas Cesar , Conde-Glez Carlos J. , Inostroza Jaime , Villanueva Sonia , Velázquez María Elena , Sánchez-Alemán Miguel Angel , Echániz Gabriela
Published: August 2010

We determined the seroprevalence of protective antibodies against Hib in Mexican children under the age of five using a standardized enzyme-linked immunosorbent assay. Hib antibodies (≥0.15 μg/ml) were present in 95.34% (±1.14% [seroprevalence ± standard error]) of samples. Fewer children aged 30 to 47 months had protective Hib antibody levels (91.45% ± 2.60%) than children from 12 to 29 and 48 to 59 months (97.3% ± 1.34% and 97.44% ± 1.80%, respectively).
Mycoplasma genitalium
PCR: Does Freezing of Specimens Affect Sensitivity?
Authors: Carlsen Katrina Hutton , Jensen Jorgen Skov
Published: August 2010

Mycoplasma genitalium
is an established cause of sexually transmitted infections. Studies of disease associations are often performed on archived specimens, but little is known about the effect of storage of specimens on the detection of
M. genitalium.
Genital swab and first-void urine specimens submitted for detection of
M. genitalium
were tested on the day of receipt. Remnants of positive original specimens as well as DNA preparations were stored at −20°C for up to 18 months. A total of 361
M. genitalium
-positive specimens were available. PCR after repeat DNA preparation was performed for 262 specimens. The sensitivity after repeat DNA preparation was 90%, and the median decrease in DNA load was 155 genome equivalents (geq) (
P
< 0.0001). For 327 specimens, PCR could be repeated on the primary DNA preparation. The sensitivity of PCR after storage was 95%, and the median decrease in DNA load was 13.5 geq (
P
< 0.0001). The specimens yielding negative results at repeat testing had a significantly lower median DNA load in the primary analysis than those with a repeat positive test (
P
< 0.0001). For 228 specimens, PCR could be performed both on the primary DNA preparation and after repeat DNA preparation. The median DNA load was lower after repeat DNA extraction than after repeat testing of the stored DNA extract (
P
< 0.0001). In conclusion, the
M. genitalium
DNA load as well as the detection rate decreased after storage. This was more pronounced in clinical specimens stored frozen than in stored DNA extracts, particularly in those with an initial low DNA load.
Strains of
Mycobacterium tuberculosis
from Western Maharashtra, India, Exhibit a High Degree of Diversity and Strain-Specific Associations with Drug Resistance, Cavitary Disease, and Treatment Failure
Authors: Chatterjee Anirvan , D'Souza Desiree , Vira Tina , Bamne Arun , Ambe Gurish T. , Nicol Mark P. , Wilkinson Robert J. , Mistry Nerges
Published: August 2010

We performed spoligotyping of
Mycobacterium tuberculosis
isolates from 833 systematically sampled pulmonary tuberculosis (TB) patients in urban Mumbai, India (723 patients), and adjacent rural areas in western India (110 patients). The urban cohort consisted of two groups of patients, new cases (646 patients) and first-time treatment failures (77 patients), while only new cases were recruited in the rural areas. The isolates from urban new cases showed 71% clustering, with 168 Manu1, 62 CAS, 22 Beijing, and 30 EAI-5 isolates. The isolates from first-time treatment failures were 69% clustered, with 14 Manu1, 8 CAS, 8 Beijing, and 6 EAI-5 isolates. The proportion of Beijing strains was higher in this group than in urban new cases (odds ratio [OR], 3.29; 95% confidence limit [95% CL], 1.29 to 8.14;
P
= 0.003). The isolates from rural new cases showed 69% clustering, with 38 Manu1, 7 CAS, and 1 EAI-5 isolate. Beijing was absent in the rural cohort. Manu1 was found to be more common in the rural cohort (OR, 0.67; 95% CL, 0.42 to 1.05;
P
= 0.06). In total, 71% of isolates were clustered into 58 spoligotypes with 4 predominant strains, Manu1 (26%), CAS (9%), EAI-5 (4%), and Beijing (4%), along with 246 unique spoligotypes. In the isolates from urban new cases, we found Beijing to be associated with multidrug resistance (MDR) (OR, 3.40; 95% CL, 1.20 to 9.62;
P
= 0.02). CAS was found to be associated with pansensitivity (OR, 1.83; 95% CL, 1.03 to 3.24;
P
= 0.03) and cavities as seen on chest radiographs (OR, 2.72; 95% CL, 1.34 to 5.53;
P
= 0.006). We recorded 239 new spoligotypes yet unreported in the global databases, suggesting that the local TB strains exhibit a high degree of diversity.
Simultaneous Genotyping and Quantification of Hepatitis B Virus for Genotypes B and C by Real-Time PCR Assay
Authors: Zhao Yao , Zhang Xiu-Yu , Guo Jin-Jun , Zeng Ai-Zhong , Hu Jie-Li , Huang Wen-Xiang , Shan You-Lan , Huang Ai-Long
Published: August 2010

Hepatitis B virus (HBV) is an important cause of human chronic liver diseases and is a major public health problem. Viral load and HBV genotype play critical roles in determining clinical outcomes and response to antiviral treatment in hepatitis B patients. Viral genotype detection and quantification assays are currently in use with different levels of effectiveness. In this study, the performance of a real-time genotyping and quantitative PCR (GQ-PCR)-based assay was evaluated. Through the use of genotype-specific primers and probes, this assay provides simultaneous identification and quantification of genotypes B and C in a single reaction. Our GQ-PCR correctly identified all predefined genotypes B and C, and no cross-reaction between genotypes B and C were observed. The GQ-PCR identified more cases of HBV infections with mixed genotypes B and C than direct sequencing did. Samples from 127 HBV-infected Chinese patients were genotyped with GQ-PCR, revealing 56.7% HBV as genotype B, 13.4% as genotype C, and 29.8% as mixed genotypes B and C. This assay provides a reliable, efficient, and cost-effective means for quantification of the B and C genotypes of HBV in single or mixed infections. This assay is suitable for sequential monitoring of viral load levels and for determining the relationship between the genotype viral load and stage of disease in Asians.
Containment of Bioaerosol Infection Risk by the Xpert MTB/RIF Assay and Its Applicability to Point-of-Care Settings
Authors: Banada Padmapriya P. , Sivasubramani Satheesh K. , Blakemore Robert , Boehme Catharina , Perkins Mark D. , Fennelly Kevin , Alland David
Published: August 2010

The recently introduced Xpert MTB/RIF assay (Xpert) has point-of-care potential, but its capacity for biohazard containment remained to be studied. We compared the bioaerosols generated by the Xpert assay to acid-fast bacillus (AFB) microscope slide smear preparation. The Xpert assay sample treatment reagent (SR) was also studied for its sterilizing capacity, stability, and effect on assay sensitivity after prolonged treatment. During the preparation of AFB smears, sputum samples spiked with
Mycobacterium bovis
BCG at 5 × 10
8
CFU/ml produced 16 and 325 CFU/m
3
air measured with an Andersen impactor or BioSampler, respectively. In contrast, neither the sample preparation steps for the Xpert assay nor its automated processing produced any culturable bioaerosols. In testing of SR sterilizing capacity, clinical sputum samples from strongly smear-positive tuberculosis patients treated with SR at a 2:1 ratio eliminated
Mycobacterium tuberculosis
growth in all but 1/39 or 3/45 samples cultured on solid or liquid medium, respectively. These few unsterilized samples had a mean 13.1-day delay in the time to positive culture. SR treatment at a 3:1 ratio eliminated growth in all samples. SR retained a greater than 6-log-unit killing capacity despite storage at temperatures spanning 4 to 45°C for at least 3 months. The effect of prolonged SR sample treatment was also studied. Spiked sputum samples could be incubated in SR for up to 3 days without affecting Xpert sensitivity for
M. tuberculosis
detection and up to 8 h without affecting specificity for rifampin resistance detection. These results suggest that benchtop use of the Xpert MTB/RIF assay limits infection risk to the user.
Performance of Version 2.0 of the Cobas AmpliPrep/Cobas TaqMan Real-Time PCR Assay for Hepatitis B Virus DNA Quantification
Authors: Chevaliez Stéphane , Bouvier-Alias Magali , Laperche Syria , Hézode Christophe , Pawlotsky Jean-Michel
Published: August 2010

The detection and quantification of hepatitis B virus (HBV) DNA are essential for the diagnosis and treatment of chronic HBV infection. The use of real-time PCR assays for HBV DNA quantification is strongly recommended. The goal of this study was to evaluate the intrinsic characteristics and clinical performance of version 2.0 (v2.0) of the Cobas AmpliPrep/Cobas TaqMan (CAP/CTM) assay, a fully automated platform for HBV DNA quantification in serum or in plasma with a claimed lower limit of detection of 20 IU/ml and a claimed upper limit of quantification of 1.7 × 10
8
IU/ml. The specificity of the assay was 99% (95% confidence interval, 94.7 to 100%). Intra-assay and interassay coefficients of variation ranged from 0.21% to 2.67% and from 0.65% to 2.25%, respectively. The calibration of the assay was found to be satisfactory. Study of blood specimens from patients infected with HBV genotypes A to F showed good correspondence between HBV DNA levels measured by the CAP/CTM v2.0 assay, version 1.0 of the same assay, and the third-generation “branched DNA” assay. The CAP/CTM v2.0 assay quantified HBV DNA levels in serum or plasma from the same patients equally. In conclusion, the new version of the CAP/CTM assay is sensitive, specific, and reproducible. It accurately quantifies HBV DNA levels in patients chronically infected with HBV genotypes A to F. Improvements made to ensure equal quantification of HBV DNA in serum and plasma have been successful. Overall, the CAP/CTM assay, version 2.0, is well suited to monitoring clinical HBV DNA levels according to current clinical practice guidelines.
Polymicrobial Bloodstream Infection with
Eggerthella lenta
and
Desulfovibrio desulfuricans
Authors: Liderot Karin , Larsson Martin , Boräng Stina , Özenci Volkan
Published: August 2010

The advancement in culture identification methods has made possible the culture and identification of slow-growing anaerobic bacteria in clinical samples. Here, we describe a case of polymicrobial bloodstream infection (BSI) caused by
Eggerthella lenta
and
Desulfovibrio desulfuricans
, identified by API 20A and Vitek 2 systems and by 16S rRNA sequencing.
Proteasome Activator REGγ Enhances Coxsackieviral Infection by Facilitating p53 Degradation
Authors: Gao Guang , Wong Jerry , Zhang Jingchun , Mao Ivy , Shravah Jayant , Wu Yan , Xiao Allen , Li Xiaotao , Luo Honglin
Published: August 2010

Coxsackievirus B3 (CVB3) is a small RNA virus associated with diseases such as myocarditis, meningitis, and pancreatitis. We have previously demonstrated that proteasome inhibition reduces CVB3 replication and attenuates virus-induced myocarditis. However, the underlying mechanisms by which the ubiquitin/proteasome system regulates CVB replication remain unclear. In this study, we investigated the role of REGγ, a member of the 11S proteasome activator, in CVB3 replication. We showed that overexpression of REGγ promoted CVB3 replication but that knockdown of REGγ led to reduced CVB3 replication. We further demonstrated that REGγ-mediated p53 proteolysis contributes, as least in part, to the proviral function of REGγ. Although total protein levels of REGγ remained unaltered after CVB3 infection, virus infection induced a redistribution of REGγ from the nucleus to the cytoplasm, rendering an opportunity for a direct interaction of REGγ with viral proteins and/or host proteins (e.g., p53), which controls viral growth and thereby enhances viral infectivity. Further analyses suggested a potential modification of REGγ by SUMO following CVB3 infection, which was verified by both
in vitro
and
in vivo
sumoylation assays. Sumoylation of REGγ may play a role in its nuclear export during CVB3 infection. Taken together, our results present the first evidence that the host REGγ pathway is utilized and modified during CVB3 infection to promote efficient viral replication.
Poxvirus Complement Control Proteins Are Expressed on the Cell Surface through an Intermolecular Disulfide Bridge with the Viral A56 Protein
Authors: DeHaven Brian C. , Girgis Natasha M. , Xiao Yuhong , Hudson Paul N. , Olson Victoria A. , Damon Inger K. , Isaacs Stuart N.
Published: August 2010

The vaccinia virus (VACV) complement control protein (VCP) is an immunomodulatory protein that is both secreted from and expressed on the surface of infected cells. Surface expression of VCP occurs though an interaction with the viral transmembrane protein A56 and is dependent on a free N-terminal cysteine of VCP. Although A56 and VCP have been shown to interact in infected cells, the mechanism remains unclear. To investigate if A56 is sufficient for surface expression, we transiently expressed VCP and A56 in eukaryotic cell lines and found that they interact on the cell surface in the absence of other viral proteins. Since A56 contains three extracellular cysteines, we hypothesized that one of the cysteines may be unpaired and could therefore form a disulfide bridge with VCP. To test this, we generated a series of A56 mutants in which each cysteine was mutated to a serine, and we found that mutation of cysteine 162 abrogated VCP cell surface expression. We also tested the ability of other poxvirus complement control proteins to bind to VACV A56. While the smallpox homolog of VCP is able to bind VACV A56, the ectromelia virus (ECTV) VCP homolog is only able to bind the ECTV homolog of A56, indicating that these proteins may have coevolved. Surface expression of poxvirus complement control proteins may have important implications in viral pathogenesis, as a virus that does not express cell surface VCP is attenuatedin vivo. This suggests that surface expression of VCP may contribute to poxvirus pathogenesis.
Culturing the Unculturable: Human Coronavirus HKU1 Infects, Replicates, and Produces Progeny Virions in Human Ciliated Airway Epithelial Cell Cultures
Authors: Pyrc Krzysztof , Sims Amy C. , Dijkman Ronald , Jebbink Maarten , Long Casey , Deming Damon , Donaldson Eric , Vabret Astrid , Baric Ralph , van der Hoek Lia , Pickles Raymond
Published: August 2010

Culturing newly identified human lung pathogens from clinical sample isolates can represent a daunting task, with problems ranging from low levels of pathogens to the presence of growth suppressive factors in the specimens, compounded by the lack of a suitable tissue culture system. However, it is critical to develop suitable
in vitro
platforms to isolate and characterize the replication kinetics and pathogenesis of recently identified human pathogens. HCoV-HKU1, a human coronavirus identified in a clinical sample from a patient with severe pneumonia, has been a major challenge for successful propagation on all immortalized cells tested to date. To determine if HCoV-HKU1 could replicate in
in vitro
models of human ciliated airway epithelial cell cultures (HAE) that recapitulate the morphology, biochemistry, and physiology of the human airway epithelium, the apical surfaces of HAE were inoculated with a clinical sample of HCoV-HKU1 (Cean1 strain). High virus yields were found for several days postinoculation and electron micrograph, Northern blot, and immunofluorescence data confirmed that HCoV-HKU1 replicated efficiently within ciliated cells, demonstrating that this cell type is infected by all human coronaviruses identified to date. Antiserum directed against human leukocyte antigen C (HLA-C) failed to attenuate HCoV-HKU1 infection and replication in HAE, suggesting that HLA-C is not required for HCoV-HKU1 infection of the human ciliated airway epithelium. We propose that the HAE model provides a ready platform for molecular studies and characterization of HCoV-HKU1 and in general serves as a robust technology for the recovery, amplification, adaptation, and characterization of novel coronaviruses and other respiratory viruses from clinical material.