bluuebunny/crossref_metadata_2025 huggingface dataset:
Solution structure of hirsutellin A – new insights into the active site and interacting interfaces of ribotoxins
Authors: Viegas Aldino , Herrero‐Galán Elias , Oñaderra Mercedes , Macedo Anjos L. , Bruix Marta
Published: March 2009

Hirsutellin (HtA) is intermediate in size between other ribotoxins and less specific microbial RNases, and thus offers a unique chance to determine the minimal structural requirements for activities unique to ribotoxins. Here, we have determined the structure of HtA by NMR methods. The structure consists of one α‐helix, a helical turn and seven β‐strands that form an N‐terminal hairpin and an anti‐parallel β‐sheet, with a characteristic α + β fold and a highly positive charged surface. Compared to its larger homolog α‐sarcin, the N‐terminal hairpin is shorter and less positively charged. The secondary structure elements are connected by large loops with root mean square deviation (rmsd) values > 1 Å, suggesting some degree of intrinsically dynamic behavior. The active site architecture of HtA is unique among ribotoxins. Compared to α‐sarcin, HtA has an aspartate group, D40, replacing a tyrosine, and the aromatic ring of F126, located in the leucine ‘environment’ close to the catalytic H113 in a similar arrangement to that found in RNase T1. This unique active site structure is discussed in terms of its novel electrostatic interactions to understand the efficient cytotoxic activity of HtA. The contributions of the N‐terminal hairpin, loop 2 and loop 5 with regard to protein functionality, protein–protein and protein–lipid interactions, are also discussed. The truncation and reduced charge of the N‐terminal hairpin in HtA may be compensated for by the extension and new orientation of its loop 5. This novel orientation of loop 5 re‐establishes a positive charge on the side of the molecule that has been shown to be important for intermolecular interactions in ribotoxins.
Predictors of the Transition From Experimental to Daily Smoking Among Adolescents in the United States
Authors: Park Sunhee , Weaver Terri E. , Romer Daniel
Published: March 2009

PURPOSE. This study examined factors affecting the transition from experimental smoking at baseline to two types of daily smoking, temporary daily smoking, and continued daily smoking, at 1‐year follow‐up.

DESIGN AND METHODS. This study analyzed data from the National Longitudinal Study of Adolescent Health (n = 4,903 U.S. adolescents). Baseline predictors were selected based on Problem Behavior Theory.

RESULTS. Important problem behavior theory‐related predictors of smoking were the number of friends who smoke, academic performance, and alcohol, marijuana, and other illicit drug use. Other significant predictors were age, gender, race, depression, perceived general health, and cigarette availability at home.

PRACTICE IMPLICATIONS. To prevent teens from progressing to daily smoking, nursing professionals should consider multifaceted factors based on multiple theories.
The contrariwise life of a parasitic, pedomorphic copepod with a non‐feeding adult: ontogenesis, ecology, and evolution
Authors: Hendler Gordon , Dojiri Masahiro
Published: March 2009

Abstract. The extraordinary parasitic metanauplius larva of Caribeopsyllus amphiodiae is sexually dimorphic, with conspicuous gonads, and elaborate lens‐bearing eyes. The parasites usually occur singly within their host, and grow for ≤5 months within the stomach of burrowing ophiuroids (Amphiodia urtica). They transform into free‐living, semelparous, non‐feeding adults that live only 2 weeks. The species' life‐history pattern, with a larval period ∼10 × longer than the adult life span, is contrariwise to that of other copepods but not for animals with non‐feeding adults of both sexes. It appears that the life cycle of C. amphiodiae is pedomorphic, and probably evolved through a delay of metamorphosis regulated by developmental hormones. We attribute the dominance of the larval phase to the greater potential for survival and growth of the enterozoic parasitic stages than of the free‐living, post‐metamorphic stages. We note that among marine invertebrates, non‐feeding adults of both sexes occur exclusively in taxa with a complex life cycle, and that non‐feeding adults of both sexes are never found in taxa that have small larvae and delayed maturation. They occur only when there is a large larva that can provide the adult stage with sufficient nutrient reserves for reproduction.
Prevalence of cardiovascular disease and risk factors in a type 2 diabetic population of the North Catalonia diabetes study
Authors: Jurado Jeronimo , Ybarra Juan , Solanas Pascual , Caula Jacint , Gich Ignasi , Pou Jose M. , Romeo June Hart
Published: March 2009

Purpose: The purpose of the study was to evaluate the prevalence of cardiovascular disease (CVD), cardiovascular risk factors (CVRFs), and their control in patients with type 2 diabetes mellitus (T2DM) at primary care settings from the North Catalonia Diabetes Study (NCDS).

Data sources: In this multicentre cross‐sectional descriptive study, data were collected from a random sample of 307 patients with T2DM. The prevalence of CVD, CVRF, metabolic syndrome (MS), coronary heart disease (CHD) risk at 10 years (Framingham Point Scores), and CVRF control was evaluated. MS and lipid profiles were established according to Adult Treatment Panel III criteria.

Conclusions: CVD prevalence was 22.0% (CHD: 18.9% and peripheral ischemia: 4.5%) and more frequent in men. The prevalence of selected CVRF was: hypertension: 74.5%; dyslipidemia: 77.7%; smoking: 14.9%; obesity 44.9%, and familial CVD: 38.4%. Three or more CVRFs, including T2DM, were observed in 91.3%. MS prevalence was 68.7%. Framingham score was 10.0%, higher in men than in women. CVD prevalence was related to: age, number of CVRFs, duration of diabetes, familial history of CVD, waist circumference, hypertension, lipid profile, kidney disease, and Framingham score, but not to MS by itself. Correct lipid profiles and blood pressure were only observed in 18.9% and 24.0%, respectively, whereas platelet aggregation inhibitors were only recorded in 16.1% of the patient cohort. MS presence was not an independent risk factor of CVD in our study.

Implications for practice: The high prevalence of CVD and an inadequate control of CVRF, which were apparent in the NCDS population, would suggest that advanced practice nurses should consider incorporating specific cardiovascular assessment in their routine care of persons with T2DM.
The advance directive prevalence in long‐term care: A comparison of relationships between a nurse practitioner healthcare model and a traditional healthcare model
Authors: Lawrence James F.
Published: March 2009

Purpose: The purpose of this study was to examine rates of completion of advance directives (ADs) among institutionalized older adults in three geographically diverse areas of the country—Arizona, Georgia, and Massachusetts. Comparisons among four variables—gender, race, education, and type of healthcare model (Evercare vs. non‐Evercare), related to AD completion rates were examined.

Data sources: This study was a secondary data analysis using deidentified data from 11,775 older adults enrolled in the Evercare healthcare model to 91,443 non‐Evercare older adults (Minimum Data Set) during the last quarter of 2004. Chi‐square analysis was used to examine any differences in gender, race, education, and healthcare model associated with the completion rates of ADs.

Conclusions: The Evercare healthcare model that used nurse practitioners (NPs) consistently had significantly higher (p < .001) completion rates of ADs compared to the non‐Evercare healthcare model that did not use NPs. Black people and white people in the Evercare healthcare model had similar rates of AD completion (p > .001), which is contrary to previous findings where black people had a lower completion rate. Males and females in the Evercare healthcare model had similar rates of AD completion (p > .001), which is also contrary to previous findings where females had a higher completion rate. Finally, older adults with a high school education or less and older adults with greater than a high school education in the Evercare healthcare model had similar rates of AD completion (p > .001), which is contrary to previous findings where individuals with increased education had a higher completion rate.

Implications for practice: With the increasing number of older adults in the general and the long‐term care population, older adults should be encouraged to complete their ADs when discussing their medical decisions with their healthcare providers. Through the use of the Evercare healthcare model, NPs are well prepared to assist their clients and families in identifying these decisions. As a result, a significantly greater proportion of ADs have been completed by individuals enrolled in the Evercare healthcare model when compared to non–Evercare individuals living in long‐term care settings. By using this model, Evercare NPs ensure that the specific medical choices of their patients are carried out.
Indicator of UV Index Intervals
Authors: Letić Milorad
Published: March 2009

A simple device for indication of intervals of UV index (UVI) values and the level of necessary protection is constructed. The indication is based on the solar elevation angle and total ozone column values. The recommended level of protection is indicated via color‐coded (red, yellow, green) intervals on the scale of the device. Comparisons of the indication of the device to the forecast of UVI for 165 days during spring, summer and fall for Belgrade, Serbia showed that for 162 days the indication about application of protective measures was in agreement with the UVI forecast. The indicator is simple, easy to use, straightforward and reliable in determining the interval of UVI values and the level of necessary protection from UV radiation.
Building Research Collaboratives Among Schools and Universities: Lessons From the Field
Authors: Kuriloff Peter , Reichert Michael , Stoudt Brett , Ravitch Sharon
Published: March 2009

ABSTRACT— In a previous issue of Mind, Brain, and Education, Hinton and Fischer (2008) argue that educational research needs to be grounded in the lived realities of school life. They advocate for research schools as a venue for accomplishing this. The Center for the Study of Boys’ and Girls’ Lives represents an alternative model—a research collaborative among independent schools and university‐based scholars. This article describes the Center’s experience with democratic, participatory action research. It discusses major roadblocks encountered doing such work, including difficulties selecting research topics collaboratively, epistemological differences in methods and design, the scarcity of time, and resistance to results when they challenge gender stereotypes or the status quo or involve student researchers. The article concludes with strategies for overcoming these roadblocks, including clearer, upfront negotiations with schools and a compact that specifies roles and responsibilities for both school and Center personnel.
Diagnosis and management of asthma in adolescents
Authors: Towns Susan Joy , Van Asperen Peter Paul
Published: March 2009

Objectives:  In this review we explore some of the issues surrounding the diagnosis and misdiagnosis of asthma in adolescents and suggest a management approach which might facilitate the provision of optimal treatment in order to minimise morbidity from asthma in this vulnerable and often difficult‐to‐manage age group.

Results:  We highlight important diagnostic traps which occur in the adolescent age group, including the misdiagnosis of asthma in young people presenting with exercise‐related symptoms or cough, and stress the importance of considering alternative diagnoses, including vocal cord dysfunction. We explore how adolescence impacts on asthma management and emphasise the importance of an understanding of normal adolescent development and an awareness of high‐risk indicators in developing a strategy to optimally manage a young person with asthma. We also illustrate how psychosocial functioning may impact on both perceived asthma severity and quality of life as well as interfere with optimal asthma control both directly and by increasing non‐adherence. A suggested management strategy for adolescents is provided which emphasises a supportive approach to help facilitate optimal control of asthma by involving the young person in management decisions about their asthma. Specific mention is made about smoking cessation assistance and the interrelationship between physical activity, obesity and asthma. Finally we discuss the importance of an appropriate transition process to prepare the young person for transfer from child‐centred care to adult‐centred care.

Conclusion:  Many different factors is of outmost importance when diagnosing adolescents with shortness of breath; furthermore, management of asthma need a supportive approach in both paediatric and adult setting.

Please cite this paper as: Towns SJ and van Asperen PP. Diagnosis and management of asthma in adolescents. The Clinical Respiratory Journal 2009; 3: 69–76.
Where Is the Unproctored Internet Testing Train Headed Now?
Authors: Tippins Nancy T.
Published: March 2009

So, where is unproctored Internet testing (UIT) now? What has happened since 2006? Is the train stalled right outside the station, waiting on answers? Is it lurching forward, making slow but sure progress? Or, is it simply chugging in circles? The original focal article (Tippins, 2009) and the 13 subsequent commentaries are not comprehensive summaries of all the research and thought about UIT and can hardly be considered definitive as a collective body of work. In total, these articles probably do not convince many of the doubters. However, the articles collectively highlight the critical issues faced in UIT, suggest new ideas, and give guidance for the future, indicating the UIT train is at least moving. In this concluding article of the UIT series, I try to summarize where I believe we are in 2009 in terms of the important points that must be taken into consideration when contemplating UIT and emerging questions that industrial and organizational (I–O) psychologists must answer.
Work Motivation: Identifying Use-Inspired Research Directions
Authors: Kanfer Ruth
Published: March 2009

The study of work motivation progresses through the inspiration that comes from creating new alignments between scientific understanding and considerations of practical use (cf. D. E. Stokes, 1997). Using the 3 C’s framework for work motivation (Kanfer, Chen, & Pritchard, 2008a, b), I coordinate 5 practical concerns related to work motivation with recent scientific trends in order to encourage the development of new research agendas in the field.
Situational Strength as a Means of Conceptualizing Context
Authors: Meyer Rustin D. , Dalal Reeshad S.
Published: March 2009

Kanfer (2009) argues that “context” is an important consideration in the study of motivated behavior, but our knowledge of contextual constructs is immature and in need of considerable development. We agree and build on her position by proposing that situational strength (Hough & Oswald, 2008; Mischel, 1977) has the potential to help conceptualize what Johns (2006) calls “discrete context” (i.e., the particular task, social, and physical variables that influence motivation, attitudes, and behavior). The present article briefly describes situational strength, discusses its operationalization and measurement, explores its implications for practice, and describes two research questions that fall within Pasteur’s Quadrant (Stokes, 1997).
Isolation and characterization of 28 new microsatellite markers for European flounder (Platichthys flesus L.)
Authors: TYSKLIND NIKLAS , NEUPARTH TERESA , ASHCROFT GREGG R. , TAYLOR MARTIN I. , LYONS BRETT P. , MCCARTHY IAN D. , CARVALHO GARY R.
Published: March 2009

The European flounder (Platichthys flesus L.) is used in ecotoxicological studies to provide detailed information on the effects of pollution on individual fish. Data on population and evolutionary level effects are, however, limited. Here, the isolation and characterization of 28 novel species specific microsatellite loci are presented. The number of alleles ranged from 8 to 38, and observed heterozygosity from 0.542 to 1.
Genetic similarity of polyploids: a new version of the computer program POPDIST (version 1.2.0) considers intraspecific genetic differentiation
Authors: TOMIUK JÜRGEN , GULDBRANDTSEN BERNT , LOESCHCKE VOLKER
Published: March 2009

For evolutionary studies of polyploid species estimates of the genetic identity between species with different degrees of ploidy are particularly required because gene counting in samples of polyploid individuals often cannot be done, e.g., in triploids the phenotype AB can be genotypically either ABB or AAB. We recently suggested a genetic distance measure that is based on phenotype counting and made available the computer program POPDIST. The program provides maximum‐likelihood estimates of the genetic identities and distances between polyploid populations, but this approach is not informative for populations within species that only differ in their allele frequencies. We now close this gap by applying the frequencies of shared ‘bands’ in both populations to Nei's identity measure. Our simulation study demonstrates the close correlation between the band‐sharing identity and the genetic identity calculated on the basis of gene frequencies for any degree of ploidy. The new extended version of POPDIST (version 1.2.0) provides the option of choosing either the maximum‐likelihood estimator or the band‐sharing measure.
Characterization of microsatellite loci In The hihi Notiomystis cincta (Notiomystidae, Aves)
Authors: BREKKE PATRICIA , DAWSON DEBORAH A. , HORSBURGH GAVIN J. , EWEN JOHN G.
Published: March 2009

We have characterized 20 polymorphic microsatellite loci in the hihi Notiomystis cincta. Loci were identified by testing loci originally isolated in other avian species and by isolating new microsatellites from a hihi genomic library. These loci were characterized in unrelated hihi from a single population on Tiritiri Matangi Island (n = 98). Each locus displayed between two and 10 alleles, and observed heterozygosities ranged between 0.29 and 0.91. Nineteen of the 20 polymorphic loci could be assigned a chromosome location in the zebra finch, Taeniopygia guttata genome based on sequence homology.
Current computational models do not reveal the importance of the nervous system in long‐term control of arterial pressure
Authors: Osborn John W. , Averina Viktoria A. , Fink Gregory D.
Published: March 2009

Arterial pressure is regulated over long periods of time by neural, hormonal and local control mechanisms, which ultimately determine the total blood volume and how it is distributed between the various vascular compartments of the circulation. A full understanding of the complex interplay of these mechanisms can be greatly facilitated by the use of mathematical models. In 1967, Guyton and Coleman published a model for long‐term control of arterial pressure that focused on renal control of body sodium and water and thus total blood volume. The central point of their model is that the long‐term level of arterial pressure is determined exclusively by the ‘renal function curve’, which relates arterial pressure to urinary excretion of salt and water. The contribution of the sympathetic nervous system to setting the long‐term level of arterial pressure in the model is limited. In light of the overwhelming evidence for a major role of the sympathetic nervous system in long‐term control of arterial pressure and the pathogenesis of hypertension, new mathematical models for long‐term control of arterial pressure may be necessary. Despite the prominence and general acceptance of the Guyton–Coleman model in the field of hypertension research, we argue here that it overestimates the importance of renal control of body fluids and total blood volume in blood pressure regulation. Furthermore, we suggest that it is possible to construct an alternative model in which sympathetic nervous system activity plays an important role in long‐term control of arterial pressure independent of its effects on total blood volume.
Comment on "Detection, Stimulation, and Inhibition of Neuronal Signals with High-Density Nanowire Transistor Arrays"
Authors: Fromherz Peter , Voelker Moritz
Published: March 2009

Patolsky
et al
. (Reports, 25 August 2006, p. 1100) used silicon nanowires to record action potentials in rat neuronal axons and found increases in conductance of about 85 nanosiemens. We point out that the data correspond to voltage changes of about –85 millivolts on the nanowire and that conceivable mechanisms of axon-nanowire interaction lead to signals that are opposite in sign or smaller by orders of magnitude.
Response to Comment on "Detection, Stimulation, and Inhibition of Neuronal Signals with High-Density Nanowire Transistor Arrays"
Authors: Timko Brian P. , Patolsky Fernando , Lieber Charles M.
Published: March 2009

Fromherz and Voelker make incorrect assumptions about our experiments that raise serious questions about the validity of their claims. We show that our calibrated signals are consistent with previously published data and a general model with biophysically relevant parameters. Additionally, the wide variation in previously published signal amplitudes suggests caution in applying and drawing conclusions from the models of Fromherz and Voelker.
Selective Erasure of a Fear Memory
Authors: Han Jin-Hee , Kushner Steven A. , Yiu Adelaide P. , Hsiang Hwa-Lin (Liz) , Buch Thorsten , Waisman Ari , Bontempi Bruno , Neve Rachael L. , Frankland Paul W. , Josselyn Sheena A.
Published: March 2009

Memories are thought to be encoded by sparsely distributed groups of neurons. However, identifying the precise neurons supporting a given memory (the memory trace) has been a long-standing challenge. We have shown previously that lateral amygdala (LA) neurons with increased cyclic adenosine monophosphate response element–binding protein (CREB) are preferentially activated by fear memory expression, which suggests that they are selectively recruited into the memory trace. We used an inducible diphtheria-toxin strategy to specifically ablate these neurons. Selectively deleting neurons overexpressing CREB (but not a similar portion of random LA neurons) after learning blocked expression of that fear memory. The resulting memory loss was robust and persistent, which suggests that the memory was permanently erased. These results establish a causal link between a specific neuronal subpopulation and memory expression, thereby identifying critical neurons within the memory trace.
Recent Changes in Phytoplankton Communities Associated with Rapid Regional Climate Change Along the Western Antarctic Peninsula
Authors: Montes-Hugo Martin , Doney Scott C. , Ducklow Hugh W. , Fraser William , Martinson Douglas , Stammerjohn Sharon E. , Schofield Oscar
Published: March 2009

The climate of the western shelf of the Antarctic Peninsula (WAP) is undergoing a transition from a cold-dry polar-type climate to a warm-humid sub-Antarctic–type climate. Using three decades of satellite and field data, we document that ocean biological productivity, inferred from chlorophyll a concentration (Chl a), has significantly changed along the WAP shelf. Summertime surface Chl a (summer integrated Chl a ∼63% of annually integrated Chl a) declined by 12% along the WAP over the past 30 years, with the largest decreases equatorward of 63°S and with substantial increases in Chl a occurring farther south. The latitudinal variation in Chl a trends reflects shifting patterns of ice cover, cloud formation, and windiness affecting water-column mixing. Regional changes in phytoplankton coincide with observed changes in krill (
Euphausia superba
) and penguin populations.
A Functional Genomics Approach Reveals CHE as a Component of the
Arabidopsis
Circadian Clock
Authors: Pruneda-Paz Jose L. , Breton Ghislain , Para Alessia , Kay Steve A.
Published: March 2009

Transcriptional feedback loops constitute the molecular circuitry of the plant circadian clock. In
Arabidopsis
, a core loop is established between CCA1 and TOC1. Although CCA1 directly represses
TOC1
, the TOC1 protein has no DNA binding domains, which suggests that it cannot directly regulate
CCA1
. We established a functional genomic strategy that led to the identification of CHE, a TCP transcription factor that binds specifically to the
CCA1
promoter. CHE is a clock component partially redundant with LHY in the repression of
CCA1
. The expression of
CHE
is regulated by CCA1, thus adding a CCA1/CHE feedback loop to the
Arabidopsis
circadian network. Because CHE and TOC1 interact, and CHE binds to the
CCA1
promoter, a molecular linkage between TOC1 and
CCA1
gene regulation is established.
Self-Repairing Oxetane-Substituted Chitosan Polyurethane Networks
Authors: Ghosh Biswajit , Urban Marek W.
Published: March 2009

Polyurethanes have many properties that qualify them as high-performance polymeric materials, but they still suffer from mechanical damage. We report the development of polyurethane networks that exhibit self-repairing characteristics upon exposure to ultraviolet light. The network consists of an oxetane-substituted chitosan precursor incorporated into a two-component polyurethane. Upon mechanical damage of the network, four-member oxetane rings open to create two reactive ends. When exposed to ultraviolet light, chitosan chain scission occurs, which forms crosslinks with the reactive oxetane ends, thus repairing the network. These materials are capable of repairing themselves in less than an hour and can be used in many coatings applications, ranging from transportation to packaging or fashion and biomedical industries.
Explosive Percolation in Random Networks
Authors: Achlioptas Dimitris , D'Souza Raissa M. , Spencer Joel
Published: March 2009

Networks in which the formation of connections is governed by a random process often undergo a percolation transition, wherein around a critical point, the addition of a small number of connections causes a sizable fraction of the network to suddenly become linked together. Typically such transitions are continuous, so that the percentage of the network linked together tends to zero right above the transition point. Whether percolation transitions could be discontinuous has been an open question. Here, we show that incorporating a limited amount of choice in the classic Erdös-Rényi network formation model causes its percolation transition to become discontinuous.
Nonporous Organic Solids Capable of Dynamically Resolving Mixtures of Diiodoperfluoroalkanes
Authors: Metrangolo Pierangelo , Carcenac Yvan , Lahtinen Manu , Pilati Tullio , Rissanen Kari , Vij Ashwani , Resnati Giuseppe
Published: March 2009

Halogen bonding has increasingly facilitated the assembly of diverse host-guest solids. Here, we show that a well-known class of organic salts, bis(trimethylammonium) alkane diiodides, can reversibly encapsulate α,ω-diiodoperfluoroalkanes (DIPFAs) through intermolecular interactions between the host's I
–
anions and the guest's terminal iodine substituents. The process is highly selective for the fluorocarbon that forms an I
–
···I(CF
2
)
m
I···I
–
superanion that is matched in length to the chosen dication. DIPFAs that are 2 to 12 carbons in length (common industrial intermediates) can thereby be isolated from mixtures by means of crystallization from solution upon addition of the dissolved size-matched ionic salt. The solid-state salts can also selectively capture the DIPFAs from the vapor phase, yielding the same product formed from solution despite a lack of porosity of the starting lattice structure. Heating liberates the DIPFAs and regenerates the original salt lattice, highlighting the practical potential for the system in separation applications.
Time-Resolved Molecular Frame Dynamics of Fixed-in-Space CS
2
Molecules
Authors: Bisgaard Christer Z. , Clarkin Owen J. , Wu Guorong , Lee Anthony M. D. , Geßner Oliver , Hayden Carl C. , Stolow Albert
Published: March 2009

Random orientation of molecules within a sample leads to blurred observations of chemical reactions studied from the laboratory perspective. Methods developed for the dynamic imaging of molecular structures and processes struggle with this, as measurements are optimally made in the molecular frame. We used laser alignment to transiently fix carbon disulfide molecules in space long enough to elucidate, in the molecular reference frame, details of ultrafast electronic-vibrational dynamics during a photochemical reaction. These three-dimensional photoelectron imaging results, combined with ongoing efforts in molecular alignment and orientation, presage a wide range of insights obtainable from time-resolved studies in the molecular frame.
Giant Ring on the Prairie
Authors: Seidel Robert W.
Published: March 2009

Fermilab
. Physics, the Frontier, and Megascience.
By Lillian Hoddeson, Adrienne W. Kolb, and Catherine Westfall
. University of Chicago Press, Chicago, 2008. 513 pp. $45, £31. ISBN 9780226346236.

This detailed history illuminates the technical, sociological, and political complexities of establishing and operating a laboratory at the frontier of big physics.
Influence of a Probiotic Strain ofEnterococcus faeciumonSalmonella entericaSerovar Typhimurium DT104 Infection in a Porcine Animal Infection Model
Authors: Szabó István , Wieler Lothar H. , Tedin Karsten , Scharek-Tedin Lydia , Taras David , Hensel Andreas , Appel Bernd , Nöckler Karsten
Published: March 2009

The beneficial effects of probioticEnterococcusspp. in different hosts, such as mice and humans, have previously been reported in several studies. However, studies of large domestic animals, as well as challenge studies with pathogenic microorganisms, are very rare. Here, we investigated the influence of oral treatment of pigs with the probiotic bacteriumEnterococcus faeciumNCIMB 10415 onSalmonella entericaserovar Typhimurium DT104 infections in weaning piglets. Clinical symptoms, fecal excretion, the organ distribution ofSalmonella, and the humoral immune response (immunoglobulin G [IgG], IgM, and IgA levels) in serum were examined. A pool of 89 piglets was randomly divided into probiotic and control groups. The probiotic group received a feed supplement containingE. faeciumstarting on day 14 postpartum prior to challenge withSalmonellaserovar Typhimurium DT104 at 28 days postpartum. After challenge withSalmonellaserovar Typhimurium DT104, piglets in both groups showed no severe clinical signs of salmonellosis. However, fecal excretion and colonization ofSalmonellain organs were significantly greater in piglets fedE. faecium. Likewise, the humoral immune response againstSalmonella(serum IgM and IgA levels) was significantly greater in the probiotic group animals than in control animals. The results of this study suggest thatE. faeciumNCIMB 10415 treatment enhanced the course of infection in weaning piglets challenged withSalmonellaserovar Typhimurium DT104. However, the probiotic treatment also appeared to result in greater production of specific antibodies againstSalmonellaserovar Typhimurium DT104.
Identification and Characterization of the Phage Gene
sav
, Involved in Sensitivity to the Lactococcal Abortive Infection Mechanism AbiV
Authors: Haaber Jakob , Rousseau Geneviève M. , Hammer Karin , Moineau Sylvain
Published: March 2009

Lactococcus lactis
phage mutants that are insensitive to the recently characterized abortive infection mechanism AbiV were isolated and analyzed in an effort to elucidate factors involved in the sensitivity to AbiV. Whole-genome sequencing of the phage mutants p2.1 and p2.2 revealed mutations in an
orf
that is transcribed early, indicating that this
orf
was responsible for AbiV sensitivity. Sequencing of the homologous regions in the genomes of other AbiV-insensitive mutants derived from p2 and six other lactococcal wild-type phages revealed point mutations in the homologous
orf
sequences. The
orf
was named
sav
(for
s
ensitivity to
A
bi
V
), and the encoded polypeptide was named SaV. The purification of a His-tagged SaV polypeptide by gel filtration suggested that the polypeptide formed a dimer in its native form. The overexpression of SaV in
L. lactis
and
Escherichia coli
led to a rapid toxic effect. Conserved, evolutionarily related regions in SaV polypeptides of different phage groups are likely to be responsible for the AbiV-sensitive phenotype and the toxicity.
Host-Directed Evolution of a Novel Lactate Oxidase in
Streptococcus iniae
Isolates from Barramundi (
Lates calcarifer
)
Authors: Nawawi Roslina A. , Baiano Justice C. F. , Kvennefors E. Charlotte E. , Barnes Andrew C.
Published: March 2009

In
Streptococcus iniae
, lactate metabolism is dependent upon two proteins, lactate permease that mediates uptake and lactate oxidase, a flavin mononucleotide-dependent enzyme that catalyzes oxidation of α-hydroxyacids. A novel variant of the lactate oxidase gene,
lctO
, in Australian isolates of
S. iniae
from diseased barramundi was found during a diagnostic screen using LOX-1 and LOX-2 primers, yielding amplicons of 920 bp instead of the expected 869 bp. Sequencing of the novel gene variant (type 2) revealed a 51-nucleotide insertion in
lctO
, resulting in a 17-amino-acid repeat in the gene product, and three-dimensional modeling indicated formation of an extra loop in the monomeric protein structure. The activities of the lactate oxidase enzyme variants expressed in
Escherichia coli
were examined, indicating that the higher-molecular-weight type 2 enzyme exhibited higher activity. Growth rates of
S. iniae
expressing the novel type 2 enzyme were not reduced at lactate concentrations of 0.3% and 0.5%, whereas a strain expressing the type 1 enzyme exhibited reduced growth rates at these lactate concentrations. During a retrospective screen of 105 isolates of
S. iniae
from Australia, the United States, Canada, Israel, Réunion Island, and Thailand, the type 2 variant arose only in isolates from a single marine farm with unusually high tidal flow in the Northern Territory, Australia. Elevated plasma lactate levels in the fish, resulting from the effort of swimming in tidal flows of up to 3 knots, may exert sufficient selective pressure to maintain the novel, high-molecular-weight enzyme variant.
Enzyme-Linked Immunosorbent Assay Using Glycoprotein and Monoclonal Antibody for Detecting Antibodies to Vesicular Stomatitis Virus Serotype New Jersey
Authors: Lee Hyang-Sim , Heo Eun-Jeong , Jeoung Hye-Young , Ko Hyo-Rim , Kweon Chang-Hee , Youn Hee-Jeong , Ko Young-Joon
Published: March 2009

In this study, an enzyme-linked immunosorbent assay (ELISA) using glycoprotein and a monoclonal antibody (MAb) was developed for the detection of antibodies to vesicular stomatitis virus (VSV) serotype New Jersey (NJ). The glycoprotein to be used as a diagnostic antigen was extracted from partially purified VSV-NJ, and a neutralizing MAb specific to VSV-NJ was incorporated to compete with antibodies in a blocking ELISA using glycoprotein (GP ELISA). The cutoff of the GP ELISA was set at 40% inhibition, which corresponded to a virus neutralization test (VNT) titer of 32. With this threshold, the GP ELISA exhibited 99.6% specificity for naïve sera (n= 3,005) from cattle (n= 1,040), pigs (n= 1,120), and horses (n= 845) from domestic farms. The GP ELISA did not cross-react with sera positive for foot-and-mouth disease virus, swine vesicular disease virus, or VSV serotype Indiana. The GP ELISA was more compatible with the VNT than was the nucleocapsid-based ELISA for VSV-NJ-positive sera (n= 19). Taken together, this GP ELISA could be a useful tool as an alternative to the VNT for detecting antibodies specific to VSV-NJ.
Potent Adjuvant Activity of Cationic Liposome-DNA Complexes for Genital Herpes Vaccines
Authors: Bernstein David I. , Cardin Rhonda D. , Bravo Fernando J. , Strasser Jane E. , Farley Nicholas , Chalk Claudia , Lay Marla , Fairman Jeff
Published: March 2009

Development of a herpes simplex virus (HSV) vaccine is a priority because these infections are common. It appears that potent adjuvants will be required to augment the immune response to subunit HSV vaccines. Therefore, we evaluated cationic liposome-DNA complexes (CLDC) as an adjuvant in a mouse model of genital herpes. Using a whole-virus vaccine (HVAC), we showed that the addition of CLDC improved antibody responses compared to vaccine alone. Most important, CLDC increased survival, reduced symptoms, and decreased vaginal virus replication compared to vaccine alone or vaccine administered with monophosphoryl lipid A (MPL) plus trehalose dicorynomycolate (TDM) following intravaginal challenge of mice. When CLDC was added to an HSV gD2 vaccine, it increased the amount of gamma interferon that was produced from splenocytes stimulated with gD2 compared to the amount produced with gD2 alone or with MPL-alum. The addition of CLDC to the gD2 vaccine also improved the outcome following vaginal HSV type 2 challenge compared to vaccine alone and was equivalent to vaccination with an MPL-alum adjuvant. CLDC appears to be a potent adjuvant for HSV vaccines and should be evaluated further.
Characterization of Protective Mucosal and Systemic Immune Responses Elicited by Pneumococcal Surface Protein PspA and PspC Nasal Vaccines against a Respiratory Pneumococcal Challenge in Mice
Authors: Ferreira D. M. , Darrieux M. , Silva D. A. , Leite L. C. C. , Ferreira J. M. C. , Ho P. L. , Miyaji E. N. , Oliveira M. L. S.
Published: March 2009

Pneumococcal surface protein A (PspA) and PspC are virulence factors that are involved in the adhesion of
Streptococcus pneumoniae
to epithelial cells and/or evasion from the immune system. Here, the immune responses induced by mucosal vaccines composed of both antigens as recombinant proteins or delivered by
Lactobacillus casei
were evaluated. None of the PspC vaccines protected mice against an invasive challenge with pneumococcal strain ATCC 6303. On the other hand, protection was observed for immunization with vaccines composed of PspA from clade 5 (PspA5 or
L. casei
expressing PspA5) through the intranasal route. The protective response was distinguished by a Th1 profile with high levels of immunoglobulin G2a production, efficient complement deposition, release of proinflammatory cytokines, and infiltration of neutrophils. Intranasal immunization with PspA5 elicited the highest level of protection, characterized by increased levels of secretion of interleukin-17 and gamma interferon by lung and spleen cells, respectively, and low levels of tumor necrosis factor alpha in the respiratory tract.
Structural Characterization of an Oligosaccharide Made by
Neisseria sicca
Authors: O'Connor Ellen T. , Zhou Hui , Bullock Kevin , Swanson Karen V. , Griffiss J. McLeod , Reinhold Vernon N. , Miller Clinton J. , Stein Daniel C.
Published: March 2009

Neisseria sicca
4320 expresses two carbohydrate-containing components with sodium dodecyl sulfate-polyacrylamide gel electrophoresis mobilities that resemble those of lipooligosaccharide and lipopolysaccharide. Using matrix-assisted laser desorption ionization—time of flight and electrospray ionization mass spectrometry, we characterized a disaccharide carbohydrate repeating unit expressed by this strain. Gas chromatography identified the sugars composing the unit as rhamnose and
N
-acetyl-
d
-glucosamine. Glycosidase digestion confirmed the identity of the nonreducing terminal sugar of the disaccharide and established its β-anomeric configuration. Mass spectrometry analysis and lectin binding were used to verify the linkages within the disaccharide repeat. The results revealed that the disaccharide repeat is [-4) β-
l
-rhamnose (1-3) β-
N
-acetyl-
d
-glucosamine (1-] with an
N
-acetyl-
d
-glucosamine nonreducing terminus. This work is the first structural characterization of a molecule that possesses rhamnose in the genus
Neisseria
.
Parvovirus B19 Infection Localized in the Intestinal Mucosa and Associated with Severe Inflammatory Bowel Disease
Authors: Pironi Loris , Bonvicini Francesca , Gionchetti Paolo , D'Errico Antonia , Rizzello Fernando , Corsini Catia , Foroni Laura , Gallinella Giorgio
Published: March 2009

Infection by human parvovirus B19 is widespread and can be associated with a wide range of different pathologies and clinical manifestations. We provide the first evidence of localization of an active parvovirus B19 infection in the intestinal mucosa and its association with a severe inflammatory bowel disease, characterized by duodenal villous atrophy with increased intraepithelial lymphocytes and inflammatory infiltrates in the colonic mucosa. Virus in the intestinal mucosa was detected in cells of the inflammatory infiltrate, identified as T lymphocytes and selectively localized in sites of active tissue degeneration.
Performance of a Polymer-Based DNA Chip Platform in Detection and Genotyping of Human Papillomavirus in Clinical Samples
Authors: Schenk T. , Brandstetter T. , zur Hausen A. , Alt-Mörbe J. , Huzly D. , Rühe J.
Published: March 2009

Human papillomavirus (HPV) plays a key role in the development of cervical and laryngeal cancers. The aim of our study was to compare the performance of a new hydrogel-based HPV genotyping biochip assay (Biochip) to a commercially available and CE-marked conventional PCR followed by reverse hybridization (GenID-PCR). One hundred twenty-three samples were available for the study. Of these samples, 101/123 were gynecological swabs, 8/123 were swabs or biopsy samples of genital warts, 7/123 were biopsy samples of otorhinolaryngeal lesions, 5/123 were samples of skin warts, and 2/123 were samples of orolabial abnormalities. These molecular methods for HPV genotyping showed comparable sensitivity and specificity. However, 19/123 of the results were discrepant. Specifically, Biochip showed better performance in the detection of multiple infections, especially when more than one high-risk genotype was present. Due to the different probe configurations used in the two assays, GenID-PCR achieves only group-specific detection of many HPV genotypes, whereas Biochip allows for specific identification. Overall, the newly developed HPV chip system (Biochip) proved to be a suitable tool for HPV detection and genotyping; it also proved to be superior for establishing HPV genotyping methods.
Improvement in Mycobacterial Yield and Reduced Time to Detection in Pediatric Samples by Use of a Nutrient Broth Growth Supplement
Authors: Brittle W. , Marais B. J. , Hesseling A. C. , Schaaf H. S. , Kidd M. , Wasserman E. , Botha T.
Published: March 2009

There is an urgent need to improve the methods used for the bacteriological diagnosis of childhood mycobacterial disease. This study compared the mycobacterial yields and the times to detection (in days) of mycobacteria in pediatric clinical specimens by using Mycobacterial Growth Indicator Tubes (MGITs) and solid Löwenstein-Jensen (LJ) slants with and without a nutrient broth supplement. A total of 801 specimens from 493 patients were processed: 82.8% were gastric aspirate specimens, 15.6% were sputum specimens, and 1.6% were fine-needle-aspiration biopsy specimens. The mycobacterial yield obtained with MGITs (with and without nutrient broth) was 11.0%, and that obtained with LJ slants was 1.6% (
P
< 0.001). Of the 88 positive cultures, 62 were detected in MGITs and 73 were detected in MGITs supplemented with nutrient broth (
P
= 0.11). The mean time to detection in MGITs (without nutrient broth) was 18.5 days, whereas it was 12.4 days in MGITs with nutrient broth (
P
< 0.001). Supplementation of standard MGITs improved the mycobacterial yield and significantly reduced the time to detection of mycobacteria in pediatric samples.
Cervid Herpesvirus 2 Causes Respiratory and Fetal Infections in Semidomesticated Reindeer
Authors: das Neves Carlos G. , Rimstad Espen , Tryland Morten
Published: March 2009

Members of the viral subfamilyAlphaherpesvirinaeestablish latency from which they can be reactivated. Bovine herpesvirus 1 causes infectious bovine rhinotracheitis and infectious pustular vulvovaginitis in cattle, as well as abortion and weak calves. Serological evidence of alphaherpesvirus infection has been reported for wild and semidomesticated reindeer (Rangifer tarandus tarandus) in Norway. To address the possibility that reindeer alphaherpesvirus (cervid herpesvirus 2 [CvHV-2]) infection might affect the respiratory system and in part explain the relatively high mortality of reindeer calves during their first year, tissue samples were obtained from reindeer and reindeer fetuses at slaughterhouses in Finnmark County, Norway. A nested pan-alphaherpesvirus PCR amplification targeting the highly conserved UL27 gene (encoding glycoprotein B) was used. Sequencing of amplicons revealed the presence of CvHV-2 DNA. The detection of CvHV-2 DNA in trigeminal ganglia (27 of 143 samples), nasal swabs (5 of 75 samples), and fetal tissues (12 of 48 samples) indicates that CvHV-2 infection is endemic in this reindeer population. Moreover, the virus is transmitted horizontally by the respiratory route, establishing latency in the trigeminal ganglion, and vertically to the fetus through the placenta. Further studies should focus on the reproductive impact of CvHV-2 infection in reindeer.
The LMP1 Promoter Can Be Transactivated Directly by NF-κB
Authors: Demetriades Constantinos , Mosialos George
Published: March 2009

A bioinformatic analysis identified two putative NF-κB binding sites in the Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) promoter. The ability of p65RelA to interact with the LMP1 promoter was shown by in vitro and in vivo assays. Using an EBV-transformed lymphoblastoid cell line as a reporter system for the activity of the +40/−328 LMP1 promoter region, the functional importance of NF-κB and other transcription factor binding sites was demonstrated. p65RelA could also induce LMP1 expression from the EBV genome in Daudi and P3HR1 Burkitt's lymphoma cell lines. Finally, it was shown that p65RelA could cooperate with EBNA2 or the aryl hydrocarbon receptor in the transactivation of the LMP1 promoter. Our study established the importance of NF-κB and several
cis
-acting elements in the regulation of the LMP1 promoter in a latency III environment and highlighted a complex interplay between NF-κB and other transcription factors in this process.
Broad Neutralization of Human Immunodeficiency Virus Type 1 (HIV-1) Elicited from Human Rhinoviruses That Display the HIV-1 gp41 ELDKWA Epitope
Authors: Arnold Gail Ferstandig , Velasco Paola K. , Holmes Andrew K. , Wrin Terri , Geisler Sheila C. , Phung Pham , Tian Yu , Resnick Dawn A. , Ma Xuejun , Mariano Thomas M. , Petropoulos Christos J. , Taylor John W. , Katinger Hermann , Arnold Eddy
Published: March 2009

In efforts to develop AIDS vaccine components, we generated combinatorial libraries of recombinant human rhinoviruses that display the well-conserved ELDKWA epitope of the membrane-proximal external region of human immunodeficiency virus type 1 (HIV-1) gp41. The broadly neutralizing human monoclonal antibody 2F5 was used to select for viruses whose ELDKWA conformations resemble those of HIV. Immunization of guinea pigs with different chimeras, some boosted with ELDKWA-based peptides, elicited antibodies capable of neutralizing HIV-1 pseudoviruses of diverse subtypes and coreceptor usages. These recombinant immunogens are the first reported that elicit broad, albeit modest, neutralization of HIV-1 using an ELDKWA-based epitope and are among the few reported that elicit broad neutralization directed against any recombinant HIV epitope, providing a critical advance in developing effective AIDS vaccine components.