bluuebunny/crossref_metadata_2025 huggingface dataset:
What are the Links Between Theory of Mind and Social Relations? Review, Reflections and New Directions for Studies of Typical and Atypical Development
Authors: Hughes Claire , Leekam Sue
Published: October 2004

Twenty years after research on children's ‘theory of mind’ began, this field continues to be a leading influence in the study of developmental psychology and psychopathology. In this review we examine the contribution of research on children's theories of mind to our understanding of children's developing social relationships. Evidence shows that for both typical and atypical populations this relationship is neither uniform nor unidirectional. Theory‐of‐mind skills are multifaceted and the nature of the developmental relationship between different aspects is not yet known, and there is evidence that theory‐of‐mind skills both transform and are transformed by interpersonal and family relationships and by language communities. To understand the multifaceted and complex nature of development in this area, we need clearer definitions of the skills under investigation, as well as research designs that capture the transactional nature of the relationship between theory of mind and social relations. Addressing these issues should help to clarify (i) the processes by which children's developing understanding of others is influenced by the social environment, and (ii) issues concerning the specificity of theory‐of‐mind impairments in atypical populations and the processes by which these impairments develop.
ECONOMIC CONSEQUENCES OF THE POSTED WORKERS DIRECTIVE
Authors: Meier Volker
Published: October 2004

The consequences of an increase in the minimum wage for foreign workers in the construction sector, implied by the EU Posted Workers Directive, are analyzed. Due to the rising price of construction services, the factor demand for both construction services and capital in the tradeable good sector falls, and the wage rate in this sector declines. While the share of domestic workers increases with respect to both foreign workers and capital in the construction sector, this need not suffice to reduce unemployment. A possible higher level of employment of natives is not sufficient to raise natives’ welfare.
The Interface Between Physical and Mental Health Problems and Medical Help Seeking in Children and Adolescents: A Research Perspective
Authors: Elena Garralda M.
Published: October 2004

This paper addresses child and adolescent psychopathology as it presents to general practitioners and paediatricians, and explores psychosocial aspects of unexplained medical symptoms in children and adolescents. High rates of psychopathology have been identified amongst children and adolescents attending general practice and paediatric services, most of it ‘‘hidden’’ at presentation and emotional in nature. It is often linked to poor physical well being and to maternal stress focused on the child. It may be of special relevance to medical help seeking in socio‐economically advantaged areas. Co‐morbid psychopathology, mainly emotional disorders, is common amongst children with unexplained medical symptoms. However, there are specific psychosocial aspects that differentiate these children from those with emotional disorders. They involve disease beliefs, illness behaviour and predicament. The latter may be characterised by special reactivity to stress in children with personality vulnerability, in a context of parents with high levels of mental distress, unexplained medical symptoms and emotional over‐involvement with the child. There is comparatively little interface work between CAMHS and primary health care. An important research priority would seem to lie in the development of interventions that can be adapted for use by primary care staff. Similarly, there are few dedicated CAMHS paediatric liaison teams. Their more extensive development should help attend in a more informed and focused way than at present to children and adolescents suffering from unexplained physical symptoms and disorders. Further research is needed into vulnerability mechanisms and maintaining factors, health beliefs, treatment engagement and interventions.
Social Services and CAMHS: A National Survey
Authors: Kerfoot Michael , Panayiotopoulos Christos , Harrington Richard
Published: October 2004

Background:  Links between Social Service Departments (SSDs) and CAMHS are explored and described through a national survey and a number of site visits.

Method:  Data were collected via (1) a national questionnaire survey yielding a response rate of 75%, and (2) pre‐selected site visits to seven SSDs.

Results:  Additional to existing CAMHS assessment and treatment provision to SSDs, there is increasing outreach consultation and support, some provided through primary mental health workers and dedicated clinical psychology sessions. Some SSDs have developed innovative in‐house provision for Tier 2 type referrals.

Conclusions:  Outreach and core CAMHS provision to SSDs need equal investment and development.
Chronic Fatigue: A Survey, Outcome Study and Proposals
Authors: Lines Sharon
Published: October 2004

Background:  Sample sizes in studies of childhood chronic fatigue remain small and there is a need to integrate research findings and establish common criteria.

Method:  This is a detailed report of 20 children with medically unexplained fatigue, referred to as Chronic Fatigue Syndrome (CFS). It includes an uncontrolled outcome study of treatment based on a pragmatic cognitive‐behavioural model adapted for children at Great Ormond Street Hospital for Children.

Results:  The data revealed psychological distress at 30 times the prevalence found in the normal child population, with specific high‐ranking symptoms, including headache, social withdrawal and decreased appetite. Onset tended to occur at 11 years of age with some earlier problems at school age, suggesting it might be possible to identify those most at risk. Follow‐up demonstrated improvement in 87% of the sample (parental report), a significant increase in school attendance and a significant decrease in symptom severity. Factors associated with good outcome were short duration of fatigue (less than a year) and behaviours that tended to combat the cycle of avoidance, such as carefully planned home‐tuition.

Conclusions:  Recording a comprehensive range of symptoms and factors facilitates comparisons within and between child studies. It is proposed that symptom severity is a particularly relevant measure for Chronic Fatigue Syndrome which is characterised by persisting symptoms.
The Impact of Primary Mental Health Workers on Referrals to CAMHS
Authors: Whitworth Denise , Ball Clare
Published: October 2004

Background:  This paper describes the impact of a new primary mental health team within the existing child and adolescent health service on referrals and attendance rates.

Method:  100 referrals to CAMHS from prior to the PMHT and 100 referrals prior to the evaluation were examined and classified as appropriate or inappropriate for CAMHS. Information was also gathered as to whether the child and/or family attended for their appointment with CAMHS.

Results and Conclusions:  a large increase in attendance and decrease in non‐attendance rates to CAMHS was seen. The referrals to CAMHS have become more appropriate for a psychiatry service. However, the service provoked a mixed response from Tier 1 professionals. Recommendations are made for the skills needed by primary mental health workers and further evaluative techniques.
Understanding Children and Young People's Rights to Confidentiality
Authors: Harbour Anthony
Published: October 2004

This article develops some themes in relation to young people and confidentiality by focusing on recent guidance from the Department of Health. The Child and Adolescent Faculty of the Royal College of Psychiatrists, Julian Sheather and Rebecca Mussell from the BMA, and Colin Hawkes and Eileen Vizard from the Young Abusers Project greatly assisted.
Decision-Making in Betting Markets
Authors: Vaughan Williams Leighton
Published: October 2004

The placing of a bet is a classic example of decision-making under uncertainty. A betting market is also an example of a simple financial market, but one which possesses the advantage that each bet is characterised by a well-defined end point at which it possesses a definite value, i.e. the amount won or lost. Leighton Vaughan Williams explains the implications for our understanding of economic decision-making of the observed tendency for the expected returns on bets to differ markedly at different odds levels.
It Shouldn't Happen to a Statistician
Authors: Clough Helen , Brown Patrick
Published: October 2004

Helen Clough and Patrick Brown are statisticians working in a cross-disciplinary veterinary epidemiology collaboration, uniting statisticians, mathematical modellers, veterinarians, molecular biologists and microbiologists. They describe the strengths that statisticians bring to the collaboration and the creative tensions that arise when both statistical and mathematical modelling techniques are used.
Career Story: Official Statistician
Authors: Rice Melissa
Published: October 2004

It was a desire to travel which first attracted Melissa Rice to a career in the Civil Service. And, even though she hasn't yet done as much travelling as she would like, she already has several job changes under her belt and, as she tells Helen Joyce, opportunities to broaden her horizons in other ways.
Improving a Food Process by Experimentation
Authors: Gilmour Steven
Published: October 2004

Like any other industry, the food industry continuously develops and improves its products and the processes that are used to make them, says Steven Gilmour. The customer is mainly interested in the sensory characteristics of the product: how does it taste; how does it smell; how does it look? However, commercial success also depends on whether or not the product can be produced economically, consistently and safely. Product development and improvement require research, which often means experimentation.
Triangle Plots of Three-Level Explanatory Variables
Authors: Alexander Neal
Published: October 2004

Thalassaemias are genetic blood disorders which cause varying degrees of anaemia. Their geographical distribution suggests a compensating protection against malaria, which kills between 0.5 and 2.5 million people per year in developing countries. Neal Alexander describes a study in Papua New Guinea which estimated this association more directly, and a triangle plot which clarified, for himself and his non-statistician colleagues, the relative risks of malaria for those with none, one or two copies of the relevant haemoglobin mutation.
Estimation of Fault Strength: Reconstruction of Stress Before the 1995 Kobe Earthquake
Authors: Yamashita Futoshi , Fukuyama Eiichi , Omura Kentaro
Published: October 2004

We have estimated the stress field before the 1995 Kobe, Japan, earthquake (moment magnitude 6.9) using in situ post-shock stress measurements obtained from hydraulic fracturing experiments near the fault. We reconstructed the pre-shock stress field using a kinematic source model inverted from seismic waveforms and geodetic deformations. We found that at the center of the fault, two sides of the fault surface coupled completely before the earthquake, with a coefficient of friction of 0.6, which is equivalent to strong crust. At the edge of the fault, a possible aseismic slip is expected to occur from the pre-shock stress orientation.
Morphological Disparity of Ammonoids and the Mark of Permian Mass Extinctions
Authors: Villier Loïc , Korn Dieter
Published: October 2004

The taxonomic diversity of ammonoids, in terms of the number of taxa preserved, provides an incomplete picture of the extinction pattern during the Permian because of a strongly biased fossil record. The analysis of morphological disparity (the variety of shell shapes) is a powerful complementary tool for testing hypotheses about the selectivity of extinction and permits the recognition of three distinct patterns. First, a trend of decreasing disparity, ranging for about 30 million years, led to a minimum disparity immediately before the Permian-Triassic boundary. Second, the strongly selective Capitanian crisis fits a model of background extinction driven by standard environmental changes. Third, the end-Permian mass extinction operated as a random, nonselective sorting of morphologies, which is consistent with a catastrophic cause.
The Scaling of Animal Space Use
Authors: Jetz Walter , Carbone Chris , Fulford Jenny , Brown James H.
Published: October 2004

Space used by animals increases with increasing body size. Energy requirements alone can explain how population density decreases, but not the steep rate at which home range area increases. We present a general mechanistic model that predicts the frequency of interaction, spatial overlap, and loss of resources to neighbors. Extensive empirical evidence supports the model, demonstrating that spatial constraints on defense cause exclusivity of home range use to decrease with increasing body size. In large mammals, over 90% of available resources may be lost to neighbors. Our model offers a general framework to understand animal space use and sociality.
Activating Mutations of
NOTCH1
in Human T Cell Acute Lymphoblastic Leukemia
Authors: Weng Andrew P. , Ferrando Adolfo A. , Lee Woojoong , Morris John P. , Silverman Lewis B. , Sanchez-Irizarry Cheryll , Blacklow Stephen C. , Look A. Thomas , Aster Jon C.
Published: October 2004

Very rare cases of human T cell acute lymphoblastic leukemia (T-ALL) harbor chromosomal translocations that involve
NOTCH1
, a gene encoding a transmembrane receptor that regulates normal T cell development. Here, we report that more than 50% of human T-ALLs, including tumors from all major molecular oncogenic subtypes, have activating mutations that involve the extracellular heterodimerization domain and/or the C-terminal PEST domain of NOTCH1. These findings greatly expand the role of activated NOTCH1 in the molecular pathogenesis of human T-ALL and provide a strong rationale for targeted therapies that interfere with NOTCH signaling.
Variable Azimuthal Anisotropy in Earth's Lowermost Mantle
Authors: Garnero Edward J. , Maupin Valérie , Lay Thorne , Fouch Matthew J.
Published: October 2004

A persistent reversal in the expected polarity of the initiation of vertically polarized shear waves that graze the D″ layer (the layer at the boundary between the outer core and the lower mantle of Earth) in some regions starts at the arrival time of horizontally polarized shear waves. Full waveform modeling of the split shear waves for paths beneath the Caribbean requires azimuthal anisotropy at the base of the mantle. Models with laterally coherent patterns of transverse isotropy with the hexagonal symmetry axis of the mineral phases tilted from the vertical by as much as 20° are consistent with the data. Small-scale convection cells within the mantle above the D″ layer may cause the observed variations by inducing laterally variable crystallographic or shape-preferred orientation in minerals in the D″ layer.
Engaged with Sequences
Authors: Winters R. Scott
Published: October 2004

Gene(sis)
Contemporary Art Explores Human Genomics. Robin Held, curator. Organized by the Henry Art Gallery, the University of Washington, Seattle, WA, in affiliation with the Berkeley Art Museum, Berkeley, CA. At the Mary and Leigh Block Museum of Art, Northwestern University, Evanston, IL, through 28 November 2004.
www.gene-sis.net
. CD-ROM catalogue, $24.95.

The nearly 60 works, by 24 contemporary artists or ensembles, in this exhibit seek to promote considerations of the potential social, emotional, and ethical implications of the Human Genome Project.
Origins of Microsatellite Diversity in the
Trichophyton rubrum
-
T. violaceum
Clade (Dermatophytes)
Authors: Ohst T. , de Hoog S. , Presber W. , Stavrakieva V. , Gräser Y.
Published: October 2004

We analyzed the population structure of the anthropophilic dermatophyte species
Trichophyton violaceum
, which mainly causes tinea capitis, and
T. rubrum
, the most frequently isolated agent of dermatophytosis worldwide. A microsatellite marker (T1) was developed by using the enrichment technique for microsatellites. The T1 marker containing a (GT)
8-10
repeat was proven to specifically amplify both species, underlining their close kinship. Four polymorphic alleles were detected within a set of about 130 strains by using polyacrylamide gel electrophoresis with this marker. An association with geographic origin of the isolates was apparent. Given the close relatedness of both species, these data suggest an African origin of the entire
T. rubrum
complex, followed by the emergence of a new genotype (B) in Asia with subsequent spread of this genotype over Europe and the United States.
Serotypes and Sequence Types of Pneumococci Causing Invasive Disease in Scotland Prior to the Introduction of Pneumococcal Conjugate Polysaccharide Vaccines
Authors: Clarke S. C. , Scott K. J. , McChlery S. M.
Published: October 2004

Pneumococcal conjugate polysaccharide (Pnc) vaccines are now available, and the need for an improved understanding of circulating pneumococcal serotypes and sequence types (STs) is recognized. Three hundred sixty-eight pneumococci isolated in cases of invasive disease in Scotland in the first 6 months of 2003 were analyzed. The isolates belonged to 30 serotypes, and there was a strong correlation between serotype and ST, although only nine serotypes consisted of a single ST. The following serotypes coexisted with the following numbers of STs: serotype 14, 10 STs, serotype 8, 8 STs; serotype 4, 6 STs; serotype 22F, 8 STs; serotype 9V, 7 STs; serotype 23F, 6 STs; serotype 6B, 6 STs; serotype 1, 3 STs; serotype 3, 3 STs; and serotype 7F, 3 STs. Our data also showed a strong association between ST and serotype, although 19 STs contained multiple serotypes. Of the 10 most common STs, 6 coexisted with a single serotype each. Vaccine coverage in all age groups was 94.9% for the 23-valent polysaccharide vaccine and 50.7, 55.4, and 64.1% for the 7-, 9-, and 11-valent Pnc vaccines, respectively. For those under the age of 2 years, 79% coverage would be provided by the 7-, 9-, and 11-valent Pnc vaccines.
Detection of U.S., Lelystad, and European-Like Porcine Reproductive and Respiratory Syndrome Viruses and Relative Quantitation in Boar Semen and Serum Samples by Real-Time PCR
Authors: Wasilk A. , Callahan J. D. , Christopher-Hennings J. , Gay T. A. , Fang Y. , Dammen M. , Reos M. E. , Torremorell M. , Polson D. , Mellencamp M. , Nelson E. , Nelson W. M.
Published: October 2004

Transmission of porcine reproductive and respiratory syndrome virus (PRRSV) via boar semen has been documented. Since semen is widely disseminated for artificial insemination and the virus can cause significant health and economic consequences, it is essential to have well-validated, rapid diagnostic techniques to detect and quantitate the virus for diagnostic and research purposes. Previously, boar semen was tested by a nested PCR (nPCR) assay which was compared to the “gold standard” swine bioassay. A correlation of 94% was observed, indicating that, most of the time, PCR detected infectious virus. Subsequently, a real-time PCR targeting the 3′ untranslated region of the PRRSV genome was compared with nPCR by testing 413 serum and semen samples from PRRSV-inoculated and control boars. There was 95% agreement between the results of the two tests, with the majority of samples with discordant results containing virus at the lower range of detection by the assays. The virus in all samples was quantitated by using a standard curve obtained by serial dilution of an in vitro transcript. By using the in vitro transcript, the lower limit of sensitivity was observed to be approximately 33 copies/ml. Reactivity with a panel of more than 100 PRRSV isolates from various geographical regions in the United States was also documented. No reactivity with nine nonrelated swine viruses was noted. A real-time PCR was also developed for the detection of the European Lelystad virus and the European-like PRRSV now found in the United States. In six of six PRRSV-inoculated boars, peak levels of viremia occurred at 5 days postinoculation (DPI) and were most consistently detectable throughout 22 DPI. In five of six boars, PRRSV was shed in semen for 0 to 2 days during the first 10 DPI; however, one of six boars shed the virus in semen through 32 DPI. Therefore, in general, the concentration and duration of PRRSV shedding in semen did not correlate with the quantity or duration of virus in serum. These differences warrant further studies into the factors that prevent viral replication in the reproductive tract.
Epidemiologic and Molecular Characterization of an Outbreak of
Candida parapsilosis
Bloodstream Infections in a Community Hospital
Authors: Clark Thomas A. , Slavinski Sally A. , Morgan Juliette , Lott Timothy , Arthington-Skaggs Beth A. , Brandt Mary E. , Webb Risa M. , Currier Mary , Flowers Richard H. , Fridkin Scott K. , Hajjeh Rana A.
Published: October 2004

Candida parapsilosis
is an important cause of bloodstream infections in the health care setting. We investigated a large
C. parapsilosis
outbreak occurring in a community hospital and conducted a case-control study to determine the risk factors for infection. We identified 22 cases of bloodstream infection with
C. parapsilosis
: 15 confirmed and 7 possible. The factors associated with an increased risk of infection included hospitalization in the intensive care unit (adjusted odds ratio, 16.4; 95% confidence interval, 1.8 to 148.1) and receipt of total parenteral nutrition (adjusted odds ratio, 9.2; 95% confidence interval, 0.9 to 98.1). Samples for surveillance cultures were obtained from health care worker hands, central venous catheter insertion sites, and medical devices. Twenty-six percent of the health care workers surveyed demonstrated hand colonization with
C. parapsilosis
, and one hand isolate was highly related to all case-patient isolates by tests with the DNA probe Cp3-13. Outbreak strain isolates also demonstrated reduced susceptibilities to fluconazole and voriconazole. This largest known reported outbreak of
C. parapsilosis
bloodstream infections in adults resulted from an interplay of host, environment, and pathogen factors. Recommendations for control measures focused on improving hand hygiene compliance.
Tropheryma whipplei
Infection of an Acellular Porcine Heart Valve Bioprosthesis in a Patient Who Did Not Have Intestinal Whipple's Disease
Authors: Dreier Jens , Szabados Florian , von Herbay Axel , Kröger Thilo , Kleesiek Knut
Published: October 2004

Rare cases of culture-negative infective endocarditis are caused by
Tropheryma whipplei
, the uncommon bacterium of Whipple's disease. We evaluated an 80-year-old woman with valvular heart disease but without intestinal Whipple's disease. The diagnosis of aortic valve xenograft culture-negative infection with
T. whipplei
was established by multiple molecular assays and by electron microscopy. First, a PCR with broad-range primers identified the complete 16S ribosomal DNA of
T. whipplei
in bioprosthesis tissue. Novel real-time reverse transcription-PCR assays were developed to detect mRNAs encoding recently identified proteins determined from the
T. whipplei
genome, specifically Whipplei surface protein (TW113) and a DNA polymerase III subunit (TW727). The positive detection of mRNAs indicated the presence of metabolically active bacteria and suggested the viability of
T. whipplei
. The quantification of
T. whipplei
genome equivalents by real-time PCR indicated a high-density bacterial colonization of the valve tissue. Additionally, an ultrastructural examination revealed numerous rod-shaped bacteria consistent in size with
T. whipplei
in the extracellular collagen matrix of the bioprosthesis. We conclude that extracellular growth of
T. whipplei
can occur in the microenvironment of biological prosthetic valve tissue and that
T. whipplei
endocarditis can occur in the absence of intestinal Whipple's disease.
Genotyping of 44 Isolates ofFusarium solani, the Main Agent of Fungal Keratitis in Brazil
Authors: Godoy Patricio , Cano Josep , Gené Josepa , Guarro Josep , Höfling-Lima Ana Luisa , Lopes Colombo Arnaldo
Published: October 2004

In a retrospective study performed over 6 years in Brazil,Fusarium solaniwas found to be the most common species causing mycotic keratitis. The genetic diversity of 44 isolates from 39 patients was assessed by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and PCR restriction fragment length polymorphism (PCR-RFLP) fingerprinting. ERIC-PCR was more discriminatory than PCR-RFLP for differentiating the strains. By combining of the results of both techniques, we identified 40 genotypes. Molecular typing revealed a high genomic heterogeneity of the strains ofF. solanistudied.
Detection of Mutations Associated with Isoniazid and Rifampin Resistance in
Mycobacterium tuberculosis
Isolates from Samara Region, Russian Federation
Authors: Nikolayevsky V. , Brown T. , Balabanova Y. , Ruddy M. , Fedorin I. , Drobniewski F.
Published: October 2004

High incidence rates of isoniazid-, rifampin-, and multiple-drug-resistant tuberculosis have been reported in countries of the former Soviet Union (FSU). Genotypic (unlike phenotypic) drug resistance assays do not require viable cultures but require accurate knowledge of both the target gene and the mutations associated with resistance. For these assays to be clinically useful, they must be able to detect the range of mutations seen in isolates from the population of tuberculosis patients to which they are applied. Two novel macroarrays were applied to detect mutations associated with rifampin (
rpoB
) and isoniazid (
katG
and
inhA
) resistance. In a sample of 233 isolates from patients in Samara, central Russia, 46.5% of isolates possessed mutations in both the
rpoB
and the
katG
(or
inhA
) genes. Combined results from the macroarrays demonstrated concordance in 95.4 and 90.4% of phenotypically defined rifampin- and isoniazid-resistant isolates, respectively. The contribution of different mutations to resistance was comparable to that reported previously for non-FSU countries, with 90% of rifampin-resistant isolates and 93% of isoniazid resistant isolates due to
rpoB531
and
katG315
mutations, respectively. The percentage of phenotypically resistant rifampin isolates with no mutations in the
rpoB
codons 509 to 536 was 4.2%, which was similar to previous reports. Novel macroarrays offer a rapid, accurate, and relatively cheap system for the identification of rifampin-, isoniazid-, and multiple-drug-resistant
Mycobacterium tuberculosis
isolates.
Novel Real-Time PCR Assay for Detection of
Helicobacter pylori
Infection and Simultaneous Clarithromycin Susceptibility Testing of Stool and Biopsy Specimens
Authors: Schabereiter-Gurtner Claudia , Hirschl Alexander M. , Dragosics Brigitte , Hufnagl Peter , Puz Sonja , Kovách Zsuzsanna , Rotter Manfred , Makristathis Athanasios
Published: October 2004

A biprobe real-time PCR protocol, followed by hybridization melting point analysis, to detect point mutations in the 23S rRNA gene of
Helicobacter pylori
associated with clarithromycin resistance was established and evaluated in a clinical study. Of 92 patients who underwent endoscopy, 45 were found to be
H. pylori
infected and invariably were also culture positive. Of the 45 isolates, 11 were shown to be resistant to clarithromycin by E-test. With respect to the detection of
H. pylori
infection, PCR showed sensitivities of 100% in biopsies and 98% in stool specimens and a specificity of 98% in both biopsy and stool samples. All clarithromycin-sensitive cases were identified as such by PCR in both biopsy and stool samples. Of the cases with a resistant strain, eight were identified as such in stool DNA and nine were identified in biopsy DNA. Failure of PCR to detect the resistant genotype in the biopsy DNA, stool DNA, or both (one case) was associated with mixed populations. In these cases, patients had not been treated for
H. pylori
infection before, and the sensitive population showed to be present in considerably higher numbers than the resistant population. In five of six cases in which infection with a resistant genotype only was identified by PCR, the patients had received clarithromycin-based eradication therapy in the past. Thus, the assay presented provides a highly accurate noninvasive method to detect
H. pylori
infection in stool and at the same time allows for culture-independent clarithromycin susceptibility testing.
Genotyping Hepatitis C Virus by Heteroduplex Mobility Analysis Using Temperature Gradient Capillary Electrophoresis
Authors: Margraf Rebecca L. , Erali Maria , Liew Michael , Wittwer Carl T.
Published: October 2004

The genotype of the infecting hepatitis C virus (HCV) helps determine the patient's prognosis and the duration of treatment. Heteroduplex mobility analysis (HMA) is a rapid, inexpensive method for genotyping of HCV that does not require sequencing. We developed an HMA that uses temperature gradient capillary electrophoresis (TGCE) to differentiate HCV genotypes. A 56-bp region of the HCV 5′ untranslated region (UTR) that was conserved within a genotype yet whose sequence differed between genotypes was amplified for HMA-TGCE analysis. HCV amplicons of types 1, 2a, 2b, 3a, 4, and 6a were hybridized in pairs and analyzed by TGCE. Amplicons hybridized to the same subtype yielded one homoduplex peak, while hybridization of different subtypes resulted in heteroduplexes and generated multiple TGCE peaks. Heteroduplexes contain thermodynamically unstable nucleotide mismatches that reduced their TGCE mobilities compared to those of homoduplexes. Three HCV subtypes (subtypes 1a, 3a, and 4) generated unique peak patterns when they were combined with each genotype analyzed and were chosen as the reference genotypes. A blinded study with 200 HCV-infected samples was 97% accurate compared to genotyping by 5′ UTR sequence analysis. The majority of discordant results were unexpected sequence variants; however, five of nine sequence variants were correctly genotyped. The assay also detected and correctly genotyped mixed HCV infections. Compared to conventional HMA, TGCE improves the resolution, with better separation of heteroduplexes and homoduplexes. All common HCV genotypes can be detected and differentiated by this HMA-TGCE assay.
Seroincidence of Coccidioidomycosis during Military Desert Training Exercises
Authors: Crum Nancy F. , Potter Mark , Pappagianis Demosthenes
Published: October 2004

Coccidioidomycosis is a common fungal infection acquired in the southwestern United States. This is the first study in over 2 decades to determine the seroincidence of
Coccidioides immitis
infections among U.S. military members performing training exercises in an area of endemicity. Only 8% of participants were aware of coccidioidomycosis, despite the majority having visited or lived previously in an area of endemicity. One (0.6%) of the 178 participants developed “definite” serologic evidence of infection over a 5-week training period; four (2.3%) additional patients developed “possible” coccidioidomycosis infections. None had complicated disease. The calculated annual incidence ranged from 6 to 32%. This study suggests that the risk of serious coccidioidomycosis is low among military personnel during desert training exercises; however, disease incidence may vary depending on specific activities and geographic factors. Due to the potential morbidity and mortality of this infection, preventative strategies, including vaccine development, are advocated.
Molecular Epidemiological Analysis of
Mycoplasma bovis
Isolates from the United Kingdom Shows Two Genetically Distinct Clusters
Authors: McAuliffe Laura , Kokotovic Branko , Ayling Roger D. , Nicholas Robin A. J.
Published: October 2004

Mycoplasma bovis
is an important veterinary pathogen causing pneumonia, arthritis, and mastitis in infected cattle. We investigated the genetic diversity of 53 isolates collected in the United Kingdom between 1996 and 2002 with pulsed-field gel electrophoresis (PFGE), amplified fragment length polymorphism (AFLP), and random amplified polymorphic DNA (RAPD) analysis. In addition, the influence of variable surface protein (Vsp) profiles on the profiles generated with molecular typing techniques was studied. Both AFLP and RAPD separated the isolates into two distinct groups, but PFGE showed less congruence with the other techniques. There was no clear relationship between the geographic origin or year of isolation of the isolates and the profiles produced. No correlation between Vsp profiles and any of the molecular typing techniques was observed. We propose that RAPD and AFLP provide valuable tools for molecular typing of
M. bovis
.
Large-Scale Comparative Genomics Meta-Analysis of
Campylobacter jejuni
Isolates Reveals Low Level of Genome Plasticity
Authors: Taboada Eduardo N. , Acedillo Rey R. , Carrillo Catherine D. , Findlay Wendy A. , Medeiros Diane T. , Mykytczuk Oksana L. , Roberts Michael J. , Valencia C. Alexander , Farber Jeffrey M. , Nash John H. E.
Published: October 2004

We have used comparative genomic hybridization (CGH) on a full-genome
Campylobacter jejuni
microarray to examine genome-wide gene conservation patterns among 51 strains isolated from food and clinical sources. These data have been integrated with data from three previous
C. jejuni
CGH studies to perform a meta-analysis that included 97 strains from the four separate data sets. Although many genes were found to be divergent across multiple strains (
n
= 350), many genes (
n
= 249) were uniquely variable in single strains. Thus, the strains in each data set comprise strains with a unique genetic diversity not found in the strains in the other data sets. Despite the large increase in the collective number of variable
C. jejuni
genes (
n
= 599) found in the meta-analysis data set, nearly half of these (
n
= 276) mapped to previously defined variable loci, and it therefore appears that large regions of the
C. jejuni
genome are genetically stable. A detailed analysis of the microarray data revealed that divergent genes could be differentiated on the basis of the amplitudes of their differential microarray signals. Of 599 variable genes, 122 could be classified as highly divergent on the basis of CGH data. Nearly all highly divergent genes (117 of 122) had divergent neighbors and showed high levels of intraspecies variability. The approach outlined here has enabled us to distinguish global trends of gene conservation in
C. jejuni
and has enabled us to define this group of genes as a robust set of variable markers that can become the cornerstone of a new generation of genotyping methods that use genome-wide
C. jejuni
gene variability data.
Clinical Evaluation of the Sensititre YeastOne Colorimetric Antifungal Plate for Antifungal Susceptibility Testing of the New Triazoles Voriconazole, Posaconazole, and Ravuconazole
Authors: Pfaller M. A. , Espinel-Ingroff A. , Jones R. N.
Published: October 2004

A commercially prepared dried colorimetric microdilution panel (Sensititre YeastOne, TREK Diagnostic Systems, Cleveland, Ohio) was compared in three different laboratories with the National Committee for Clinical Laboratory Standards (NCCLS) reference microdilution method by testing two quality control strains and 300 clinical isolates of
Candida
spp. against fluconazole, voriconazole, posaconazole, and ravuconazole. Reference MIC endpoints were established after 48 h of incubation and YeastOne colorimetric endpoints were established after 24 h of incubation. YeastOne endpoints were determined to be the lowest concentration at which the color in the well changed from red (indicating growth) to purple (indicating growth inhibition) or blue (indicating no growth). Excellent agreement (within two dilutions) between the reference and colorimetric MICs was observed. Overall agreement was 95.4%. Agreement ranged from 92.3% with posaconazole to 98.0% with fluconazole. The YeastOne colorimetric method appears to be comparable to the NCCLS reference method for testing the susceptibility of
Candida
spp to the new triazoles voriconazole, posaconazole, and ravuconazole.
Performance Assessment of a Nested-PCR Assay (the RAPID BAP-MTB) and the BD ProbeTec ET System for Detection of
Mycobacterium tuberculosis
in Clinical Specimens
Authors: Wang Jann-Yuan , Lee Li-Na , Chou Chin-Sheng , Huang Chung-Yi , Wang Shu-Kuan , Lai Hsin-Chih , Hsueh Po-Ren , Luh Kwen-Tay
Published: October 2004

The performance of a nested PCR-based assay (the RAPID BAP-MTB; AsiaGen, Taichung, Taiwan) and the BD ProbeTec ET (DTB) system (Becton Dickinson, Sparks, Md.) for detection of
Mycobacterium tuberculosis
was evaluated with 600 consecutive clinical samples. These samples, including 552 respiratory specimens and 48 nonrespiratory specimens, were collected from 333 patients treated at National Taiwan University Hospital from September to October 2003. The results of both assays were compared to the gold standard of combined culture results and clinical diagnosis. The overall sensitivity and specificity of the RAPID BAP-MTB assay for respiratory specimens were 66.7% and 97.2%, respectively, and for the DTB assay they were 56.7% and 95.3%, respectively. The positive and negative predictive values for the RAPID BAP-MTB were 74.1% and 96.0%, respectively, and for the DTB assay they were 59.6% and 94.7%, respectively. For smear-negative samples, the sensitivity of the RAPID BAP-MTB and DTB assays was 57.1% and 40.5%, respectively. The RAPID BAP-MTB assay produced 14 false-positive results in 14 samples, including one of the six samples yielding
Mycobacterium abscessus
, one of the six samples yielding
Mycobacterium avium intracellulare
, one sample from a patient with a history of pulmonary tuberculosis with complete treatment, and three samples from three patients with a previous diagnosis of tuberculosis who were under treatment at the time of specimen collection. Among the 48 nonrespiratory specimens, the RAPID BAP-MTB assay was positive in one biopsy sample from a patient with lumbar tuberculous spondylitis and one pus sample from a patient with tuberculous cervical lymphadenopathy. Our results showed that the RAPID BAP-MTB assay is better than the DTB assay for both respiratory specimens and nonrespiratory specimens. The overall time for processing this assay is only 5 h. In addition, its diagnostic accuracy in smear-negative samples is as high as in smear-positive samples.
Relative Importance of Nasopharyngeal versus Oropharyngeal Sampling for Isolation of
Streptococcus pneumoniae
and
Haemophilus influenzae
from Healthy and Sick Individuals Varies with Age
Authors: Greenberg David , Broides Arnon , Blancovich Irena , Peled Nechama , Givon-Lavi Noga , Dagan Ron
Published: October 2004

Streptococcus pneumoniae
and
Haemophilus influenzae
carriage is a useful index for measuring the emergence of resistance and outcome in vaccination trials. We performed a study to determine which sampling site, nasopharynx (NP) or oropharynx (OP), yields the highest rate of
S. pneumoniae
and
H. influenzae
isolation at different ages. Both NP and OP cultures were obtained from 216 children aged <60 months and their mothers. The total
S. pneumoniae
carriage rate was 68% among children and 15% among mothers (
P
< 0.001). Using NP alone for the isolation of
S. pneumoniae
would have missed 2, 2, and 42% and using OP alone would have missed 77, 66, and 45% of
S. pneumoniae
in children aged 0 to 23 months, 24 to 59 months, and mothers, respectively. Using NP cultures alone for
H. influenzae
would have missed 23, 24, and 81% of the isolates, respectively. The respective figures for
H. influenzae
isolation from OP alone are 38, 29, and 9%. In children,
S. pneumoniae
was carried mainly in the NP while
H. influenzae
was equally carried in the NP and OP. In mothers,
S. pneumoniae
was carried equally in the NP and OP while
H. influenzae
was carried significantly more often in the OP. In children,
H. influenzae
colonization increased during illness, mainly in the NP. Culturing only one site significantly reduced the recovery of
H. influenzae
at all ages. NP cultures for
S. pneumoniae
detected close to 100% of isolates in children but only 58% of isolates in mothers.
Evidence of an Antigenic Shift among Palyam Serogroup Orbiviruses
Authors: Ohashi Seiichi , Matsumori Youichi , Yanase Tohru , Yamakawa Makoto , Kato Tomoko , Tsuda Tomoyuki
Published: October 2004

The Japanese isolates of Palyam serogroup viruses isolated from 1985 to 2001 were investigated for the genome sequence of segments 2 and 7 and were phylogenetically analyzed in comparison with Australian and African isolates of the same serogroup. The nucleotide sequences of segment 7 were highly conserved within Japanese isolates (95.1 to 100%) and between Japanese and Taiwanese isolates (96.0 to 100%), whereas the identities between Japanese and Taiwanese isolates and Australian and African isolates were fairly conserved (84.2 to 92.0%). Phylogenetic analysis based on segment 7 revealed three clusters according to geographical origin. As a result of the nucleotide sequence analysis of segment 2, which encodes a serotype-specific antigen, Japanese isolates were classified into two groups by genome length and nucleotide identities. Four of the nine Japanese isolates were categorized into the same group as prototype strain K-47 of the Chuzan virus, and the remaining isolates were categorized into the same group as the D'Aguilar virus and Nyabira virus. Phylogenetic analysis based on segment 2 revealed two clusters, the cluster containing Chuzan virus and the cluster containing the D'Aguilar and Nyabira viruses. To examine the antigenic relationship among viruses categorized in different clusters, we conducted a cross-neutralization test. KSB-29/E/01, isolated in 2001 in Japan, was neutralized by antiserum not only to strain B8112 of D'Aguilar virus but also to Chuzan virus. These results indicated that genetically and antigenically unique characteristics of KSB-29/E/01 were attributed to genetic reassortment of segment 2 between Chuzan virus and D'Aguilar virus.
Evaluation of the Immunoglobulin G Avidity Test for Diagnosis of Toxoplasmic Lymphadenopathy
Authors: Montoya Jose G. , Huffman Heather B. , Remington J. S.
Published: October 2004

Toxoplasmic lymphadenopathy (TL) is the most common clinical manifestation of acute acquired toxoplasma infection in normal individuals. The diagnosis is established by serologic methods and lymph node biopsy. Recently, tests for avidity of toxoplasma immunoglobulin G (IgG) antibodies have been introduced to help discriminate between recently acquired and distant infection with the parasite. We studied an avidity test to define the usefulness of this method and to determine the evolution of the IgG avidity in TL. Seventy-three consecutive patients diagnosed as having TL were studied. IgG avidity test titers were noted to be time dependent from the clinical onset of lymphadenopathy. Low IgG avidity test results were observed in patients who had developed lymphadenopathy from <1 month to 17 months prior to the sampling of sera, emphasizing that low IgG avidity test results are not reliable for diagnosis of recently acquired infection. In contrast, high IgG avidity test results were observed only in patients who had developed lymphadenopathy at least 4 months earlier. Thus, a high IgG avidity test result in an individual who has recent onset of lymphadenopathy (e.g., within 2 to 3 months of sera sampling) suggests a cause other than toxoplasmosis. In such cases, further workup is warranted in order to determine the cause of the lymphadenopathy.
High Prevalence of Genital Mycoplasmas among Sexually Active Young Adults with Urethritis or Cervicitis Symptoms in La Crosse, Wisconsin
Authors: Schlicht Michael J. , Lovrich Steven D. , Sartin Jeffrey S. , Karpinsky Patricia , Callister Steven M. , Agger William A.
Published: October 2004

Sexually active young adults in the small college town of La Crosse, Wisconsin, were evaluated for conventional sexually transmitted pathogens and tested for infections with mycoplasmas. The prevalence in 65 symptomatic men or women and 137 healthy volunteers (67 men and 70 women) was compared. Urine specimens from both cohorts were tested by ligase chain reaction for
Chlamydia trachomatis
or
Neisseria gonorrhoeae
. In addition, the urethral or cervical swabs from the symptomatic subjects were tested by PCR for
Mycoplasma genitalium
and cultured for
Mycoplasma hominis
and the ureaplasmas. The results confirmed a relatively low prevalence of gonorrhea among symptomatic men (12%) and chlamydia among symptomatic men (15%) and normal women (3%). In contrast, infections with mycoplasmas, especially the ureaplasmas (57%), were common and the organisms were the only potential sexually transmitted pathogen detected in 40 (62%) symptomatic subjects. Because of the high prevalence, we also evaluated urethral swabs from an additional 25 normal female volunteers and recovered ureaplasmas from 4 (16%) subjects. Additionally, the participants rarely used protection during sexual intercourse and some symptomatic subjects apparently acquired their infections despite using condoms regularly. The findings demonstrate a strong association between abnormal urogenital findings and detection of myoplasmas, particularly ureaplasmas, and suggest the infections will remain common.
Tracking Methicillin-Resistant
Staphylococcus aureus
Clones during a 5-Year Period (1998 to 2002) in a Spanish Hospital
Authors: Pérez-Roth Eduardo , Lorenzo-Díaz Fabián , Batista Ninivé , Moreno Antonio , Méndez-Álvarez Sebastián
Published: October 2004

Three hundred seventy-five consecutive methicillin-resistant
Staphylococcus aureus
(MRSA) clinical isolates recovered between 1998 and 2002 at the Nuestra Señora de Candelaria University Hospital in Tenerife, Spain, were analyzed by molecular fingerprinting techniques to determine MRSA clonal types and their prevalence over time. After determining antibiotic susceptibility, we used SmaI-digested genomic DNA separated by pulsed-field gel electrophoresis (PFGE) to characterize MRSA isolates and to establish PFGE types. Additionally, several selected isolates representative of each major PFGE type were tested by multilocus sequence typing (MLST) and by a multiplex PCR method capable of identifying the structural type of the staphylococcal cassette chromosome
mec
(SCC
mec
), generating the corresponding sequence type (ST)-SCC
mec
types. Results of PFGE, supported by those of MLST and SCC
mec
typing, allowed us to identify six MRSA clones within the five major PFGE types and document temporal shifts in the prevalence of these MRSA clones from 1998 to 2002. Four of the clones were the pandemic “Iberian” (designated ST247-MRSA-IA), EMRSA-15 (ST22-MRSA-IV), EMRSA-16 (ST36-MRSA-II), and the so-called pediatric (ST5-MRSA-IV) clones, while the other two (ST125-MRSA-IVA and ST146-MRSA-IVA) clones could be derived from the pediatric one. The most striking temporal shift in the dominance of MRSA clones was the replacement of the multidrug-resistant and highly epidemic Iberian clone by the so-called British EMRSA-16 clone during the 5-year surveillance period. Our results are in accordance with previously stated findings showing the worldwide hospital dominance of relatively few pandemic and presumably virulent MRSA clones. We report for the first time the detection in Spain of the British EMRSA-15 and pediatric clones, as well as the abrupt replacement of the Iberian by the EMRSA-16 as the major MRSA clone.