bluuebunny/crossref_metadata_2025 huggingface dataset:
Bedside assessment of fibrinogen level in postpartum haemorrhage by thrombelastometry
Authors: Huissoud C , Carrabin N , Audibert F , Levrat A , Massignon D , Berland M , Rudigoz R‐C
Published: May 2009

Objectives  To establish whether reagent‐supported thrombelastometry with the rotation thrombelastometry system (ROTEM®) point‐of‐care device correlated with fibrinogen level in postpartum haemorrhages.

Design  Prospective observational study.

Population and setting  Ninety‐one women at the third trimester of pregnancy: 37 with postpartum haemorrhage (study group) and 54 without abnormal bleeding (control group).

Methods  Standard laboratory test results were compared with those obtained at bedside from the ROTEM with the FIBTEM® test (54 tests in the control group and 51 in the study group).

Main outcome measures  Analysis of correlations between fibrinogen levels and FIBTEM test results: clotting time (CT), clot amplitude at 5 and 15 minutes (CA5; CA15) and maximal amplitude [maximum clot firmness (MCF)].

Results  Median fibrinogen level was significantly lower in the haemorrhage group than in the control group (3.4 and 5.1 g/l, respectively, P < 0.0001). Median CT was higher in the haemorrhage group than in the control group (P = 0.05). CA5, CA15 and MCF were significantly lower in the haemorrhage group than in controls (P < 0.0001) and strongly correlated with fibrinogen levels in both groups (r = 0.84–0.87, P < 0.0001). A cut‐off value of CA5 at 5 mm and CA15 at 6 mm presented an excellent sensitivity (100% for both parameters) and a good specificity (respectively 85 and 88%) to detect fibrinogen levels <1.5 g/l in postpartum haemorrhage.

Conclusions  The early parameters obtained from the FIBTEM test correlated well with fibrinogen levels. ROTEM might be helpful in guiding fibrinogen transfusion during postpartum haemorrhage.
Priorities in emergency obstetric care in Bolivia––maternal mortality and near‐miss morbidity in metropolitan La Paz
Authors: Rööst M , Altamirano VC , Liljestrand J , Essén B
Published: May 2009

Objective  To document the frequency and causes of maternal mortality and severe (near‐miss) morbidity in metropolitan La Paz, Bolivia.

Design  Facility‐based cross‐sectional study.

Setting  Four maternity hospitals in La Paz and El Alto, Bolivia, where free maternal health care is provided through a government‐subsidised programme.

Population  All maternal deaths and women with near‐miss morbidity.

Methods  Inclusion of near‐miss using clinical and management‐based criteria.

Main outcome measures  Maternal mortality ratio (MMR), severe morbidity ratio (SMR), mortality indices and proportion of near‐miss cases at hospital admission.

Results  MMR was 187/100 000 live births and SMR was 50/1000 live births, with a relatively low mortality index of 3.6%. Severe haemorrhage and severe hypertensive disorders were the main causes of near‐miss, with 26% of severe haemorrhages occurring in early pregnancy. Sepsis was the most common cause of death. The majority of near‐miss cases (74%) were in critical condition at hospital admission and differed from those fulfilling the criteria after admission as to diagnostic categories and socio‐demographic variables.

Conclusions  Pre‐hospital barriers remain to be of great importance in a setting of this type, where there is wide availability of free maternal health care. Such barriers, together with haemorrhage in early pregnancy, pre‐eclampsia detection and referral patterns, should be priority areas for future research and interventions to improve maternal health. Near‐miss upon arrival and near‐miss after arrival at hospital should be analysed separately as that provides additional information about factors that contribute to maternal ill‐health.
Effects of female genital mutilation on birth outcomes in Switzerland
Authors: Wuest S , Raio L , Wyssmueller D , Mueller MD , Stadlmayr W , Surbek DV , Kuhn A
Published: May 2009

Objective  The primary aim of this study was to determine the desires and wishes of pregnant patients vis‐à‐vis their external genital anatomy after female genital mutilation (FGM) in the context of antenatal care and delivery in a teaching hospital setting in Switzerland.

Our secondary aim was to determine whether women with FGM and non‐mutilated women have different fetal and maternal outcomes.

Design  A retrospective case–control study.

Setting  A teaching hospital.

Population  One hundred and twenty‐two patients after FGM who gave consent to participate in this study and who delivered in the Department of Obstetrics and Gynaecology in the University Hospital of Berne and 110 controls.

Methods  Data for patients’ wishes concerning their FGM management, their satisfaction with the postpartum outcome and intrapartum and postpartum maternal and fetal data. As a control group, we used a group of pregnant women without FGM who delivered at the same time and who were matched for maternal age.

Main outcome measures  Patients’ satisfaction after delivery and defibulation after FGM, maternal and fetal delivery data and postpartum outcome measures.

Results  Six percent of patients wished to have their FGM defibulated antenatally, 43% requested a defibulation during labour, 34% desired a defibulation during labour only if considered necessary by the medical staff and 17% were unable to express their expectations. There were no differences for FGM patients and controls regarding fetal outcome, maternal blood loss or duration of delivery. FGM patients had significantly more often an emergency Caesarean section and third‐degree vaginal tears, and significantly less first‐degree and second‐degree tears.

Conclusion  An interdisciplinary approach may support optimal antenatal and intrapartum management and also the prevention of FGM in newborn daughters.
Intensity of Nordic Walking in young females with different peak O2 consumption
Authors: Jürimäe Toivo , Meema Krislin , Karelson Kalle , Purge Priit , Jürimäe Jaak
Published: May 2009

The purpose of this cross‐sectional study was to determine the physiological reaction to the different intensity Nordic Walking exercise in young females with different aerobic capacity values. Twenty‐eight 19–24‐year‐old female university students participated in the study. Their peak O2 consumption (VO2 peak kg−1) and individual ventilatory threshold (IVT) were measured using a continuous incremental protocol until volitional exhaustion on treadmill. The subjects were analysed as a whole group (n = 28) and were also divided into three groups based on the measured VO2 peak kg−1 (Difference between groups is 1 SD) as follows: 1. >46 ml min−1 kg−1 (n = 8), 2. 41–46 ml min−1 kg−1 (n = 12) and 3. <41 ml min−1 kg−1 (n = 8). The second test consisted of four times 1 km Nordic Walking with increasing speed on the 200 m indoor track, performed as a continuous study (Step 1 – slow walking, Step 2 – usual speed walking, Step 3 – faster speed walking and Step 4 – maximal speed walking). During the walking test expired gas was sampled breath‐by‐breath and heart rate (HR) was recorded continuously. Ratings of perceived exertion (RPE) were asked using the Borg RPE scale separately for every 1 km of the walking test. No significant differences emerged between groups in HR of IVT (172·4 ± 10·3–176·4 ± 4·9 beats min−1) or maximal HR (190·1 ± 7·3–191·6 ± 7·8 beats min−1) during the treadmill test. During maximal speed walking the speed (7·4 ± 0·4–7·5 ± 0·6 km h−1) and O2 consumption (30·4 ± 3·9–34·0 ± 4·5 ml min−1 kg−1) were relatively similar between groups (P > 0·05). However, during maximal speed walking, the O2 consumption in the second and third groups was similar with the IVT (94·9 ± 17·5% and 99·4 ± 15·5%, respectively) but in the first group it was only 75·5 ± 8·0% from IVT. Mean HR during the maximal speed walking was in the first group 151·6 ± 12·5 beats min−1, in the second (169·7 ± 10·3 beats min−1) and the third (173·1 ± 15·8 beats min−1) groups it was comparable with the calculated IVT level. The Borg RPE was very low in every group (11·9 ± 2·0–14·4 ± 2·3) and the relationship with VO2and HR was not significant during maximal speed Nordic Walking. In summary, the present study indicated that walking is an acceptable exercise for young females independent of their initial VO2 peak level. However, females with low initial VO2 peak can be recommended to exercise with the subjective ‘faster speed walking’. In contrast, females with high initial VO2 peak should exercise with maximal speed.
Geographic variation in sterilizing parasite species and the Red Queen
Authors: King Kayla C. , Lively Curtis M.
Published: May 2009

The Red Queen hypothesis predicts that sexual reproduction should be favoured in locations where the risk of infection by virulent parasites is consistently high. When hosts are exposed to multiple parasites over their geographic range, the coevolving parasite species may vary among host populations. We surveyed 26 streams on the South Island of New Zealand to determine whether the frequency of snails (Potamopyrgus antipodarum) infected by various sterilizing trematode parasite species was correlated with the frequency of sexual individuals. We compared the results with a survey conducted over 20 years ago to determine whether the associations were consistent. We also evaluated different measures of parasite‐mediated selection among populations, including prevalence of the most common local parasite (MCLP) species and parasite diversity to assess the best predictor of sexual reproduction among stream populations. The results showed that the relationship between male frequency and parasite infection is more geographically widespread than previously recorded. Additionally, we found that the prevalence of the MCLP was the best predictor of sex in habitats where hosts populations are infected with multiple parasites (approximately 15 trematode species). This study provides evidence that sexual snails occur more often in environments with high infection levels, and that the pattern of parasite‐imposed selection is geographically variable. Support for the Red Queen may be strengthened by focussing on the MCLP, which may vary among host populations.
Intron 4 polymorphism of the endothelial nitric oxide synthase eNOS gene and early microangiopathy in type 1 diabetes
Authors: Mamoulakis D. , Bitsori M. , Galanakis E. , Vazgiourakis V. , Panierakis C. , Goulielmos G. N.
Published: May 2009

Nitric oxide (NO) is an endogenous vasodilator involved in inflammatory and autoimmune response, and in the pathophysiology of diabetic vascular disease. Endothelium‐derived NO is formed from l‐arginine by endothelial NO synthase (eNOS), and earlier studies have provided evidence for altered NO metabolism and impaired endothelial function in diabetes, probably due to polymorphisms in eNOS gene. In the present study we investigated the association of the eNOS gene intron 4 a/b VNTR polymorphism with diabetic microangiopathy in 61 young individuals with type 1 diabetes (T1D), 35 male and 26 female, aged 5.0–29.1 (mean 15.6) years, and followed up for 3.24–11.4 (mean 7.44) years. Ten patients (16.4%) had developed microalbuminuria, three hypertension and two retinopathy. Wild‐type b/b homozygosity for eNOS gene intron 4 VNTR was found in 37 (60.7%) and a/b polymorphism in 24 (39.3%). No significant relationship was demonstrated between eNOS gene intron 4 polymorphisms and microalbuminuria, hypertension or retinopathy in these young individuals. Our findings suggest that a/b polymorphism of the intron 4 eNOS gene is not associated with early onset diabetic microangiopathy.
Comparative analysis of the impact of a free cysteine in tapasin on the maturation and surface expression of murine MHC class I allotypes
Authors: Wang X. , Simone L. C. , Tuli A. , Solheim J. C.
Published: May 2009

Tapasin is a key molecule in the major histocompatibility complex (MHC) class I peptide‐loading complex, interacting with several other proteins in the complex. An amino acid substitution at a free cysteine position in tapasin has been shown to disrupt the covalent association of tapasin with ERp57. In this study, we mutated the free cysteine in mouse tapasin, and analysed the effects on the cell surface expression of the mouse MHC class I molecules Kd and Kb. The C95S substitution in mouse tapasin increased the proportion of open forms relative to folded forms for both types of MHC class I molecules at the cell surface. Furthermore, the C95S substitution resulted in increased association of tapasin with folded Kd. Overall, our studies with these mouse MHC class I allotypes have revealed that the free cysteine 95 in mouse tapasin influences stable expression at the plasma membrane for both MHC class I allotypes, and have shown that tapasin's interaction with folded Kd is elevated by the C95S substitution in tapasin.
HLA class I and class II polymorphisms in Saudi patients with myasthenia gravis
Authors: Hajeer A. H. , Sawidan F. Al , Bohlega S. , Saleh S. , Sutton P. , Shubaili A. , Tahan A. Al , Al Jumah M.
Published: May 2009

Myasthenia gravis (MG) is a rare autoimmune disease of the neuromuscular junction. MG has been shown to be associated with many HLA antigens in different populations. Here we have analysed the frequency of HLA‐A, B, DR and DQ in a group of Saudi MG patients and compared their results to a group of healthy controls. MG in Saudi patients is found to be associated with HLA‐A*23, B*08, B*18, DRB1*16 and DRB1*13. The strongest association was with HLA‐B*08, which was associated with young age at onset and female gender. Our results are in line with other published results from around the world and warrant fine mapping of the area using microsatellite to map the disease gene.
The CTLA4 +49 A/G polymorphism is not associated with susceptibility to type 1 diabetes mellitus in the Portuguese population
Authors: Lemos M. C. , Coutinho E. , Gomes L. , Bastos M. , Fagulha A. , Barros L. , Carrilho F. , Geraldes E. , Regateiro F. J. , Carvalheiro M.
Published: May 2009

CTLA4 genetic polymorphisms have been associated with type 1 diabetes. We genotyped 207 patients and 249 controls for the most frequently investigated polymorphism of the CTLA4 gene (+49A/G (rs231775)). No significant differences were observed, suggesting that this polymorphism is not strongly associated with type 1 diabetes in the Portuguese population.
Full‐length sequence and expression analysis of a myeloid differentiation factor 88 (MyD88) in half‐smooth tongue sole Cynoglossus semilaevis
Authors: Yu Y. , Zhong Q. W. , Zhang Q. Q. , Wang Z. G. , Li C. M. , Yan F. S. , Jiang L. M.
Published: May 2009

Myeloid differentiation factor 88 (MyD88) is a universal and crucial adaptor protein, which plays an essential role in the intracellular signalling elicited by IL‐1R/TLR superfamily. In the present study, we report the full‐length sequence of MyD88 gene in half‐smooth tongue sole (Cynoglossus semilaevis). In the 2855 bp genomic sequence, five exons and four introns were identified. The cloned cDNA exhibited 110 bp of 5′ UTR, 576 bp of 3′ UTR and 858 bp of the entire open‐reading frame encoding a polypeptide of 285 amino acids. The protein sequence included a typical conserved cytosolic Toll/interleukin‐1 receptor (TIR) domain, an intermediate domain (ID) and a death domain (DD), and shared greater than 70% identity with Japanese flounder Paralichthys olivaceu ortholog. Real‐time polymerase chain reaction (RT‐PCR) analysis indicated a broad expression of csMyD88, especially in ovary and spleen. Quantitative RT‐PCR analysis indicated that the csMyD88 mRNA levels were significantly increased in the spleen and head kidney after inactive Vibrio anguillarum challenge and the expression of csMyD88 appeared to be developmentally regulated during C. semilaevis ontogeny. Although, species‐specific differences were present, the similarity between mammalian and piscine MyD88s suggested that the main function of MyD88 might be conserved across vertebrates.
Effects of disturbance by Siberian marmots (Marmota sibirica) on spatial heterogeneity of vegetation at multiple spatial scales
Authors: Yoshihara Yu , Ohkuro Toshiya , Bayarbaatar Buuveibaatar , Takeuchi Kazuhiko
Published: May 2009

An understanding of the relationship between vegetation spatial heterogeneity and disturbance and its application to the management are important for maintaining biodiversity and functions of ecosystems. We examined the effects of disturbance by Siberian marmots on the spatial heterogeneity of vegetation at three spatial scales (fine, intermediate and coarse) in a Mongolian grassland. We established a 50 m × 50 m plot around five marmot mounds and another plot in an area with no mounds. Each plot was subdivided into 625 adjacent 4‐m2 square quadrats and the plants in them were surveyed. Spatial heterogeneity was calculated as the mean dissimilarity in species composition among sample quadrats in each plot. The off‐colony plant community was dominated by graminoids, but a variety of plants, including graminoids, forbs and shrubs, were present in the on‐colony plot. The slightly greater fine‐scale heterogeneity in the on‐colony plot compared to the off‐colony plot reflected the presence of either disturbed or undisturbed patch within a single measurement unit, whereas the lower coarse‐scale heterogeneity in the on‐colony plot reflected the presence of both disturbed and undisturbed patches within a single measurement unit. Our results demonstrate the possibility of using the marmot as a conservation tool in our study site as far as a plant biodiversity is targeted.
Effects of land‐use change on chemical composition of soil organic matter in tropical lowland Bolivia
Authors: Abe Susumu S. , Mueller Carsten W. , Steffens Markus , Koelbl Angelika , Knicker Heike , Koegel‐Knabner Ingrid
Published: May 2009

Land‐use change affects not only the amount of soil organic matter (SOM) but also its composition. We performed cross‐polarization magic angle spinning (CPMAS)13C and15N nuclear magnetic resonance (NMR) spectroscopy to investigate the chemical composition of bulk SOM in topsoils (0–15 cm) under different land use, namely native forest (NF), 27‐year cropland with wheat/soybean rotation (CL) and 27‐year rangelands with guineagrass (RG) and with bahiagrass (RB), in south‐east Bolivia. The findings of this study showed only a subtle alteration of composition of bulk SOM despite the large changes in carbon (C) content. Nevertheless, NF and RB showed a slightly lower abundance of aromatic C but a higher proportion of alkyl C compared to CL and RG where the loss of organic matter was substantial. This suggests that relatively stable components dominated by aromatic structures had relatively enriched during SOM decomposition under agricultural practices. A slight disparity of SOM composition observed between RG and RB (less O‐alkyl C but more aromatic C in RG than RB) suggests that grass species influenced SOM quality even under the same land use, namely the rangeland. On the other hand, organic N composition was less affected by land use or management practice than C forms.
Drugs Used in the Treatment of Rheumatoid Arthritis: Relationship between Current Use and Cardiovascular Risk Factors
Authors: Rho Young Hee , Oeser Annette , Chung Cecilia P. , Milne Ginger L. , Stein C. Michael
Published: May 2009

Objectives.  Drugs used for the treatment of rheumatoid arthritis (RA) have the potential to affect cardiovascular risk factors. There is concern that corticosteroids, non‐steroidal anti‐inflammatory drugs (NSAIDs) and COX‐2 inhibitors could affect cardiovascular risk adversely, while drugs such as the antimalarial, hydroxychloroquine, may have beneficial effects. However, there is limited information about cardiovascular risk factors in patients with RA receiving different drugs.

Methods.  We measured cardiovascular risk factors including systolic and diastolic blood pressure, serum HDL and LDL cholesterol, glucose and homocysteine concentrations and urinary F2‐isoprostane excretion in 169 patients with RA. Risk factors were compared according to current use of corticosteroids, methotrexate, antimalarials, NSAIDs, COX‐2 inhibitors, leflunomide and TNF‐α blockers. Comparisons were adjusted for age, sex, race, disease activity (DAS28 score), current hypertension, diabetes, smoking status and statin use.

Results.  No cardiovascular risk factor differed significantly among current users and non‐users of NSAIDs, COX‐2 inhibitors, methotrexate and TNF‐α blockers. Serum HDL cholesterol concentrations were significantly higher in patients currently receiving corticosteroids (42.2 ± 10.5 vs. 50.2 ± 15.3 mg/dL, adjusted P < 0.001). Diastolic blood pressure (75.9 ± 11.2 vs. 72.0 ± 9.1 mm Hg, adjusted P = 0.02), serum LDL cholesterol (115.6 ± 34.7 vs. 103.7 ± 27.8 mg/dL, adjusted P = 0.03) and triglyceride concentrations (157.7 ± 202.6 vs. 105.5 ± 50.5 mg/dL, adjusted P = 0.03) were significantly lower in patients taking antimalarial drugs. Plasma glucose was significantly lower in current lefunomide users (93.0 ± 19.2 vs. 83.6 ± 13.4 mg/dL, adjusted P = 0.006).

Conclusions.  In a cross‐sectional setting drugs used to treat RA did not have major adverse effects on cardiovascular risk factors and use of antimalarials was associated with beneficial lipid profiles.
Scaffolding Collaborative Exchanges Between Expert and Novice Language Teachers in Threaded Discussions
Authors: Lee Lina
Published: May 2009

Abstract: This article presents a two‐semester study that examines the effectiveness of threaded discussions for foreign language teaching methods courses. In particular, this study focuses on how students view the role of asynchronous computer‐mediated communication in the development of language pedagogy through an exchange with expert teachers from secondary schools. The author gathered data from 32 students who participated in weekly discussion boards, wrote monthly reflective logs, filled out surveys, and joined group interviews. The findings demonstrated that online discussions promoted scaffolding by which expert teachers assisted students in creating reflective messages. In addition, threaded discussions fostered learners' autonomy and accountability while supporting collaborative learning through social interaction. The study suggests three essential ingredients to maximize the potential benefits of discussion board for language teacher training: (1) use of carefully designed tasks that engage critical thinking, (2) scaffolding strategies for monitoring group discussions, and (3) inclusion of online etiquette to avoid confusion and reduce personal conflicts.
Heritage Language Learners in First‐Year Foreign Language Courses: A Report of General Data Across Learner Subtypes
Authors: Reynolds Rachel R. , Howard Kathryn M. , Deák Julia
Published: May 2009

Abstract: This article reports on a survey of language study motivations for first‐year language learners at two large private East Coast American universities. Surveys were completed by 401 students in courses for 19 different foreign languages. Forty percent of respondents were designated as heritage language learners (HLLs), and they were divided into two subclasses: narrowly defined HLLs, who indicated that the target language was regularly spoken in their home and who constituted 34% of all HLLs (55/162); and broadly defined HLLs, who indicated familial or heritage connections to the language without regular prior exposure to the target language and who constituted 66% (107/162) of all HLLs. This article reports students' prior exposure to the language, reasons for studying it, and goals and attitudes among these subclasses, and outlines avenues for further investigation.
The Language Learning Motivation of Early Adolescent French Immersion Graduates
Authors: Wesely Pamela M.
Published: May 2009

Abstract: This interpretive multiple case study examines the motivation to learn a second language among sixth grade students who attended a French immersion school for grades K–5. Parent surveys, student surveys based on Gardner's Attitude/Motivation Test Battery, and individual and group interviews with students were the data sources used to identify students' second language (L2) learning motivating factors. Survey data indicated that the students shared home background characteristics and had high motivation and similar motivational orientations. Cross‐case theme analyses of interviews suggested that, although students appeared to be motivated by both instrumental and integrative factors, relationships with teachers and peers were also influential. Consideration of individual cases indicated that other theories in motivation and second language acquisition could shed more light on L2 learning motivation.
An International Comparison of Socially Constructed Language Learning Motivation and Beliefs
Authors: Kouritzin Sandra G. , Piquemal Nathalie A. , Renaud Robert D.
Published: May 2009

Abstract: In our global economy, it is important to understand all factors influencing successful language learning. A survey of more than 6,000 university students in Canada, Japan, and France revealed differences in language learning beliefs, attitudes, and motivations in the three countries. Learners in Canada and France exhibited primarily instrumental and integrative motivation, respectively, whereas learners from Japan displayed a different form, social capital motivation, in which knowledge of a foreign language carries value in and of itself. Knowledge of these different forms of motivation has pedagogical and political implications for language teachers.
Reassessment of the Potential Sea-Level Rise from a Collapse of the West Antarctic Ice Sheet
Authors: Bamber Jonathan L. , Riva Riccardo E. M. , Vermeersen Bert L. A. , LeBrocq Anne M.
Published: May 2009

Collapse and Rise

The West Antarctic Ice Sheet (WAIS) is thought to be inherently unstable and susceptible to rapid collapse if it reaches a certain warming threshold. Although such an event is considered unlikely, to predict the consequences of collapse it is important to know how much sea level would rise in such a case. The WAIS is thought to contain enough ice to raise sea level by 5 to 7 meters were it to collapse.

Bamber
et al.

(p.
901
, see the cover; see the Perspective by

Ivins

) have reassessed that number, on the basis of better data on the geometry of the WAIS, and conclude that its sudden collapse would raise sea level by about 3.2 meters, on average, with large and important regional variations. Although this is only about half as much as previously thought, its impact on coastal areas would still be devastating.
Photovoltaics Power Up
Authors: Swanson Richard M.
Published: May 2009

The power-generating capacity of solar cells, while currently small relative to other sources, is increasing exponentially.
Seasons and Life Cycles
Authors: Steltzer Heidi , Post Eric
Published: May 2009

A conceptual framework explains how individual species' responses to climate warming affect the length of the growing season.
Ice Sheet Stability and Sea Level
Authors: Ivins Erik R.
Published: May 2009

How much will sea levels rise if the West Antarctic Ice Sheet becomes unstable?
Far from a Rational Crowd
Animal Spirits
How Human Psychology Drives the Economy, and Why It Matters for Global Capitalism

by George A. Akerlof and Robert J. Shiller

Princeton University Press, Princeton, NJ, 2009. 246 pp. $24.95, £14.95. ISBN 9780691142333.
Authors: Baddeley Michelle
Published: May 2009

Arguing for a "behaviorally informed Keynesianism," Akerlof and Shiller discuss a variety of macroeconomic topics including business cycles, inflation, unemployment, financial and real estate booms and busts, poverty, and monetary policy.
Austria's Possible CERN Withdrawal Rattles Physicists
Authors: Clery Daniel
Published: May 2009

Austrian science minister Johannes Hahn announced on 7 May that he intends to withdraw the country from CERN membership by the end of 2010 because his ministry thinks its €16 million annual contribution would be better spent on smaller research projects.
Cytometric Approach for Detection ofEncephalitozoon intestinalis, an Emergent Agent
Authors: Barbosa Joana , Rodrigues Acácio Gonçalves , Pina-Vaz Cidália
Published: May 2009

Encephalitozoon intestinalisis responsible for intestinal disease in patients with AIDS and immunocompetent patients. The infectious form is a small spore that is resistant to water treatment procedures. Its detection is very important, but detection is very cumbersome and time-consuming. Our main objective was to develop and optimize a specific flow cytometric (FC) protocol for the detection ofE. intestinalisin hospital tap water and human feces. To determine the optimal specific antibody (Microspor-FA) concentration, a known concentration ofE. intestinalisspores (Waterborne, Inc.) was suspended in hospital tap water and stool specimens with different concentrations of Microspor-FA, and the tap water and stool specimens were incubated under different conditions. The sensitivity limit and specificity were also evaluated. To study spore infectivity, double staining with propidium iodide (PI) and Microspor-FA was undertaken. Distinct approaches for filtration and centrifugation of the stool specimens were used.E. intestinalisspores stained with 10 μg/ml of Microspor-FA at 25°C overnight provided the best results. The detection limit was 5 × 104spores/ml, and good specificity was demonstrated. Simultaneous staining with Microspor-FA and PI ensured that theE. intestinalisspores were dead and therefore noninfectious. With the stool specimens, better spore recovery was observed with a saturated solution of NaCl and centrifugation at 1,500 ×gfor 15 min. A new approach for the detection ofE. intestinalisfrom tap water or human feces that ensures that the spores are not viable is now available and represents an important step for the prevention of this threat to public health.
Contributions of Immunoblotting, Real-Time PCR, and the Goldmann-Witmer Coefficient to Diagnosis of Atypical Toxoplasmic Retinochoroiditis
Authors: Talabani H. , Asseraf M. , Yera H. , Delair E. , Ancelle T. , Thulliez P. , Brézin A. P. , Dupouy-Camet J.
Published: May 2009

Ocular toxoplasmosis is a major cause of posterior uveitis worldwide. The diagnosis is based mainly on ophthalmological examination. Biological diagnosis is necessary in atypical cases, and this requires aqueous humor sampling by anterior chamber paracentesis. We evaluated real-time PCR targeting the
Toxoplasma gondii
529-bp repeat element, the Goldmann-Witmer coefficient (GWC), and immunoblotting for the diagnosis of toxoplasmic retinochoroiditis in 54 patients with atypical uveitis. The results of these biological tests, applied to paired aqueous humor-serum samples, were compared to the clinical findings. Combining either PCR or the GWC with immunoblotting increased the sensitivity to 73% or 70%, respectively. Together, PCR and the GWC had 80% sensitivity. If feasible, sensitivity can be increased by combining the three methods (85% sensitivity). The interval between symptom onset and anterior chamber paracentesis strongly influenced the detection of specific intraocular antibody synthesis. The sensitivity of the GWC increased from 45% to 56% when sampling was performed 10 days after symptom onset, and that of immunoblotting increased from 53% to 72% when puncture was performed 30 days after symptom onset. PCR analysis of aqueous humor samples detected toxoplasmic DNA in 55% of patients. In contrast to the results of immunoblotting and the GWC, the results of PCR were not influenced by the interval between symptom onset and paracentesis. PCR was more informative than the GWC and immunoblotting for immunocompromised patients. Acute necrotizing retinal lesions were significantly larger in PCR-positive patients, with a mean of 3.5 optic disc diameters, than in PCR-negative patients, with a mean of 1.5 optic disc diameters.
Diversity of Group B Streptococcus Serotypes Causing Urinary Tract Infection in Adults
Authors: Ulett Kimberly B. , Benjamin William H. , Zhuo Fenglin , Xiao Meng , Kong Fanrong , Gilbert Gwendolyn L. , Schembri Mark A. , Ulett Glen C.
Published: May 2009

Serotypes of group B streptococcus (GBS) that cause urinary tract infection (UTI) are poorly characterized. We conducted a prospective study of GBS UTI in adults to define the clinical and microbiological characteristics of these infections, including which serotypes cause disease. Patients who had GBS cultured from urine over a 1-year period were grouped according to symptoms, bacteriuria, and urinalysis. Demographic data were obtained by reviewing medical records. Isolates were serotyped by latex agglutination and multiplex PCR-reverse line blotting (mPCR/RLB). Antibiotic susceptibilities were determined by disc diffusion. GBS was cultured from 387/34,367 consecutive urine samples (1.1%): 62 patients had bacteriuria of >10
7
CFU/liter and at least one UTI symptom; of these patients, 31 had urinary leukocyte esterase and pyuria (others not tested), 50 (81%) had symptoms consistent with cystitis, and 12 (19%) had symptoms of pyelonephritis. Compared with controls (who had GBS isolated without symptoms), a prior history of UTI was an independent risk factor for disease. Increased age was also significantly associated with acute infection. Serotyping results were consistent between latex agglutination and mPCR/RLB for 331/387 (85.5%) isolates; 22 (5.7%) and 7 (1.8%) isolates were nontypeable with antisera and by mPCR/RLB, respectively; and 45/56 (80.4%) isolates with discrepant results were typed by mPCR/RLB as belonging to serotype V. Serotypes V, Ia, and III caused the most UTIs; serotypes II, Ib, and IV were less common. Nontypeable GBS was not associated with UTI. Erythromycin (39.5%) and clindamycin (26.4%) resistance was common. We conclude that a more diverse spectrum of GBS serotypes causes UTI than previously recognized, with the exception of nontypeable GBS.
Meningitis Due to a “
Bartonella washoensis
”-Like Human Pathogen
Authors: Probert Will , Louie Janice K. , Tucker James R. , Longoria Rose , Hogue Robin , Moler Silvia , Graves Margot , Palmer Heather J. , Cassady Joseph , Fritz Curtis L.
Published: May 2009

We report the second human case of infection caused by an organism identified as the proposed
Bartonella
species, “
B
.
washoensis
.” The organism was isolated from a blood sample from a patient presenting with meningitis and early sepsis.
Oropsylla montana
fleas were implicated as the vector for disease transmission in this case.
Molecular Determination of
Mycobacterium leprae
Viability by Use of Real-Time PCR
Authors: Martinez Alejandra N. , Lahiri Ramanuj , Pittman Tana L. , Scollard David , Truman Richard , Moraes Milton O. , Williams Diana L.
Published: May 2009

Mycobacterium leprae
, the etiological agent of leprosy, is noncultivable on axenic media. Therefore, the viability of
M. leprae
for clinical or experimental applications is often unknown. To provide new tools for
M. leprae
viability determination, two quantitative reverse transcriptase PCR (RT-PCR) assays were developed and characterized.
M. leprae sodA
mRNA and 16S rRNA were used as RNA targets, and
M. leprae
repetitive element (RLEP) DNA was used to determine relative bacterial numbers in the same purified bacterial preparations or from crude biological specimens. Results demonstrated that both assays were good predictors of
M. leprae
viability during short-term experiments (48 h) involving rifampin (rifampicin) treatment in axenic medium, within rifampin-treated murine macrophages (MΦ), or within immune-activated MΦ. Moreover, these results strongly correlated those of other
M. leprae
viability assays, including radiorespirometry-based and Live/Dead BacLight viability assays. The 16S rRNA/RLEP assay consistently identified the presence of
M. leprae
in eight multibacillary leprosy patient biopsy specimens prior to multidrug therapy (MDT) and demonstrated a decline in viability during the course of MDT. In contrast, the
sodA
/RLEP assay was able to detect the presence of
M. leprae
in only 25% of pretreatment biopsy specimens. In conclusion, new tools for
M. leprae
viability determination were developed. The 16S rRNA/RLEP RT-PCR
M. leprae
viability assay should be useful both for short-term experimental purposes and for predicting
M. leprae
viability in biopsy specimens to monitor treatment efficacy, whereas the
sodA
/RLEP RT-PCR
M. leprae
viability assay should be limited to short-term experimental research purposes.
Single Nucleotide Polymorphisms in the IS
900
Sequence of
Mycobacterium avium
subsp.
paratuberculosis
Are Strain Type Specific
Authors: Castellanos Elena , Aranaz Alicia , de Juan Lucia , Álvarez Julio , Rodríguez Sabrina , Romero Beatriz , Bezos Javier , Stevenson Karen , Mateos Ana , Domínguez Lucas
Published: May 2009

Insertion sequence IS
900
is used as a target for the identification of
Mycobacterium avium
subsp.
paratuberculosis.
Previous reports have revealed single nucleotide polymorphisms within IS
900
. This study, which analyzed the IS
900
sequences of a panel of isolates representing
M. avium
subsp.
paratuberculosis
strain types I, II, and III, revealed conserved type-specific polymorphisms that could be utilized as a tool for diagnostic and epidemiological purposes.
Evaluation of CHROMagar
Acinetobacter
for Detection of Enteric Carriage of Multidrug-Resistant
Acinetobacter baumannii
in Samples from Critically Ill Patients
Authors: Gordon N. C. , Wareham D. W.
Published: May 2009

CHROMagar
Acinetobacter
was used to screen stool and perineal swabs for enteric carriage of multidrug-resistant
Acinetobacter baumannii
in samples from critically ill patients. Results were compared with a molecular assay resulting in sensitivity and specificity of culture compared to PCR of 91.7% and 89.6%, respectively.
Rapid and Sensitive Detection of Shiga Toxin-Producing
Escherichia coli
from Nonenriched Stool Specimens by Real-Time PCR in Comparison to Enzyme Immunoassay and Culture
Authors: Grys Thomas E. , Sloan Lynne M. , Rosenblatt Jon E. , Patel Robin
Published: May 2009

Shiga toxin (Stx)-producing
Escherichia coli
(STEC) bacteria are a frequent cause of food-borne gastroenteritis, hemorrhagic colitis, and hemolytic uremic syndrome. Because antimicrobial agents are generally contraindicated in patients infected with STEC, a sensitive and specific diagnostic test with rapid turnaround is essential. Current culture methods may fail to detect non-O157 STEC. We evaluated a Stx gene real-time PCR assay using hybridization probes and the LightCycler instrument with 204 prospectively collected stool specimens, which were also tested for Stx by enzyme immunoassay (EIA) (ProSpecT STEC; Remel, Lenexa, KS) and by culturing on chromogenic agar (Chromagar O157; BD BBL, Sparks, MD). In addition, 85 archived stool specimens previously tested for Stx (by EIA) and/or
E. coli
O157:H7 (by culture) were tested by PCR. Sample preparation for PCR included mixing the stool in sterile water and extraction of nucleic acid using the MagNA Pure LC instrument (Roche Diagnostics). The PCR assay had 100% sensitivity and specificity compared to EIA and culture for specimens collected prospectively (4 of 204 specimens were positive) and compared to culture and/or EIA for archival specimens (42 of 85 specimens were positive). Both the EIA and PCR produced positive results from a specimen containing an O103 serotype STEC in the prospective specimens, and the PCR test detected three positive specimens that contained nonviable STEC in the archived specimens. The PCR assay demonstrated 100% sensitivity and specificity compared to EIA and/or culture and more rapid turnaround than either EIA or culture.
Comparison of Chromogenic Media to BD GeneOhm Methicillin-Resistant
Staphylococcus aureus
(MRSA) PCR for Detection of MRSA in Nasal Swabs
Authors: Bischof Larry J. , Lapsley Linda , Fontecchio Karen , Jacosalem Dollie , Young Carol , Hankerd Rosemary , Newton Duane W.
Published: May 2009

To select a method for detecting methicillin-resistant
Staphylococcus aureus
(MRSA) in nasal swabs, we compared BD GeneOhm MRSA PCR and various culture media (mannitol salt agar with cefoxitin, MRSA
Select
, CHROMagar MRSA, and Spectra MRSA). While PCR detection of MRSA was more rapid, MRSA
Select
and Spectra MRSA demonstrated performance equivalent to that of PCR with maximal detection at 24 h.
Evidence that the Linker between the Methyltransferase and Helicase Domains of Potato Virus X Replicase Is Involved in Homologous RNA Recombination
Authors: Draghici Heidrun-Katharina , Varrelmann Mark
Published: May 2009

Recombination in RNA viruses, one of the main factors contributing to their genetic variability and evolution, is a widespread phenomenon. In this study, an in vivo assay to characterize RNA recombination in potato virus X (PVX), under high selection pressure, was established.Agrobacterium tumefacienswas used to express inNicotiana benthamianaleaf tissue both a PVX isolate labeled with green fluorescent protein (GFP) containing a coat protein deletion mutation (ΔCP) and a transcript encoding a functional coat protein +3′-ntr. Coexpression of the constructs led to virus movement and systemic infection; reconstituted recombinants were observed in 92% of inoculated plants. Similar results were obtained using particle bombardment, demonstrating that recombination mediated byA. tumefacienswas not responsible for the occurrence of PXC recombinants. The speed of recombination could be estimated by agroinfection of two PVX mutants lacking the 3′ and 5′ halves of the genome, respectively, with an overlap in the triple gene block 1 gene, allowing GFP expression only in the case of recombination. Ten different pentapeptide insertion scanning replicase mutants with replication abilities comparable to wild-type virus were applied in the different recombination assays. Two neighboring mutants affecting the linker between the methyltransferase and helicase domains were shown to be strongly debilitated in their ability to recombine. The possible functional separation of replication and recombination in the replicase molecule supports the model that RNA recombination represents a distinct function of this protein, although the underlying mechanism still needs to be investigated.
The M33 Chemokine Receptor Homolog of Murine Cytomegalovirus Exhibits a Differential Tissue-Specific Role during In Vivo Replication and Latency
Authors: Cardin Rhonda D. , Schaefer Gregory C. , Allen Janelle R. , Davis-Poynter Nicholas J. , Farrell Helen E.
Published: May 2009

M33, encoded by murine cytomegalovirus (MCMV), is a member of the UL33 homolog G-protein-coupled receptor (GPCR) family and is conserved across all the betaherpesviruses. Infection of mice with recombinant viruses lacking M33 or containing specific signaling domain mutations in M33 results in significantly diminished MCMV infection of the salivary glands. To determine the role of M33 in viral dissemination and/or infection in other tissues, viral infection with wild-type K181 virus and an M33 mutant virus, ΔM33B
T2
, was characterized using two different routes of inoculation. Following both intraperitoneal (i.p.) and intranasal (i.n.) inoculation, M33 was attenuated for infection of the spleen and pancreas as early as 7 days after infection. Following i.p. inoculation, ΔM33B
T2
exhibited a severe defect in latency as measured by a diminished capacity to reactivate from spleens and lungs in reactivation assays (
P
< 0.001). Subsequent PCR analysis revealed markedly reduced ΔM33B
T2
viral DNA levels in the latently infected spleens, lungs, and bone marrow. Following i.n. inoculation, latent ΔM33B
T2
viral DNA was significantly reduced in the spleen and, in agreement with results from i.p. inoculation, did not reactivate from the spleen (
P
< 0.001). Furthermore, in vivo complementation of ΔM33B
T2
virus replication and/or dissemination to the salivary glands and pancreas was achieved by coinfection with wild-type virus. Overall, our data suggest a critical tissue-specific role for M33 during infection in the salivary glands, spleen, and pancreas but not the lungs. Our data suggest that M33 contributes to the efficient establishment or maintenance of long-term latent MCMV infection.
Latency-Associated Nuclear Antigen of Kaposi's Sarcoma-Associated Herpesvirus (KSHV) Upregulates Survivin Expression in KSHV-Associated B-Lymphoma Cells and Contributes to Their Proliferation
Authors: Lu Jie , Verma Subhash C. , Murakami Masanao , Cai Qiliang , Kumar Pankaj , Xiao Bingyi , Robertson Erle S.
Published: May 2009

Survivin is a master regulator of cell proliferation and cell viability and is highly expressed in most human tumors. The molecular network linked to survivin expression in tumors has not been completely elucidated. In this study, we show that latency-associated nuclear antigen (LANA), a multifunctional protein of Kaposi's sarcoma-associated herpesvirus (KSHV) that is found in Kaposi's sarcoma tumors, upregulates survivin expression and increases the proliferation of KSHV-infected B cells. Analysis of pathway-specific gene arrays showed that survivin expression was highly upregulated in BJAB cells expressing LANA. The mRNA levels of survivin were also upregulated in HEK 293 and BJAB cells expressing LANA. Similarly, protein levels of survivin were significantly higher in LANA-expressing, as well as KSHV-infected, cells. Survivin promoter activity assays identified GC/Sp1 and p53
cis
-acting elements within the core promoter region as being important for LANA activity. Gel mobility shift assays revealed that LANA forms a complex with Sp1 or Sp1-like proteins bound to the GC/Sp1 box of the survivin promoter. In addition, a LANA/p53 complex bound to the p53
cis
-acting element within the survivin promoter, indicating that upregulation of survivin expression can also occur through suppression of p53 function. Furthermore, immunohistochemistry analyses revealed that survivin expression was upregulated in KSHV-associated Kaposi's sarcoma tissue, suggesting that LANA plays an important role in the upregulation of survivin expression in KSHV-infected endothelial cells. Knockdown of survivin expression by lentivirus-delivered small hairpin RNA resulted in loss of cell proliferation in KSHV-infected cells. Therefore, upregulation of survivin expression in KSHV-associated human cells contributes to their proliferation.
The Fowlpox Virus BCL-2 Homologue, FPV039, Interacts with Activated Bax and a Discrete Subset of BH3-Only Proteins To Inhibit Apoptosis
Authors: Banadyga Logan , Veugelers Kirstin , Campbell Stephanie , Barry Michele
Published: May 2009

Apoptosis is a potent immune barrier against viral infection, and many viruses, including poxviruses, encode proteins to overcome this defense. Interestingly, the avipoxviruses, which include fowlpox and canarypox virus, are the only poxviruses known to encode proteins with obvious Bcl-2 sequence homology. We previously characterized the fowlpox virus protein FPV039 as a Bcl-2-like antiapoptotic protein that inhibits apoptosis by interacting with and inactivating the proapoptotic cellular protein Bak. However, both Bak and Bax can independently trigger cell death. Thus, to effectively inhibit apoptosis, a number of viruses also inhibit Bax. Here we show that FPV039 inhibited apoptosis induced by Bax overexpression and prevented both the conformational activation of Bax and the subsequent formation of Bax oligomers at the mitochondria, two critical steps in the induction of apoptosis. Additionally, FPV039 interacted with activated Bax in the context of Bax overexpression and virus infection. Importantly, the ability of FPV039 to interact with active Bax and inhibit Bax activity was dependent on the structurally conserved BH3 domain of FPV039, even though this domain possesses little sequence homology to other BH3 domains. FPV039 also inhibited apoptosis induced by the BH3-only proteins, upstream activators of Bak and Bax, despite interacting detectably with only two: BimL and Bik. Collectively, our data suggest that FPV039 inhibits apoptosis by sequestering and inactivating multiple proapoptotic Bcl-2 proteins, including certain BH3-only proteins and both of the critical “gatekeepers” of apoptosis, Bak and Bax.