bluuebunny/crossref_metadata_2025 huggingface dataset:
Biotribological and biomechanical changes after experimental haemarthrosis in the rabbit knee
Authors: RAVANBOD R. , TORKAMAN G. , ESTEKI A.
Published: September 2010

Summary.  Changes in articular cartilage after haemarthrosis have not been completely elucidated in haemophilic arthropathy. Insights into the pathophysiological mechanisms of blood‐induced joint damage mainly derived from histological, inflammatory and biochemical investigations. A structure–function relationship is another reasonable way to determine the joint overall health status. Cartilage, a viscoelastic connective tissue, is at least a biphasic material that should also work under minimal friction. Pendulum friction tester measures the mechanical aspects of joint lubrication and quantifies the biotribological properties of the joint. Indentation test is an in situ method characterizing the biomechanical properties of the cartilage. Gross, biotribological and biomechanical properties were determined in a rabbit model of experimental haemarthrosis. A sample of 1 mL of fresh autologous blood was injected in the left knee of rabbit’s joint twice weekly for four consecutive weeks. The right knee and animals in the control group were left untreated. After 8 days, joint perimeter, biotribological and biomechanical tests were performed. In a consistent manner, all data showed detrimental effects of the blood on the overall cartilage function under loading. Non‐weight bearing and early blood aspiration seem wise to be considered after haemarthrosis.
A cytoskeletal actin gene in the mosquito Anopheles gambiae
Authors: Salazar C. E. , Hamm D. Mills , Wesson D. M. , Beard C. B. , Kumar V. , Collins F. H.
Published: September 2010

Five actin genes have been identified in the mosquito Anopheles gambiae, and a constitutively expressed actin gene has been chosen for detailed analysis. We have physically mapped and sequenced this gene and six associated cDNAs, including translated coding regions, as well as the 5 and 3 flanking sequences. Analysis of stage‐specific RNA shows this gene to be present in all stages of mosquito development and in an established A. gambiae cell line, thus indicating a cytoskeietal actin. In the sequence of the translated coding region and in pattern of expression, this gene is very similar to the cytoskeietal actin genes of Droso‐phila melanogaster, and in sequence, equally similar to the Artemia cytoskeietal actin gene 403 (99.2% identity among the three amino acid sequences). Sequencing of this A. gambiae actin gene (designated actWior its location in chromosome division 1D) and selected cDNAs shows that it possesses three alternative leader sequences; thus the gene appears to have three alternative promoters. These promoters should ultimately prove useful in the production of transgenic constructs for constitutive expression.
Tissue specific distribution of Drosophila sarcomeric myosin heavy‐chain protein isoforms
Authors: Crough E. M. , Kazzaz J. A. , Rozek C. E.
Published: September 2010

The sarcomeric myosin heavy‐chain (sMHC) gene of Drosophila is single‐copy and RNA transcription from this gene is developmentally regulated. Numerous sMHC mRNAs that differ in exon composition can be formed by alternate RNA processing. These transcriptional events result in the presence of multiple sMHC isoforms in the developing organism. We have developed and characterized two antibodies which are specific for different types of sarcomeric myosin heavy‐chain protein isoforms in Drosophila and have begun to examine the tissue distribution and function of these various protein isoforms. One of the antibodies (anti‐A) is capable of distinguishing between two classes of sMHC protein isoforms which differ in their carboxy terminal amino acid sequences. The second antibody (anti‐MHC) recognizes a separate and distinct domain in sMHC protein isoforms. We demonstrate the specificity and the utility of these antibodies in examining the developmental and tissue‐specific expression of sMHC protein isoforms in the developing fly.
A cDNA encoding an ADP/ATP carrier from the mosquito Anopheles gamblae
Authors: Beard C. B. , Crews‐Oyen A. E. , Kumar V. K. , Collins F. H.
Published: September 2010

Two cDNAs are described from Anopheles gamblae that correspond to the ADP/ATP carrier or translocase. The clones are identical exceptfor minor differences in the 5 non‐coding region and in the lengths of the poly‐A tails. They code for mRNAs of 1261 and 1263 bp and contain one open reading frame of 906 bp. A probe made from the 1263 bp cDNA hybridized to bands of approximately 1260 and 1700 bp on developmental Northern blots. The putative 300 amino acid peptide sequence shows from 53.4–78.5% identity to AAC peptide sequences from a range of organisms from Zea maysto human. Both clones mapped to region 26a on the left arm of chromosome 2 in An. gamblae.
20‐Hydroxyecdysone regulation of two isoforms of the Ets transcription factor E74 gene in programmed cell death in the silkworm anterior silk gland
Authors: Sekimoto T. , Iwami M. , Sakurai S.
Published: September 2010

Programmed cell death of larval‐specific tissues in insects is under the control of 20‐hydroxyecdysone (20E). The ecdysteroid‐regulated early genes are conserved in the programmed cell death of anterior silk glands (ASGs) in Bombyx mori and salivary glands in Drosophila melanogaster. We identified and characterized two isoforms of the Ets transcription factor E74 gene in B. mori (BmE74). In ASGs of B. mori last instar larvae, the Bm74A mRNA level increased concomitantly with an increase in haemolymph ecdysteroid titre after gut purge. The optimal 20E concentration for stimulation of Bm74A in ASGs was 4 µM, a similar value to the peak haemolymph ecdysteroid concentration after gut purge. In contrast, BmE74B expression peaked on day 5 of the feeding period, after which it did not increase again. These findings suggest that the BmE74 isoforms play different roles in the regulation of programmed cell death in B. mori ASGs.
Germ‐line transformation of the Mexican fruit fly
Authors: Condon K. C. , Condon G. C. , Dafa’alla T. H. , Forrester O. T. , Phillips C. E. , Scaife S. , Alphey L.
Published: September 2010

Germ‐line transformation of a major agricultural pest, the Mexican fruit fly (Anastrepha ludens Loew, Mexfly), was achieved using composite piggyBac transposable elements marked with green, yellow and red fluorescent proteins (CopGreen, PhiYFP and J‐Red). We also investigated the possibility of generating transposon‐free insertions, in order to address potential concerns relating to proposed field use of transgenic Mexfly. We describe a highly efficient method for transforming Mexfly, compare efficiency of piggyBac terminal sequences for transformation and also describe the derivation of a transposon‐free insertion line. The development of an efficient transformation system for Mexfly holds great promise for improved applications of the sterile insect technique, a major component of the present control measures for this economically important pest species.
Targeting ie‐1 gene by RNAi induces baculoviral resistance in lepidopteran cell lines and in transgenic silkworms
Authors: Kanginakudru S. , Royer C. , Edupalli S. V. , Jalabert A. , Mauchamp B. ,   Chandrashekaraiah, Prasad S. V. , Chavancy G. , Couble P. , Nagaraju J.
Published: September 2010

RNA interference (RNAi)‐mediated viral inhibition has been used in a few organisms for eliciting viral resistance. In the present study, we report the use of RNAi in preventing baculovirus infection in a lepidopteran. We targeted the baculoviral immediate early‐1 (ie‐1) gene in both a transformed lepidopteran cell line and in the transgenic silkworm Bombyx mori L. Constitutive expression of double‐stranded RNA was achieved by piggyBac‐mediated transformation of Sf9 cell line with a transgene encoding double‐stranded ie‐1 RNA (dsie‐1). Strong viral repression was seen at early stages of infection but subsequent recovery of viral proliferation was observed. In contrast, the same transgene inserted into the chromosomes of transgenic silkworms induced long‐term inhibition of B. mori nucleopolyhedrovirus infection, with nearly 40% protection compared with nontransgenic animals. Protection was efficient at larval stages after oral infection with occlusion bodies or hemocoel injection of budded viruses. Virus injected pupae also displayed resistance. These results show that heritable RNAi can be used to protect silkworm strains from baculovirus infection.
Tetracycline treatment influences mitochondrial metabolism and mtDNA density two generations after treatment in Drosophila
Authors: Ballard J. W. O. , Melvin R. G.
Published: September 2010

Tetracycline is commonly used to clear Wolbachia from infected insects. Studies then compare specific biochemical and/or life‐history traits between infected and uninfected individuals with the same genetic background. We investigated the potential for tetracycline to influence mitochondrial efficiency and mitochondrial (mt)DNA density two generations after treatment in Drosophila simulans. We observed that antibiotic treatment resulted in a decline in inorganic phosphate incorporated into ATP per mole of oxygen consumed (ADP:O ratio). Further, tetracycline treatment caused a significant increase in mtDNA density in naturally Wolbachia‐uninfected but not in naturally Wolbachia‐infected lines suggesting a dosage effect. These data suggest that the current practice of comparing Wolbachia‐infected and Wolbachia‐uninfected insects two generations after tetracycline treatment needs to be re‐evaluated.
Diverse Mariner‐like elements in fig wasps
Authors: Haine E. R. , Kabat P. , Cook J. M.
Published: September 2010

Mariner transposable elements are widespread and diverse in insects. We screened 10 species of fig wasps (Hymenoptera: Agaonidae) for mariner elements. All 10 species harbour a large diversity of mariner elements, most of which have interrupted reading frames in the transposase gene region, suggesting that they are inactive and ancient. We sequenced two full‐length mariner elements and found evidence to suggest that they are inserted in the genome at a conserved region shared by other hymenopteran taxa. The association between mariner elements and fig wasps is old and dominated by vertical transmission, suggesting that these ‘selfish genetic elements’ have evolved to impart only very low costs to their hosts.
Expression analysis of the FoxP homologue in the brain of the honeybee, Apis mellifera
Authors: Kiya T. , Itoh Y. , Kubo T.
Published: September 2010

The transcription factor FoxP2 is related to acoustic communication in vertebrates and, although widely expressed in various tissues, its mutations cause a speech disorder in humans and disrupt vocalization in mice. In honeybee colonies, workers transmit information about a food location using ‘dance communication’, which is a form of acoustic communication. We identified a honeybee FoxP2‐homologue, AmFoxP, and investigated its expression in the honeybee brain to elucidate its possible role in dance communication. The relative abundance of AmFoxP mRNA in the worker brain increased during the first 4 days of adult life. In situ hybridization revealed AmFoxP expression around the optic lobes, central complex, dorsal lobes, and protocerebral lobes, which was not dependent on the caste or division of labour.
Over expression of a Cytochrome P450 (CYP6P9) in a Major African Malaria Vector, Anopheles Funestus, Resistant to Pyrethroids
Authors: Amenya D. A. , Naguran R. , Lo T.‐C. M. , Ranson H. , Spillings B. L. , Wood O. R. , Brooke B. D. , Coetzee M. , Koekemoer L. L.
Published: September 2010

Anopheles funestus Giles is one of the major African malaria vectors. It has previously been implicated in a major outbreak of malaria in KwaZulu/Natal, South Africa, during the period 1996 to 2000. The re‐emergence of this vector was associated with monooxygenase‐based resistance to pyrethroid insecticides. We have identified a gene from the monooxygenase CYP6 family, CYP6P9, which is over expressed in a pyrethroid resistant strain originating from Mozambique. Quantitative Real‐Time PCR shows that this gene is highly over expressed in the egg and adult stages of the resistant strain relative to the susceptible strain but the larval stages showed almost no difference in expression between strains. This gene is genetically linked to a major locus associated with pyrethroid resistance in this A. funestus population.
The Gram‐Negative Bacteria‐Binding Protein gene family: Its role in the innate immune system of Anopheles gambiae and in anti‐Plasmodium defence
Authors: Warr E. , Das S. , Dong Y. , Dimopoulos G.
Published: September 2010

Gram‐negative bacteria‐binding proteins (GNBPs) are pattern recognition receptors which contribute to the defensive response against Plasmodium infection in Anopheles. We have characterized the GNBP gene family in Anopheles gambiae at the molecular level, and show that they are functionally diverse components of the A. gambiae innate immune system. GNBPB4 is a major factor in the defence against a broad range of pathogens, while the other GNBPs have narrower defence specificities. GNBPB4 is associated with the regulation of immune signalling pathways and was found to interact with the Gram‐negative Escherichia coli and weakly co‐localized with Plasmodium berghei ookinetes in the mosquito midgut epithelium.
piggyBac‐like elements in cotton bollworm, Helicoverpa armigera (Hübner)
Authors: Sun Z. C. , Wu M. , Miller T. A. , Han Z. J.
Published: September 2010

Two piggyBac‐like elements (PLEs) were identified in the cotton bollworm, Helicoverpa armigera, and were designated as HaPLE1 and HaPLE2. HaPLE1 is flanked by 16 bp inverted terminal repeats (ITRs) and the duplicated TTAA tetranucleotide, and contains an open reading frame (ORF) of 1794 bp with the presumed DDD domain, indicating that this element may be an active autonomously mobile element. HaPLE2 was found with the same ITRs, but lacks the majority of an ORF‐encoding transposase. Thus, this element was thought to be a non‐autonomous element. Transposable element displays and distribution of the two PLEs in individuals from three different H. armigera populations suggest that transmobilization of HaPLE2 by the transposase of HaPLE1 may be likely, and mobilization of HaPLE1 might occur not only within the same individual, but also among different individuals. In addition, horizontal transfer was probably involved in the evolution of PLEs between H. armigera and Trichoplusia ni.
Low‐voltage‐activated calcium channels are not involved in capacitation and biological response to progesterone in human sperm
Authors: Bonaccorsi Lorella , Forti Gianni , Baldi Elisabetta
Published: September 2010

The involvement of voltage‐dependent calcium channels in the biological effects exerted by progesterone (P) on human spermatozoa is still a controversial issue. We have investigated the involvement of T‐type calcium channels [voltage‐operated calcium channels (VOCCT)] in two biological functions of human sperm, responsiveness to P and capacitation, by employing three different pharmacological antagonists of VOCCT, namely mibefradil (Ro 5967), pimozide and amiloride. Intracellular calcium [Ca2+]iincrease in response to P was essentially unaffected by pre‐treatment with mibefradil and pimozide at concentrations previously shown to prevent [Ca2+]iincrease in response to zona proteins. Amiloride could not be tested in these experiments because it was found to interfere with fura‐2 fluorescence. The increase in tyrosine phosphorylation stimulated by P in a protein of about 97 kDa was unaffected by the three antagonists. Acrosome reaction (AR) induced by P was also unaffected by mibefradil or pimozide but was significantly inhibited by amiloride at high concentrations (100 and 500 but not 10 μM). At 100 and 500 μMamiloride also inhibited Na/H exchanger as assessed by a fluorimetric method. We conclude that VOCCT are not involved in calcium increase and AR stimulated by P in human sperm.

We next investigated the effect of the three VOCCT inhibitors on sperm capacitation by evaluating tyrosine phosphorylation and AR in basal conditions and in response to P. We found that the presence of pimozide and amiloride during capacitation stimulated a higher increase of tyrosine phosphorylation, whereas mibefradil was less effective. The ability of P to induce the AR, considered an index of occurrence of capacitation, was not affected by pimozide and mibefradil, whereas was inhibited by amiloride at concentrations that inhibit Na/H exchanger. In conclusion, our results do not support a major role of low‐voltage‐activated calcium channels in capacitation and response to P of human spermatozoa.
Combination of subcutaneous levonorgestrel implants and transdermal dihydrotestosterone gel for male hormonal contraception
Authors: Pöllänen Pasi , Nikkanen Vesa , Huhtaniemi Ilpo
Published: September 2010

The primary aim of the present study was to determine the therapeutic dose of subcutaneous levonorgestrel (LNG) to induce azoospermia or severe oligozoospermia (<3 × 106/mL) in normal men requiring contraception. Transdermal 5α‐dihydrotestosterone (DHT) was combined to the treatments to maintain peripheral androgen level. Forty‐three 21–45‐year‐old healthy men were enrolled in this phase II randomised and comparative clinical performance study. The subjects were allocated to five groups to receive: (1) transdermal DHT (AndractimR, Besins Iscovesco, Paris, France) and one subdermal LNG implant (Jadelle, Leiras, Turku, Finland); (2) transdermal DHT and two subdermal LNG implants; (3) transdermal DHT and four subdermal LNG implants; (4) transdermal DHT and oral LNG (Microluton, Schering, Germany); or (5) transdermal DHT only. A total of 27 men completed the suppression phase. None of them reached azoospermia. One subject with oral LNG and transdermal DHT reached <3 million/mL at 5 months of suppression, but not repeatedly. Together 2/27 (7%) subjects, one with oral LNG and DHT and the other with four subdermal LNG implants and DHT reached <5 million/mL temporarily. Altogether 9/27 (33%) subjects reached <20 million/mL. Serum testosterone concentrations (S‐T) decreased significantly during the first 3 months of treatment with one, two and four LNG implants and DHT and during the next 3 months S‐T remained significantly lower with two or four implants. Serum oestradiol concentrations (S‐E2) decreased significantly during the first 3 months only with four implants, but at 6 months S‐E2 was lower also in the group with two implants. Serum luteinizing hormone (LH) decreased significantly only with two LNG implants and DHT gel at 5 and 6 months. Serum FSH did not decrease in any of the groups. None of the subjects filled the criteria to continue to the efficacy phase. A total of 16 men discontinued for various reasons. Of the 27 men completing the suppression phase, all have recovered to sperm levels >20 million/mL. There were no changes in blood count, lipid profile, liver function tests, prostate‐specific antigen (PSA), sex hormone binding globulin (SHBG), prolactin or cortisol. The mixed antiglobulin reaction (MAR)‐IgG, MAR‐IgA or tray agglutination test (TAT) did not change during any of the treatments. The present study shows that the LNG implants themselves are well‐tolerated by men and safe, and might be suitable for replacing part of the testosterone used as injections to reduce the androgen dose during male hormonal contraception. The DHT gel was considered as quite or very uncomfortable by 66% of the men because of feeling cold during the time it was on the skin, but uncompliance in using the gel was not reported by the men.
Comparison of sperm preparation methods: effect on chromatin and morphology recovery rates and their consequences on the clinical outcome after in vitro fertilization embryo transfer
Authors: Hammadeh M. E. , Kühnen A. , Amer A. S. , Rosenbaum P. , Schmidt W.
Published: September 2010

The aim of this study was to evaluate the efficacy of swim‐up, PureSperm® gradient centrifugation and glass–wool filtration methods for semen preparation and to assess the possible enhancement of the quality of the subpopulation of spermatozoa in terms of sperm concentration, morphology and chromatin condensation. Moreover, to determine the effect of this semen processing technique on the clinical outcome after in vitro fertilization embryo transfer (IVF‐ET).

A total of 180 semen samples of patients’ husbands who were undergoing IVF therapy were prepared by swim‐up (G1, n= 60), PureSperm® gradient centrifugation (G2, n=60) or glass–wool (G3, n=60) methods. Chromatin condensation was assessed by Chromomycin (CMA3), whereas sperm morphology was evaluated according to strict criteria. In all three semen processing methods, the percentage of chromatin condensed and morphologically normal spermatozoa was higher after semen processing in comparison with native semen samples. The proportion of normal chromatin condensed spermatozoa prepared in glass–wool filtration was significantly higher than that in swim‐up (G.I, p=0.02) or PureSperm® (G.II, p=0.001). In addition semen processing with PureSperm® yields significantly a higher percentage of morphologically normal spermatozoa than swim‐up (p < 0.001) or glass–wool method (p < 0.002). However, the fertilization, implantation and pregnancy rates, in turn were similar in all semen preparation methods.

In conclusion, PureSperm® gradient centrifugation yields a higher percentage of morphologically normal spermatozoa than shown in traditional swim‐up or glass–wool filtration. However, the percentage of chromatin condensed spermatozoa was significantly higher after semen processing via glass–wool in comparison with the other two methods.

Nevertheless, there were no significant difference in the fertilization, implantation and pregnancy rates of sperm prepared by means of swim‐up, PureSperm® or glass–wool filtration.

Therefore, glass–wool filtration should be recommended as the first choice for semen preparation for Intracytoplasmic sperm injection (ICSI) technique as the natural selection is bypassed. Whereas, swim‐up and PureSperm should be used for semen processing in IVF programme.
Validation of biological collections as a source of phenological data for use in climate change studies: a case study with the orchid Ophrys sphegodes
Authors: Robbirt Karen M. , Davy Anthony J. , Hutchings Michael J. , Roberts David L.
Published: September 2010

1. The scarcity of reliable long‐term phenological data has severely hindered the study of the responses of species to climate change. Biological collections in herbaria and museums are potential sources of long‐term data for such study, but their use for this purpose needs independent validation. Here we report a rigorous test of the validity of using herbarium specimens for phenological studies, by comparing relationships between climate and time of peak flowering derived from herbarium records and from direct field‐based observations, for the terrestrial orchid Ophrys sphegodes.

2. We examined herbarium specimens of O. sphegodes collected between 1848 and 1958, and recorded peak flowering time directly in one population of O. sphegodes between 1975 and 2006. The response of flowering time to variation in mean spring temperature (March–May) was virtually identical in both sets of data, even though they covered different periods of time which differ in extent of anthropogenic temperature change. In both cases flowering was advanced by c. 6 days per °C rise in average spring temperature.

3. The proportion of variation in flowering time explained by spring temperature was lower in the herbarium record than in direct field observations. It is likely that some of the additional variation was due to geographical variation in collection site, as flowering was significantly earlier at more westerly sites, which have had warmer springs, over their range of 3.44° of longitude.

4. Predictions of peak flowering time based on the herbarium data corresponded closely with observed peak flowering times in the field, indicating that flowering response to temperature had not altered between the two separate periods over which the herbarium and field data were collected.

5. Synthesis. These results provide the first direct validation of the use of herbarium collections to examine the relationships between phenology and climate when field‐based observational data are not available.
Bayesian and frequentist models: legitimate choices for different purposes of clinical research
Authors: Berger Zackary
Published: September 2010

Objective  Bayesian and frequentist approaches to statistical modelling in epidemiology are often pitted against each other as if they represented diametrically opposing philosophies. However, both approaches have a role to play in clinical epidemiology and the evaluation of clinical practice.

Methods  Here I present an overview of the philosophical underpinnings of the Bayesian and frequentist approaches, showing that each model has its place depending on the philosophical and evaluative needs of the user.

Results  If the user's approach to a clinical problem places an emphasis on identifying causal relationships, a frequentist approach might be best suited. On the other hand, if the user takes an approach in which estimating a priori probabilities is appropriate, a Bayesian approach might be more appropriate. One could imagine both approaches used for the same study.

Conclusions  Bayesian and frequentist approaches are complementary tools in the clinical evaluator's toolkit.
Effectiveness of unattended ambulatory sleep studies for the diagnosis and treatment of OSAS
Authors: Domingo Christian , Vigil Laura
Published: September 2010

Background  Unattended home sleep studies have been implemented to minimize the cost and delay in establishing obstructive sleep apnoea syndrome (OSAS) diagnosis and therapy. However, their accuracy and validity remain disputed. The purpose of our study is to assess their effectiveness (i.e. their technical reliability in real life conditions).

Setting  Private sleep unit attending to private and social security patients.

Population  Consecutively admitted patients, aged over 18 years, referred to our clinic for confirmation or treatment of OSAS without co‐morbidities.

Methods  A clinical history was obtained for each patient with special attention to sleep, smoking habit, educational level and type of insurance. Chest X‐ray and forced spirometry were performed. For the portable respiratory recording device an eight‐channel polygraph was used and for continuous positive airway pressure (CPAP) titration an automatic positive airway pressure device was applied; both instruments were previously validated. Patients were instructed how to use the devices at the clinic and then applied them at home by themselves.

Results  For diagnostic sleep studies, 145 out of 150 (96.7%: CI: 92.4–98.9) and for CPAP titration 42 out of the 61 patients (69%: CI: 56.4–81.3) performed the sleep study correctly according to the pre‐established criteria. No statistically significant differences were found between correct performance and educational level, sex, smoking habit or type of health insurance.

Conclusions  (1) Unattended home sleep studies appear as effective as in‐hospital studies for diagnosis of OSAS without co‐morbidities; (2) for CPAP titration, the effectiveness was slightly lower; and (3) factors such as patients' educational level or type of health insurance do not appear to influence outcome.
Focusing the focus group: impact of the awareness of major factors contributing to non‐adherence to acute paediatric asthma guidelines
Authors: Bhogal Sanjit Kaur , McGillivray David , Bourbeau Jean , Plotnick Laurie H. , Bartlett Susan Joan , Benedetti Andrea , Ducharme Francine Monique
Published: September 2010

Rationale  The administration of oral corticosteroids within the first hour in the emergency department is associated with reduced hospitalization rates in children with moderate and severe asthma, yet less than half of patients benefit from this recommendation. To ensure patients receive recommended treatment, a clear understanding of what is causing suboptimal care management is needed. The assessment of barriers and solutions to optimal care is often done without a thorough examination of the factors associated with non‐adherence.

Objective  To evaluate whether knowledge of factors associated with delayed administration of systemic corticosteroids modifies the focus and prioritization of barriers and solutions identified by focus groups.

Methods  We conducted two parallel focus groups of emergency health care professionals – one group informed and the other non‐informed of key factors. Both groups received a presentation on the acute asthma guidelines, the evidence supporting its recommendations, and current practice. In addition, the informed group was provided with the factors associated and not associated with delayed administration. The groups were given 20 minutes to discuss barriers and solutions, with 5 minutes each for voting for the main barriers and solutions. Group difference in the misdirection of discussion was measured as time spent discussing barriers that were shown not to be associated with systemic corticosteroids. Prioritization of barriers and solutions was based on group endorsement.

Results  The non‐informed group spent more time discussing barriers not associated with delayed administration (15 vs. 2 minutes, P = 0.05). Although the non‐informed group proposed more solutions, most were to overcome barriers not associated with delayed administration. Of the main barriers and solutions identified by each group, only one barrier and solution were similar between the two groups: emergency department overcrowding and administrating corticosteroids at triage.

Conclusion  The awareness of objective factors of non‐adherence enabled a more directed discussion on relevant barriers and solutions, affecting prioritization of each. The administration of oral corticosteroids at triage appears to be the best solution to overcome delayed administration.
Indications for hip and knee replacement in Sweden
Authors: Löfvendahl Sofia , Bizjajeva Svetlana , Ranstam Jonas , Lidgren Lars
Published: September 2010

Objectives  The aim of this paper was to compare selected indication parameters for patients scheduled for hip and knee replacement at orthopaedic units in Sweden.

Methods  Swedish orthopaedic clinics performing joint replacement were invited to enrol in the study. The study time was set to 2 years (from June 2006 to June 2008). The study subjects were patients undergoing hip or knee replacement for osteoarthritis (OA). For data collection, we used a Swedish priority criteria tool based on a translation from a form used in Canada with minor changes. The reliability and validity of the Swedish tool were investigated, with good reproducibility. The questionnaires (one for the doctor and one for the patient) were completed during decision making for surgery.

Results  Eleven hospitals enrolled in the study. In total, 2961 patients were included during the study period. Among these, 1662 were hip replacement patients and 1299 were knee replacement patients. The vast majority of patients undergoing hip or knee replacement had findings indicating severe OA, both clinically and radiologically according to the clinical priority tool. Statistically significant self‐reported problems with pain at rest, walking and impaired activities of daily living were also observed. There were statistically significant differences in reported indications between the hospitals, both for hip OA patients and for knee OA patients.

Conclusions  A clinical priority criteria tool is a useful means of following changes in indications for certain procedures. It could also contribute to explaining differences in case mix when evaluating clinical outcome and patient satisfaction.
Measuring perceptions of safety climate in primary care: a cross‐sectional study
Authors: de Wet Carl , Johnson Paul , Mash Robert , McConnachie Alex , Bowie Paul
Published: September 2010

Rationale, aims and objectives  Building a strong and positive safety culture in health care teams and organizations is essential for patient safety. Measuring individual perceptions of safety climate is an integral part of this process. Evidence of the successful application and potential usefulness of this approach is increasingly available for secondary care settings but little is known about the safety climate in UK primary care. We therefore aimed to measure perceptions of safety climate in primary care. Further aims were to determine whether perceptions varied significantly between practice teams and according to specific participant and practice characteristics.

Method  We undertook a cross‐sectional, anonymous postal questionnaire survey of randomly selected west of Scotland primary care teams. Safety climate mean scores with standard deviations were calculated for respondents, practice teams and the region.

Results  A total of 563 (84%) team members from 49 practices (24.5%) returned questionnaires. The overall safety climate mean score was 5.48 (0.78). Significant differences in safety climate perceptions were found at the practice team level (P < 0.001) and for specific characteristics: respondents' years of experience, whether they were community or practice based, their professional roles and practices' training status. Practice managers and general practitioners perceived the safety climate more positive than other respondents (P < 0.001).

Conclusion  This was the first known attempt to measure perceptions of safety climate in UK primary care with a validated instrument specifically developed for that purpose. Reported perceptions of the prevailing safety climate were generally positive. This may reflect ongoing efforts to build a strong safety culture in primary care or alternatively point to an overestimation of the effectiveness of local safety systems. The significant variation in perception between certain staff groups has potential safety implications and may have to be aligned for a positive and strong safety culture to be built. While safety climate measurement has various benefits at the individual, practice team and regional level, further research of its association with specific safety outcomes is required.
Is the SAPS II score valid in surgical intensive care unit patients?
Authors: Sakr Yasser , Marques Juliana , Mortsch Stefan , Gonsalves Matheus Demarchi , Hekmat Khosro , Kabisch Bjorn , Kohl Matthias , Reinhart Konrad
Published: September 2010

Aims and objectives  We investigated the performance of the simplified acute physiology score II (SAPS II) in a large cohort of surgical intensive care unit (ICU) patients and tested the hypothesis that customization of the score would improve the uniformity of fit in subgroups of surgical ICU patients.

Methods  Retrospective analysis of prospectively collected data from all 12 938 patients admitted to a postoperative ICU between January 2004 and January 2009. Probabilities of hospital death were calculated for original and customized (C1‐SAPS II and C2‐SAPS II) scores. A priori subgroups were defined according to age, probability of death according to the SAPS II score, ICU length of stay (LOS), surgical procedures and type of admission.

Results  The median ICU LOS was 1 (1–3) day. ICU and hospital mortality rates were 5.8% and 10.3%, respectively. Discrimination of the SAPS II was moderate [area under receiver operating characteristic curve (aROC) = 0.76 (0.75–0.78)], but calibration was poor. This model markedly overestimated hospital mortality rates [standardized mortality rate: 0.35 (0.33–0.37)]. First‐level customization (C1‐SAPS II) did not improve discrimination in the whole cohort or the subgroups, but calibration improved in some subgroups. Second‐level customization (C2‐SAPS II) improved discrimination in the whole cohort [aROC = 0.82 (0.79–0.85)] and most of the subgroups (aROC range 0.65–86). Calibration in this model (C2‐SAPS II) improved in the whole cohort and in subgroups except in patients with ICU LOS 4–14 days and those undergoing neuro‐ or gastrointestinal surgery.

Conclusions  In this large cohort of surgical ICU patients, performance of the original SAPS II model was generally poor. Although second‐level customization improved discrimination and calibration in the whole cohort and most of the subgroups, it failed to simultaneously improve calibration in the subgroups stratified according to the type of surgery, age or ICU LOS.
Independent evolutionary origins of landlocked alewife populations and rapid parallel evolution of phenotypic traits
Authors: PALKOVACS ERIC P. , DION KIRSTIN B. , POST DAVID M. , CACCONE ADALGISA
Published: September 2010

Alewife,Alosa pseudoharengus, populations occur in two discrete life‐history variants, an anadromous form and a landlocked (freshwater resident) form. Landlocked populations display a consistent pattern of life‐history divergence from anadromous populations, including earlier age at maturity, smaller adult body size, and reduced fecundity. In Connecticut (USA), dams constructed on coastal streams separate anadromous spawning runs from lake‐resident landlocked populations. Here, we used sequence data from the mtDNA control region and allele frequency data from five microsatellite loci to ask whether coastal Connecticut landlocked alewife populations are independently evolved from anadromous populations or whether they share a common freshwater ancestor. We then used microsatellite data to estimate the timing of the divergence between anadromous and landlocked populations. Finally, we examined anadromous and landlocked populations for divergence in foraging morphology and used divergence time estimates to calculate the rate of evolution for foraging traits. Our results indicate that landlocked populations have evolved multiple times independently. Tests of population divergence and estimates of gene flow show that landlocked populations are genetically isolated, whereas anadromous populations exchange genes. These results support a ‘phylogenetic raceme’ model of landlocked alewife divergence, with anadromous populations forming an ancestral core from which landlocked populations independently diverged. Divergence time estimates suggest that landlocked populations diverged from a common anadromous ancestor no longer than 5000 years ago and perhaps as recently as 300 years ago, depending on the microsatellite mutation rate assumed. Examination of foraging traits reveals landlocked populations to have significantly narrower gapes and smaller gill raker spacings than anadromous populations, suggesting that they are adapted to foraging on smaller prey items. Estimates of evolutionary rates (in haldanes) indicate rapid evolution of foraging traits, possibly in response to changes in available resources.
Seasonal change in the opportunity for sexual selection
Authors: REICHARD M. , SMITH C. , BRYJA J.
Published: September 2010

Environmental and population parameters that influence the strength of sexual selection may vary considerably over the course of the reproductive season. However, the potential for sexual selection frequently fails to translate into variation in reproductive success among individuals. We investigated seasonal changes in variation in reproductive success, measured as the opportunity for sexual selection, using parentage analysis in 20 experimental populations of the European bitterling (Rhodeus amarus, Cyprinidae), a small freshwater fish with a promiscuous, resource‐based mating system. We showed that although the largest males sired most offspring over the entire reproductive season, variation in reproductive success and hence the opportunity for sexual selection was low at the start of the season but increased significantly at its end. This seasonal difference probably arose from the superior competitive endurance of large males and from a higher temporal clustering of reproductively active females at the start of the breeding season than later in the season. The spatial distribution of oviposition sites had a negligible effect on the variation in reproductive success. We discuss the potential implications of our results for the importance and strength of sexual selection in natural populations.
Insight into the routes of Wolbachia invasion: high levels of horizontal transfer in the spider genus Agelenopsis revealed by Wolbachia strain and mitochondrial DNA diversity
Authors: BALDO LAURA , AYOUB NADIA A. , HAYASHI CHERYL Y. , RUSSELL JACOB A. , STAHLHUT JULIE K. , WERREN JOHN H.
Published: September 2010

The pandemic distribution of Wolbachia (alpha‐proteobacteria) across arthropods is largely due to the ability of these maternally inherited endosymbionts to successfully shift hosts across species boundaries. Yet it remains unclear whether Wolbachia has preferential routes of transfer among species. Here, we examined populations of eight species of the North American funnel‐web spider genus Agelenopsis to evaluate whether Wolbachia show evidence for host specificity and the relative contribution of horizontal vs. vertical transmission of strains within and among related host species. Wolbachia strains were characterized by multilocus sequence typing (MLST) and Wolbachia surface protein (WSP) sequences, and analysed in relation to host phylogeny, mitochondrial diversity and geographical range. Results indicate that at least three sets of divergent Wolbachia strains invaded the genus Agelenopsis. After each invasion, the Wolbachia strains preferentially shuffled across species of this host genus by horizontal transfer rather than cospeciation. Decoupling of Wolbachia and host mitochondrial haplotype (mitotypes) evolutionary histories within single species reveals an extensive contribution of horizontal transfer also in the rapid dispersal of Wolbachia among conspecific host populations. These findings provide some of the strongest evidence to support the association of related Wolbachia strains with related hosts by means of both vertical and horizontal strain transmission. Similar analyses across a broader range of invertebrate taxa are needed, using sensitive methods for strain typing such as MLST, to determine if this pattern of Wolbachia dispersal is peculiar to Agelenopsis (or spiders), or is in fact a general pattern in arthropods.
Insight into the genetic bases of climatic adaptation in experimentally evolving wheat populations
Authors: RHONÉ BÉNÉDICTE , REMOUÉ CARINE , GALIC NATHALIE , GOLDRINGER ISABELLE , BONNIN ISABELLE
Published: September 2010

Experimental populations evolving under natural selection represent an interesting tool to study genetic bases of adaptation. Evolution of genes possibly involved in adaptive response can be followed together with the corresponding phenotypic traits. Using experimental populations of hexaploid wheat, we studied the evolution of flowering time, a major adaptive trait that synchronizes the initiation of reproduction and the occurrence of favourable environmental conditions. During 12 generations, three populations were grown in contrasted environments (Vervins North France, Le Moulon near Paris, Toulouse South France) under the influence of natural selection, drift, mutation and recombination. Evolution of diversity at the major gene VRN‐1 involved in wheat vernalization response has been analysed jointly with earliness estimated in controlled conditions. Whatever the population, rapid phenotypic changes as well as parallel genotypic variations were observed in the first seven generations, probably as the result of selection acting on this major gene which explains 80% of the trait variation overall. Different allelic combinations at physically unlinked copies of VRN‐1 located on distinct genomes (A, B and D) were selected between populations. As theoretically expected, due to population differentiation, a high level of genetic diversity was maintained overall in generation 12. Surprisingly, in two populations out of three, the emergence of new alleles by mutation or migration, coupled with temporal variable selection or frequency‐dependent selection, allowed to maintain within‐population diversity despite local genetic drift and natural selection. This result may plead for an evolutionary approach of wheat genetic resource conservation.
Soft selective sweep near a gene that increases plant height in wheat
Authors: RAQUIN A.‐L. , BRABANT P. , RHONÉ B. , BALFOURIER F. , LEROY P. , GOLDRINGER I.
Published: September 2010

Strong selection within a given population locally reduces genetic variability not only in the selected gene itself but also in neighbouring loci. This so‐called hitch‐hiking effect is related to the initial linkage disequilibrium between markers and the selected gene, and depends mainly on the number of copies of the beneficial allele at the start of the selection phase. Contrary to the classical case, in which selection acts on a single, newly arisen beneficial mutation, we considered selection from standing variation (soft selective sweeps) on a gene (Rht‐B1) with a major effect on plant height, a selected trait in an experimental wheat population grown for 17 generations, and we documented the evolution of gene diversity and linkage disequilibrium near this gene. As expected,Rht‐B1was found to be under strong selection (s = 0.15) and its variation in frequency accounted for 15% of the total trait evolution. This led to a smaller genetic effective population size atRht‐B1(Neg = 18) compared to the whole genome estimation (Neg = 167). When compared with expectations under genetic drift only, no significant decrease in gene diversity was found at the closest loci. We computed expected di‐locus frequencies for any linked marker–Rht‐B1pair due to hitch‐hiking effects. We found that hitch‐hiking was expected to affect the two most closely linked loci, but expected reduction in gene diversity was not greater than that due to genetic drift, which was consistent with the observations. Such limited effect was attributed to the low level of linkage disequilibrium (0.16) estimated after parental intercrosses, together with a relatively high initial frequency of the gene. This situation is favourable to candidate gene approaches where small linkage disequilibrium around selected genes is expected.
Hotspots of diversity in a clonal world — the Mediterranean moss Pleurochaete squarrosa in Central Europe
Authors: GRUNDMANN MICHAEL , ANSELL STEPHEN W. , RUSSELL STEPHEN J. , KOCH MARCUS A. , VOGEL JOHANNES C.
Published: September 2010

Diversity patterns of the dioecious haploid Mediterranean moss Pleurochaete squarrosa were analysed from Central and Northwest Europe using nuclear and chloroplast DNA sequencing and enzyme electrophoresis. Across 69 populations, 38 distinct haploid multilocus genotypes (MLGs) were detected, but nearly all populations were clonal. Only five MLGs occurred in more than two regions, and two diversity hotspots were detected. The Kaiserstuhl mountains in Southwest Germany harboured 34 MLGs, 25 being endemic within Central Europe. Levels of linkage disequilibrium and population structure in Kaiserstuhl populations were similar to levels and structure in sexually reproducing populations in the Mediterranean Basin. In the Moselle‐Nahe area, some 250 km north, a comparably high allelic diversity, but no evidence of recombination, was detected. Genetic diversity measures were significantly lower than estimates obtained in the Mediterranean Basin and a GST of 0.89 signified extreme population differentiation. Mantel tests identified a positive correlation on genetic and geographical distance for distances up to 50 km. Seven nrITS and three cpDNA haplotypes were detected, their geographical structure mirroring enzyme data set results. Comparative analysis with Mediterranean data demonstrated multiple recolonization of Central Europe from both the Iberian Peninsula and the Balkans. A suture zone of genotypes was detected along the border of Belgium/France and Germany. Despite P. squarrosa having haploid spore and/or vegetative propagules dispersal, we found patterns of postglacial recolonization of Central Europe comparable with those reported in flowering plants and animals. This study demonstrates the importance of comparative research on population genetics and phylogeography of a diverse range of organisms.