Overstating the evidence - double counting in meta-analysis and related problems

Background: The problem of missing studies in meta-analysis has received much attention. Less attention has been paid to the more serious problem of double counting of evidence. Methods: Various problems in overstating the precision of results from meta-analyses are described and illustrated with examples, including papers from leading medical journals. These problems include, but are not limited to, simple double-counting of the same studies, double counting of some aspects of the studies, inappropriate imputation of results, and assigning spurious precision to individual studies. Results: Some suggestions are made as to how the quality and reliability of meta-analysis can be improved. It is proposed that the key to quality in meta-analysis lies in the results being transparent and checkable. Conclusions: Existing quality check lists for meta-analysis do little to encourage an appropriate attitude to combining evidence and to statistical analysis. Journals and other relevant organisations should encourage authors to make data available and make methods explicit. They should also act promptly to withdraw meta-analyses when mistakes are found

Esparsette (Onobrychis viciifolia) als mögliche Futterpflanze zur Kontrolle von Magen-Darm-Strongyliden bei Schafen

In der Schaf- und Ziegenhaltung stellt der Befall mit Magen-Darm-Strongyliden (MDS) einen wesentlichen Problemfaktor für die Tiergesundheit dar. Klinische und subklinische Erkrankungen können von erheblicher wirtschaftlicher Relevanz sein. Eine zunehmende Resistenzentwicklung gegen seit langem eingesetzte Anthelminthika hat derzeit einen intensivierten Einsatz dieser Wirkstoffe zur Folge und verschärft das Problem somit zusätzlich. Die Entwicklung und Bereitstellung komplementärer Kontrollstrategien könnte zu einer erheblichen Reduktion des Anthelminthika-Einsatzes beitragen. Eine mögliche Strategie stellt der Einsatz tanninhaltiger Futterpflanzen dar. Wir beschreiben Ergebnisse aus einem in vivo Versuch zur antiparasitischen Wirkung von Esparsettenheu und Esparsettensilage (6,2 % und 4,4 % Tannine / Trockensubstanz) gegen Haemonchus contortus und Cooperia curticei in experimentell infizierten Lämmern. Nach 16-tägiger Fütterung mit Esparsettenheu war die H. contortus Wurmbürde im Vergleich zu einer isoproteisch und isoenergetisch gefütterten Kontrollgruppe (0,1 % Tannine / Trockensubstanz) um 53 % reduziert (P < 0,05). Es konnte gezeigt werden, dass Lämmer, die mit Esparsettenheu gefüttert wurden, bereits 10 Tage nach Versuchsbeginn 44 % (P < 0,05) weniger H. contortus Eier pro Gramm Kot (EpG) ausschieden als die Kontrolltiere. Bei Versuchsende (16 Tage nach Fütterungsbeginn) war die H. contortus Eiausscheidung im Vergleich zur Kontrolle um 58 % reduziert (P < 0,01). Die Tagesgewichtszunahme der Lämmer in der Esparsettenheu-Gruppe war im Vergleich zur Kontrollgruppe tendenziell besser (163 g/Tag und 96 g/Tag; P = 0,07). Die Fütterung mit Esparsettensilage reduzierte die H. contortus Eiausscheidung verglichen mit der Kontrolle um 48 %. Dieses Resultat war allerdings nicht signifikant (P = 0,075). Im Vergleich zu den Kontrollgruppen wurde die C. curticei Eiausscheidung durch Esparsettenheu um 74 % (P < 0,001) und durch Esparsettensilage um 81 % (P < 0,001) reduziert. Es handelt sich bei dieser Studie um die erste, welche die für den Praxiseinsatz wichtige Verwendung einer konservierten, tanninhaltigen Futterpflanze gezielt gegen H. contortus und C. curticei untersucht

Unterschiede im Leistungsverhalten von Gesunden zwischen Fahrrad- und Laufbandergometrie

Problemstellung: Zur Überprüfung des Ausdauertrainingseffektes während einer Terrainkur ist die Laufbandergometrie der Belastung auf dem Fahrrad vorzuziehen, da hierbei dieselben Muskeln beansprucht werden wie während des Trainings. Allerdings gelingt es wegen der frühzeitigen subjektiven Erschöpfung der Kurpatienten nur selten, Milchsäurewerte oberhalb der aeroben/anaeroben Schwelle zu erzielen. Gegenstand: Die vorliegende Studie mit Gesunden befaßt sich mit den Fragen, warum sich Kurpatienten auf dem Laufband metabolisch betrachtet nicht genügend ausbelasten lassen, und welches die entscheidenden Unterschiede in der Art der Belastung zwischen Laufband- und Fahrradergometer sind. Versuchsplan: 27 gesunde Testpersonen wurden auf dem Fahrrad- und Laufbandergometer submaximal belastet. Die Belastung wurde nach einem standardisierten Verfahren in 0,33 W/kg-Schritten gesteigert. Wichtigste Meßparameter waren Herzfrequenz und Milchsäurespiegel. Ergebnisse: Während der Laufbandergometrie lagen Herzfrequenz und Sauerstoffbedarf des Myokards zum Erreichen der gleichen Wattstufe signifikant höher (1,0 W/kg:p≤0,001; 1,33 W/kg: p≤0,01; 2,0 W/kg: n.s.; Erholung: p≤0,001) als auf dem Fahrrad. Dagegen zeigte sich bei gegebener Herzfrequenz von 130/min und bei derselben höchsten erreichten Wattstufe auf dem Laufband ein um 0,7 mmol/1 bzw. l,0mmol/l signifikant (p≤0,01) niedrigerer Laktatanstieg als auf dem Fahrradergometer. Schlußfolgerung: Die Ergebnisse zeigten somit eine stärkere kardiale Ausschöpfung auf dem Laufbandergometer. Die aeroben/anaerobe Schwelle wird dagegen auf dem Laufband später als auf dem Fahrrad erreicht

Development of a monitoring instrument to assess the performance of the Swiss primary care system.

The Swiss health system is customer-driven with fee-for-service paiement scheme and universal coverage. It is highly performing but expensive and health information systems are scarcely implemented. The Swiss Primary Care Active Monitoring (SPAM) program aims to develop an instrument able to describe the performance and effectiveness of the Swiss PC system. Based on a Literature review we developed a conceptual framework and selected indicators according to their ability to reflect the Swiss PC system. A two round modified RAND method with 24 inter-/national experts took place to select primary/secondary indicators (validity, clarity, agreement). A limited set of priority indicators was selected (importance, priority) in a third round. A conceptual framework covering three domains (structure, process, outcome) subdivided into twelve sections (funding, access, organisation/ workflow of resources, (Para-)Medical training, management of knowledge, clinical-/interpersonal care, health status, satisfaction of PC providers/ consumers, equity) was generated. 365 indicators were pre-selected and 335 were finally retained. 56 were kept as priority indicators.- Among the remaining, 199 were identified as primary and 80 as secondary indicators. All domains and sections are represented. The development of the SPAM program allowed the construction of a consensual instrument in a traditionally unregulated health system through a modified RAND method. The selected 56 priority indicators render the SPAM instrument a comprehensive tool supporting a better understanding of the Swiss PC system's performance and effectiveness as well as in identifying potential ways to improve quality of care. Further challenges will be to update indicators regularly and to assess validity and sensitivity-to-change over time

Hybridisation between red deer (Cervus elaphus) and Japanese sika (C. nippon) on the Kintyre Peninsula, Scotland

Hybridisation between introduced and endemic species causes conservation concerns, but also provides us with an opportunity to study the dynamics of gene flow between two species as they first meet. Japanese sika deer (Cervus nippon) were introduced to the British Isles at a number of locations at the beginning of the 20th century. In the intervening time, sika have spread and their range now extends across approximately 40% of Scotland, where they overlap with that of native red deer (C. elaphus), with which they hybridise. In this study we focus on the consequences of one particular introduction that took place at Carradale, on the Kintyre Peninsula in 1893. First, I assessed the current state of hybridisation using a sample of 735 red and sika deer samples collected in 2006/7 from forestry blocks throughout the Kintyre Peninsula. Genetic analysis was conducted with a panel of 22 highly differentiated microsatellite loci and one mtDNA marker. Population admixture analysis of the microsatellite data was conducted with the Bayesian clustering programme STRUCTURE. Over most of the study area, levels of introgression into red and sika deer were low and were consistent with a scenario of very occasional F1 hybridisation followed by backcrossing. There was, however, one forestry block where 43% of individuals could be defined as hybrids. Second, I developed a branching process model of introgression via backcrossing, to assess whether variation in introgression across microsatellite loci could be interpreted as a signature of selection, or could in fact be attributed to stochastic processes. If only a few hybridisation events have contributed to the hybridising population, the pattern of introgression, even with a large number of genetic markers, will be highly stochastic. This pattern of neutral variation in introgression can have high enough variance that it could be mistaken for selection. Therefore, even if strong selection is acting, it may not be possible to distinguish its effects from neutral variation. Third, I analysed trends in hybridisation and introgression over 15 years on the peninsula, through analysis of a dataset of 1513 red and sika deer samples at 20 microsatellite and a mtDNA marker. There was little evidence of change in the extent of hybridisation and introgression over time. MtDNA introgression was predominantly from red deer into sika. Recent introgression into sika on the peninsula can be explained by a very small number of F1 hybridisation events (~10) via analysis of the number of alleles that have introgressed from polymorphic red deer into the genetically homogenous sika population (a similar analysis cannot be conducted for introgression into red deer). Finally, I conducted a regression analysis of genetic hybrid scores against phenotypic traits to assess the effect of hybridisation on phenotype. Hybridisation has caused changes in the weight of sika-like deer and red-like females. Hybridisation has caused changes in incisor arcade breadth of both populations and jaw length (a proxy for skeletal size) in sika-like females. However, there is no evidence that hybridisation has caused changes in kidney fat (a measure of condition) or pregnancy rates in either population. In conclusion, even a small number of F1 hybridisation events can lead to extensive introgression and the timing and spatial distribution of these events is likely to have a large impact on the structure of a recently hybridising population - stochastic factors dominate both the distribution of hybrid individuals and the distribution of the genes that introgress following a hybridisation event. In red deer and sika deer, increasing phenotypic similarities of the two populations caused by hybridisation are likely to facilitate further breakdown between the two species. It is possible that breakdown in assortative mating between the two species could occur across their range

Development and evaluation of double locus sequence typing for molecular epidemiological investigations of Clostridium difficile.

Despite the development of novel typing methods based on whole genome sequencing, most laboratories still rely on classical molecular methods for outbreak investigation or surveillance. Reference methods for Clostridium difficile include ribotyping and pulsed-field gel electrophoresis, which are band-comparing methods often difficult to establish and which require reference strain collections. Here, we present the double locus sequence typing (DLST) scheme as a tool to analyse C. difficile isolates. Using a collection of clinical C. difficile isolates recovered during a 1-year period, we evaluated the performance of DLST and compared the results to multilocus sequence typing (MLST), a sequence-based method that has been used to study the structure of bacterial populations and highlight major clones. DLST had a higher discriminatory power compared to MLST (Simpson's index of diversity of 0.979 versus 0.965) and successfully identified all isolates of the study (100 % typeability). Previous studies showed that the discriminatory power of ribotyping was comparable to that of MLST; thus, DLST might be more discriminatory than ribotyping. DLST is easy to establish and provides several advantages, including absence of DNA extraction [polymerase chain reaction (PCR) is performed on colonies], no specific instrumentation, low cost and unambiguous definition of types. Moreover, the implementation of a DLST typing scheme on an Internet database, such as that previously done for Staphylococcus aureus and Pseudomonas aeruginosa ( http://www.dlst.org ), will allow users to easily obtain the DLST type by submitting directly sequencing files and will avoid problems associated with multiple databases

Increased genetic marker density reveals high levels of admixture between red deer and introduced Japanese sika in Kintyre, Scotland

Hybridization is a natural process at species range boundaries, but increasing numbers of species are hybridizing due to direct or indirect human activities. In such cases of anthropogenic hybridization, subsequent introgression can threaten the survival of native species. To date, many such systems have been studied with too few genetic markers to assess the level of threat resulting from advanced backcrossing. Here, we use 44,999 single nucleotide polymorphisms (SNPs) and the ADMIXTURE program to study two areas of Scotland where a panel of 22 diagnostic microsatellites previously identified introgression between native red deer (Cervus elaphus) and introduced Japanese sika (Cervus nippon). In Kintyre, we reclassify 26% of deer from the pure species categories to the hybrid category whereas in the NW Highlands we only reclassify 2%. As expected, the reclassified individuals are mostly advanced backcrosses. We also investigate the ability of marker panels selected on different posterior allele frequency criteria to find hybrids assigned by the full marker set and show that in our data, ancestry informative markers (i.e. those that are highly differentiated between the species, but not fixed) are better than diagnostic markers (those markers that are fixed between the species) because they are more evenly distributed in the genome. Diagnostic loci are concentrated on the X chromosome to the detriment of autosomal coverage