bluuebunny/crossref_metadata_2025 huggingface dataset:
Microspatial genetic heterogeneity and gene flow in stray cats (Felis catus L.): a comparison of coat colour and microsatellite loci
Authors: Say Ludovic , Bonhomme François , Desmarais Eric , Pontier Dominique
Published: May 2003

We analysed levels of genetic differentiation between nine local urban colonies of stray cats using eight coat colour and nine microsatellite loci. Both types of markers revealed a strong differentiation between colonies (FST = 0.15 and 0.09 for coat colour and microsatellite loci, respectively). Three coat colour loci showed extreme levels of genetic differentiation comparatively to other loci and are strongly suspected to be under divergent selective pressures. Microsatellite loci showed significant heterozygote deficiency within colonies (FIS = 0.14), suggesting that coat colour loci are not appropriate to investigate genetic structure at a fine scale because coat colour allele frequencies are based on Hardy–Weinberg equilibrium. The reported pattern conformed to that predicted from the social structuring of cat colonies: aggressive exclusion of immigrants, inbreeding and very low dispersal rate.
How much effort is required to isolate nuclear microsatellites from plants?
Authors: Squirrell J. , Hollingsworth P. M. , Woodhead M. , Russell J. , Lowe A. J. , Gibby M. , Powell W.
Published: May 2003

The attributes of codominance, reproducibility and high resolution have all contributed towards the current popularity of nuclear microsatellites as genetic markers in molecular ecological studies. One of their major drawbacks, however, is the development phase required to obtain working primers for a given study species. To facilitate project planning, we have reviewed the literature to quantify the workload involved in isolating nuclear microsatellites from plants. We highlight the attrition of loci at each stage in the process, and the average effort required to obtain 10 working microsatellite primer pairs.
Fluorescencein situhybridization: a new method for determining primary sex ratio in ants
Authors: De Menten L. , Niculita H. , Gilbert M. , Delneste D. , Aron S.
Published: May 2003

The haplodiploid sex determining system in Hymenoptera, whereby males develop from haploid eggs and females from diploid eggs, allows females to control the primary sex ratio (the proportion of each sex at oviposition) in response to ecological and/or genetic conditions. Surprisingly, primary sex ratio adjustment by queens in eusocial Hymenoptera has been poorly studied, because of difficulties in sexing the eggs laid. Here, we show that fluorescencein situhybridization (FISH) can be used to accurately determine the sex (haploid or diploid) of eggs, and hence the primary sex ratio, in ants. We first isolated the homologue coding sequences of theabdominal‐Agene from 10 species of 8 subfamilies of Formicidae. Our data show that the nucleotide sequence of this gene is highly conserved among the different subfamilies. Second, we used a sequence of 4.5 kbp from this gene as a DNA probe for primary sex ratio determination by FISH. Our results show that this DNA probe hybridizes successfully with its complementary DNA sequence in all ant species tested, and allows reliable determination of the sex of eggs. Our proposed method should greatly facilitate empirical tests of primary sex ratio in ants.
Genetic consequences of the ice ages on nurseries of the bat Myotis myotis: a mitochondrial and nuclear survey
Authors: Ruedi M. , Castella V.
Published: May 2003

Analyses of mitochondrial DNA (mtDNA) control region polymorphism and of variation at 10 nuclear microsatellite loci were used to investigate the mechanisms and genetic consequences of postglacial expansion of Myotis myotis in Europe. Initial sampling consisted of 480 bats genotyped in 24 nursery colonies arranged along a transect of ≈ 3000 km. The phylogeographical survey based on mtDNA sequences revealed the existence of major genetic subdivisions across this area, with several suture zones between haplogroups. Such zones of secondary contact were found in the Alps and Rhodopes, whereas other potential barriers to gene flow, like the Pyrenees, did not coincide with genetic discontinuities. Areas of population admixture increased locally the genetic diversity of colonies, which confounded the northward decrease in nucleotide diversity predicted using classical models of postglacial range expansion. However, when analyses were restricted to a subset of 15 nurseries originating from a single presumed glacial refugium, mtDNA polymorphism did indeed support a northwards decrease in diversity. Populations were also highly structured (ΦST = 0.384). Conversely, the same subset of colonies showed no significant latitudinal decrease in microsatellite diversity and much less population structure (FST = 0.010), but pairwise genetic differentiation at these nuclear markers was strongly correlated with increasing geographical distance. Together, this evidence suggests that alleles carried via male bats have maintained enough nuclear gene flow to counteract the effects of recurrent bottlenecks generally associated with recolonization processes. As females are highly philopatric, we argue that the maternally transmitted mtDNA marker better reflects the situation of past, historical gene flow, whereas current levels of gene flow are better reflected by microsatellite markers.
Population genetic structure of the toadBufo woodhousii: an empirical assessment of the effects of haplotype extinction on nested cladistic analysis
Authors: Masta S. E. , Laurent N. M. , Routman E. J.
Published: May 2003

Nested cladistic analysis (NCA) is increasingly being used to infer historical population‐level processes, including population fragmentation, range expansion and long‐distance colonization. However, the effects on interpretation of NCA inferences of stochastic extinction of haplotypes due to genetic drift (lineage sorting), or of haplotype loss via localized biotic or climatic influences, have not been thoroughly explored. We provide empirical evidence suggesting that NCA may misinterpret population history when haplotypes or haplotype groups from one clade are replaced by those of another clade. We do so by using NCA to analyse mitochondrial sequences from the toadBufo woodhousiifrom 45 locations spanning the Great Plains and southwestern USA. Portions of this region were glaciated and/or desertified in the late Pleistocene and early Holocene, and hence uninhabitable for plains‐dwelling organisms. Although NCA inferences of isolation‐by‐distance and gradual range expansion inB. woodhousiiare compatible with expectations based on climatic data and toad biology, NCA also detected several instances of long‐distance movement. Such movement seems unlikely, given the low vagility of this species. We conclude that inferences of long‐distance colonization likely result from extinction of haplotypes in intervening areas. We suggest using additional methods to look for congruent inferences, and amending the NCA inference key, to help avoid misinterpretations resulting from haplotype extinction.
Early male reproductive advantage, multiple paternity and sperm storage in an amphibian aggregate breeder
Authors: Tennessen J. A. , Zamudio K. R.
Published: May 2003

We tested whether the order in which males encounter females affects reproductive fitness in spotted salamanders (Ambystoma maculatum). Using mating chambers in the field, we allowed one male access to a female before a second male. We then used four microsatellite markers in paternity analyses of the resulting larvae. First males sired a significantly larger number of offspring than second males, suggesting that male reproductive success is greatly enhanced by early arrival at breeding ponds. Multiple paternity was common among clutches, and frequently larvae were assigned to unidentified males that had not been in the chambers. Sperm from these males had either been stored by females for a year or obtained more recently at other breeding sites.
Estimation of pairwise relatedness between individuals and characterization of isolation‐by‐distance processes using dominant genetic markers
Authors: Hardy Olivier J.
Published: May 2003

A new estimator of the pairwise relatedness coefficient between individuals adapted to dominant genetic markers is developed. This estimator does not assume genotypes to be in Hardy–Weinberg proportions but requires a knowledge of the departure from these proportions (i.e. the inbreeding coefficient). Simulations show that the estimator provides accurate estimates, except for some particular types of individual pairs such as full‐sibs, and performs better than a previously developed estimator. When comparing marker‐based relatedness estimates with pedigree expectations, a new approach to account for the change of the reference population is developed and shown to perform satisfactorily. Simulations also illustrate that this new relatedness estimator can be used to characterize isolation by distance within populations, leading to essentially unbiased estimates of the neighbourhood size. In this context, the estimator appears fairly robust to moderate errors made on the assumed inbreeding coefficient. The analysis of real data sets suggests that dominant markers (random amplified polymorphic DNA, amplified fragment length polymorphism) may be as valuable as co‐dominant markers (microsatellites) in studying microgeographic isolation‐by‐distance processes. It is argued that the estimators developed should find major applications, notably for conservation biology.
Checking the geographical origin of oak wood: molecular and statistical tools
Authors: Deguilloux M. F. , Pemonge M. H. , Bertel L. , Kremer A. , Petit R. J.
Published: May 2003

New methods for better identification of timber geographical origin would constitute an important technical element in the forest industry, for phytosanitary certification procedures or in the chain of custody developed for the certification of timber from sustainably managed forests. In the case of the European white oaks, a detailed reference map of chloroplast (cp) DNA variation across the range exists, and we propose here to use the strong geographical structure, characterized by a differentiation of western vs. eastern populations, for the purpose of oak wood traceability. We first developed cpDNA markers permitting the characterization of haplotype on degraded DNA obtained from wood samples. The techniques were subsequently validated by confirming the full correspondence between genotypes obtained from living tissues (buds) and from wood collected from the same individual oak. Finally, a statistical procedure was used to test if the haplotype composition of a lot of wood samples is consistent with its presumed geographical origin. Clearly, the technique cannot permit the unambiguous identification of wood products of unknown origin but can be used to check the conformity of genetic composition of wood samples with the region of alleged origin. This could lead to major applications not only in the forest industry but also in archaeology or in palaeobotany.
Characterization of Pacific golden chanterelle (Cantharellus formosus) genet size using co‐dominant microsatellite markers
Authors: Dunham S. M. , Kretzer A. , Pfrender M. E.
Published: May 2003

We characterized five co‐dominant microsatellite markers and used them to study Pacific golden chanterelle (Cantharellus formosus) genet size and its relation to forest age and disturbance. Fruit‐bodies were mapped in and collected from nine replicate study plots in old‐growth, recently thinned, and unthinned 40–60‐year‐old second‐growth stands dominated by Douglas fir (Pseudotsuga menziesii). Information from microsatellite loci, combined with random fragment length polymorphism analysis of the nuclear DNA internal transcribed spacer indicates that putative ‘C. formosus’ fruit‐body collections may include a cryptic chanterelle species. Small genets were characterized for both genetic types with mean maximum widths of 3.2 ± 3.6 m for C. formosus and 1.5 ± 1.7 m for the alternative genetic group. Variance in genet size was high and some multilocus genotypes were observed on multiple plots separated by 0.3 km or more, indicating that genets were not fully resolved by the loci described here. There was no evidence that genet size differed across the three disturbance treatments.
Queen control over reproductive decisions — no sexual deception in the ant Lasius niger
Authors: Jemielity Stephanie , Keller Laurent
Published: May 2003

Queen–worker conflicts in social insect societies have received much attention in the past decade. In many species workers modify the colony sex ratio to their own advantage or produce their own male offspring. In some other species, however, queens seem to be able to prevent workers from making selfish reproductive decisions. So far, little effort has been made to find out how queens may keep control over sex ratio and male parentage. In this study we use a Lasius niger population under apparent queen control to show that sexual deception cannot explain queen dominance in this population. The sexual deception hypothesis postulates that queens should prevent workers from discriminating against males by disguising male brood as females. Contrary to the predictions of this hypothesis, we found that workers are able to distinguish male and female larvae early in their development: in early spring workers generally placed only either female or male larvae in the uppermost chambers of the nest, although both types of larvae must have been present. At this time males were only at 11% of their final dry weight, a developmental stage at which (according to two models) workers would still have benefited from replacing queen‐produced males by females or worker‐produced males. This study thus demonstrates that sexual deception cannot account for the apparent queen control over colony sex ratio and male parentage in L. niger.
The influence of the Miocene Mediterranean desiccation on the geographical expansion and genetic variation of Androcymbium gramineum (Cav.) McBride (Colchicaceae)
Authors: Caujapé‐Castells Juli , Jansen Robert K.
Published: May 2003

Chloroplast DNA (cpDNA) restriction site and isozyme data were combined to explore the spatial–temporal influence of the Messinian desiccation in the Mediterranean on the disjunct distribution of Androcymbium gramineum in Almería and Morocco (north and south of the straits of Gibraltar, respectively). Lack of evidence for different selective pressures, divergence time estimates based on the calibration of the isozyme molecular clock with the cpDNA data, the basal position of Almerian populations in the A. gramineum clade, and the much higher isozyme polymorphism in Almería suggest that (i) only a southern European range of A. gramineum existed before the Messinian [≈ 11.2 million years ago (Ma), in the middle Miocene] and (ii) the desiccation of the Mediterranean basin about 5.5–4.5 Ma induced the migration of A. gramineum from Almería to Morocco (between 4.9 and 4.6 Ma, according to our time estimates). After the split into two allopatric units following the refilling of the Mediterranean, the major influence of drift associated with Plio‐Pleistocene recurrent glaciation cycles and range expansions/contractions probably fostered the substantial interpopulation genetic differentiation observed within Almería (CGST = 0.41, average DNei = 0.185) and, to a lesser extent, within Morocco (CGST = 0.24, average DNei = 0.089), but did not hinder the maintenance of considerable levels of genetic variation in either geographical area (A = 2.14, HE = 0.230 and A = 1.90, HE = 0.213, respectively).
Queen‐controlled sex ratios and worker reproduction in the bumble bee Bombus hypnorum, as revealed by microsatellites
Authors: Brown M. J. F. , Schmid‐Hempel R. , Schmid‐Hempel P.
Published: May 2003

Social insect colonies provide model systems for the examination of conflicts among parties with different genetic interests. As such, they have provided the best tests of inclusive fitness theory. However, much remains unknown about in which party's favour such conflicts are resolved, partly as a result of the only recent advent of the molecular tools needed to examine the outcome of these conflicts. Two key conflicts in social insect colonies are over control of the reproductive sex ratio and the production of male offspring. Most studies have examined only one of these conflicts but in reality they occur in tandem and may influence each other. Using microsatellite analyses, the outcome of conflict over sex ratios and male production was examined in the bumble bee, Bombus hypnorum. The genotypes were determined for mother queens, their mates and males for each of 10 colonies. In contrast to other reports of mating frequency in this species, all of the queens were singly mated. The population sex ratio was consistent with queen control, suggesting that queens are winning this conflict. In contrast, workers produced over 20% of all males in queen‐right colonies, suggesting that they are more effective in competing over male‐production. Combining these results with previous work, it is suggested that worker reproduction is a labile trait that may well impose only small costs on queen fitness.
Was there a second adaptive radiation of giant tortoises in the Indian Ocean? Using mitochondrial DNA to investigate speciation and biogeography of Aldabrachelys (Reptilia, Testudinidae)
Authors: Austin Jeremy J. , Arnold E. Nicholas , Bour Roger
Published: May 2003

A radiation of five species of giant tortoises (Cylindraspis) existed in the southwest Indian Ocean, on the Mascarene islands, and another (of Aldabrachelys) has been postulated on small islands north of Madagascar, from where at least eight nominal species have been named and up to five have been recently recognized. Of 37 specimens of Madagascan and small‐island Aldabrachelys investigated by us, 23 yielded significant portions of a 428‐base‐pair (bp) fragment of mitochondrial (cytochrome b and tRNA‐Glu), including type material of seven nominal species (A. arnoldi, A. dussumieri, A. hololissa, A. daudinii, A. sumierei, A. ponderosa and A. gouffei). These and nearly all the remaining specimens, including 15 additional captive individuals sequenced previously, show little variation. Thirty‐three exhibit no differences and the remainder diverge by only 1–4 bp (0.23–0.93%). This contrasts with more widely accepted tortoise species which show much greater inter‐ and intraspecific differences. The non‐Madagascan material examined may therefore only represent a single species and all specimens may come from Aldabra where the common haplotype is known to occur. The present study provides no evidence against the Madagascan origin for Aldabra tortoises suggested by a previous molecular phylogenetic analysis, the direction of marine currents and phylogeography of other reptiles in the area. Ancient mitochondrial DNA from the extinct subfossil A. grandidieri of Madagascar differs at 25 sites (5.8%) from all other Aldabrachelys samples examined here.
Edaphic races and phylogenetic taxa in the Lasthenia californica complex (Asteraceae: Heliantheae): an hypothesis of parallel evolution
Authors: Rajakaruna Nishanta , Baldwin Bruce G. , Chan Raymund , Desrochers Andrée M. , Bohm Bruce A. , Whitton Jeannette
Published: May 2003

Lasthenia californica sensu Ornduff consists of two races that differ in their flavonoid pigments and edaphic tolerances. Recent phylogenetic studies of Lasthenia have revealed that members of L. californica sensu Ornduff belong to two phylogenetic species. The relationship of the edaphic races to these new species and to each other is the focus of this study. Characterization of flavonoid profiles and phylogenetic placement of 33 populations demonstrates that races and phylogenetic taxa are not concordant, suggesting that one or both edaphic races evolved in parallel in the two clades. We hypothesize an edaphically linked ecological role for flavonoid differences that first revealed the existence of two races.
High multiple paternity and low last‐male sperm precedence in a hermaphroditic planarian flatworm: consequences for reciprocity patterns
Authors: Pongratz N. , Michiels N. K.
Published: May 2003

It is difficult to predict a priori how mating success translates into fertilization success in simultaneous hermaphrodites with internal fertilization. Whereas insemination decisions will be determined by male interests, fertilization will depend on female interests, possibly leading to discrepancies between insemination and fertilization patterns. The planarian flatworm Schmidtea polychroa, a simultaneous hermaphrodite in which mating partners trade sperm was studied. Sperm can be stored for months yet individuals mate frequently. Using microsatellites, maternity and paternity data were obtained from 748 offspring produced in six groups of 10 individuals during four weeks. Adults produced young from four mates on average. Reciprocal fertilization between two mates was found in only 41 out of 110 registered mate combinations, which is clearly less than what is predicted from insemination patterns. Multiple paternity was high: > 80% of all cocoons had two to five fathers for only three to five offspring per cocoon. Because animals were collected from a natural population, 28% of all hatchlings were sired by unknown sperm donors in the field, despite a 10‐day period of acclimatization and within‐group mating. This percentage decreased only moderately throughout the experiment, showing that sperm can be stored and used for at least a month, despite frequent mating and sperm digestion. The immediate paternity a sperm donor could expect to obtain was only about 25%. Male reproductive success increased linearly with the number of female partners, providing support for Bateman's principle in hermaphrodites. Our results suggest that hermaphrodites do not trade fertilizations when trading sperm during insemination, lending support to the view that such conditional sperm exchange is driven by exchange of resources.
Measuring gene flow from two birdsfoot trefoil (Lotus corniculatus) field trials using transgenes as tracer markers
Authors: De Marchis F. , Bellucci M. , Arcioni S.
Published: May 2003

Genetic engineering is becoming a useful tool in the improvement of plants but concern has been expressed about the potential environmental risks of releasing genetically modified (GM) organisms into the environment. Attention has focused on pollen dispersal as a major issue in the risk assessment of transgenic crop plants. In this study, pollen‐mediated dispersal of transgenes via cross‐fertilization was examined. Plants of Lotus corniculatus L. transformed with either the Escherichia coli asparagine synthetase gene asnA or the β‐glucuronidase gene uidA, were used as the pollen donor. Nontransgenic plants belonging to the species L. corniculatus L., L. tenuis Waldst. and Kit. ex Willd, and L. pedunculatus Cav., were utilized as recipients. Two experimental fields were established in two areas of central Italy. Plants carrying the uidA gene were partially sterile, therefore only the asnA gene was used as a tracer marker. No transgene flow between L. corniculatus transformants and the nontransgenic L. tenuis and L. pedunculatus plants was detected. As regards nontransgenic L. corniculatus plants, in one location flow of asnA transgene was detected up to 18 m from the 1.8 m2 donor plot. In the other location, pollen dispersal occurred up to 120 m from the 14 m2 pollinating plot.
Demographic expansion of parasitic nematodes of livestock based on mitochondrial DNA regions that conflict with the infinite‐sites model
Authors: Mes T. H. M.
Published: May 2003

Mitochondrial ND4 sequences of populations of four species of parasitic nematodes of livestock were subjected to demographic analyses. Deviation from selective neutrality was detectable using the frequency spectrum of segregating sites and highly negative neutrality statistics. However, the mitochondrial data sets do not comply with the infinite‐sites model that underlies these tests, and as a consequence, it was not established whether these features are solely a result of population expansion, or whether aspects of the molecular evolution of these mitochondrial regions are also involved. Coalescent analyses based on Fu's Fs neutrality test, which incorporated estimates of rate heterogeneity, the transition–transversion ratio and nucleotide bias, as well as analyses that are fairly robust to deviations from the infinite‐sites model supported population expansion. Also analyses that do not depend on the infinite‐sites model suggested historical population expansion of these nematodes. The very similar time since expansion, the absence of signatures of positive selection in ND4 and the logical association with human demography imply that selective sweeps of mitochondrial variants are less probable, and that expansion is the most likely scenario for the parasitic nematodes of livestock. The methods used to characterize the expansion have different assumptions and emphasize different aspects of expansions. The resulting restrictions on the interpretation of expansions are outlined.
Evidence for interspecies transmission of viruses in natural populations of filamentous fungi in the genus Cryphonectria
Authors: Liu Y.‐C. , Linder‐Basso D. , Hillman B. I. , Kaneko S. , Milgroom M. G.
Published: May 2003

Interspecies transmission is a significant evolutionary event that has allowed a variety of pathogens to invade new host species. We investigated interspecies transmission of viruses between the chestnut blight fungus, Cryphonectria parasitica, and a sympatric unidentified Cryphonectria species in Japan. Two isolates of Cryphonectria sp. were found to contain Cryphonectria hypovirus 1 (CHV‐1), which has been typically found in C. parasitica. Three lines of evidence support the hypothesis of interspecies transmission of CHV‐1. First, host species occur sympatrically and therefore have the opportunity to come into physical contact. Second, we transmitted CHV‐1 between species experimentally in the laboratory. Third, phylogenetic analysis of 476 bp of the ORF B region of CHV‐1 showed that sequences from Cryphonectria sp. were more closely related to those from C. parasitica than to each other. Local geographical subdivision of virus sequences from both host species argues against the alternative hypothesis of independent evolution of CHV‐1 since speciation of their hosts. Based on these findings, we rule out the hypotheses that CHV‐1 diverged from viruses in a common ancestor of the hosts, or that ancestral polymorphisms in CHV‐1 persisted in the two host taxa. Estimating the direction and frequency of interspecies transmission in nature will require more extensive samples of CHV‐1 from both host species.
Reliable microsatellite genotyping of the Eurasian badger (Meles meles) using faecal DNA
Authors: Frantz A. C. , Pope L. C. , Carpenter P. J. , Roper T. J. , Wilson G. J. , Delahay R. J. , Burke T.
Published: May 2003

The potential link between badgers and bovine tuberculosis has made it vital to develop accurate techniques to census badgers. Here we investigate the potential of using genetic profiles obtained from faecal DNA as a basis for population size estimation. After trialling several methods we obtained a high amplification success rate (89%) by storing faeces in 70% ethanol and using the guanidine thiocyanate/silica method for extraction. Using 70% ethanol as a storage agent had the advantage of it being an antiseptic. In order to obtain reliable genotypes with fewer amplification reactions than the standard multiple‐tubes approach, we devised a comparative approach in which genetic profiles were compared and replication directed at similar, but not identical, genotypes. This modified method achieved a reduction in polymerase chain reactions comparable with the maximum‐likelihood model when just using reliability criteria, and was slightly better when using reliability criteria with the additional proviso that alleles must be observed twice to be considered reliable. Our comparative approach would be best suited for studies that include multiple faeces from each individual. We utilized our approach in a well‐studied population of badgers from which individuals had been sampled and reliable genotypes obtained. In a study of 53 faeces sampled from three social groups over 10 days, we found that direct enumeration could not be used to estimate population size, but that the application of mark–recapture models has the potential to provide more accurate results.
Frontotemporal and motor neurone degeneration with neurofilament inclusion bodies: additional evidence for overlap between FTD and ALS
Authors: Bigio E. H. , Lipton A. M. , White C. L. , Dickson D. W. , Hirano A.
Published: May 2003

We present the case of a patient who had clinical frontal lobe dementia without apparent motor neurone disease (MND), with pathologic findings not typical of any single currently classified frontotemporal degeneration (FTD). At autopsy, the brain had frontal and temporal atrophy with neuronal loss, gliosis, and superficial spongiosis, typical of all FTDs. There were at least three different morphologic types of intracytoplasmic neuronal inclusions in a variety of brain and brainstem regions, including the hippocampal dentate gyrus and pyramidal neurones, the neocortex (in particular, the motor cortex), basal ganglia, thalamus, subthalamic nucleus, basis pontis, and inferior olivary nuclei. Inclusions had the morphologies of Pick‐like bodies, pleomorphic inclusions, and hyaline conglomerate (HC)‐like inclusions. None of these were positive with tau immunostains. Pick‐like bodies in the dentate gyrus were labelled with ubiquitin. The pleomorphic inclusions in the neocortex and dentate gyrus and the HC‐like inclusions in the motor and parietal cortex were strongly positive with immunostains for neurofilament. We discuss the differential diagnosis and compare this case with those disorders to which it is most similar. In particular, we compare the unique neurofilament‐positive inclusions to the inclusions of FTD‐MND, to Pick bodies, and to the basophilic and HC inclusions that are occasionally seen in amytrophic lateral sclerosis (ALS). Although FTD‐MND may be found in ALS, the findings in this case may have additional implications for a link between FTD and ALS.
The practice of antenatal care: comparing four study sites in different parts of the world participating in the WHO Antenatal Care Randomised Controlled Trial
Authors: Piaggio G. , Ba’aqeel H. , Bergsjø P. , Carroli G. , Farnot U. , Lumbiganon P. , Pinol A. , Villar J.
Published: May 2003

In the preparation of a randomised controlled trial to evaluate a new programme of antenatal care (ANC) in different parts of the world, we conducted a baseline survey of the ANC procedures in all 53 clinics participating in the trial. There were two components of this survey: (1) description of clinic characteristics and services offered: the staff of each clinic was interviewed and direct observation was made by field supervisors, and (2) the actual use of services by pregnant women attending these clinics: we reviewed a random sample of 2913 clinical histories.

  The clinical units surveyed were offering most of the activities, screening, laboratory tests and interventions recommended as effective according to the Cochrane Pregnancy and Childbirth Database (PCD), although some of these were not available in some sites. On the other hand, some tests and interventions that are considered not effective according to these criteria are reportedly offered. There was a difference across sites in the availability and offer to low‐risk women of vaginal examination, evaluation of pelvic size, dental examination, external version for breech presentation and formal risk score classification, and a notable difference in the type of principal provider of ANC. There was a large variation in the actual use of screening and laboratory tests and interventions that should be offered to all women according to Cochrane PCD criteria: some of these are simply not available in a site; others are available, but only a fraction of women attending the clinics are receiving them. The participating sites all purport to follow the traditional ‘Western’ schedule for ANC, but in three sites we found that a high percentage of women initiate their ANC after the first trimester, and therefore do not have either the recommended minimum number of visits during pregnancy or the minimum first trimester evaluation. It is concluded that the variability and heterogeneity of ANC services provided in the four study sites are disturbing to the profession and cast doubts on the rationale of routine ANC.
Managing data for a randomised controlled clinical trial: experience from the WHO Antenatal Care Trial
Authors: Pinol A. , Bergel E. , Chaisiri K. , Diaz E. , Gandeh M.
Published: May 2003

The World Health Organisation, in collaboration with four developing countries, is conducting a randomised controlled clinical trial to evaluate a new programme of antenatal care. In a city or region in Argentina, Cuba, Saudi Arabia and Thailand, 53 clinical units were randomly allocated to provide either the new programme or the programme currently in use. This paper describes the organisation of the data management system used to collect the data.

  Each woman participating in the trial is uniquely identified, and information such as her name, address and expected delivery date is recorded in the trial ‘subject number list’. If the clinic belongs to the intervention group, information about the woman's eligibility is recorded on the classification form. Details of the outcome of the pregnancy are indicated on two additional case report forms: the antenatal hospital admission form and the summary form. When forms are completed by the investigators, they are submitted to the country data coordinating centre (CDCC).

  The CDCCs are responsible for the processing of the country study forms. This includes verification of the batch of forms, data capture into computer files, data verification, data validation, production of query sheets for data problems, maintenance and updating of study master files. All operations on data such as additions or modifications are performed using transaction processing. At monthly intervals, recruitment reports and transaction files are sent to the trial coordinating centre in Geneva. All transaction files are processed to accumulate data on the trial's consolidated master files. A monthly report including number of women recruited in the trial, adverse events reported by the countries, recruitment charts by clinic and analyses on eligible women in the intervention group is prepared and submitted to the data safety and monitoring committee.

  A workshop was organised in 1995, before the start of the trial, to introduce the data management system to the four participating countries. Annual site visits were made to each CDCC to monitor progress. Additional visits were made when major or critical problems could not be solved by the CDCC. At the closure of data collection, a visit is made to review and assess all data management procedures including form filling, maintenance of registers, computer files, query sheets, data modifications. In addition, final cleaning of the data is performed, and an analysis file is produced for inclusion in the centralised trial analyses and in the country‐specific analyses.

  Based on the experience gained in this trial, the decentralised data management model can be advocated only if CDCCs that will be involved in the trial are already in place with competent and experienced staff. Uniformity of the data management system and of standard operating procedures across countries is also a crucial issue for the effective management of the data collection phase.
Monitoring data and safety in the WHO Antenatal Care Trial
Authors: Bergsjø P. , Bréart G. , Morabia A.
Published: May 2003

A committee to monitor data and safety in a large clinical trial should have members with expertise in biostatistics, epidemiology and the clinical field relevant to the trial. Its mandate should cover both logistics and the safety of study subjects, issues which to some extent overlap. While a steering committee and field staff members ideally should be blinded to the experimental and control arms of a randomised clinical trial, the data and safety monitoring committee (DSMC) should have full access to interim trial data to fulfil its role as watchdog. One initial question to be resolved concerns if and when to advise stopping a trial because of danger to study subjects, or on the other hand obvious benefits, in one trial arm. The DSMC of the WHO Antenatal Care Trial decided not to establish any definite stopping rules before implementation. After a scrutiny of the adopted procedures for data collection and handling, the DSMC received monthly reports of recruitment, and individual summary reports of three adverse events by site and trial arm: maternal deaths, fetal deaths and cases of eclampsia. At the time of writing (December 1997) recruitment to the trial is almost complete, but data collection will continue throughout most of 1998, until every index pregnancy has ended in birth or miscarriage. So far, the balance of untoward events between the intervention and control arms have not given cause for alarm.
The WHO Antenatal Care Randomised Controlled Trial: rationale and study design
Authors: Villar J. , Bakketeig L. , Donner A. , Al‐Mazrou Y. , Ba’aqeel H. , Belizán J.M. , Carroli G. , Farnot U. , Lumbiganon P. , Piaggio G. , Berendes H.
Published: May 2003

The World Health Organisation and collaborating institutions in developing countries are conducting a multicentre randomised controlled trial to evaluate a new antenatal care (ANC) programme, consisting of tests, clinical procedures and follow‐up actions scientifically demonstrated to be effective in improving maternal and newborn outcomes. These activities are distributed, for practical reasons, over four visits during the course of pregnancy and are aimed at achieving predetermined goals. The study is taking place in four countries, Argentina, Cuba, Saudi Arabia and Thailand. Recruitment of study subjects started on 1 May 1996. All 53 ANC clinical units had been enrolled by December 1996. Clinics in each country were randomly allocated (cluster randomisation) to provide either the new programme or the traditional programme currently in use. Approximately 24 000 women presenting for ANC at these clinics over an average period of 18 months will have been recruited. As women attending the control clinics receive the ‘best standard treatment’ as currently offered in these clinics, individual informed consent is requested only from women attending the intervention clinics. Authorities of the corresponding health districts and all participating clinics have provided written institutional informed consent before randomisation.

  The primary outcome of the trial in relation to maternal conditions is the rate of a morbidity indicator index, defined as the presence of at least one of the following conditions for which ANC is relevant: (a) pre‐eclampsia or eclampsia during pregnancy or within 24 h of delivery; (b) postpartum anaemia (haemoglobin < 90 g/L); or (c) severe urinary tract infection/pyelonephritis, defined as an episode requiring antibiotic treatment and/or hospitalisation. The primary fetal outcome is the rate of low birthweight (< 2500 g). Adverse maternal and fetal outcomes are expected for ≈ 10% of the control group. Several maternal and perinatal secondary outcomes are also considered. A comprehensive cost‐effective‐ness analysis and women's and providers’ satisfaction evaluation are performed concurrently with the trial. Health‐care programmes should be rigorously evaluated by randomised controlled trials, which are feasible in developing countries and should be conducted before introducing new treatments or health interventions.
Methodological considerations in the design of the WHO Antenatal Care Randomised Controlled Trial
Authors: Donner A. , Piaggio G. , Villar J. , Pinol A. , Al‐Mazrou Y. , Ba’aqeel H. , Bakketeig L. , Belizán J.M. , Berendes H. , Carroli G. , Farnot U. , Lumbiganon P.
Published: May 2003

We discuss methodological issues arising in a recent evaluation trial of a new antenatal care programme, as sponsored by the Special Programme of Research, Development and Research Training in Human Reproduction, and WHO's Division of Reproductive Health (Technical Support). The randomisation unit for the trial is the antenatal care clinic, with 53 clinics located in four countries randomly allocated to provide either the new programme or the traditional programme currently in use. Approximately 24 000 women presenting for antenatal care over an average period of 18 months will have been recruited.
Paper title with footnote if needed
Authors: Mugford M. , Hutton G. , Fox‐Rushby J.
Published: May 2003

The WHO is testing a new rationalised programme of antenatal care in a multicentre randomised trial. The motivation for this trial arose from the current uncertainty about the effectiveness of different approaches to provision of routine antenatal care. Decision makers also lack information about the costs of providing routine antenatal care and the cost‐effectiveness of one programme over another. Such information will be needed before the final choice of programme can be made. The WHO trial provides an ideal opportunity to estimate and compare the incremental costs and cost‐effectiveness of the new programme in four countries (Argentina, Cuba, Saudi Arabia, Thailand). A separate economic component has been organised to measure the costs of antenatal care. Methods for cost identification and measurement, and methods for economic analysis in the context of an international study are based on current recommendations for the conduct of economic evaluations alongside trials. However, several aspects require further development. In particular, this includes defining standard methods for costing in different countries; measuring women's costs of access to care; and making comparisons across international settings. The economic evaluation will also inform similar multicentre international trials and investigate issues of generalisability beyond trial settings.
Conceptual bases and methodology for the evaluation of women's and providers’ perception of the quality of antenatal care in the WHO Antenatal Care Randomised Controlled Trial
Authors: Langer A. , Nigenda G. , Romero M. , Rojas G. , Kuchaisit C. , Al‐Osimi M. , Orozco E.
Published: May 2003

In this paper, we describe the conceptual bases and methodology used to assess women's and providers’ perception of the quality of antenatal care, as part of a large randomised trial in four developing countries. Information has been obtained by applying both qualitative and quantitative methodologies. The focus group discussions and in‐depth interviews have contributed useful insights into the cultural milieu in which care is provided, users’ and providers’ expectations, and their concept of quality. Based on these findings, we developed two standardised questionnaires, one being administered to a representative sample of pregnant women (n = 1600) and the other for all care providers. In this paper we present some of the findings of the focus group discussions and in‐depth interviews with women in one country as an example of the kind of information we have obtained. Women expressed their point of view concerning a reduced number of visits, type of provider, information that they get during clinical encounters and interpersonal relations with health professionals. The qualitative information, together with the data we obtain from the surveys, will highlight the aspects that will have to be considered if the new model of care is to be introduced on a routine basis.
Forty years on: Professor Neville Butler and the British Birth Cohort studies
Authors: Ferri Elsa
Published: May 2003

This paper traces the major contribution made by Professor Neville Butler, over a period of 40 years, to the foundation and subsequent development of two of Britain's three national birth cohort studies: the National Child Development Study, which has monitored the lives of all those in Britain born in the week 3–9 March 1958, and the British Cohort Study 1970, which has similarly followed the development of those born in the week 5–11 April 1970. Some recent findings from the two studies in the areas of health and health behaviour are briefly summarised, as are the plans for their future development.
Intergenerational influences affecting birth outcome. I. Birthweight for gestational age in the children of the 1958 British Birth Cohort
Authors: Hennessy Enid , Alberman Eva
Published: May 2003

There is considerable literature on intergenerational influences on birthweight. Few studies have been able to investigate such influences on the more basic measures of birthweight for gestational age and gestational age itself. This paper considers fetal growth. The investigations are derived from the 1958 British birth cohort followed from birth to age 33 years. Included were questions on physical and social characteristics of each parent and the grandparents, and birth details of parent and first child. In the present study, fetal growth in non‐preterm babies, after adjustment for the known effects of smoking and sex of the child, is explained best by factors relating to the parent's own growth, primarily in utero, but also to adulthood. There are small additional effects of education or social class but not of parent's gestational age. Only 15% of the variability in the child's fetal growth can be explained by the mother's characteristics and ≈ 7% by the father’s. Parent's own fetal growth accounts for nearly half of the variability if unadjusted for other factors and nearly a third after adjustment for sex of child, smoking, parental height and weight, maternal age at menarche and paternal age at first birth. Parental fetal growth makes the greatest anthropometric contribution.
Intergenerational influences affecting birth outcome. II. Preterm delivery and gestational age in the children of the 1958 British birth cohort
Authors: Hennessy Enid , Alberman Eva
Published: May 2003

The 1958 British cohort study has data to investigate intergenerational effects on preterm delivery and on gestational age in non‐preterm births, allowing for many confounders that may differ in the more pathological preterm babies. Previous results for all gestational ages have been inconsistent. The strongest and only likely independent intergenerational effect on non‐preterm gestational age found is parental gestational age (adjusted regression coefficient = 0.067 weeks per week in mothers and 0.045 in fathers). The preterm analysis has low power; however, reported history of hypertension in mothers (any), in fathers and in the maternal grandmother (measured in the 1958 pregnancy) all significantly and independently increased the risk of preterm birth [OR = 1.7, 2.0, 1.5 respectively]. The absolute risk was particularly high in hypertensive mothers who had been preterm themselves (21%). Other possible intergenerational influences of height, weight, fetal growth and gestation were not significant enough and/or consistent enough between parents to speculate whether they are truly intergenerational or confounded by other factors acting during the pregnancy. Excepting mother's weight for height, no genetic or environmental influence studied affects both gestational age and fetal growth in term births. However, many maternal factors that reduce either fetal growth or gestation in term births are associated with increased risk of preterm birth.