bluuebunny/crossref_metadata_2025 huggingface dataset:
DNA sequence of the porcine α‐lactalbumin 5' flanking region and single‐base polymorphisms within this region
Authors: Bleck G T , Johnson‐Roberts D L , Jiménez‐Flores R , Wheeler M B
Published: July 2010

The 5′ flanking region of the α‐lactalbumin (α‐LA) gene was sequenced for the Duroc, Yorkshire and Meishan breeds of swine to identify potential sequence variants within this regulatory region of the porcine α‐LA gene. The sequenced region of the gene encompasses 391 bp 5′ of the translation start site to 11 bp 3′ of the translation start site. Within this sequence of the porcine α‐LA gene two single‐base pair differences were detected. One variant occurs at position – 178 and the other at position – 235 from the translation start site. Each of the variations can be detected by a restriction fragment length polymorphism within a polymerase chain reaction amplified product. The polymorphisms at the – 178 and – 235 positions appear to be genetically linked in the animals that have been analysed.
Genetic polymorphism of the sixth component of complement (C6) in dogs
Authors: Shibata T , Abe T , Tanabe Y
Published: July 2010

Using agarose gel isoelectric focusing and immunoblotting with rabbit anti‐rabbit C6, a genetic polymorphism was found in the sixth component of complement (C6) in 18 Asian native breeds or populations and three European breeds of dog. The C6 locus was highly polymorphic. The phenotype distribution data indicated that dog C6 phenotypes were controlled by seven codominant alleles, C6A,C6B,C6C,C6D,C6E,C6F and C6G, at a single autosomal locus. Breed differences were observed among the gene frequencies, especially between Asian and European breeds. Two gene flows from the adjacent areas into Japanese native dogs were postulated.
Random amplified polymorphic DNA in cattle and sheep: application for detecting genetic variation
Authors: Kantanen J , Vilkki J , Elo K , Mäki‐Tanila A
Published: July 2010

The present study investigated the use of the random amplified polymorphic DNA (RAPD) method to detect genetic variation in cattle and sheep. The animals studied consisted of samples from five Finnish cattle breeds: native Eastern (18 animals), Northern (24), Western Finncattle (24), Finnish Ayrshire (24), and Finnish Friesian (18); as well as a white (6 animals) and a grey (9) colour type of Finnsheep. The cattle and sheep populations were analysed with 11 and 13 RAPD primers demonstrating the most repeatable amplification pattern. Two out of ten RAPD fragments tested by cross hybridization showed homology between the two species. The RAPD method did not prove efficient for finding new polymorphisms in either species, because we found only three polymorphic RAPD markers for cattle and seven markers for sheep with different allele frequencies between the breeds. Although there is a greater presence of polymorphic RAPD markers in sheep, according to the similarity indices the sheep populations showed a higher degree of homogeneity than the cattle breeds. However, the interbreed and intrabreed similarity indices for cattle did not suggest any significant differentiation of the Finnish breeds, contrary to earlier results based on blood group and protein polymorphism.
Polymorphic CAC/T repetitive sequences in the pig genome 1
Authors: Coppieters W , Van de Weghe A , Depicker A , Coppieters J , Peelman L , VAN Zeveren A , Bouquet Y
Published: July 2010

Three genomic clones were isolated from a sizeselected pig DNA library by hybridization with a DNA‐fingerprint probe. Analysis at the sequence level revealed that all three clones contain interrupted stretches of triplet repeats mainly composed of CAC and CAT triplets. Evaluation of the corresponding loci for polymorphism by Southern blot hybridization showed considerable length variation. For two loci the polymorphism was also demonstrated by polymerase chain reaction (PCR) amplification. The PiGMaP reference pedigree was typed for all three loci.
Assignment of the growth hormone receptor gene to bovine chromosome 20 using linkage analysis and somatic cell mapping
Authors: Moody D E , Pomp D , Barendse W , Womack J E
Published: July 2010

A polymorphism was identified in the bovine growth hormone receptor (GHR) gene by digesting polymerase chain reaction (PCR) products with the restriction enzyme Alul. Two alleles were segregating in cattle of Bos indicus descent, but one allele appears to be fixed in Bos taurus cattle. GHR was localized to bovine chromosome 20 using bovine‐rodent hybrid cell lines and linkage analysis.
Identification and characterization of new BoLA‐DRB3 alleles by heteroduplex analysis and direct sequencing
Authors: Sitte K , East I J , Lavin M F , Jazwinska E C
Published: July 2010

A sample of 52 mixed‐breed dairy cattle (Holstein Friesian and Jersey) and 51 beef cattle (Hereford) from south‐east Queensland was studied. The second exon of BoLA‐DRB3 was amplified by polymerase chain reaction (PCR), and polymorphisms were detected by heteroduplex analysis. A large number of different heteroduplex patterns indicated extensive sequence polymorphism. Direct sequencing of PCR products from 17 homozygotes and cloning and sequencing of PCR product from two heterozygotes resulted in the identification and characterization of four novel alleles. The previously described allele BoLA‐DRB3* 2A is characterized by an amino acid deletion at position 65. We have identified three animals that are homozygous for this amino acid deletion, indicating that the deletion is unlikely to result in loss of function. Two of these animals had allele BoLA‐DRB3* 2A, and one had a novel allele with codon 65 deleted but differing from BoLA‐DRB3* 2A at a number of other amino acid positions. In conclusion, heteroduplex analysis allows rapid discrimination between homozygotes and heterozygotes, and enables rapid identification of new BoLA‐DRB3 alleles.
Prediction of informativeness for microsatellite markers among progeny of sires used for detection of economic trait loci in dairy cattle
Authors: Ron M , Lewin H , Da Y , Band M , Yanai A , Blank Y , Feldmesser E , Weller J I
Published: July 2010

Individual loci affecting economic traits can be located using genetic linkage. Application of either daughter or granddaughter designs requires determination of allele origin in the progeny. If only the sires and their progeny are genotyped, the paternal allele origin of progeny having the same genotype as the sire cannot be determined. The expected frequency of informative sons can be predicted for each sire and genetic marker from the allele frequencies in the population. The accuracy of a predictor of the frequency of informative progeny was tested on 103 grandsire x microsatellite combinations. Number of sons per grandsire varied from 24 to 129. Allele frequencies in the population were estimated by genotyping seven sires. The regression of the frequency of informative sons on the predicted frequency was 1.04 with a zero intercept model. Thus, considering the large number of genetic markers available for analysis, predicted informative frequency is a useful criterion for selection of genetic markers.
Organization of the sex‐linked late‐feathering haplotype in chickens
Authors: Iraqi F , Smith E J
Published: July 2010

Nucleotide sequence analysis of polymerase chain reaction products confirmed that ev21 integrated into one of two large homologous elements on the Z chromosome of late‐feathering (LF) White Leghorn chickens. Southern blots of NotI‐, NaeI‐, KspI‐ and BamHI‐digested DNA from early‐feathering (EF) and LF White Leghorns, that had been hybridized with a probe that flanks ev21, indicated a 180 kb duplication of an unoccupied repeat in the LF genotype of White Leghorns. A KspI fragment that carries ev21 was about 32 kb smaller than the KspI fragment found in EF DNA. In the evolution of LF, retroviral insertion into one of two large repeats and a 32 kb deletion may have generated LF.
Sequence and PCR‐RFLP analysis of 14 novel BoLA‐DRB3 alleles
Authors: Gelhaus A , Schnittger L , Mehlitz D , Horstmann R D , Meyer C G
Published: July 2010

The genetic diversity of the bovine class IIDRB3 locus was investigated by polymerase chain reaction (PCR) amplification and DNA sequencing of the first domain exon. Studying 34 animals of various cattle breeds, 14 previously unrecognized DRB3 alleles were identified. In three alleles, amino acid substitutions were observed that had not been previously found in bovine DRB3, but occurred at the same position in bovine DQB and in the DRB alleles of other mammals. For all newly identified alleles, the restriction fragment length polymorphism (RFLP) patterns of PCR products obtained with the enzymes RsaI, BstYI, and HaeIII were compared with patterns of 38 previously described alleles. Altogether, eleven novel PCR‐RFLP types were defined. Twelve out of the 42 PCR‐RFLP types identified so far were not found to be fully informative because they corresponded to more than one allelic sequence. PCR‐RFLP may therefore be a rapid and useful method for DRB3 typing in cattle families, but for studies on outbred populations, sequencing and hybridization techniques are required.
Probability of random sire exclusion using microsatellite markers for parentage verification
Authors: Usha A. P. , Simpson S. P. , Williams J. L.
Published: July 2010

Many microsatellite sequences have been described in the bovine genome. Being highly polymorphic these have been suggested as markers for parentage verification and individual identification in cattle. We have evaluated the use of five highly polymorphic microsatellite markers for parentage verification in 14 breeds of cattle in the UK. Three of the microsatellite loci occur within introns in genes: BoLA DRB3, steroid 21‐hydroxylase, and the beta subunit of the follicle‐stimulating hormone. The other two are anonymous sites ETH131 and HEL6. Results were analysed by a statistical approach that takes in to account deviations from Hardy‐Wienberg equilibrium and linkage disequilibrium for multiple loci. The method of determining the probability of random sire exclusion uses observed genotype frequencies instead of allele frequencies. Independently, the markers used have a probability of between 0.72 and 0.62 of identifying a parentage error, while used together the five markers give, on average across breeds, a probability of 0.99 of excluding an incorrect sire.
Extensive mtDNA diversity in horses revealed by PCR–SSCP analysis
Authors: Marklund S , Chaudhary R , Marklund L , Sandberg K , Andersson L
Published: July 2010

The hypervariable D‐loop region of mitochondrial DNA (mtDNA) was amplified with the polymerase chain reaction using total horse DNA samples. Analysis of single strand conformation polymorphism (SSCP) of denatured amplification products was carried out by native polyacrylamide (8%) gel electrophoresis followed by silver staining. As many as 15 distinct SSCP variants were revealed when screening a total of 78 maternally unrelated horses representing five different breeds. All breeds showed a high degree of polymorphism and the estimated probability (PImt) that two maternally unrelated individuals have, by chance, identical SSCP variants varied between 0.14 and 0.30. We detected no heteroplasmy or deviations from strict and stable maternal inheritance when examining four maternal lineages, each represented by six to eight horses, separated by up to five generations from a common ancestral mare. The study establishes a simple screening method for detecting equine mtDNA types, which can be applied for tracing maternal genealogies and for association studies.
Report of the First Workshop on the Genetic Map of Bovine Chromosome 23
Authors: Beever J. E. , Lewin H. A. , Barendse W. , Andersson L. , Armitage S. M. , Beattie C. W. , Burns B. M. , Davis S. K. , Kappes S. M. , Kirkpatrick B. W. , Ma R. Z. , McGraw R. A. , Stone R. T. , Taylor J. F.
Published: July 2010

A report of the first workshop on the genetic map of bovine chromosome 23 (BTA23) is given. Five laboratories contributed data from 29 loci, including a total 11586 informative genotypes. The combined pedigrees represented 1930 potentially informative meioses. Eighteen of the 29 loci were common to two or more data sets and were used to construct a framework linkage map of BTA23. Twelve of the 18 could be ordered on the linkage map with a likelihood ratio of greater than 1000:1. Thus, a low resolution consensus map was constructed with a high level of support for order. The sex‐averaged, female and male maps span 54.5, 52.7 and 55.8 cM, respectively. Sex‐specific differences in recombination frequency were identified for eight pairs of framework loci. Average genetic distance between framework loci on the sex‐averaged map is 5.0 cM.
Development and mapping of polymorphic microsatellite markers derived from a chicken brain cDNA library
Authors: Ruyter‐Spira C. P. , Crooijmans RPMA , Dijkhof R. J. M. , van Oers P. A. M. , Strijk J. A. , van der Poel J. J. , Groenen M. A. M.
Published: July 2010

Until now the genetic linkage map in chicken has been based mainly on random genomic markers. The addition of expressed sequence tags (ESTs) to the genetic linkage maps is becoming more important because ESTs can form the basis for comparative mapping studies. This may be helpful for the detection of candidate genes for quantitative trait loci (QTLs).

In our study we used a (TG)13repeat as probe for the detection of microsatellites in a chicken brain cDNA library. After hybridization 0.15% of the cDNA clones gave a positive signal. The cDNA complexity of the library was high; of the 90 cDNA clones that were sequenced 60 occurred only once. For 29 clones primer sets for the polymerase chain reaction could be developed. Twenty‐one microsatellites were polymorphic on one or more of the test panels and 15 markers could be mapped on either or both of the international reference families. Because sequence homology between chicken and mammalian cDNAs is sometimes low it was difficult to assess the level of sequence homology that indicated a true homologous transcript. In our study seven cDNA clones, of which three could be mapped, showed a relatively high percentage of sequence homology with sequences found in other species. Because sequencing and mapping of expressed sequence tags in human and mouse is progressing very rapidly, it is predicted that further information will soon be readily available. Therefore, increasing the number of expressed sequences on the chicken genetic linkage map will be of value for comparative mapping studies in the near future.
Mapping economic trait loci for somatic cell score in Holstein cattle using microsatellite markers and selective genotyping
Authors: Ashwell M. S. , Rexroad C. E. , Miller R. H. , Van Raden P. M.
Published: July 2010

Marker‐assisted selection (MAS) uses genetic marker genotypes to predict an animal's production potential and will provide additional selection information for progeny testing. With the discovery of highly polymorphic microsatellite markers, the tools now exist to begin the search for economic trait loci (ETL), which is the first step toward MAS. The objective of this study was to identify ETL for somatic cell score in an existing Holstein population. Using the granddaughter design, sons from seven grandsire families were genotyped with 20 autosomal microsatellites from five chromosomes (4, 8, 13, 17, 23), with an emphasis on chromosome 23, which is the location of the bovine major histocompatibility complex (BoLA). Selective genotyping was used to reduce the number of genotypes required, in which the 10 highest and 10 lowest sons from the phenotypic distribution curve were tested (140 sons in seven families). One marker (513), located near BoLA, showed evidence of an ETL in three of five polymorphic families. Additional sons were genotyped from the five families to estimate the effect and to compare selective and ‘complete’ genotyping. Both methods detected an ETL at marker 513, but in different families. This study provides evidence of the usefulness of microsatellite markers and the granddaughter design in the detection of ETL; however, additional markers need to be evaluated to determine the usefulness of selective genotyping. Based on the results from the 20 studied markers, the most likely position of a somatic cell score ETL lies near marker 513, located on chromosome 23.
Evidence for a single pedigree source of the hyperkalemic periodic paralysis susceptibility gene in Quarter Horses
Authors: Bowling A. T. , Byrns G. , Spier S.
Published: July 2010

The pedigree origin of a base pair substitution in the horse muscle sodium channel gene that confers susceptibility to the muscle disease hyperkalemic periodic paralysis (HYPP) was investigated with a set of 978 Quarter Horses. The horses were chosen at random, based on a collection of blood samples taken between 1989 and 1991 to meet parentage testing requirements, primarily but not exclusively from breeding stallions. The frequency of Quarter Horses positive for the base pair substitution, all heterozygotes, was 4‐4%, which corresponds to an allelic frequency of 0.02. All horses positive for the gene traced to a single previously identified stallion as first, second or third generation descendants. A higher frequency of the HYPP susceptibility trait than expected by random occurrence was found among his descendants in this study.
Comparison of protein markers and microsatellites in differentiation of cattle populations
Authors: Arranz J J , Bayón Y , Primitivo F San
Published: July 2010

Five cattle populations, representing four breeds, were analysed for 14 protein markers and five microsatellite loci. The breeds studied were Brown Swiss and three autochthonous Spanish cattle: Avileña‐Negra Ibérica (A‐NI), two populations (A‐NI 1 and A‐NI 2) from different, reproductively isolated, locations; Sayaguesa; and Morucha. A total of 752 animals were examined for biochemical polymorphisms, of which 488 were also DNA typed. Genetic parameters and phylogenetic trees were obtained separately for each group of markers and results were compared. Estimates of heterozygosity and genetic distances from microsatellites were greater than those obtained using protein markers. The overall topology of the two dendrograms was similar. A‐NI 1 and A‐NI 2 populations were grouped together, related to Morucha, and the three of them related to Sayaguesa. Brown Swiss appeared in a separate branch from Spanish cattle. These results support the usefulness of microsatellites in the study of genetic relationships among closely related populations and breeds.
Introgression of the avian naked neck gene assisted by DNA fingerprints
Authors: Yancovich A , Levin I , Cahaner A , Hillel J
Published: July 2010

Theoretical predictions suggest that DNA markers can be useful tools for genomic selection in gene introgression programmes. An experiment was carried out to evaluate the efficiency of using multi‐locus DNA markers in an introgression programme designed to transfer the naked neck gene from a donor to a recipient chicken line. The donor line was a commercial egg layer chicken stock heterozygous at the naked neck locus (Na/na+), while the recipients were from a Cornish broiler line. These two lines differ markedly in their average body weight, a quantitative trait that can also represent the comprehensive differences between the genomes of the two lines involved. Three groups of naked neck BC1 individuals were selected according to the following criteria: (i) low band‐sharing with their donor grandsires evaluated by multi‐locus DNA markers, (ii) high body weight at six weeks of age, and (iii) selection at random as a control group. Birds from each of these groups were mated at random to individuals from the heavier Cornish line to produce three groups of BC2 individuals whose body weights were recorded weekly from three to seven weeks of age. Results indicated that BC2 birds obtained from BC1 parents selected for band‐sharing levels and those selected for body weight, performed equally well at 4–7 weeks of age; both were 3.1–3.9% heavier than birds from the randomly selected group. The additional genome recovery of the heavier broiler line, obtained by DNA markers, was found to be in agreement with theoretically predicted values.
A search for quantitative trait loci for ovulation rate in cattle
Authors: Blattman A N , Kirkpatrick B W , Gregory K E
Published: July 2010

Seventy‐seven polymorphic microsatellites were analysed in offspring of three elite sires that were part of the foundation of an experimental population selected for twinning rate at the US Meat Animal Research Center, Clay Center, Nebraska. All females were assessed for ovulation rate by rectal palpation of corpora lutea over 8–10 consecutive oestrous cycles from approximately 12 to 18 months of age, and associations between ovulation rate and sire allele were examined in each of the three sire groups. A preliminary analysis was performed using selectively genotyped daughters of each sire. Markers found significant or approaching significance were also genotyped in all daughters, sons and granddaughters of these sires. A test of marker associations limited to the granddaughter data provided an independent confirmation of marker effect and significance relative to the initial test with daughter data. Putative ovulation rate quantitative trait loci were detected on chromosomes 7 and 23. Marker UWCA20 on chromosome 7 was associated with an effect in excess of one phenotypic standard deviation and accounted for approximately 10% of phenotypic variation ovulation rate. Marker CYP21 (steroid 21‐hydroxylase) on chromosome 23 was associated with an effect of slightly less than half a phenotypic standard deviation and accounted for approximately 4% of phenotypic variation.
Integration of the PiGMaP and USD A maps for porcine chromosome 14
Authors: Kapke P , Wang L , Helm J , Rothschild M F
Published: July 2010

In order to align two previously published genetic linkage maps, a set of four of the United States Department of Agriculture (USDA) microsatellite linkage markers was mapped in the International Pig Gene Mapping Project (PiGMaP) reference families. Two‐point linkage analysis was used between these USDA markers and the set of genes and markers previously mapped on the PiGMaP chromosome 14 map‐Markers with threshold lod scores of three or greater were used for multipoint map construction. The USDA and PigGMaP linkage maps of chromosome 14 were aligned using the four USDA microsatellite markers along with three markers that are common to both maps. The PiGMaP genetic linkage map order for chromosome 14 was confirmed and the map was expanded to 193 cM with addition of the new markers.
Linkage between the loci for serum albumin and vitamin D binding protein (GC) in the Japanese quail
Authors: Shibata T , Abe T
Published: July 2010

Vitamin D binding protein (GC) and serum protease inhibitor (PI) have been added to genetic markers in the Japanese quail. Both loci were controlled by autosomal codominant alleles named GCA, GCB and PIA, PIB, PIC, respectively. Close linkage between the loci for serum albumin (ALB) and GC protein is reported. Two recombinants were observed in 145 informative offspring of 14 families. The recombination frequency between the loci was estimated as 0.014±0.006. Thus, GC was assigned to linkage group II in the Japanese quail. No signs of linkage were observed among the loci for the ALB‐GC complex, PI. serum prealbumin 2 (PA2), phosphoglucose isomerase (PG1), 6‐phosphogluconate dehydrogenase (PGD) and esterase‐D (ESD).
Simultaneous analysis of bovine K‐casein and BLAD alleles by multiplex PCR followed by parallel digestion with two restriction enzymes
Authors: Zsolnai A , Fésüs L
Published: July 2010

An improved and simplified method allowing simultaneous genetic typing of K‐casein and CD 18 (bovine leucocyte adhesion deficiency; BLAD) loci has been developed. The method is based on the simultaneous amplification of fragments of the two groups of alleles by multiplex PCR, and on a concurrent, parallel digestion of the products by two restriction enzymes (PstI andHaeIII) in the same incubation buffer. Digestion withPstI distinguishes K‐casein A and B alleles and does not cut within any of the BLAD alleles, while digestion withHaeIII allows the differentiation between normal and mutant allele variants of theCD28locus. All combinations of the known mutants of the two alleles, characterized to the regions amplified and resulting in phenotypic effect, could be detected by electrophoretic separation performed on the same agarose gel owing to the vast differences in the length of the restriction fragments.
Molecular genetic characterization of new bovine kappa‐casein alleles CSN3F and CSN3G and genotyping by PCR‐RFLP
Authors: Prinzenberg E‐M , Hiendleder S , Ikonen T , Erhardt G , Prinzenberg E‐M , Hiendleder S , Erhardt G , Ikonen T
Published: July 2010

In order to characterize the two new kappa‐casein variants F and G (CSN3F and CSN3G) recently detected in Ayrshire and Pinzgauer cattle, exon IV of CSN3 from heterozygous animals was amplified by polymerase chain reaction (PCR), cloned and sequenced. The sequencing data revealed single point mutations at nucleotide positions 10530 (G→A) for CSN3F and 10790 (C→T) for CSN3G, corresponding to amino acid exchanges in positions 10 (Arg→His) and 97 (Arg→Cys) respectively. These mutations alter recognition sites for the restriction enzymes HhaI and MaeII, which were subsequently used to confirm these polymorphisms in cattle carrying CSN3F or CSN3G. A PCR‐restriction fragment length polymorphism (RFLP) genotyping procedure for all currently known CSN3 alleles (CSN3A, CSN3B, CSN3C, CSNJE, CSNF, CSN3G) was developed.
Characterization of 42 highly polymorphic bovine microsatellite markers
Authors: Hirano T , Nakane S , Mizoshitat K , Yamakuchit H , Inoue‐Murayama M , Watanabe T , Sugimoto Y , Barendse W
Published: July 2010

We have isolated 42 highly polymorphic microsatellite (GT/CA)n markers from Japanese black cattle Wagyu (Bos taurus). Forty‐one of the markers were assigned to bovine autosomes with lod scores > 6, through linkage analyses performed on the International Bovine Reference Family Panel (IBRP). The remaining marker showed X‐linked inheritance. These markers exhibited an average heterozygosity value of 0.67 with between four and 17 alleles on the IBRP.
Polymerase chain reaction‐based polymorphisms in the porcine cholecystokinin (CCK) gene and assignment to chromosome 13
Authors: Clutter A C , Sasaski S , Pomp D
Published: July 2010

Polymorphisms were identified in the porcine cholecystokinin (CCK) gene by digestion of products from polymerase chain reaction (PCR) with the restriction enzyme DpnII. Individuals from the European pig gene mapping project (PiGMaP) consortium reference families (eight full‐sib families, 91 total progeny) were genotype to determine linkage relationships between the CCK gene and previously mapped loci. Linkage analysis revealed that the CCK gene is located on porcine chromosome 13.
Genetic markers linked to quantitative traits in poultry
Authors: Lamont S. J. , Lakshmanan N. , Plotsky Y. , Kaiser M. G. , Kuhn M. , Arthur J. A. , Beck N. J. , O'Sullivan N. P.
Published: July 2010

This study utilized DNA fingerprints and crosses of two genetically distinct lines of layer‐type chickens to identify genetic markers linked to quantitative trait loci (QTL). In phase I, back‐cross (BC1) hens were separately ranked for each of eight traits and then blood pools were produced in groups along each phenotypic distribution. The DNA was isolated from the blood pools and used in a gradient analysis to screen for DNA fingerprint bands that exhibited intensity gradients associated with the phenotypic traits. To identify linkage of bands with QTL and to estimate band effects, F2 progeny were produced in phase II from the phase I BC, population. A single‐trait animal model was used for analysis of associations of all individual DNA fingerprint bands of sires and their progeny phenotypic performance. Twenty fingerprint bands, only two of which had shown trait‐associated gradients in phase I, were identified by the animal model analysis of the progeny test as QTL linked (P≤005) to specific traits of growth, reproduction and egg quality. These 20 bands warrant further study as potentially valuable molecular markers for QTL.
Efficient resolution of parentage in dogs by amplification of microsatellites
Authors: Fredholm M. , Winterø A. K.
Published: July 2010

Parentage control has been performed for 15 litters from 12 different dog breeds by amplification of microsatellites. As it was possible to include all putative parents in all cases, they were solved by exclusion. Discrimination between parents/non‐parents was made after genotyping of 6–9 microsatellite loci. In 12 of the cases all but one of the alleged fathers were excluded while in three cases it was unambiguously shown that superfecundation had taken place. Furthermore, one inclusion case concerning disputed maternity has been investigated. Maternity indices were calculated for 12 loci and probability of maternity was estimated to be 99‐99%. These results testify that microsatellites can be applied very efficiently for resolution of parentage in dogs.
Isolation and characterization of 45 polymorphic microsatellites from the bovine genome
Authors: Ma R. Z. , Russ I. , Park C. , Heyen D. W. , Beever J. E. , Green C. A. , Lewin H. A.
Published: July 2010

A small‐insert bovine genomic library was constructed in pBluescript II SK(+) and enriched for microsatellites by selective rescue of single‐stranded pBluescript DNA carrying (CA)n/(TG) n tandem repeats. Approximately 50% of the clones in the enriched library contained (CA) nrepeats or CA‐rich sequences. Sequencing of clones selected for (CA) n repeats resulted in the identification and characterization of 45 (CA) npolymorphic microsatellites. Genotyping in 9 large paternal half‐sib families indicated that 40 of these microsatellite markers exhibit autosomal Mendelian inheritance. The numbers of alleles range from 2 to 18, with an average of 6‐3 per locus. The polymorphic microsatellite markers we have identified and characterized will contribute to the construction of a high‐resolution linkage map of bovine genome.
‘More than Words’: touch practices in child and family social work
Authors: Lynch Rachel , Garrett Paul Michael
Published: July 2010

This paper explored the subject of physical touch occurring between social workers and children in the Republic of Ireland. Here, it was maintained that touch practices occupy an ambiguous area for social work practitioners. The empirical component was based on interviews that were conducted with a small group of practitioners in one particular Irish county. The study provided insight into the social workers' personal experiences of, and opinions on, touch with children in the context of their practice. Despite a lack of prescriptive ‘do's and don'ts’ regarding physical touch and social work, the findings suggested that social workers do not consciously engage in physical touch with children. Rather, touch occurs as a result of practicality and safety concerns for a child. Recognized as beneficial in terms of communication, reassurance and conveying empathy, touch practices are also guided by a fear of misinterpretation, allegations and concern for causing harm to the child. The place of physical touch with children is regarded as being outside the remit of the social work role. Despite this, a majority of those interviewed spoke of a desire to have more discussion and guidance on the subject of physical touch with children within their work locations.
Factor XIII deficiency
Authors: HSIEH L. , NUGENT D.
Published: July 2010

Summary.  Inherited factor XIII (FXIII) deficiency is a rare bleeding disorder that can present with umbilical bleeding during the neonatal period, delayed soft tissue bruising, mucosal bleeding and life‐threatening intracranial haemorrhage. FXIII deficiency has also been associated with poor wound healing and recurrent miscarriages. FXIII plays an integral role in haemostasis by catalysing the cross‐linking of fibrin, platelet membrane and matrix proteins throughout thrombus formation, thus stabilizing the blood clot. The molecular basis of FXIII deficiency is characterized by a high degree of heterogeneity, which contributes to the different clinical manifestations of the disease. There have been more than 60 FXIII mutations identified in the current literature. In addition, single nucleotide polymorphisms have been described, some of which have been shown to affect FXIII activity, contributing further to the heterogeneity in patient presentation and severity of clinical symptoms. Although there is a lifelong risk of bleeding, the prognosis is excellent when current prophylactic treatment is available using cryoprecipitate or plasma‐derived FXIII concentrate.
Self‐reported drug allergies and the diagnostic work‐up in the surgical population
Authors: Tamayo Eduardo , Álvarez F. Javier , Castrodeza Javier , Yánez Javier , Arnaiz Pilar , Lajo Carmen , Soria Susana
Published: July 2010

Objective  The diagnostic work‐up of a drug hypersensitivity reaction is indeed difficult. In general, medical documentation of allergic reactions in medical reports is usually highly deficient or non‐existent. The aim of this study was to analyse the prevalence of self‐reported drug allergies in the surgical population as well as the criteria used in the diagnosis of drug hypersensitivity reactions.

Methods  A prospective study with the consecutive participation of 1439 patients, following surgical intervention, attended the Post‐Operative Care Unit. Previously, as a routine process during the pre‐anesthesia consultation, all patients were questioned about whether they had any drug allergies to report and diagnostic work‐up.

Results  The prevalence of self‐reported drug allergies was 8.3% (119/1439): 3.6% considered themselves allergic to β‐lactams and 2.4% to non‐steroidal anti‐inflammatory drugs. Approximately one‐third of the subjects (40 out of the 119) had not been subjected to any allergy diagnostic procedure and with 79 (66.4%), the only diagnostic test used by the Allergy Unit had been the skin prick‐test. None of those participating in the study had tryptase, methylhistamine, specific IgE or intradermal tests carried out to characterize the diagnosis of the allergic reaction.

Conclusions  These results show that self‐reported drug allergies are highly prevalent and as yet little explored. It is an important prevalence which should bring about modifications to the prescription of certain medicaments. The medical personnel must be made aware of the need to make an accurate diagnosis of allergies to medicaments.
The improper use of research placebos
Authors: Kottow Miguel
Published: July 2010

Rationale  Reasons given for the routine use of placebos in Phase III clinical research are not convincing. Ethically inadequate strategies such as using placebos and recruiting the mentally incompetent for non‐therapeutic research are allegedly permissible because research is purportedly aimed at benefiting the common weal.

Method and results  A review of current literature concludes that placebos are not equivalent to inactive substances, as has been amply shown in clinical use; in research settings they may actually be harmful, for patients recruited as research subjects often develop expectations that are all the more frustrated if they receive a control substance that is not therapeutic but may show unexpected side effects. Supposedly inert control substances have met with strong opposition, for patients are placed at risk of becoming therapeutic orphans when their current medication is withdrawn and they have a 50% chance of being randomized to the inactive control arm. Using placebos, it is claimed, increase the internal validity of a study, but this concern appears misplaced when scant attention to external validity is given.

Conclusions  Although rejected and disregarded by many investigators, it is an accepted practice that control groups should receive the best treatment in existence, not what is locally available nor a placebo. Inactive comparators should only be used when no effective medications exists for the disease under scrutiny. Bioethics should be wary of the tenuous justifications presented, as researchers rarely resort to placebos in their own sponsor countries, while actively endorsing them in Third World nations where most of their trials are being located.
FabAct®: a decision‐making tool for the anticipation of the preparation of anticancer drugs
Authors: Bonan Brigitte , Martelli Nicolas , Berhoune Malik , Vidal Ludovic‐Alexandre , Sahin Evren , Prognon Patrice
Published: July 2010

Rationale  Due to the increase of new cancer cases, our chemotherapy compounding unit must face with ever‐growing production needs. To support this increasing workload, we decided to anticipate the preparation of several anti‐cancer drugs.

Aims and objectives  To help us in the decision making, we needed a modern tool able to combine several criteria for selecting appropriate medications for an anticipated preparation. The aim of this study was to assess the decision‐making software, FabAct® (Version 1.0).

Methods  FabAct® ranked all of the anti‐cancer drugs used in our chemotherapy compounding unit according to price, chemical stability, compounding difficulties, dosage and production per year. Then, we started to anticipate currently the preparation of four medications and conducted a follow‐up of destroyed preparations between January and May 2007. We tried to identify the destruction causes and calculated the time saved for the patients and for the pharmacy technicians.

Results  According to the decision‐making software, the first four drugs for an anticipated preparation were: fluorouracil, cisplatin, carboplatin and paclitaxel. A total of 3913 (50.2%) anticipated preparations were performed and among those, 470 (12%) were destroyed. The main cause of destruction was due to the preparation expiration. Finally, the mean waiting time per patient was reduced from 118 minutes to 68 minutes after the application of the anticipated model.

Conclusion  According to this 5‐month follow‐up, FabAct® helped us to select appropriate anti‐cancer drugs to anticipate the compounding. Most of the anticipated preparations were administrated to patients and the patient waiting time was significantly reduced.
Doctors' perceptions of laboratory monitoring in office practice
Authors: Goldman Roberta E. , Soran Christine S. , Hayward Geoffrey L. , Simon Steven R.
Published: July 2010

Background  Laboratory monitoring has been increasingly recognized as an important area for improving patient safety in ambulatory care. Little is known about doctors' attitudes towards laboratory monitoring and potential ways to improve it.

Methods  Six focus groups and one individual interview with 20 primary care doctors and nine specialists from three Massachusetts communities.

Results  Participants viewed laboratory monitoring as a critical, time‐consuming task integral to their practice of medicine. Most believed they commit few laboratory monitoring errors and were surprised at the error rates reported in the literature. They listed various barriers to monitoring, including not knowing which doctor was responsible for ensuring the completion of laboratory monitoring, uncertainty regarding the necessity of monitoring, lack of alerts/reminders and patient non‐adherence with recommended monitoring. The primary facilitator of monitoring was ordering laboratory tests while the patient is in the office. Primary care doctors felt more strongly than specialists that computerized alerts could improve laboratory monitoring. Participants wanted to individualize alerts for their practices and warned that alerts must not interrupt work flow or require too many clicks.

Conclusions  Doctors in community practice recognized the potential of computerized alerts to enhance their monitoring protocols for some medications. They viewed patient non‐adherence as a barrier to optimal monitoring. Interventions to improve laboratory monitoring should address doctor workflow issues, in addition to patients' awareness of the importance of fulfilling recommended therapeutic monitoring to prevent adverse drug events.
Pitfalls in double‐blind, placebo‐controlled oral food challenges
Authors: Niggemann B. , Beyer K.
Published: July 2010

Although controlled oral food challenges are considered to be the gold‐standard in the diagnosis of food related symptoms, especially if performed in a double‐blind, placebo‐controlled food challenges (DBPCFC) manner, there are still many unanswered questions and newer aspects, which may explain some pitfalls encountered during oral food challenges. For stopping an oral food challenge and declaring a challenge as positive or negative, symptoms should be objective and/or repetitive. The time interval between administering the food and observing the clinical reaction is an ambivalent factor. Possible reasons for false negative assessments include inadvertent drug use during oral challenges, and the fact that a short‐term specific oral tolerance induction (SOTI) may be induced as increasing amounts of the offended food are administered during a titrated oral food challenge. Possible reasons for false positive assessments are the difficulty to maintain an appropriate strict diet throughout the oral challenge procedure, and that the elimination diet implemented before the oral food challenge in children with atopic eczema and suspected food related symptoms may itself be responsible for immediate type clinical symptoms, which had not been reported by the parents before. Finally augmentation factors are among the most plausible explanations for the inadequate reproducibility of an oral food challenge. Although a 100% standardization of the challenge procedure does not seem realistic, efforts should be made to improve the methodology used so far. On the contrary, the possible relation of DBPCFC and SOTI may offer potential advantages for future therapeutic approaches of food allergy.
Association of prostaglandin‐endoperoxide synthase 2 gene polymorphisms with asthma and atopy in Chinese children
Authors: Chan I. H. S. , Tang N. L. S. , Leung T. F. , Ma S. L. , Zhang Y. P. , Wong G. W. K. , Wong C. K. , Lam C. W. K.
Published: July 2010

Background:  Cyclooxygenase‐2 (COX‐2) plays essential roles in inflammation. Previous studies have suggested associations between prostaglandin‐endoperoxide synthase 2 (PTGS2) polymorphisms and prostaglandins production in asthma.

Objective:  We have investigated the effects of Chinese tagging single nucleotide polymorphisms (SNPs) of PTGS2 on asthma traits in 299 Chinese asthmatic children and 175 controls.

Methods:  Plasma total and allergen‐specific IgE were measured by enzyme immunoassay. PTGS2.8473T→C in the 3′‐untranslated region of exon 10 and three tag SNPs covering most of the variations in PTGS2 haplotypes in Chinese were genotyped by restriction fragment length polymorphism.

Results:  Among the four SNPs, only PTGS2.8473 showed significant association with asthma (P = 0.034) and atopy (P = 0.005 when compared with non‐atopic controls; P = 0.023 with all controls). Carriers of the C allele had a 1.5‐fold (95% confidence interval: 1.01–2.30) risk of developing asthma than those homozygous for the T allele. Multivariate regression revealed significant correlations between PTGS2.8473 and forced expiratory volume in 1 s (FEV1; P = 0.002) and peak expiratory flow rate (PEFR; P = 0.001) with age and gender adjusted. Patients with the C allele of PTGS2.8473 had significantly lower FEV1 (median: 90.0%vs 98.0%; P = 0.0047) and PEFR (70.0%vs 73.5%; P = 0.0065) than those homozygous for the T allele. No significant association between plasma total and allergen‐specific IgE and these SNPs or with their haplotypes was found.

Conclusions:  PTGS2.8473 polymorphism is associated with asthma, atopy and lung function but not plasma IgE in Chinese children. This may help to explore the pharmacogenetics of COX‐2 inhibitors.
Factors associated with severity of occupational asthma with a latency period at diagnosis
Authors: Descatha A. , Leproust H. , Choudat D. , Garnier R. , Pairon J.‐C. , Ameille J.
Published: July 2010

Background:  Severity of occupational asthma at diagnosis is an important prognostic factor. The aim of this study was to determine which factors affect the severity of occupational asthma with a latency period at diagnosis.

Methods:  The study population consisted of 229 consecutive subjects with occupational asthma with a latency period recruited by four occupational health departments and divided into two groups according to the severity of the disease at diagnosis. The moderate‐severe (FEV1 <70% predicted, or PD20 methacholine ≤300 μg; n = 101) and mild (FEV1≥70% predicted and PD20 methacholine >300 μg, n = 128) groups were compared in terms of clinical and demographic parameters. Multivariate analysis using logistic regressions was performed to examine factors associated with asthma severity.

Results:  Duration of symptoms before diagnosis was significantly longer in the moderate‐severe group (mean ± SD: 6.3 ± 6.8 years vs 3.4 ± 4.4 years, P < 0.001). Sex ratio, age, atopy, smoking habits, duration of exposure before symptoms, and molecular weight of the causal agent were not significantly different between the two groups. On multivariate analysis, only duration of symptoms before diagnosis was associated with asthma severity (aOR = 1.12, 95% CI 1.05–1.18, P < 0.001).

Conclusions:  Severity of occupational asthma with a latency period at diagnosis was associated with duration of symptoms before diagnosis, but not with the type of causal agent. This finding emphasizes the need for early diagnosis and avoidance of exposure.
Disseminated Mycobacterium genavense with pulmonary nodules in a kidney transplant recipient: case report and review of the literature
Authors: Doggett J.S. , Strasfeld L.
Published: July 2010

J.S. Doggett, L. Strasfeld. Disseminated Mycobacterium genavense with pulmonary nodules in a kidney transplant recipient: case report and review of the literature. Transpl Infect Dis 2011: 13: 38–43. All rights reserved

Abstract: We report a case of disseminated Mycobacterium genavense with pulmonary nodules in a kidney transplant recipient and review the associated literature. Disease caused by M. genavense has been recognized in acquired immunodeficiency syndrome (AIDS) patients since 1990, with subsequent case reports in other immunocompromised host populations. In AIDS patients, pulmonary lesions are an uncommon finding. This is the first report to our knowledge of a human immunodeficiency virus (HIV)‐negative patient with pulmonary nodules as a feature of disseminated M. genavense. Diagnosis of M. genavense is often challenging and frequently requires nucleic acid‐based identification techniques. Because of limitations in culture and drug susceptibility testing, treatment regimens rely on reported clinical experience. This case report and literature review illustrates a successful approach to the diagnosis and treatment of disseminated M. genavense and summarizes the reports of M. genavense infection in HIV‐negative patients.
Regulation of Nucleolar RNA Metabolism by Hydrocortisone
Authors: Jacob S. T. , Sajdel E. M. , Munro H. N.
Published: July 2010

Purified rat liver nucleoli were examined for RNA polymerase activity in the presence of Mg2+ and of Mn2+ with added ammonium sulfate. Both types of activity were found to be present in this organelle.

A single injection of hydrocortisone 21‐phosphate into adrenalectomized rats resulted, after 3.5 hours, in a threefold increase in the Mg2+‐dependent polymerase activity while the Mn2+‐dependent polymerase activity was not significantly altered. When rats were injected with [14C]orotic acid hydrocortisone induced a 60% increase in the synthesis of 45 S RNA isolated from nucleoli. Pulse‐chase experiments using low doses of actinomycin D suggested that the hormone enhances the turnover of newly synthesized 45 S RNA.

Similar studies on extra‐nucleolar RNA produced smaller changes in the synthesis of this RNA fraction following hydrocortisone treatment and the pattern of RNA turnover in the chase experiments was not affected by the hormone.