bluuebunny/crossref_metadata_2025 huggingface dataset:
Is It What We Do or How We Do It? New Evidence on Agglomeration Economies and Metropolitan Growth
Authors: Malpezzi Stephen , Seah Kiat‐Ying , Shilling James D.
Published: May 2004

This paper uses data on U.S. metropolitan areas from 1970 to 1999 to examine the extent to which recent increases in earnings are attributable to agglomeration economies. We decompose the total change in earnings over the 1970–1999 period into components attributable to changes in relative growth, industry mix and interaction effects. We find strong evidence that relative growth matters more than industry mix. In addition, we find that relative growth effects are related to both localization and urbanization economies.
The Alpine Fault biogeographic hypothesis revisited
Authors: Heads Michael , Craw Robin
Published: May 2004

The New Zealand Alpine Fault is a major tectonic feature located on the mutual boundary of the Australian and Pacific Plates which is hypothesized to have undergone some 470 km of right‐lateral displacement. The Nelson and Westland provinces have moved north‐east relative to the rest of the South Island. Many plant and animal taxa show a conspicuous distribution gap in the central South Island, and traditionally this has been explained by glacial extirpation of the central populations. However, the gap is usually filled by a closely related taxon. Furthermore, the taxa involved occupy a wide ecological range, and include intertidal algae, shorefishes, lowland trees, and alpine herbs and insects that thrive around glaciers. The Alpine Fault biogeographic hypothesis [Heads, M., 1998a. Biogeographic disjunction along the Alpine Fault, New Zealand. Biol. J. Linn. Soc. 63, 161–176;Heads, M., 1998b.Coprosma decurva(Rubiaceae), a new species from New Zealand. NZ J. Bot. 36, 65–69.], based on track and phylogenetic analyses, proposes that the disjunction has instead been caused by movement on the fault pulling apart plant and animal populations.Wallis and Trewick (2001) [Wallis, G.P., Trewick, S.A., 2001. Finding fault with vicariance: a critique of Heads (1998). Syst. Biol. 50, 602–609.] provided a critique of this idea and pointed out what they felt were nine problems with it. These problems are answered here and shown to result from Wallis and Trewick's misunderstanding of aspects of geology and biology involved in the hypothesis. In particular, they have confused the age of inception of the fault with the age of movement along it, and, by neglecting the related central taxa in the “gap”, have assumed the biogeographic hypothesis to be an uninformative two‐area statement, whereas in fact it is an informative three‐ or four‐area statement.
Phylogeny of the Myllaenini and related taxa (Coleoptera: Staphylinidae: Aleocharinae)
Authors: Ahn Kee‐Jeong , Ashe James S.
Published: May 2004

A cladistic analysis of the tribe Myllaenini Ganglbauer and related genera is presented. Monophyly of the Myllaenini is tested, and the tribe is hypothesized to be a monophyletic group consisting of nine genera (Myllaena Erichson, Amazonopora Pace, Dimonomera Cameron, Bryothinusa Casey, Philomina Blackwelder, Polypea Fauvel, Brachypronomaea Sawada, Rothium Moore and Legner, and Lautaea Sawada), based on the synapomorphy of antero‐lateral angles of mentum prolonged into spinose processes. A history of the classification of the Myllaenini is discussed. The data set for phylogenetic analysis comprised 99 characters representing 297 character states derived from adult morphology. The analysis agrees on the monophyly of the Myllaenini and the monophyly of the Pronomaeini Ganglbauer (Pronomaea Erichson, Pseudomniophila Pace, Nopromaea Cameron and Tomoxelia Bernhauer). The tribe Dimonomerini (Dimonomera Cameron) is confirmed to be a member of the Myllaenini. Masuriini is a possible sister group of the Myllaenini. Stylopalpus Cameron shows a sister group relationship to the Pronomaeini. Several other clades are also consistently recovered. However, the phylogenetic relationships of the genus Dysacrita are ambiguous. The rogue genus Diglotta Champion is not recovered as a member of the Myllaenini or Pronomaeini. On the contrary, it forms a monophyletic clade with the liparocephaline genera Halorhadinus Sawada and Amblopusa Casey. Evolution of the defensive gland on abdominal tergite VII among aleocharine lineages is reconsidered, and the origin of an intertidal habitat in the Myllaenini is discussed.
Do larvae evolve the same way as adults in Tingidae (Insecta: Heteroptera)?
Authors: Guilbert Eric
Published: May 2004

A recent cladistic analysis showed that adult traits of Tingidae, which exhibit a great variety of shapes, evolved homoplastically from simple to complex (Guilbert, 2001). These complex traits, often exaggerated, were hypothesized to be adaptive. However, this study, as well as another by Lis (1999), both based on adult morphology, contradict the traditional classification of Tingidae. A new analysis is performed here, that includes larval characters, which, like those of adults, have a great variety of shapes. The results corroborate the traditional classification of the Tingidae. No clear divisions among Tinginae are drawn from the analysis, but an evolutionary pattern of shapes among Tingidae emerges from this study. There is a global tendency for larval traits to evolve convergently from simple to complex, as suggested for adults. The pattern seen in adult and larval traits is independent, but consistent. These traits can be used in the same anti‐predation context, but with different roles.
Phylogeny of the Brachytheciaceae (Bryophyta) based on morphology and sequence level data
Authors: Huttunen Sanna , Ignatov Michael S.
Published: May 2004

Brachytheciaceae is often considered a taxonomically difficult group of mosses. For example, morphological variation has led to difficulty in generic delimitation. We used DNA sequence data (chloroplast psbT‐H and trnL‐F and nuclear ITS2) together with morphology (63 characters) to examine the relationships within this family. The combined unaligned length of the DNA sequences used in the phylogenetic analyses varied between 1277 and 1343 bp. For phylogeny reconstruction we performed direct optimization, as implemented in POY. Analyses were performed with three different gap costs and the morphological data partition was weighted both: (1) equal to gap cost, and (2) with a weight of one. The utility of sensitivity analysis has recently been cast into doubt; hence in this study it was performed only to explore the effects of weighting on homology statements and topologies and to enable more detailed comparisons between earlier studies utilizing the direct optimization method. The wide sequence length variation of non‐coding ITS2 sequences resulted in character optimizations (i.e., “alignments”) of very different lengths when various gap costs were applied. Despite this variation, the topologies of equally parsimonious trees remained fairly stable. The inclusion of several outgroups, instead of only one, was observed to increase the congruence between data sets and to slightly increase the resolution. An inversion event in the 9 bp loop region in the chloroplast psbT‐N spacer in mosses has been postulated to include only uninformative variation, thus possibly negatively impacting the phylogeny reconstruction. Despite this inversion, its variation within Brachytheciaceae was clearly congruent with information from other sources, but inclusion of these 9 bp in the analysis had only a minor effect on the phylogenetic results. In the most parsimonious topology, which was obtained with equal weighting of all data, Meteoriaceae and Brachytheciaceae were resolved as monophyletic sister groups, which had recently been suggested based on a few shared morphological characters. Our study revealed some new generic relationships within the Brachytheciaceae, which are discussed in light of the morphological characters traditionally used for generic delimitation.
Phylogenetic relationships of the genus Cheilosia and the tribe Rhingiini (Diptera, Syrphidae) based on morphological and molecular characters
Authors: Ståhls Gunilla , Stuke Jens‐Hermann , Vujic Ante , Doczkal Dieter , Muona Jyrki
Published: May 2004

The phylogenetic relationships of the tribe Rhingiini and the genus Cheilosia (Diptera, Syrphidae) were investigated using morphological and molecular characters. The genus Cheilosia is one of the most diverse lineages of hoverflies (Syrphidae). The mitochondrial protein coding gene cytochrome c oxidase subunit I (COI), and the D2‐3 region of the nuclear 28S rRNA gene were chosen for sequencing, and morphological characters were scored for both adults and immature stages. The combined dataset included 56 ingroup taxa. The datasets were analyzed separately and in conjunction, using both static and dynamic alignment under the parsimony criterion. The aim of the study was to assess the phylogenetic relationships of the tribe Rhingiini, and to explore if the subgenera of Cheilosia were supported as monophyletic clades. Results showed that the monophyly of subtribes of Rhingiini remained ambiguous, especially due to unstable phylogenetic placements of the genera Portevinia and Rhingia. We recovered most subgenera of Cheilosia as monophyletic clades. Dynamic alignment, using the optimization alignment program POY, always recovered more parsimonious topologies under all parameter weighting schemes, than did parsimony analyses using static alignment and analyzed with NONA.
Close linkage of genes encoding receptors for subgroups A and C of avian sarcoma/leucosis virus on chicken chromosome 28
Authors: Elleder D. , Plachý J. , Hejnar J. , Geryk J. , Svoboda J.
Published: May 2004

Avian sarcoma and leucosis viruses (ASLV) are classified into six major subgroups (A to E and J) according to the properties of the viral envelope proteins and the usage of cellular receptors for virus entry. Subgroup A and B receptors are identified molecularly and their genomic positions TVA and TVB are mapped. The subgroup C receptor is unknown, its genomic locus TVC is reported to be genetically linked to TVA, which resides on chicken chromosome 28. In this study, we used two chicken inbred lines that carry different alleles coding for resistance (TVCR) and sensitivity (TVCS) to infection by subgroup C viruses. A backross population of these lines was tested for susceptibility to subgroup C infection and genotyped for markers from chicken chromosome 28. We confirmed the close linkage between TVA and TVC loci. Further, we have described the position of TVC on chromosome 28 relative to markers from the consensus map of the chicken genome.
cDNA cloning, genomic structure and polymorphism of the porcine FHL3 gene
Authors: Zuo B. , Xiong Y. Z. , Deng C. Y. , Su Y. H. , Wang J. , Lei M. G. , Li F. E. , Jiang S. W. , Zheng R.
Published: May 2004

LIM domain proteins are important regulators of the growth, determination and differentiation of cells. Four‐and‐a‐half LIM‐only protein 3 (FHL3) is a type of LIM‐only protein that contains four tandemly repeated LIM motifs with an N‐terminal single zinc finger (half LIM motif). In this study, we have determined the complete coding sequence of pig FHL3 which encodes a 280 amino acid protein. The coding region of the pig FHL3 gene is organized in five exons and spans an approximately 2.1‐kb genomic region. Comparative sequencing of six pig breeds revealed three single nucleotide polymorphisms (SNPs) within exon 2 of which an A→G substitution at position 313 changes a codon for arginine into a codon for glycine. The substitution was situated within a PstI recognition site and developed as a PCR‐RFLP marker for further use in population variation investigations and association analysis. The A/G polymorphism was segregating only in Landrace pigs. Association studies of the FHL3 polymorphism with carcass traits provided preliminary evidence that the PstI PCR‐restriction fragment length polymorphism (RFLP) genotype may be associated with variation in several carcass traits of interest for pig breeding. Further investigations in more Landrace pigs are needed to confirm this.
Mapping and transcription profiling of CASP1, 3, 6, 7 and 8 in relation to caspase activity in the bovine cumulus–oocyte complex
Authors: Yuan Y. Q. , Peelman L. J. , Williams J. L. , Van Zeveren A. , de Kruif A. , Law A. , Van Soom A.
Published: May 2004

So far 12 caspases have been described in mouse and human while only one (CASP13) is known in cattle. The aim of this study was to (1) search for other bovine caspases by reverse transcription and polymerase chain reaction (RT–PCR) and (2) examine the presence of bovine caspase mRNA and active protein in the cumulus–oocyte complex. Five caspases (1, 3, 6, 7 and 8) were identified, partially cloned and sequenced. Four of them were mapped. Differential transcription of the caspase genes was detected, but no active caspase protein was found in diverse bovine oocytes. Cumulus granulosa cells (CGC) contain CASP1, 6, 7 and 8 mRNA and active caspase protein. The presence of caspase mRNA and active caspase proteins in CGCs suggests the occurrence of apoptosis in cumulus–oocyte complex, while caspase activation is blocked in fresh oocytes and therefore caspase transcription cannot be used to predict the oocyte developmental capacity.
The chicken proinflammatory cytokines interleukin‐1β and interleukin‐6: differences in gene structure and genetic location compared with their mammalian orthologues
Authors: Kaiser P. , Rothwell L. , Goodchild M. , Bumstead N.
Published: May 2004

The genes encoding the chicken proinflammatory cytokines interleukin (IL)‐1B and IL‐6 were cloned, sequenced and mapped. The exon:intron structure of the coding region of chicken IL1B corresponds almost exactly to those of mammalian IL1B. As yet, we have no evidence for a 5′‐UTR non‐coding exon equivalent to that found in mammalian IL1B. The exon:intron structure of chicken IL6 differs from those of mammalian IL6, having one exon fewer (the first two exons in mammalian IL6 genes appear to be fused in the chicken gene). We were unable to clone or sequence the promoter of chicken IL1B. The chicken IL6 promoter shares a number of potential regulatory sequences similar to those found in the human IL6 promoter. These putative elements include (5′–3′) a glucocorticoid response element (GRE), an AP‐1 binding site, an NF‐IL‐6 binding site (albeit in the reverse orientation), an NF‐κB binding site, a second AP‐1 binding site and a TATAAA box. A further GRE, a cAMP response element and regions with homology to c‐fos serum responsive elements or retinoblastoma control elements were absent. Promoter sequence polymorphisms were not identified in eight different inbred chicken lines. A restriction single‐stranded conformational polymorphism was identified which enabled chicken IL1B to be genetically mapped to one end of chromosome 2. Chicken IL6 was mapped by fluorescent in situ hybridization also to chromosome 2, at an FLpter of 0.26.
Genetic polymorphisms and antiviral activity in the bovine MX1 gene
Authors: Nakatsu Y. , Yamada K. , Ueda J. , Onogi A. , Ables G. P. , Nishibori M. , Hata H. , Takada A. , Sawai K. , Tanabe Y. , Morita M. , Daikohara M. , Watanabe T.
Published: May 2004

Bovine MX1 cDNAs consisting of 2280 bp from 11 animals of five breeds and from a cultured cell line were sequenced and compared with previously reported data. Ten nucleotide substitutions were synonymous mutations, and a single nucleotide substitution at 458 resulted in an amino acid exchange of Ile (ATT) and Met (ATG). A 13‐bp deletion–insertion mutation was also found in the 3′‐UTR. Based on the nucleotide substitutions found in this study, bovine MX1 cDNA was classified into 11 genotypes. A phylogenetic tree of the 11 genotypes suggested that the genotypes observed in Brahman were a great genetic distance from other genotypes. An 18‐bp deletion–insertion variation at position 171 was found to be the result of alternative splicing. The 18‐bp deletion–insertion is located at the boundary between exon  3 and intron 3. Permanently transfected 3T3 cell lines expressing bovine MX1 mRNA were established to analyse the antiviral potential against VSVΔG*‐G infection. Transfected cell clones expressing bovine MX1 mRNA showed a significantly smaller number of cells infected with VSVΔG*‐G compared with the control cells. These results indicate that the bovine MX1 protein has potent antiviral activity.
Radiation hybrid map assignments of 11 ESTs obtained from a 28‐day‐old swine embryo cDNA library to the IMpRH map
Authors: Sarker N. , Kiuchi S. , Yasue H. , Mitsuhashi T.
Published: May 2004

In order to improve the map resolution and to locate more genes on the porcine radiation hybrid map, expressed sequence tags (ESTs) were isolated from a 28‐day‐old normal pig embryo cDNA library. The ESTs were sequenced from the 5′‐end and similarities were checked with sequences registered in the NCBI DNA database (http://www.ncbi.nlm.nih.gov/blast/). The ESTs sequences which have high identity scores (>80%) against human genes or ESTs were further sequenced from the 3′ untranslated region. The ESTs which were sequenced successfully were used to design primers for PCR analysis of the radiation hybrid panel. Eleven ESTs were physically mapped to porcine chromosomes 2, 4, 8, 10, 13, 14 and X. The localizations are in agreement with the comparative mapping data between human and pig. The results will provide unique information to the comparative map of human and pig.
Genetic mapping of quantitative trait loci affecting body weight, egg character and egg production in F2 intercross chickens
Authors: Sasaki O. , Odawara S. , Takahashi H. , Nirasawa K. , Oyamada Y. , Yamamoto R. , Ishii K. , Nagamine Y. , Takeda H. , Kobayashi E. , Furukawa T.
Published: May 2004

Phenotypic measurements of chicken egg character and production traits are restricted to mature females only. Marker assisted selection of immature chickens using quantitative trait loci (QTL) has the potential to accelerate the genetic improvement of these traits in the chicken population. The QTL for 12 traits (i.e. body weight (BW), six for egg character, three for egg shell colour and two for egg production) of chickens were identified. An F2 population comprising 265 female chickens obtained by crossing White Leghorn and Rhode Island Red breeds and genotyped for 123 microsatellite markers was used for detecting QTL. Ninety‐six markers were mapped on 25 autosomal linkage groups, and 13 markers were mapped on one Z chromosomal linkage group. Eight previous unmapped markers were assigned to their respective chromosomes in this study. Significant QTL were detected for BW on chromosomes 4 and 27, egg weight on chromosome 4, the short length of egg on chromosome 4, and redness of egg shell colour (using the L*a*b* colour system) on chromosome 11. A significant QTL on the Z chromosome was linked with age at first egg. Significant QTL could account for 6–19% of the phenotypic variance in the F2 population.
A first‐generation microsatellite linkage map of the Japanese quail
Authors: Kayang B. B. , Vignal A. , Inoue‐Murayama M. , Miwa M. , Monvoisin J. L. , Ito S. , Minvielle F.
Published: May 2004

A linkage map of the Japanese quail (Coturnix japonica) genome was constructed based upon segregation analysis of 72 microsatellite loci in 433 F2 progeny of 10 half‐sib families obtained from a cross between two quail lines of different genetic origins. One line was selected for long duration of tonic immobility, a behavioural trait related to fearfulness, while the other was selected based on early egg production. Fifty‐eight of the markers were resolved into 12 autosomal linkage groups and a Z chromosome‐specific linkage group, while the remaining 14 markers were unlinked. The linkage groups range from 8 cM (two markers) to 206 cM (16 markers) and cover a total map distance of 576 cM with an average spacing of 10 cM between loci. Through comparative mapping with chicken (Gallus gallus) using orthologous markers, we were able to assign linkage groups CJA01, CJA02, CJA05, CJA06, CJA14 and CJA27 to chromosomes. This map, which is the first in quail based solely on microsatellites, is a major step towards the development of a quality molecular genetic map for this valuable species. It will provide an important framework for further genetic mapping and the identification of quantitative trait loci controlling egg production and fear‐related behavioural traits in quail.
Verification of selective DNA pooling methodology through identification and estimation of the DGAT1 effect
Authors: Fisher P. J. , Spelman R. J.
Published: May 2004

The discovery of markers linked to genes that are responsible for traits of interest to the dairy industry might prove useful because they could aid in selection and breeding decisions. We have developed a selective DNA pooling methodology to allow us to efficiently screen the bovine genome in order to find genes responsible for production traits. Using markers on chromosome 14 as a test case, we identified a gene (DGAT1) previously known to affect three traits (fat yield, protein yield and total milk yield). Furthermore, we predicted similar effects to those previously shown for DGAT1 in a New Zealand Holstein‐Friesian herd. Additionally, we showed a low error rate (1.6%) for the pooling procedure. Hence we are confident that we can apply this procedure to an entire genome scan in the search for quantitative trait loci (QTL).
Evidence for quantitative trait loci affecting twinning rate in North American Holstein cattle
Authors: Cruickshank J. , Dentine M. R. , Berger P. J. , Kirkpatrick B. W.
Published: May 2004

Twinning in dairy cattle has been associated with many negative health and reproductive events that cause economic loss to the producer. Reports have suggested that twinning rates are increasing and that there may be a positive relationship between milk production and twinning frequency. Putative quantitative trait loci (QTL) for twinning and ovulation rate on bovine chromosomes 5, 7, 19 and 23 have been previously identified in other populations. The objective of this study was to detect and possibly confirm the existence and effects of these QTL in the North American Holstein population. Half‐sib families of 20 North American Holstein sires with above average twinning rate predicted transmitting abilities (PTA) comprised the sample population under investigation. Twinning rate PTA values had been estimated from calving data. DNA extracted from semen samples was analysed using 45–61 microsatellite markers across the four chromosomes. Marker heterozygosity of the patriarchs averaged 62%. Evidence of twinning QTL was found in multiple families on chromosomes 5, 7 and 23 and in one family on chromosome 19. Four of the sires formed one three‐generation family: one sire and three half‐sib sons with sons of their own. This extended family was analysed with additional markers confirming a twinning QTL of significant size on chromosome 5.
Mapping of the porcine oestrogen receptor 2 gene and association study with litter size in Iberian pigs
Authors: Muñoz G. , Óvilo C. , Amills M. , Rodríguez C.
Published: May 2004

Genes encoding the oestrogen receptors (ESR) are considered candidate genes for prolificacy traits due to the key role these molecules play in the regulation of reproductive physiology. In this paper, we report the assignment of the pig ESR2 gene to porcine chromosome 1 by radiation hybrid mapping. Most of the ESR2 cDNA was sequenced from Iberian pig ovarian RNA samples and one A/G single nucleotide polymorphism (SNP) was found at exon  5, being associated with a Met/Val substitution at position 949. This SNP was genotyped using a PCR‐RFLP (Hsp92II) protocol and its potential effect on litter size was evaluated in two Iberian pig populations. However, no statistically significant association between the ESR2 polymorphism and litter size was found.
Impact of candidate sire number and sire relatedness on DNA polymorphism‐based measures of exclusion probability and probability of unambiguous parentage
Authors: Sherman G. B. , Kachman S. D. , Hungerford L. L. , Rupp G. P. , Fox C. P. , Brown M. D. , Feuz B. M. , Holm T. R.
Published: May 2004

Genetic paternity testing can provide sire identity data for offspring when females have been exposed to multiple males. However, correct paternity assignment can be influenced by factors determined in the laboratory and by size and genetic composition of breeding groups. In the present study, DNA samples from 26 commingled beef bulls and their calves from the Nebraska Reference Herd‐1 (NRH1), along with previously reported Illinois Reference/Resource Families data, were used to estimate the impact of sire number and sire relatedness on microsatellite‐based paternity testing. Assay performance was measured by exclusion probabilities and probabilities of unambiguous parentage (PUP) were derived. Proportion of calves with unambiguous parentage (PCUP) was also calculated to provide a readily understandable whole‐herd measure of unambiguous paternity assignment. For NRH1, theoretical and observed PCUP values were in close agreement (85.3 and 85.8%, respectively) indicating good predictive value. While the qualitative effects on PUP values of altering sire number and sire relatedness were generally predictable, we demonstrate that the impacts of these variables, and their interaction effects, can be large, are non‐linear, and are quantitatively distinct for different combinations of sire number and degree of sire relatedness. In view of the potentially complex dynamics and practical consequences of these relationships in both research and animal production settings, we suggest that a priori estimation of the quantitative impact of a given set of interacting breeding group‐specific and assay‐specific parameters on PUP may be indicated, particularly when candidate sire pools are large, sire relatedness may be high, and/or loci numbers or heterozygosity values may be limiting.
Subclinical alteration of taste sensitivity induced by candesartan in healthy subjects
Authors: Tsuruoka Shuichi , Wakaumi Michi , Nishiki Kenta , Araki Nobutaka , Harada Kenichi , Sugimoto Koichi , Fujimura Akio
Published: May 2004

Aims

There have been case reports of taste disturbance for the angiotensin II receptor blockers losartan and valsartan, but not for candesartan. This study was undertaken to examine whether candesartan causes taste disturbance.

Methods

Candesartan cilexetil (4 mg day−1) or vehicle was given to healthy volunteers (n = 8) for 7 days in a randomized, double‐blind, placebo‐controlled, cross‐over design with a 2‐week washout period. Clinical gustometry using the filter‐paper disc test and electrogustometry were sequentially performed before and at the end of each trial. Serum and salivary zinc concentrations were also measured.

Results

Detection thresholds of four basic tastes (sweet, salty, sour and bitter) determined by the paper disc test were significantly (P < 0.05 in all tests) worsened (i.e. score of test increased) after repeated dosing of the drug, although the subjects did not notice such changes. The mean ± SEM (and 95% CI) scores of the four tastes at just before the seventh dosing of candesartan or vehicle was 3.38 ± 0.32 (3.02, 3.74) and 2.63 ± 0.18 (2.18, 3.08) for sweetness, 3.63 ± 0.38 (4.49, 2.77) and 2.63 ± 0.26 (3.27, 1.98) for salt, 4.01 ± 0.42 (3.04, 4.98) and 2.61 ± 0.32 (3.35, 1.87) for sourness, 4.01 ± 0.38 (3.22, 4.80) and 2.99 ± 0.33 (2.24, 3.74) for bitterness, for candesartan and placebo, respectively. Electrogustometry confirmed the candesartan‐related taste disturbance. Serum and salivary zinc concentrations were not influenced by candesartan.

Conclusions

These data suggest that candesartan subclinically reduces taste sensitivity after repeated dosing in healthy subjects. Because similar events are reported for losartan and valsartan in case reports, this adverse effect might be a class effect of angiotensin‐II receptor blockers (ARBs).
Population pharmacokinetics of pyrimethamine and sulfadoxine in children with congenital toxoplasmosis
Authors: Trenque T. , Simon N. , Villena I. , Chemla C. , Quereux C. , Leroux B. , Jaussaud R. , Rémy G. , Dupouy D. , Millart H. , Pinon J.‐M. , Urien S.
Published: May 2004

Aims

To develop a population pharmacokinetic model for pyrimethamine (PYR) and sulfadoxine (SDX) in children with congenital toxoplasmosis.

Methods

Children were treated with PYR (1.25 mg kg−1) and SDX (25 mg kg−1) (Fansidar®) plus folinic acid (Lederfoline® 5 mg). Plasma concentrations, available from a therapeutic drug monitoring database, were determined by high‐performance liquid chromatogrphy. Population pharmacokinetic analysis was performed using a nonlinear mixed effects model.

Results

Eighty‐nine children, aged 1 week to 14 years and weighing 2.9–59 kg, were available for evaluation. Both PYR and SDX concentration‐time profiles were best described by a one‐compartment open model. Volume of plasma distribution (V) and clearance (CL) were significantly related to body weight (BW) using an allometric function. Typical CL and V estimates (95% confidence interval), for a child weighing 11 kg were 5.50 (5.28, 5.73) l day−1 and 36 (33, 39) l for PYR and 0.26 (0.25, 0.27) l day−1 and 2.1 (1.9, 2.3) l for SDX. For BW between 3.5 and 60 kg, plasma half‐lives were predicted to vary from 4.0 to 5.2 days for PYR, and from 5.0 to 7.5 days for SDX.

Conclusion

This study indicated that body weight influences PYR and SDX pharmacokinetics in children. To optimize PYR/SDX combination treatment in congenital toxoplasmosis, short dosing intervals in very young low‐wight children are probably appropriate.
Purging iron from the heart
Authors: Hershko Chaim , Cappellini Maria D , Galanello Renzo , Piga Antonio , Tognoni Gianni , Masera Giuseppe
Published: May 2004

Methods are now available to measure the magnitude of iron accumulation in the heart. Their validation currently relies on indirect evidence and not on chemical estimation in cardiac biopsies. All patients with symptomatic heart disease appear to have abnormal T2* values, but many patients without symptomatic heart disease also have evidence of increased myocardial iron. Although there is no proof to date that increased myocardial iron, as evidenced by abnormal magnetic resonance imaging, carries an adverse prognosis, it is likely that such new information will affect the chelating programme of patients. In these cases, there are a number of options available: (i) ongoing treatment with either desferrioxamine (DFO) or deferiprone may be intensified; (ii) the patient may be switched to the alternative chelator or (iii) combined chelation with both DFO and deferiprone may be started, which is more effective than using either chelator alone. For patients with symptomatic heart disease, continuous intravenous DFO with, or without deferiprone, remains the currently recommended treatment, in view of its documented ability to salvage these patients.
Genetic variants of the extra‐large stimulatory Gs protein alpha‐subunit and risk of thrombotic and haemorrhagic disorders
Authors: González‐Conejero R. , Corral J. , Guerrero J. A. , Iniesta J. A. , Rivera J. , De Arriba F. , Vicente V.
Published: May 2004

A polymorphism of the gene encoding the extra‐large stimulatory G‐protein α‐subunit (XLαs), originally identified in three patients with a bleeding tendency, involved a 36‐bp insertion and two missense changes. A paternally‐inherited insertion displayed a moderate platelet Gsα over‐expression, which lead to platelet hypo‐reactivity. These data prompted us to investigate the genetic, functional and clinical relevance of this polymorphism in the Mediterranean population. We included 414 healthy subjects and three case/control studies: 263 consecutive patients with a first episode of primary intracerebral haemorrhage, 195 patients with deep venous thrombosis, and 104 patients with cerebrovascular disease. Controls were selected by approximating criteria to match selected risk factors to patients. Moreover, we performed studies of platelet function. We developed a simple method to determine the methylated allele, by digestion of genomic DNA with Sma I before polymerase chain reaction amplification. We identified two new rare variants, resulting from the loss of repeat units 7 and 5. The AB genotype was present in 3·6% of healthy population and the prevalence of the B allele was similar among cases and controls. Accordingly, the non‐methylated B allele did not modify either the expression of platelet Gsα or the platelet response to Gs‐agonists. Thus, our study suggests a minor functional role of XLαs polymorphism in thrombotic or in haemorrhagic disorders.
Characterisation of Mcl‐1 cleavage during apoptosis of haematopoietic cells
Authors: Clohessy John G. , Zhuang Jianguo , Brady Hugh J. M.
Published: May 2004

Mcl‐1 is essential for normal haematopoiesis, being required for lymphocyte development and maintenance. Its role in haematopoietic differentiation and development is associated with its function as an anti‐apoptotic member of the Bcl‐2 family of proteins although the underlining mechanism is poorly understood. We have characterized caspase cleavage of the Mcl‐1 protein during apoptosis. Caspase cleavage resulted in the removal of the PEST regions from the protein and generation of a fragment containing the BH‐1, ‐2 and ‐3 homology domains. Removal of the PEST regions did not appear to alter Mcl‐1 stability, suggesting that these regions are not responsible for Mcl‐1's short half‐life. In addition, unlike cleavage of Bcl‐2 and Bcl‐XL, which resulted in pro‐apoptotic fragments, cleaved forms of Mcl‐1 were unable to induce apoptosis. This novel regulation of Mcl‐1 may have important implications not only for its role in apoptosis but also for the essential role it plays in the differentiation and development of haematopoietic cells.
Early onset hereditary hemochromatosis resulting from a novel TFR2 gene nonsense mutation (R105X) in two siblings of north French descent
Authors: Le Gac G. , Mons F. , Jacolot S. , Scotet V. , Férec C. , Frébourg T.
Published: May 2004

The molecular basis of hereditary hemochromatosis (HH) is more complex than previously expected. More than 80% of hemochromatosis probands of Northern European descent are homozygous for the C282Y HFE gene mutation. However, five novel non‐related‐HFE HH forms have now been identified. The transferrin receptor(TFR2)‐linked form is inherited in an autosomal recessive pattern and is considered to be an adult‐onset syndrome. Until now, it has been associated with five mutations that have only been detected in Japanese and southern European patients. Here, we report the identification of a novel TFR2 nonsense mutation in two related French adolescents. We discuss the phenotype of this sibling pair from precedent biological and clinical findings as well as the expected role of TFR2 in iron homeostasis. Finally, we suggest that iron overload phenotypes associated with mutations in TFR2 may be intermediate between those related to mutations in HFE and those related to mutations in juvenile hemochromatosis genes.
Infused CD34+ cell dose, but not tumour cell content of peripheral blood progenitor cell grafts, predicts clinical outcome in patients with diffuse large B‐cell lymphoma and follicular lymphoma grade 3 treated with high‐dose therapy
Authors: Blystad A. K. , Delabie J. , Kvaløy S. , Holte H. , Vålerhaugen H. , Ikonomou I. , Kvalheim G.
Published: May 2004

Previously, we have shown that patients with diffuse large B‐cell lymphoma (DLBCL) transplanted with contaminated bone marrow (BM) generally have a poor outcome. Whether this is also the case when peripheral blood progenitor cell (PBPC) grafts are used is not known. Forty‐three patients with chemosensitive DLBCL or follicular lymphoma grade 3 (FLgr3) were treated with high‐dose therapy (HDT) and autologous stem cell support. Nine patients received purged grafts. Quantitative real‐time polymerase chain reaction (QRT‐PCR) for either the BCL2/IgH translocation or allele specific oligonucleotide (ASO) QRT‐PCR for the immunoglobulin heavy chain (IgH) complementarity‐determining region 3 were used. Nine of 25 (36%) PBPC grafts contained tumour cells as tested by QRT‐PCR, including two grafts purged by CD34+ cell enrichment combined with B‐cell depletion. The level of contamination of the PBPC/CD34+ cells ranged from 0 to 8·28%. No relationship could be shown between the total number of tumour cells infused and relapse. Patients receiving PCR‐positive or PCR‐negative PBPC grafts had similar progression‐free survival (PFS) (P = 0·49). However, a significant difference was seen in PFS and overall survival (OS) for the patients given ≥6·1 × 106 CD34+ cells/kg compared with those given <6·1 × 106 CD34+ cells/kg (P = 0·01 and P < 0·05 respectively).
WT1 gene expression: an excellent tool for monitoring minimal residual disease in 70% of acute myeloid leukaemia patients – results from a single‐centre study
Authors: Østergaard Mette , Olesen Lene Hyldahl , Hasle Henrik , Kjeldsen Eigil , Hokland Peter
Published: May 2004

Following induction chemotherapy for acute myeloid leukaemia (AML), sensitive determination of minimal residual disease (MRD) in patients achieving complete remission (CR) should enable the detection of early relapse and allow intervention at a more favourable stage than at overt relapse. We have determined the expression levels of the Wilms’ tumour gene (WT1) by real‐time quantitative polymerase chain reaction (RQ‐PCR) in peripheral blood and bone marrow in 133 newly diagnosed AML patients and compared them with those in healthy volunteers. At diagnosis, the WT1 level exceeded normal expression in 118 of 133 (89%) patients, and was high enough to allow for detection of a WT1 decrease of least 1000‐fold in 98 of 133 (74%) patients following induction therapy. Concomitant monitoring of fusion transcripts (PML‐RARα, AML1‐ETO, MLL‐MLL, CBFβ‐MYH11, or DEK‐CAN) in 38 patients identified different relationships between WT1 and fusion transcript levels, the AML1‐ETO group showing remarkably low levels of WT1 compared with fusion transcript. In 32 patients analysed longitudinally there was close concordance between relapse and increased WT1 levels. Parallel longitudinal monitoring of WT1 and fusion transcript showed close correlation in 18 of 18 patients. We conclude that WT1 expression by RQ‐PCR may be employed as a tool to detect MRD in the majority of fusion transcript‐negative AML patients.
E4BP4 expression is regulated by the t(17;19)‐associated oncoprotein E2A‐HLF in pro‐B cells
Authors: Yeung Jenny , O'Sullivan Elaine , Hubank Mike , Brady Hugh J. M.
Published: May 2004

The E4BP4 basic leucine zipper (bZIP) transcription factor is regulated by interleukin‐3 (IL‐3) in pro‐B cells and has been reported to promote survival of the murine IL‐3‐dependent pro‐B cell lines, FL5.12 and Baf‐3. The E2A‐HLF oncoprotein arises from a t(17;19) translocation in childhood pro‐B cell acute lymphoblastic leukaemia and acts as an anti‐apoptotic factor in FL5.12 and Baf‐3 cells. To assess the functions of E2A‐HLF and E4BP4 in cell survival, a tetracycline‐inducible system was established in Baf‐3 cells to express E4BP4 or E2A‐HLF. Upon IL‐3 withdrawal, expression of E2A‐HLF conferred resistance to apoptosis whereas overexpression of E4BP4 did not. E4BP4 and E2A‐HLF both recognized the same DNA sequence in reporter gene assays, but had opposite effects on transcription. E2A‐HLF acts as a transcriptional activator and E4BP4 as a transcriptional repressor. Furthermore, E4BP4 is a downstream transcriptional target of E2A‐HLF. Our data suggests that the overexpression of E4BP4 is unable to block apoptosis induced by IL‐3 withdrawal and that the expression of E2A‐HLF does not replace the function of E4BP4 in mediating survival.
Validation and extension of the EBMT Risk Score for patients with chronic myeloid leukaemia (CML) receiving allogeneic haematopoietic stem cell transplants
Authors: Passweg Jakob R. , Walker Irwin , Sobocinski Kathleen A. , Klein John P. , Horowitz Mary M. , Giralt Sergio A. ,   Chronic Leukemia Study Writing Committee of the International Bone Marrow Transplant Registry
Published: May 2004

The European Group for Blood and Marrow Transplantation (EBMT) devised a scoring system to predict survival after allogeneic haematopoietic stem cell transplantation (HSCT) for chronic myeloid leukaemia (CML). The present International Bone Marrow Transplant Registry study of 3211 patients tested the EBMT Risk Score in a independent population, investigated the value of adding other variables, evaluated a new risk score specifically for chronic phase and compared the allograft risk scores with risk scores established by Sokal in 1984 and Hasford in 1998 for survival with non‐transplant treatments. The primary outcome was 5‐year survival after HSCT; survival curves, regression models and measurements of explained variation were used to compare scores. Using the EBMT scoring system, survival in the independent dataset was almost identical to those in the original EBMT publication, thus validating the EBMT Risk Score. Adding one extra variable, performance status, or designing a score specifically for early chronic phase by using the original five variables with different breakpoints gave results only slightly better than the original EBMT Score. Sokal and Hasford Scores did not predict survival after HSCT. We concluded that the EBMT Risk Score does not currently require modification.
Depletion and impaired interferon‐α‐producing capacity of blood plasmacytoid dendritic cells in human T‐cell leukaemia virus type I‐infected individuals
Authors: Hishizawa Masakatsu , Imada Kazunori , Kitawaki Toshio , Ueda Maki , Kadowaki Norimitsu , Uchiyama Takashi
Published: May 2004

Dendritic cells (DCs) play an important role in innate and adaptive immunity. There are two major populations of blood DCs, myeloid DCs (myDCs) and plasmacytoid DCs (pcDCs). pcDCs are particularly important in antiviral as well as in general host defence, as they are the principal producers of type I interferons (IFNs). In this study, we analysed myDCs and pcDCs in healthy controls, human T‐cell leukaemia virus type I (HTLV‐I)‐infected asymptomatic carriers (ACs), and patients with adult T‐cell leukaemia (ATL). ATL patients had significantly decreased number of pcDCs and myDCs compared with controls. IFN‐α production by peripheral blood mononuclear cells (PBMCs) was markedly reduced in ATL patients. Purified pcDCs from ACs were found to have impaired IFN‐α‐producing capacity, suggesting a functional defect in pcDCs in HTLV‐I‐infected individuals. Interestingly, pcDCs were shown to be susceptible to HTLV‐I infection. Thus, impaired IFN‐α production by pcDCs may contribute to the immunodeficiency observed in ATL. Furthermore, IFN‐α‐producing capacity was inversely correlated with HTLV‐I proviral load in PBMCs from ACs, suggesting a role for pcDCs in maintaining the carrier state. Taken together, we hypothesize that the depletion and impaired IFN‐α‐producing capacity of blood DCs may contribute to the immunodeficiency in ATL and/or the development of ATL.
Secretion of functional plasma haemostasis proteins in long‐term primary cultures of human hepatocytes
Authors: Biron‐Andréani Christine , Bezat‐Bouchahda Corinne , Raulet Edith , Pichard‐Garcia Lydiane , Fabre Jean‐Michel , Saric Jean , Baulieux Jacques , Schved Jean‐François , Maurel Patrick
Published: May 2004

This study was designed to investigate the ability of long‐term primary cultures of adult human hepatocytes to secrete the main haemostasis proteins. Factors II, V, VII, VIII, PIVKA‐II (protein induced by vitamin K 1 absence or antagonist II), fibrinogen and antithrombin were quantified in culture medium by immunological methods and by measuring the coagulant activity of factors II, V and VII. All the haemostasis protein antigens except the factor VIII antigen (FVIII:Ag) were found in the culture medium throughout the culture period. The clotting activity of each factor correlated well with antigen level. In addition, fibrinogen and fibrin were detected in the fibrillar material following incubation of the culture medium with thromboplastin. Moreover, adding vitamin K 1 to the culture medium resulted in a significant increase of factors II and VII and a reciprocal decrease of the PIVKA‐II, and adding von Willebrand factor resulted in a drastic increase of the level of FVIII:Ag. We conclude that, in our culture system, normal adult human hepatocytes retain their capacity to secrete haemostasis proteins for at least 30 days.
Gene expression profiling in the myelodysplastic syndromes using cDNA microarray technology
Authors: Pellagatti Andrea , Esoof Noor , Watkins Fiona , Langford Cordelia F. , Vetrie David , Campbell Lisa J. , Fidler Carrie , Cavenagh James D. , Eagleton Helen , Gordon Peter , Woodcock Barrie , Pushkaran Beena , Kwan Mark , Wainscoat James S. , Boultwood Jacqueline
Published: May 2004

The myelodysplastic syndromes (MDS) comprise a heterogeneous group of clonal disorders of the haematopoietic stem cell and primarily involve cells of the myeloid lineage. Using cDNA microarrays comprising 6000 human genes, we studied the gene expression profiles in the neutrophils of 21 MDS patients, seven of which had the 5q‐ syndrome, and two acute myeloid leukaemia (AML) patients when compared with the neutrophils from pooled healthy controls. Data analysis showed a high level of heterogeneity of gene expression between MDS patients, most probably reflecting the underlying karyotypic and genetic heterogeneity. Nevertheless, several genes were commonly up or down‐regulated in MDS. The most up‐regulated genes included RAB20, ARG1, ZNF183 and ACPL. The RAB20 gene is a member of the Ras gene superfamily and ARG1 promotes cellular proliferation. The most down‐regulated genes include COX2, CD18, FOS and IL7R. COX2 is anti‐apoptotic and promotes cell survival. Many genes were identified that are differentially expressed in the different MDS subtypes and AML. A subset of genes was able to discriminate patients with the 5q‐ syndrome from patients with refractory anaemia and a normal karyotype. The microarray expression results for several genes were confirmed by real‐time quantitative polymerase chain reaction. The MDS‐specific expression changes identified are likely to be biologically important in the pathophysiology of this disorder.
Variant MYST4‐CBP gene fusion in a t(10;16) acute myeloid leukaemia
Authors: Murati Anne , Adélaïde José , Mozziconacci Marie‐Joëlle , Popovici Cornel , Carbuccia Nadine , Letessier Anne , Birg Françoise , Birnbaum Daniel , Chaffanet Max
Published: May 2004

We report a novel fusion of the MYST4 and CBP genes in an acute myeloid leukaemia (AML)‐M4 patient exhibiting t(10;16)(q22;p13) and t(11;17)(q23;q21). The t(10;16)(q22;p13) resulted in a rearrangement, where MYST4‐CBP and CBP‐MYST4 chimaeric transcripts were products of in‐frame fusions of MYST4 exon  17 to CBP exon  6 and CBP exon  4 to MYST4 exon  18 respectively. The potential resulting chimaeric proteins showed similarities with MYST3‐CBP, MYST3‐P300 and MYST3‐NCOA2 putative fusion proteins found in other cases of AML.