bluuebunny/crossref_metadata_2025 huggingface dataset:
The role of calcium/calmodulin-dependent protein kinase II on the inactivation of MAP kinase and p34cdc2 kinase during fertilization and activation in pig oocytes
Authors: Ito Junya , Kawano Natsuko , Hirabayashi Masumi , Shimada Masayuki
Published: September 2004

The objective of this study was to investigate the role of calmodulin-dependent protein kinase II (CaMKII) during fertilization in the pig. Since it has been reported that CaMKII is involved in the capacitation and acrosome reaction of spermatozoa, we tested whether supplementation with the CaMKII inhibitor, KN-93, in the fertilization medium affected sperm penetration. The results showed that the addition of KN-93 in the fertilization medium significantly reduced the rate of sperm penetration into oocytes. However, pre-treatment with KN-93 beforein vitrofertilization (IVF) did not significantly affect sperm penetration, but it did affect pronuclear formation in a dose-dependent manner. In the oocytes pre-treated with KN-93 before IVF and then co-cultured with spermatozoa without the drug, the decrease in p34cdc2kinase and the cyclin B1 level were significantly suppressed as compared with those in penetrated oocytes without treatment with KN-93. However, the decrease in MAP kinase activity was not affected by KN-93. Additional treatment with KN-93 after Ca2+ionophore treatment also inhibited the reduction in p34cdc2kinase activity and the cyclin B1 level, but not MAP kinase activity. Treatment with KN-92, an inactive KN-93 analogue, did not significantly affect sperm penetration and pronuclear formation. In conclusion, the activation of CaMKII by artificial stimuli or sperm stimulated the disruption of cyclin B1 and the inactivation of p34cdc2kinase, but did not affect MAP kinase inactivation during oocyte activation in pigs.
Fyn kinase–tubulin interaction during meiosis of rat eggs
Authors: Talmor-Cohen A , Tomashov-Matar R , Tsai W B , Kinsey W H , Shalgi R
Published: September 2004

Prior to fertilization, the spindle of vertebrate eggs must remain stable and well organized during the second meiotic meta-phase arrest (MII). In a previous study we have determined that the completion of meiosis is a Src family kinase (SFK)-dependent event. In the current study we have used the SFK inhibitors, SU6656 and PP2, and demonstrated that inhibition of SFKs caused the formation of a disorganized spindle. The observation that proper organization of an MII spindle is an SFK-dependent process, combined with our previous finding that Fyn kinase is localized at the microtubules (MTs), prompted us to examine the potential role of Fyn in MT signaling. Our results show an association between Fyn and tubulin, the ability of Fyn to phosphorylate tubulinin vitroand stimulation of meiosis completion by injection of a constitutively active form of Fyn (CAF).

We suggested that SFKs mediate significant functions during the organization of the MII spindle. In view of CAF injection experiments, and of the pronounced concentration of Fyn kinase at the spindle, we propose that Fyn may play an important role in some aspects of the spindle functions, possibly those involving the MTs.
Development of mouse-specific contraceptive vaccines: infertility in mice immunized with peptide and polyepitope antigens
Authors: Hardy Christopher M , Clydesdale Gavin , Mobbs Karen J
Published: September 2004

Mouse-specific immunocontraceptive peptides have been identified in mouse proteins with key roles in reproduction from sequence comparisons to other species and tested for efficacy as immunocontraceptive antigens. Peptides were derived from granulocyte-macrophage colony-stimulating factor (GMCSF), the placental 27 kDa heat-shock protein (HSP), leukemia inhibitory factor receptor (LIFR), oviduct glycoprotein (OGP), proliferin (PLF), prolactin (PRL), sperm protein SP56 and mouse zona pellucida subunits 1 and 3 (ZP1, ZP3). Fertility of female BALB/c mice was reduced after immunization with several peptides either conjugated to a carrier protein or in the form of recombinant polyepitopes. The most effective conjugated peptides (SP56, GMCSF and PRL) induced peptide-specific serum antibodies and reduced fertility by 50%. Fertility of mice was also reduced after immunization with polyepitope antigens containing up to five different peptides fused to maltose-binding protein, but antibodies were not produced against all the encoded peptides. The most effective polyepitope antigen (containing PLF, SP56, ZP1 and ZP3 peptides) reduced fertility by 50% but induced only SP56 and ZP1 antibodies. We demonstrate that lack of antibody response to a given peptide epitope (ZP3) can be overcome if repeated copies are used in the polyepitope antigen construct, but the effect varies between mouse strains. We conclude that infertility induced in mice with a range of peptide-based vaccines is dependent on antigen formulation and genetic factors but does not necessarily correlate with peptide-specific antibody levels. In light of these results, strategies to improve the efficacy of peptide-based antifertility vaccines are discussed.
A Microsatellite Genetic Linkage Map for XiphophorusSequence data from this article have been deposited with the EMBL/GenBank Data Libraries under accession nos. AY258640, AY258896.
Authors: Walter R B , Rains J D , Russell J E , Guerra T M , Daniels C , Johnston Dennis A , Kumar Jay , Wheeler A , Kelnar K , Khanolkar V A , Williams E L , Hornecker J L , Hollek L , Mamerow M M , Pedroza A , Kazianis S
Published: September 2004

Interspecies hybrids between distinct species of the genus Xiphophorus are often used in varied research investigations to identify genomic regions associated with the inheritance of complex traits. There are 24 described Xiphophorus species and a greater number of pedigreed strains; thus, the number of potential interspecies hybrid cross combinations is quite large. Previously, select Xiphophorus experimental crosses have been shown to exhibit differing characteristics between parental species and among the hybrid fishes derived from crossing them, such as widely differing susceptibilities to chemical or physical agents. For instance, genomic regions harboring tumor suppressor and oncogenes have been identified via linkage association of these loci with a small set of established genetic markers. The power of this experimental strategy is related to the number of genetic markers available in the Xiphophorus interspecies cross of interest. Thus, we have undertaken the task of expanding the suite of easily scored markers by characterization of Xiphophorus microsatellite sequences. Using a cross between Xiphophorus maculatus and X. andersi, we report a linkage map predominantly composed of microsatellite markers. All 24 acrocentric chromosome sets of Xiphophorus are represented in the assembled linkage map with an average intergenomic distance of 7.5 cM. Since both male and female F1 hybrids were used to produce backcross progeny, these recombination rates were compared between “male” and “female” maps. Although several genomic regions exhibit differences in map length, male- and female-derived maps are similar. Thus Xiphophorus, in contrast to zebrafish, Danio rerio, and several other vertebrate species, does not show sex-specific differences in recombination. The microsatellite markers we report can be easily adapted to any Xiphophorus interspecies and some intraspecies crosses, and thus provide a means to directly compare results derived from independent experiments.
Radiation Hybrid Mapping of the Species Cytoplasm-Specific (scsae) Gene in Wheat
Authors: Hossain Khwaja G , Riera-Lizarazu Oscar , Kalavacharla Venugopal , Vales M Isabel , Maan Schivcharan S , Kianian Shahryar F
Published: September 2004

Radiation hybrid (RH) mapping is based on radiation-induced chromosome breakage and analysis of chromosome segment retention or loss using molecular markers. In durum wheat (Triticum turgidum L., AABB), an alloplasmic durum line [(lo) durum] has been identified with chromosome 1D of T. aestivum L. (AABBDD) carrying the species cytoplasm-specific (scsae) gene. The chromosome 1D of this line segregates as a whole without recombination, precluding the use of conventional genome mapping. A radiation hybrid mapping population was developed from a hemizygous (lo) scsae− line using 35 krad gamma rays. The analysis of 87 individuals of this population with 39 molecular markers mapped on chromosome 1D revealed 88 radiation-induced breaks in this chromosome. This number of chromosome 1D breaks is eight times higher than the number of previously identified breaks and should result in a 10-fold increase in mapping resolution compared to what was previously possible. The analysis of molecular marker retention in our radiation hybrid mapping panel allowed the localization of scsae and 8 linked markers on the long arm of chromosome 1D. This constitutes the first report of using RH mapping to localize a gene in wheat and illustrates that this approach is feasible in a species with a large complex genome.
Population Genetic Evidence for Rapid Changes in Intraspecific Diversity and Allelic Cycling of a Specialist Defense Gene in ZeaSequence data from this article have been deposited with the EMBL/GenBank Data Libraries under accession nos. AY320258–320280, AY52550, AY52551, AY52552, AY52553, AY52554, AY52555, AY52556, AY52557, AY52558, AY52559, and AY549598, AY549599, AY549600, AY549601, AY549602, AY549603, AY549604, AY549605, AY549606, AY549607, AY549608, AY549609, AY549610, AY549611, AY549612, AY549613, AY549614, AY549615, AY549616, AY549617, AY549618, AY549619, AY549620, AY549621, AY549622, AY549623, AY549624, AY549625, AY549626, AY549627, AY549628, AY549629, AY549630, AY549631, AY549632, AY549633, AY549634, AY549635, AY549636, AY549637, AY549638.
Authors: Tiffin Peter , Hacker Robert , Gaut Brandon S
Published: September 2004

Two patterns of plant defense gene evolution are emerging from molecular population genetic surveys. One is that specialist defenses experience stronger selection than generalist defenses. The second is that specialist defenses are more likely to be subject to balancing selection, i.e., evolve in a manner consistent with balanced-polymorphism or trench-warfare models of host-parasite coevolution. Because most of the data of specialist defenses come from Arabidopsis thaliana, we examined the genetic diversity and evolutionary history of three defense genes in two outcrossing species, the autotetraploid Zea perennis and its most closely related extant relative the diploid Z. diploperennis. Intraspecific diversity at two generalist defenses, the protease inhibitors wip1 and mpi, were consistent with a neutral model. Like previously studied genes in these taxa, wip1 and mpi harbored similar levels of diversity in Z. diploperennis and Z. perennis. In contrast, the specialist defense hm2 showed strong although distinctly different departures from a neutral model in the two species. Z. diploperennis appears to have experienced a strong and recent selective sweep. Using a rejection-sampling coalescent method, we estimate the strength of selection on Z. diploperennis hm2 to be ∼3.0%, which is approximately equal to the strength of selection on tb1 during maize domestication. Z. perennis hm2 harbors three highly diverged alleles, two of which are found at high frequency. The distinctly different patterns of diversity may be due to differences in the phase of host-parasite coevolutionary cycles, although higher hm2 diversity in Z. perennis may also reflect reduced efficacy of selection in the autotetraploid relative to its diploid relative.
Quantitative Trait Loci Affecting Life Span in Replicated Populations of Drosophila melanogaster. I. Composite Interval Mapping
Authors: Forbes Scott N , Valenzuela Robert K , Keim Paul , Service Philip M
Published: September 2004

Composite interval mapping was used to identify life-span QTL in F2 progeny of three crosses between different pairs of inbred lines. Each inbred line was derived from a different outbred population that had undergone long-term selection for either long or short life span. Microsatellite loci were used as genetic markers, and confidence intervals for QTL location were estimated by bootstrapping. A minimum of 10 QTL were detected, nine of which were located on the two major autosomes. Five QTL were present in at least two crosses and five were present in both sexes. Observation of the same QTL in more than one cross was consistent with the hypothesis that genetic variation for life span is maintained by balancing selection. For all QTL except one, allelic effects were in the direction predicted on the basis of outbred source population. Alleles that conferred longer life were always at least partially dominant.
Estimating the Degree of Saturation in Mutant Screens
Authors: Pollock David D , Larkin John C
Published: September 2004

Large-scale screens for loss-of-function mutants have played a significant role in recent advances in developmental biology and other fields. In such mutant screens, it is desirable to estimate the degree of “saturation” of the screen (i.e., what fraction of the possible target genes has been identified). We applied Bayesian and maximum-likelihood methods for estimating the number of loci remaining undetected in large-scale screens and produced credibility intervals to assess the uncertainty of these estimates. Since different loci may mutate to alleles with detectable phenotypes at different rates, we also incorporated variation in the degree of mutability among genes, using either gamma-distributed mutation rates or multiple discrete mutation rate classes. We examined eight published data sets from large-scale mutant screens and found that credibility intervals are much broader than implied by previous assumptions about the degree of saturation of screens. The likelihood methods presented here are a significantly better fit to data from published experiments than estimates based on the Poisson distribution, which implicitly assumes a single mutation rate for all loci. The results are reasonably robust to different models of variation in the mutability of genes. We tested our methods against mutant allele data from a region of the Drosophila melanogaster genome for which there is an independent genomics-based estimate of the number of undetected loci and found that the number of such loci falls within the predicted credibility interval for our models. The methods we have developed may also be useful for estimating the degree of saturation in other types of genetic screens in addition to classical screens for simple loss-of-function mutants, including genetic modifier screens and screens for protein-protein interactions using the yeast two-hybrid method.
A Genetic Screen for Dominant Modifiers of a cyclin E Hypomorphic Mutation Identifies Novel Regulators of S-Phase Entry in Drosophila
Authors: Brumby Anthony , Secombe Julie , Horsfield Julie , Coombe Michelle , Amin Nancy , Coates Deborah , Saint Robert , Richardson Helena
Published: September 2004

Cyclin E together with its kinase partner Cdk2 is a critical regulator of entry into S phase. To identify novel genes that regulate the G1- to S-phase transition within a whole animal we made use of a hypomorphic cyclin E mutation, DmcycEJP, which results in a rough eye phenotype. We screened the X and third chromosome deficiencies, tested candidate genes, and carried out a genetic screen of 55,000 EMS or X-ray-mutagenized flies for second or third chromosome mutations that dominantly modified the DmcycEJP rough eye phenotype. We have focused on the DmcycEJP suppressors, S(DmcycEJP), to identify novel negative regulators of S-phase entry. There are 18 suppressor gene groups with more than one allele and several genes that are represented by only a single allele. All S(DmcycEJP) tested suppress the DmcycEJP rough eye phenotype by increasing the number of S phases in the postmorphogenetic furrow S-phase band. By testing candidates we have identified several modifier genes from the mutagenic screen as well as from the deficiency screen. DmcycEJP suppressor genes fall into the classes of: (1) chromatin remodeling or transcription factors; (2) signaling pathways; and (3) cytoskeletal, (4) cell adhesion, and (5) cytoarchitectural tumor suppressors. The cytoarchitectural tumor suppressors include scribble, lethal-2-giant-larvae (lgl), and discs-large (dlg), loss of function of which leads to neoplastic tumors and disruption of apical-basal cell polarity. We further explored the genetic interactions of scribble with S(DmcycEJP) genes and show that hypomorphic scribble mutants exhibit genetic interactions with lgl, scab (αPS3-integrin—cell adhesion), phyllopod (signaling), dEB1 (microtubule-binding protein—cytoskeletal), and moira (chromatin remodeling). These interactions of the cytoarchitectural suppressor gene, scribble, with cell adhesion, signaling, cytoskeletal, and chromatin remodeling genes, suggest that these genes may act in a common pathway to negatively regulate cyclin E or S-phase entry.
Linkage Maps of Microsatellite DNA Markers for the Pacific Oyster Crassostrea gigasWe dedicate this study to the memory of Will Borgenson, who reared and cared for the parents of the mapping families.
Authors: Hubert Sophie , Hedgecock Dennis
Published: September 2004

We constructed male and female consensus linkage maps for the Pacific oyster Crassostrea gigas, using a total of 102 microsatellite DNA markers typed in 11-day-old larvae from three families. We identified 11 and 12 linkage groups in the male and female consensus maps, respectively. Alignment of these separate maps, however, suggests 10 linkage groups, which agrees with the haploid chromosome number. The male linkage map comprises 88 loci and spans 616.1 cM, while the female map comprises 86 loci and spans 770.5 cM. The male and the female maps share 74 loci; 2 markers remain unlinked. The estimated coverages for the consensus linkage maps are 79% for the male and 70–75% for the female, on the basis of two estimates of genome length. Ninety-five percent of the genome is expected to lie within 16 and 21 cM of markers on the male and female maps, respectively, while 95% of simulated minimum distances to the male and female maps are within 10.1 and 13.6 cM, respectively. Females have significantly more recombination than males, across 118 pairs of linked markers in common to the parents of the three families. Significant differences in recombination and orders of markers are also evident among same-sex parents of different families as well as sibling parents of opposite sex. These observations suggest that polymorphism for chromosomal rearrangements may exist in natural populations, which could have profound implications for interpreting the evolutionary genetics of the oyster. These are the first linkage maps for a bivalve mollusc that use microsatellite DNA markers, which should enable them to be transferred to other families and to be useful for further genetic analyses such as QTL mapping.
Gene Conversion and Crossing Over Along the 405-kb Left Arm of Saccharomyces cerevisiae Chromosome VII
Authors: Malkova Anna , Swanson Johanna , German Miriam , McCusker John H , Housworth Elizabeth A , Stahl Franklin W , Haber James E
Published: September 2004

Gene conversions and crossing over were analyzed along 10 intervals in a 405-kb region comprising nearly all of the left arm of chromosome VII in Saccharomyces cerevisiae. Crossover interference was detected in all intervals as measured by a reduced number of nonparental ditypes. We have evaluated interference between crossovers in adjacent intervals by methods that retain the information contained in tetrads as opposed to single segregants. Interference was seen between intervals when the distance in the region adjacent to a crossover was <∼35 cM (90 kb). At the met13 locus, which exhibits ∼9% gene conversions, those gene conversions accompanied by crossing over exerted interference in exchanges in an adjacent interval, whereas met13 gene conversions without an accompanying exchange did not show interference. The pattern of exchanges along this chromosome arm can be represented by a counting model in which there are three nonexchange events between adjacent exchanges; however, maximum-likelihood analysis suggests that ∼8–12% of the crossovers on chromosome VII arise by a separate, noninterfering mechanism.
Comparative Mapping in the Pinaceae
Authors: Krutovsky Konstantin V , Troggio Michela , Brown Garth R , Jermstad Kathleen D , Neale David B
Published: September 2004

A comparative genetic map was constructed between two important genera of the family Pinaceae. Ten homologous linkage groups in loblolly pine (Pinus taeda L.) and Douglas fir (Pseudotsuga menziesii [Mirb.] Franco) were identified using orthologous expressed sequence tag polymorphism (ESTP) and restriction fragment length polymorphism (RFLP) markers. The comparative mapping revealed extensive synteny and colinearity between genomes of the Pinaceae, consistent with the hypothesis of conservative chromosomal evolution in this important plant family. This study reports the first comparative map in forest trees at the family taxonomic level and establishes a framework for comparative genomics in Pinaceae.
Stress and Survival of Aging Escherichia coli rpoS Colonies
Authors: Saint-Ruf Claude , Taddei François , Matic Ivan
Published: September 2004

In Escherichia coli, the expression of the RpoS regulon is known to be crucial for survival in liquid cultures during stationary phase. By measuring cell viability and by transcriptome analysis, here we show that rpoS cells as well as wild-type cells survive when they form colonies on solid media.
Analysis of Deinococcus radiodurans's Transcriptional Response to Ionizing Radiation and Desiccation Reveals Novel Proteins That Contribute to Extreme Radioresistance
Authors: Tanaka Masashi , Earl Ashlee M , Howell Heather A , Park Mie-Jung , Eisen Jonathan A , Peterson Scott N , Battista John R
Published: September 2004

During the first hour after a sublethal dose of ionizing radiation, 72 genes were upregulated threefold or higher in D. radiodurans R1. Thirty-three of these loci were also among a set of 73 genes expressed in R1 cultures recovering from desiccation. The five transcripts most highly induced in response to each stress are the same and encode proteins of unknown function. The genes (ddrA, ddrB, ddrC, ddrD, and pprA) corresponding to these transcripts were deleted, both alone and in all possible two-way combinations. Characterization of the mutant strains defines three epistasis groups that reflect different cellular responses to ionizing radiation-induced damage. The ddrA and ddrB gene products have complementary activities and inactivating both loci generates a strain that is more sensitive to ionizing radiation than strains in which either single gene has been deleted. These proteins appear to mediate efficient RecA-independent processes connected to ionizing radiation resistance. The pprA gene product is not necessary for homologous recombination during natural transformation, but nevertheless may participate in a RecA-dependent process during recovery from radiation damage. These characterizations clearly demonstrate that novel mechanisms significantly contribute to the ionizing radiation resistance in D. radiodurans.
Functional Distinction Between Cln1p and Cln2p Cyclins in the Control of the Saccharomyces cerevisiae Mitotic Cycle
Authors: Queralt Ethel , Igual J Carlos
Published: September 2004

Cln1p and Cln2p are considered as equivalent cyclins on the basis of sequence homology, regulation, and functional studies. Here we describe a functional distinction between the Cln1p and Cln2p cyclins in the control of the G1/S transition. Inactivation of CLN2, but not of CLN1, leads to a larger-than-normal cell size, whereas overexpression of CLN2, but not of CLN1, results in smaller-than-normal cells. Furthermore, mild ectopic expression of CLN2, but not of CLN1, suppresses the lethality of swi4swi6 and cdc28 mutant strains. In the absence of Cln1p, the kinetics of budding, initiation of DNA replication, and activation of the Start-transcription program are not affected; by contrast, loss of Cln2p causes a delay in bud emergence. A primary role for Cln2p but not for Cln1p in budding is reinforced by the observation that only the cln2 mutation is synthetic lethal with a cdc42 mutation, and only the cln2 mutant strain is hypersensitive to latrunculin B. In addition, we found that Cln1p showed a more prominent nuclear staining than Cln2p. Finally, chimeric proteins composed of Cln1p and Cln2p revealed that Cln2p integrity is required for its functional specificity.
stall-Mediated Extrinsic Control of Ovarian Follicle Formation in Drosophila
Authors: Willard Stacey S , Ozdowski Emily F , Jones Natasha A , Cronmiller Claire
Published: September 2004

Complex patterns of morphogenesis require intricate coordination of multiple, regulatory processes that control cellular identities, shapes, and behaviors, both locally and over vast distances in the developing organism or tissue. Studying Drosophila oogenesis as a model for tissue morphogenesis, we have discovered extraovarian regulation of follicle formation. Clonal analysis and ovary transplantation have demonstrated that long-range control of follicle individualization requires stall gene function in cells outside of the ovary. Although tissue nonautonomous regulation has been shown to govern follicle maturation and survival, this is the first report of an extraovarian pathway involved in normal follicle formation.
Interactions Between Imprinting Effects in the Mouse
Authors: Cattanach Bruce M , Beechey Colin V , Peters Josephine
Published: September 2004

Mice with uniparental partial or complete disomies for any one of 11 identified chromosomes show abnormal phenotypes. The abnormalities, or imprinting effects, can be attributable to an incorrect dosage of maternal or paternal copies of imprinted gene(s) located within the regions involved. Here we show that combinations of partial disomies may result in interactions between imprinting effects that seemingly independently affect fetal and/or placental growth in different ways or modify neonatal and postnatal imprinting effects. Candidate genes within the regions have been identified. The findings are generally in accord with the “conflict hypothesis” for the evolution of genomic imprinting but do not clearly demonstrate common growth axes within which imprinted genes may interact. Instead, it would seem that any gene that represses or limits embryonic/fetal growth to the advantage of the mother—by any developmental means—will have been subject to evolutionary selection for paternal allele repression. Likewise, any gene that favors embryonic/fetal development at consequent cost to the mother—by any developmental means—will have faced selection for maternal allele repression. The classical Igf2-Igf2r axis may therefore be unique. The findings involve reinterpretation of older imprinting data and consequently revision of the mouse imprinting map.
A Novel Yeast Mutation, rad52-L89F, Causes a Specific Defect in Rad51-Independent Recombination That Correlates With a Reduced Ability of Rad52-L89F to Interact With Rad59
Authors: Cortés-Ledesma Felipe , Malagón Francisco , Aguilera Andrés
Published: September 2004

We isolated a novel rad52 mutation, rad52-L89F, which specifically impairs recombination in rad51Δ cells. rad52-L89F displays phenotypes similar to rad59Δ and encodes a mutant protein impaired in its ability to interact with Rad59. These results support the idea that Rad59 acts in homologous recombination via physical interaction with Rad52.
Near‐fatal uterine hemorrhage during induction chemotherapy for acute myeloid leukemia: A case report of bilateral uterine artery embolization
Authors: Phelan John T. , Broder Jonathan , Kouides Peter A.
Published: September 2004

Severe transfusion‐dependent uterine hemorrhage is a relatively uncommon complication of induction chemotherapy for acute myeloid leukemia (AML). Even less common is the failure of systemic conjugated estrogens in this setting. We report a case of life‐threatening uterine hemorrhage in a 38‐year‐old woman in the setting of transfusion‐refractory thrombocytopenia after completing induction chemotherapy for AML. She experienced dramatic breakthrough uterine hemorrhage despite multiple platelet transfusions, conjugated estrogens, recombinant factor VIIa, ϵ‐aminocaproic acid, and intracavitary thrombin‐soaked gauze tamponade. At the point of near‐exsanguination in the setting of hypotension, hematocrit of 14%, and a platelet count of 3,000/μL, she underwent bilateral uterine artery embolization which proved immediately successful. We review the literature and indications for this procedure in the oncologic patient care setting. Am. J. Hematol. 77:151–155, 2004. © 2004 Wiley‐Liss Inc. © 2004 Wiley‐Liss, Inc.
Second case of CML with aberrant BCR‐ABL fusion transcript (e8/a2) with insertion of an inverted ABL intron 1b sequence
Authors: Sugimoto Takeshi , Ijima Kentaro , Hisatomi Hisashi , Murayama Tohru , Mizuno Ishikazu , Hato Akio , Imoto Shion , Nishimura Ryuichiro , Koizumi Tamio
Published: September 2004

We found a case of Ph‐positive chronic myelogenous leukemia (CML) patient with an atypical BCR‐ABL transcript that was undetectable by a routine reverse transcription polymerase chain reaction (RT‐PCR) for major BCR‐ABL. Additional RT‐PCR and sequence analyses have demonstrated that the aberrant transcript consists of a fusion of BCR exon 8 (e8) and ABL exon 2 (a2) with an insertion of a 55‐bp inverted sequence of intron 1b between them. The nucleotide sequences of the aberrant transcript were identical to those of a previously reported CML patient. These are the only two CML cases in the literature with identical aberrant BCR‐ABL transcripts. Am. J. Hematol. 77:164–166, 2004. © 2004 Wiley‐Liss, Inc.
Dysplastic changes in the peripheral blood of children with microdeletion 22q11.2
Authors: Özbek Namık , Derbent Murat , Olcay Lale , Yılmaz Zerrin , Tokel Kürşat
Published: September 2004

Myelodysplasia refers to abnormal morphology of the bone marrow and/or peripheral blood. This condition is frequently due to myelodysplastic syndrome; however, myelodysplasia is caused by a variety of factors and is not always a sign of pre‐malignant disease. We observed myelodysplastic changes in peripheral blood smears and bone marrow specimens from a patient who had chromosomal microdeletion 22q11.2 (del22q11.2). We then investigated such changes in several other patients who were newly diagnosed with del22q11.2 (n = 5 total, including the index case) and compared the findings to those in four sets of controls without this chromosomal abnormality. Specifically, the controls were children with conotruncal heart defects (n = 3); otherwise healthy children with bacterial (n = 4) or viral infection (n = 4); and healthy children (n = 4). The myelodysplasia scores in the myeloid cells and eosinophils of the children with del22q11 were higher than those in all four of the control groups. Myelodysplastic changes in the peripheral blood of children with del22q11.2 have not been reported previously. We believe that certain gene(s) in the deleted region may be responsible for the myelodysplastic changes that we observed in our patients with this chromosomal abnormality. Am. J. Hematol. 77:126–131, 2004. © 2004 Wiley‐Liss, Inc.
Microvascular blood flow and stasis in transgenic sickle mice: Utility of a dorsal skin fold chamber for intravital microscopy
Authors: Kalambur Venkatasubramaniam S. , Mahaseth Hemchandra , Bischof John C. , Kielbik Miroslaw C. , Welch Thomas E. , Vilbäck Åsa , Swanlund David J. , Hebbel Robert P. , Belcher John D. , Vercellotti Gregory M.
Published: September 2004

Vascular inflammation, secondary to ischemia–reperfusion injury, may play an essential role in vaso‐occlusion in sickle cell disease (SCD). To investigate this hypothesis, dorsal skin fold chambers (DSFCs) were implanted on normal and transgenic sickle mice expressing human α and βs/βs‐Antilles globin chains. Microvessels in the DSFC were visualized by intravital microscopy at baseline in ambient air and after exposure to hypoxia–reoxygenation. The mean venule diameter decreased 9% (P < 0.01) in sickle mice after hypoxia–reoxygenation but remained constant in normal mice. The mean RBC velocity and wall shear rate decreased 55% (P < 0.001) in sickle but not normal mice after hypoxia–reoxygenation. None of the venules in normal mice became static at any time during hypoxia–reoxygenation; however, after 1 hr of hypoxia and 1 hr of reoxygenation, 11.9% of the venules in sickle mice became static (P < 0.001). After 1 hr of hypoxia and 4 hr of reoxygenation, most of the stasis had resolved; only 3.6% of the subcutaneous venules in sickle mice remained static (P = 0.01). All of the venules were flowing again after 24 hr of reoxygenation. Vascular stasis could not be induced in the subcutaneous venules of sickle mice by tumor necrosis factor alpha (TNF‐α). Leukocyte rolling flux and firm adhesion, manifestations of vascular inflammation, were significantly higher at baseline in sickle mice compared to normal (P < 0.01) and increased 3‐fold in sickle (P < 0.01), but not in normal mice, after hypoxia–reoxygenation. Plugs of adherent leukocytes were seen at bifurcations at the beginning of static venules. Misshapen RBCs were also seen in subcutaneous venules. Am. J. Hematol. 77:117–125, 2004. © 2004 Wiley‐Liss, Inc.
Evaluation of kidney damage in patients with acute lymphoblastic leukemia in long‐term follow‐up: Value of renal scan
Authors: Yetgin Sevgi , Olgar Şeref , Aras Tülin , Çetin Mualla , Düzova Ali , Beylergil Volkan , Akhan Okan , Oĝuz Oĝuzhan , Saraçbaşı Osman
Published: September 2004

In order to evaluate potential long‐term kidney damage of childhood leukemia and risk factors affecting renal damage, we studied 116 children treated for acute lymphoblastic leukemia (ALL) using the St. Jude Total XI and XIII protocols in 1991–1998. The median follow‐up period after the completion of treatment was 35 months. The following parameters were examined: urinalysis, urinary creatinine (Cr), calcium (Ca), phosphorus, β2‐microglobulin, glomerular filtration rate (GFR), tubular phosphorus reabsorption (TPR), and renal function tests. Radiological evaluation included renal ultrasonography (US), and renal scans with DMSA or MAG‐3 were performed. Blood chemistry and renal US patients were normal in all patients except two. GFR, TPR, urinary Ca/Cr, β2‐microglobulin, and renal scan were abnormal in 19.0%, 16.4%, 13.8%, 6.0%, and 40.5% of patients, respectively. The abnormality rate in GFR was significantly higher in patients <2 years of age. TPR abnormality was found to be significantly higher in patients who did not have G‐CSF. An abnormal renal scan was associated with Hb < 10 g/dL, kidney infiltration, or hypertension at presentation and also occurred patients who underwent methotrexate treatment with frequent intervals during the follow‐up period. Patients should be followed‐up after cessation of therapy with the conventional tests mentioned above. In case of any abnormality, further detailed tests should be performed; renal scan seems to be more predictive for renal damage. Am. J. Hematol. 77:132–139, 2004. © 2004 Wiley‐Liss, Inc.
Genetic variability of platelet glycoprotein Ibα gene
Authors: Ozelo Margareth C. , Costa Devanira S.P. , Siqueira Lucia H. , Machado Tania M.F. , Castro Vagner , Gonçalves Marilda S. , Menezes Raimundo C. , Soares Manoel , Annichino‐Bizzacchi Joyce M. , Costa Fernando F. , Arruda Valder R.
Published: September 2004

Platelet membrane glycoprotein (GP) Ibα is a critical component of platelet adhesion complex to subendothelium structures following tissue injury or pathological surfaces, such as atherosclerotic plaques. Polymorphisms of the GPIbα gene have been associated with a high risk for occlusive vascular disease, and its distribution varies considerably among distinct populations. These polymorphisms comprise the human platelet antigen (HPA)‐2 system, the −5C/T dimorphism of the Kozak sequence, and the variable number of tandem 39‐bp repeats (VNTR). Here we report the prevalence of the GPIbα gene polymorphisms among Brazilians, a highly ethnically diverse population. We analyzed 492 subjects of European, African, or Indigenous origin. It was possible to determine ten distinct haplotypes. The most common (∽40%) haplotype was the Kozak‐TT/HPA‐2aa/VNTR‐CC for both Caucasian and African descent. However, among Indigenous, Kozak‐TT/HPA‐2aa/VNTR‐CC and Kozak‐TC/HPA‐2aa/VNTR‐CC were equally present. Although a strong linkage disequilibrium between VNTR and HPA‐2 polymorphism had also been observed, here we determined incomplete linkage disequilibrium in 10% of subjects from all ethnic groups. VNTR‐E, a rare variant lacking the 39‐bp repeat, was identified in two unrelated subjects, and functional platelet studies revealed no abnormalities. The VNTR‐A allele, the largest variant containing four copies of the repeats, was not identified in this population. However, homozygosity for the VNTR‐A allele (Kozak‐TT/HPA‐2aa/VNTR‐AA) was determined in two distinct species of nonhuman primates. These results suggest a greater complex evolutionary mechanism in the macroglycoprotein region of the GPIbα gene and may be useful in the design of gene–disease association studies for vascular disease. Am. J. Hematol. 77:107–116, 2004. © 2004 Wiley‐Liss, Inc.
Monoclonal gammopathy in extranodal marginal zone lymphoma (ENMZL) correlates with advanced disease and bone marrow involvement
Authors: Asatiani Ekatherine , Cohen Philip , Ozdemirli Metin , Kessler Craig M. , Mavromatis Blanche , Cheson Bruce D.
Published: September 2004

Monoclonal gammopathy is a well‐recognized occurrence in splenic marginal zone lymphoma (MZL); however, its prevalence has never been reported in extranodal MZL (ENMZL). We present results of a retrospective analysis of 26 patients with newly diagnosed as well as previously treated ENMZL. Monoclonal gammopathy was detected in 7 out of 26 (27%) patients. All patients with a monoclonal gammopathy had stage IV disease, and most of them had involvement of lymph nodes and/or bone marrow (n = 6). Only 1 out of 7 patients with a monoclonal spike had stage IV disease diagnosed based on multifocal mucosal involvement. There was also a strong correlation between the involvement of bone marrow and the presence of a monoclonal spike (Fisher's exact test, P = 0.0007). Prevalence of monoclonal gammopathy is higher than previously recognized and indicates advanced disease. However, the prognostic significance of the presence of monoclonal gammopathy in this population is unknown. Am. J. Hematol. 77:144–146, 2004. © 2004 Wiley‐Liss, Inc.
Presentation and pattern of symptoms in 382 patients with Glanzmann thrombasthenia in Iran
Authors: Toogeh G. , Sharifian R. , Lak M. , Safaee R. , Artoni A. , Peyvandi F.
Published: September 2004

Glanzmann thrombasthenia (GT) is a rare autosomal recessive disease characterized by prolonged bleeding time with normal platelet count and morphology. It is caused by the quantitative or qualitative deficiency of the platelet glycoprotein IIb–IIIa. In 382 Iranian patients with GT diagnosed at a single center during the period 1969–2001, consanguinity between parents was 86.6%, in accord with the high frequency of intrafamilial marriages in Iran. Almost all patients had had abnormal mucocutaneous bleeding (epistaxis and gum bleeding); at follow‐up, 4/5 of the patients had been transfused at least once to control hemorrhagic episodes. As expected, almost all the patients had a normal platelet count while the leukocyte count was increased in 19.3%. Among women, an unexpected low rate of pregnancies was observed. Am. J. Hematol. 77:198–199, 2004. © 2004 Wiley‐Liss, Inc.
Squamous cell carcinoma of the tongue complicating chronic oral mucosal graft‐versus‐host disease after allogeneic hematopoietic stem cell transplantation
Authors: Szeto Ching H. , Shek Tony W.H. , Lie Albert K.W. , Au Wing Y. , Yuen Anthony P.W. , Kwong Yok L.
Published: September 2004

Two patients underwent allogeneic hematopoietic stem cell transplantation (HSCT) for acute myeloid leukemia. Chronic graft‐versus‐host disease (GVHD) developed, with persistent symptomatic oral lesions. At 2 and 6 years post‐HSCT, both patients developed squamous cell carcinoma (SCC) of the tongue in areas previously involved by chronic GVHD. None had any known risk factor for SCC. Histologically, moderate to severe dysplasia was present in noncancerous oral mucosa. Oral SCC is rarely described after HSCT, and a review of the reported cases showed chronic GVHD to be a common risk, suggesting that the chronic inflammation associated with GVHD might be of pathogenetic significance. Am. J. Hematol. 77:200–202, 2004. © 2004 Wiley‐Liss, Inc.
Home prothrombin time monitoring: A literature analysis
Authors: Yang David T. , Robetorye Ryan S. , Rodgers George M.
Published: September 2004

The anticoagulant activity of warfarin sodium is monitored by the prothrombin time (PT) using the international normalized ratio (INR). Standard oral anticoagulant therapy monitoring requires frequent patient visits to physicians' offices and/or laboratories to optimize warfarin dosage. Home PT monitoring by patients can increase testing frequency and may thus decrease complications associated with oral anticoagulant therapy. Clinical studies suggest that home PT monitoring is more effective than uncoordinated management and is as effective as care through specialized anticoagulation clinics for keeping INRs within a therapeutic range. There are accurate and reliable instruments available, but paramount to the success of home PT monitoring is sound patient selection, appropriate patient training, and consistent quality control. Am. J. Hematol. 77:177–186, 2004. © 2004 Wiley‐Liss, Inc.
Prevention of bleeds in hemophilia patients with inhibitors: Emerging data and clinical direction
Authors: Leissinger Cindy A.
Published: September 2004

In patients with hemophilia, the development of high‐responding inhibitors to factor VIII prevents adequate replacement therapy and results in increased risk of serious bleeding episodes, poor control of joint bleeding, and progressive, debilitating joint disease. Immune tolerance therapy can eradicate inhibitors, but it is not uniformly successful. Emerging data suggest that prophylaxis using activated prothrombin complex concentrates may be effective and safe in reducing the incidence of joint bleeding during immune tolerance therapy and for patients in whom immune tolerance induction fails. However, only controlled clinical trials will ultimately demonstrate whether prophylaxis can prevent joint bleeding and damage, and improve quality of life in patients with inhibitors. Am. J. Hematol. 77:187–193, 2004. © 2004 Wiley‐Liss, Inc.
Acute lymphoblastic leukemia with the phenotype of a putative B‐cell/T‐cell bipotential precursor
Authors: Lau Lee Gong , Tan Lip Kun , Koay Evelyn S.C. , Ee Melvin H.L. , Tan Suat Hoon , Liu Te Chih
Published: September 2004

Biphenotypic acute leukemias (BALs) are uncommon. Most are of myeloid‐B‐cell or myeloid‐T‐cell lineage. We report herein a 70‐year‐old man with an unusual acute leukemia where the blasts expressed both B‐ and T‐lymphoid markers. He presented to us with an enlarging cutaneous tumor. The presenting peripheral blood and bone marrow aspirate showed 40% and 90% blasts, respectively, which were negative for the usual cytochemical stains. The flow cytometric analysis revealed that the blasts were positive for CD19, CD20, CD22, cytoplasmic (Cyt) CD79a, CD10, Cyt CD3, CD5, CD7, CD4, HLA‐DR, TdT, and were negative for myeloid markers. According to the scoring system from the European Group for the Immunological Characterization of Acute Leukaemias (EGIL), this case was an unequivocal B‐cell/T‐cell BAL. Conventional cytogenetic analysis revealed 46XY [t(4;11)(q31;q13), add(8)(q24), der(9)del(9)(p21)del(9)(q32q34), −13, +mar] in all 25 metaphases analyzed. Fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR) for 11q23 rearrangements as well as t(9;22) were negative. PCR for both TCR‐γ and IgH gene analyses revealed polyclonal rearrangements. We postulate that this case of BAL might have arisen from the putative common lymphoid progenitor cell. Am. J. Hematol. 77:156–160, 2004. © 2004 Wiley‐Liss, Inc.
Case of chronic‐phase chronic myelogenous leukemia with an abdominal hematopoietic tumor of leukemic clone origin
Authors: Sakakura Miho , Ohishi Kohshi , Nomura Kenichi , Katayama Naoyuki , Nishii Kazuhiro , Masuya Masahiro , Nakase Kazunori , Shiku Hiroshi
Published: September 2004

We report a 59‐year‐old man with chronic myelogenous leukemia (CML) in chronic phase who presented with a large abdominal tumor. Biopsy revealed proliferation of granulocytic‐, erythroid‐, and megakaryocytic‐lineage cells in a retroperitoneal lymph node. The BCR/ABL fusion gene was detected on a paraffin‐embedded tissue section of the lymph node by double‐color fluorescence in situ hybridization, indicating an extramedullary hematopoietic tumor of CML origin. This patient has achieved a complete cytogenetic response for 19 months with imatinib mesylate (STI571; Gleevec), in association with the regression of the tumor. However, the development of an extramedullary tumor in chronic‐phase CML generally indicates a poor prognosis, because it commonly consists of blast proliferation and is followed by blast crisis in the marrow within a few months. This case, therefore, points to the importance of histological examination of extramedullary tumors in CML for evaluation of disease status and for therapeutic decisions. Am. J. Hematol. 77:167–170, 2004. © 2004 Wiley‐Liss, Inc.
Role of ELISA D‐dimer test in patients with unstable angina pectoris presenting at the emergency department with a normal electrocardiogram
Authors: Shitrit David , Bar‐Gil Shitrit Ariella , Rudensky Bernard , Sulkes Jaqueline , Gutterer Natan , Zviony Dan
Published: September 2004

Patients with unstable angina pectoris and acute myocardial infarction have higher than normal D‐dimer levels. The aim of the study was to determine the value of the D‐dimer test in patients with unstable angina pectoris and a normal electrocardiogram on presentation at the emergency department. The study sample included 81 patients who met these criteria. Blood samples collected at admission were subjected to ELISA D‐dimer. Findings were correlated with coronary risk factors, use of cardiac medications, blood levels of acute phase reactants (fibrinogen and C‐reactive protein), cardiac enzymes levels, length of hospital stay, and catheterization findings. ELISA D‐dimer levels were statistically significantly correlated with cardiac risk factors, namely male sex, older age, smoking, and hypertension (r = 0.25, P = 0.02; r = 0.43, P = 0.0001; r = 0.26, P = 0.03; r = 0.35, P = 0.002, respectively), in addition to use of cardiac medications (beta blockers, aspirin, nitrates), levels of acute phase reactants, length of stay, and catheterization findings. On multivariate analysis, only D‐dimer level, age, and sex were predictors of length of stay (P = 0.018). The study suggests that D‐dimer levels at admission to the emergency department may serve as an additional tool to predict the magnitude of unstable angina pectoris in patients with a normal electrocardiogram. Am. J. Hematol. 77:147–150, 2004. © 2004 Wiley‐Liss, Inc.
Response of myasthenia gravis to rituximab in a patient with non‐Hodgkin lymphoma
Authors: Gajra Ajeet , Vajpayee Neerja , Grethlein Sara J.
Published: September 2004

Myasthenia gravis is a B‐cell‐mediated autoimmune neuromuscular disorder characterized by weakness and fatigability of skeletal muscles. The underlying defect is an autoantibody‐mediated attack on the acetylcholine receptors (AchRs) at the neuromuscular junction. Rituximab is a genetically engineered chimeric murine/human monoclonal antibody indicated for treatment of patients with low‐grade or follicular, CD20‐positive, B‐cell non‐Hodgkin lymphoma. Based on its potential for elimination of auto‐reactive B‐cell clones, rituximab may have a role in the management of some autoimmune disorders. We report a patient with B‐cell, follicular non‐Hodgkin lymphoma and a long‐standing history of myasthenia gravis and the favorable impact of rituximab on both disorders. Am. J. Hematol. 77:196–197, 2004. © 2004 Wiley‐Liss, Inc.