bluuebunny/crossref_metadata_2025 huggingface dataset:
Development and use of a microdissected swine chromosome 6 DNA library
Authors: Ambady S. , Mendiola J.R. , Louis C.F. , Janzen M.A. , Schook L.B. , Buoen L. , Lunney J.K. , Grimm D.R. , Ponce de León F.A.
Published: May 2008

To facilitate the identification of microsatellite genetic markers from a single swine chromosome, chromosome microisolation and microcloning have been used to generate a swine chromosome 6-specific DNA library. Ten copies of swine chromosome 6 were scraped from metaphase spreads, ligated to custom-prepared adaptors, and amplified by PCR. The purity of the amplified product was verified by fluorescent in situ hybridization. The utility of the chromosome painting probe for heterologous painting was demonstrated and confirmed that swine chromosome 6 is syntenic to human chromosomes p and 19q. A small insert genomic library of 1.39 × 106 clones was generated from the PCR-amplified chromosome 6 genomic DNA and screened for (GT)n microsatellite genetic markers. Nine (GT)n microsatellite markers were developed and genotyped on a Yorkshire × Meishan swine reference family. All nine markers genetically mapped to chromosome 6, confirming the purity of the microisolation method. The method used here should be adaptable to the microdissection of sub-chromosomal regions of not only the swine genome but also other livestock genomes.
FISH localization of the human Y-homolog of protein kinase PRKX (PRKY) to Yp11.2 and two pseudogenes to 15q26 and Xq12→q13
Authors: Schiebel K. , Mertz A. , Winkelmann M. , Gläser B. , Schempp W. , Rappold G.
Published: May 2008

Recently, we reported the isolation of a new subfamily of serine-threonine protein kinases. This subfamily was shown to consist of at least four members. Sequencing and FISH mapping of all 4 members now reveals that the Y-homolog (PRKY) of the previously mapped PRKX gene (Xp22.3) is located in Yp11.2, in close vicinity to AMELY. The other two copies reside on Xq112→q13(PRKXP2) and 15q26(PRKXP1, containing CA repeat STS D15S87) and represent pseudogenes.
The gene cluster containing the LCAT gene is conserved between human and pig
Authors: Frengen E. , Thomsen P.D. , Brede G. , Solheim J. , de Jong P.J. , Prydz H.
Published: May 2008

A pooled DNA probe from P1artificial chromosome clones (PACs) containing the human lecithin:cholesterol acyl transferase (LCAT) gene cluster was used in fluorescence in situ hybridization (FISH) experiments assigning the genes to pig chromosome 6p13. In addition, probes derived from the coding regions in the human gene cluster were used in long range mapping experiments to show that the overall structures of the human and porcine LCAT gene clusters are identical. Both the linear order and the close physical distance of five apparently unrelated genes have been maintained throughout 90 million years of divergent evolution between human and pig. The extremely dense clustering of the genes in the LCAT gene cluster suggests that this gene organization has biological significance. The conservation of the gene cluster between human and pig supports this suggestion.
Instability of CHO chromosomes containing interstitial telomeric sequences originating from Chinese hamster chromosome 10
Authors: Slijepcevic P. , Xiao Y. , Natarajan A.T. , Bryant P.E.
Published: May 2008

We identified a marker chromosome in the CHO K1 cell line containing amplifications of interstitial telomeric sequences originating from Chinese hamster chromosome 10. Analysis of the progression of this chromosome in two sub-clones of CHO K1 revealed sensitivity of one amplicon to chromosome breakage, resulting in telomere function at the break site. In addition, two more marker chromosomes, both containing amplifications of interstitial telomeric sequences from chromosome 10, were formed during karyotypic evolution of the CHO K1 subclones. The presence of some of the marker chromosomes was also identified in the radiosensitive xrs 5 cell line derived from CHO K1. These results indicate instability of CHO K1 chromosomes containing interstitial telomeric sequences originating from Chinese hamster chromosome 10.
Characterization of a de novo unbalanced chromosome rearrangement by comparative genomic hybridization and fluorescence in situ hybridization
Authors: Levy B. , Gershin I.F. , Desnick R.J. , Babu A. , Gelb B.D. , Hirschhorn K. , Cotter P.D.
Published: May 2008

Fluorescence in situ hybridization (FISH) has proven useful for the identification of chromosomal material of unknown origin. More recently, comparative genomic hybridization (CGH) has been used to identify deletions and amplifications, particularly in neoplastic samples. Here, we describe the combined use of CGH and FISH to identify the origin of a de novo unbalanced translocation in a newborn with multiple congenital anomalies. GTG banding of metaphases from cultured lymphocytes showed an unbalanced karyotype, with extra material on a chromosome 5: 46, XX, add(5)(q35). Parental karyotypes were both normal. CGH revealed the additional material was from distal 1 1q (11q23→qter). This finding was confirmed by FISH with a whole chromosome paint for chromosome 11. Based on the CGH and FISH analyses, the pro-band’s karyotype was therefore 46, XX, der(5)t(5;11)(q35.2; q23.2).ish der(5)(wcp11+). This case demonstrates the efficient use of CGH and confirmatory FISH for the identification of chromosomal material of unknown origin.
Construction of chicken × human microcell hybrids for human gene targeting
Authors: Koi M. , Lamb P.W. , Filatov L. , Feinberg A.P. , Barrett J.C.
Published: May 2008

Human chromosomes 1, 2, 3, and 11 were tagged with pSV2 neo and transferred via microcell fusion from mouse A9 human monochromosomal hybrids to a chicken pre-B cell line, DT40, proficient for homologous recombination. Hybrids containing two copies of human chromosome 11 were transfected with targeting vectors containing a mammalian selectable gene with either the Dl 1S16 or HRAS genomic sequences corresponding to two different chromosome 11 loci. Analysis of stable transfectants showed a high frequency (∼80%) of targeted integration of these constructs into each of the homologous loci of human chromosome 11 in DT40 hybrids. The results suggest that any human genomic sequences on human chromosomes transferred into DT40 cells could be targeted at high frequency, thereby allowing for subsequent modification of human genes and chromosomes.
Mapping in pig of genes involved in sexual differentiation: AMH, WT1 FTZF1, SOX2, SOX9, AHC, and placental and embryonic CYP19
Authors: Lahbib-Mansais Y. , Barbosa A. , Yerle M. , Parma P. , Milan D. , Pailhoux E. , Gellin J. , Cotinot C.
Published: May 2008

Intersexuality has been reported in pigs and investigations are in progress to identify and physically map the genes involved in sex-reversal. In this study we have mapped on porcine chromosomes seven genes which might be implicated in this developmental pathway. Four genes were mapped by radioactive in situ hybridization: AMH (Anti-Müllerian Hormone) and WT1 (Wilms’ Tumor gene 1) were both mapped to pig chromosome 2 (SSC2) in the q14→q21 and p14→q11 regions, placental CYP19 (cytochrome P450, subfamily XIX) and FTZF1 (fushi tarazu factor [Drosophila] homolog 1, alias SF1 [steroidogenic factor 1]) to pig chromosome 1 (SSC1) in the q14→q17 and q210→q211 regions respectively. Four other genes were regionally located by PCR analysis on a cytogenetically characterized porcine somatic cell hybrid panel: a second CYP19 gene (expressed in embryo) was mapped to porcine 1q12→q17, AHC (alias DAX1 adrenal hypoplasia congenital) to porcine Xp24, SOX2 and SOX9 (SRY sex determining region Y-box 2 and 9) to 13q23→q41 and 12p13→p11 respectively. These results are in global agreement with mapping data available in other mammalian species.
Microdissection-mediated selection of chromosome region-specific cDNAs
Authors: Yokoyama Y. , Ohsugi K. , Kozaki T. , Sakuragawa N.
Published: May 2008

K562 is a cell line with two acrocentric marker chromosomes containing abnormally banded regions (ABRs), derived from a Ph-positive chronic myelogenous leukemia (CML) patient. Using reverse and forward chromosome painting FISH analysis, we found that 9q34, 13q31, and 22q11 regions co-amplified in the ABRs-bearing acrocentric marker chromosomes of K562. Utilizing the ABRs of the cell line as target DNA for cDNA selection, we established a simple procedure for chromosome region-specific cDNA isolation. After first strand cDNA synthesis from fetal brain mRNAs, short fragment cDNAs (sf-cDNAs) were synthesized with a two-step amplification system by use of our modified Degenerate Oligonucleotide Primed Shuttle Polymerase Chain Reaction (DOP-Shuttle-PCR) method. The sf-cDNAs were hybridized onto RNase A treated metaphases from K562, and the ABRs were microdissected and reamplified with DOP-Shuttle-PCR primer-II. The reamplified sf-cDNAs were cloned into a pBluescript vector. Twenty randomly chosen clones were sequenced and classified into 8 groups. Three out of the 8 grouped clones had been mapped to the long arm of chromosome 22 (22q11), whereas the other 5 were novel cDNAs. Quantitative Southern blot analysis indicated that 7 out of the 8 grouped clones (87.5%) were derived from the co-amplified regions.
Regional assignment of 64 human gene transcripts on chromosome 8
Authors: Ugolini F. , Chaffanet M. , Houlgatte R. , Patterson D. , Wells D. , Auffray C. , Birnbaum D. , Pébusque M.-J.
Published: May 2008

A panel of somatic cell hybrids specific for human chromosome 8 was used to localize 64 expressed sequence-tagged site (ESTS) markers to six individual regions by PCR amplification. Nine ESTS correspond to 8 known human genes and 6 others show similarities with vertebrate genes, whereas the remaining 49 ESTS markers correspond to novel genes with no database similarities. These gene transcript markers will contribute to the developing physical and expression maps of chromosome 8, and to the search for candidate genes for various human diseases.
Conservation of human vs. feline genome organization revealed by reciprocal chromosome painting
Authors: Wienberg J. , Stanyon R. , Nash W.G. , O’Brien P.C.M. , Yang F. , O’Brien S.J. , Ferguson-Smith M.A.
Published: May 2008

We employed fluorescence in situ hybridization (FISH) with probes established by flow sorting metaphase chromosomes of the domestic cat (Felis cattus, 2n = 38) to “paint” homologous segments on human chromosomes and, reciprocally, using human chromosome paints on feline metaphase preparations. The results revealed, by direct microscopic observation, widespread conservation of genome organization between the two mammalian orders and confirmed 90% of the homologous genes mapped to both species. Fourteen of 23 human chomosomes were hybridized with single cat probes, and 9 of 19 cat chromosomes were entirely labeled by a single human probe. All other chromosomes were labeled with only two or, at most, three probes of the respective species. Y-chromosome probes gave no signals. Approximately 30 syntenic segments were identified, and the number of translocations could be estimated to be on the order of one new translocation per 10 million years in the phylogenetic lines leading to human and cat. Using the principle of maximum parsimony, the primitive vs. derived human chromosome segments were identified by comparison to the feline, cattle, and pig genomes, a first step in reconstructing the evolutionary heritage of the mammalian radiations. The results suggest that reciprocal chromosome painting will help reconstruct the history of genomic changes by determining the polarity of chromosomal rearrangements and establishing the ancestral karyotype for each principle branching point in mammalian evolution.
Cloning and chromosome assignment to 1q32 of a human cDNA (RAB7L1) encoding a small GTP-binding protein, a member of the RAS superfamily
Authors: Shimizu F. , Katagiri T. , Suzuki M. , Watanabe T.K. , Okuno S. , Kuga Y. , Nagata M. , Fujiwara T. , Nakamura Y. , Takahashi E.
Published: May 2008

A full-length cDNA homologous to RAB7, a member of the RAB-related GTP-binding protein subfamily, was isolated from a human placenta cDNA library. This cDNA, designated RAB7L1, has an open reading frame of 609 nucleotides encoding 203 amino acids. Northern analysis showed that the mRNA is ubiquitously expressed in human tissues, although signal intensities were different among the various organs examined. This gene was located on chromosome band 1q32 by fluorescence in situ hybridization.
Localization of the 18S, 5.8S and 28S rRNA genes and the 5S rRNA genes in the babirusa and the white-lipped peccary
Authors: Zijlstra C. , Mellink C.H.M. , de Haan N.A. , Bosma A.A.
Published: May 2008

The locations of the genes encoding 18S, 5.8S and 28S rRNA and 5S rRNA were studied in two relatives of the domestic pig, the babirusa (Babyrousa babyrussa) and the white-lipped peccary (Tayassu pecari). In the babirusa, the 18S, 5.8S and 28S rDNA is located on chromosomes 6, 8 and 10. The genes on chromosomes 8 and 10 are actively transcribed, in contrast to those on chromosomes 6. In the white-lipped peccary, this rDNA was found to be located on chromosomes 4 and 8. The genes on both of these pairs of chromosomes are actively transcribed. The 5S rDNA was physically mapped to chromosome 16 in the babirusa, and to chromosome 11 in the white-lipped peccary. These data are compared to similar data obtained for the domestic pig, and confirm previously recognized chromosome homologies.
Assignment of the canalicular multispecific organic anion transporter gene (CMOAT) to human chromosome 10q24 and mouse chromosome 19D2 by fluorescent in situ hybridization
Authors: van Kuijck M.A. , Kool M. , Merkx G.F.M. , Geurts van Kessel A. , Bindels R.J.M. , Deen P.M.T. , van Os C.H.
Published: May 2008

Rabbit epithelial basolateral chloride conductance regulator (EBCR) and rat canalicular multispecific organic anion transporter (Cmoat) are found to be homologues based on protein sequence comparison and Northern blot analysis. EBCRis, therefore, renamed as rabbit Cmoat. The gene encoding CMOAT, a transporter possibly involved in Dubin-Johnson syndrome in humans, is mapped on human chromosome 10q24 and mouse chromosome 19D2.
Undermethylation of Alu sequences in ICF syndrome: molecular and in situ analysis
Authors: Miniou P. , Bourc’his D. , Molina Gomes D. , Jeanpierre M. , Viegas-Péquignot E.
Published: May 2008

The methylation status of young Alu sequences was investigated in four ICF patients. In fibroblast and leukocyte DNAs, Alu repeats were either undermethylated (HhaI and HpaII digestion) or demethylated (BstUl digestion), in contrast with the methylated status of Alus in control subjects. The methylation profile exhibited in ICF patients reproduces the normal profile of placental or sperm DNA. High-sensitivity immunocytochemical detection of HhaI and HpaII restriction sites on metaphase chromosomes provided further evidence of this undermethylation. The DNA methylation defect in ICF patients, first detected in satellite DNAs (constitutive heterochromatin) and CpG islands of genes on the inactive X chromosome (facultative heterochromatin), thus includes Alu sequences that are widely distributed throughout the human genome.
Physical and linkage mapping of the bovine bone morphogenetic protein 1 on the evolutionary break region of BTA 8
Authors: Martín-Burriel I. , Goldammer T. , Elduque C. , Lundin M. , Barendse W. , Zaragoza P. , Olsaker I.
Published: May 2008

Bone morphogenetic protein 1 (BMP1) has been proposed as a regulatory molecule involved in the binding or activation of other BMP molecules. It has been described as being identical to the enzyme C-proteinase that processes pro-collagens to fibrillar collagens. A fragment containing an exon of the bovine BMP1 gene was amplified from an adapter-ligated genomic DNA library. The isolated bovine BMP1 fragment was assigned by fluorescence in situ hybridization (FISH) to bovine (BTA) chromosome 8q21, a region containing an evolutionary breakpoint between homology to human (HSA) chromosomes 8 and 9. The assignment to BTA 8 was confirmed by using a hybrid somatic cell panel. The 3’ intron region of the amplified genomic BMP1 fragment showed a SSCP polymorphism, and linkage analysis in the International Bovine Reference Panel (IBRP) of families confirmed the boundary position of this gene. The BMP1 gene presents total linkage (Θ = 0.00, Z = 3.61) with the lipoprotein lipase gene on BTA 8 and HSA 8 and a larger recombination fraction (Θ = 0.11 Z = 5.03) with the marker GGTB2 on BTA 8 and HSA 9. Physical and genetic mapping of the bovine BMP1 gene contribute to narrow the boundary between the HSA 8 and HSA 9 homology segments on BTA 8 and to the comparative mapping between BTA 8, murine chromosome MMU 14, and HSA 8.
Characteristics of two salmonid repetitive DNA families in rainbow trout, Oncorhynchus mykiss
Authors: Reed K.M. , Dorschner M.O. , Phillips R.B.
Published: May 2008

DNA sequence and genomic location of two repetitive DNA families in rainbow trout (Oncorhynchus mykiss) were investigated to develop molecular markers for chromosome identification. DNA fragments with sequences similar to the tandem and interspersed elements described in other salmonids were isolated. One clone showed differential hybridization to 12 pairs of chromosomes and should be a useful marker for physical mapping.
Rapid isolation of genomic clones for individual members of human multigene families: identification and localisation of UBE2L4, a novel member of a ubiquitin conjugating enzyme dispersed gene family
Authors: Ardley H.C. , Moynihan T.P. , Thompson J. , Leek J.P. , Markham A.F. , Robinson P.A.
Published: May 2008

We describe a rapid, PCR-based, screening procedure for the isolation of human genomic clones in λ bacteriophage, containing sequences coding for individual homologous members of a multigene family. The approach is based upon the identification, by dilution, of sub-pools of the genomic library that contain members of the gene family, prior to phage isolation. The presence of specific genes is established by PCR of aliquots of individually amplified library pools, using consensus primers and subsequent sequencing. We have used the approach to isolate a fourth member of the UBE2L gene family, UBE2L4, and located it on chromosome 19q13.1→q13.2. This PCR-based approach to library screening has wider applicability in that it could be used to isolate alternate-spliced products from cDNA libraries.
Assignment of STK6 to human chromosome 20q13.2→q13.3 and a pseudogene STK6P to 1q41→q42
Authors: Kimura M. , Matsuda Y. , Eki T. , Yoshioka T. , Okumura K. , Hanaoka F. , Okano Y.
Published: May 2008

Fluorescence in situ hybridization analysis of human STK6 encoding a mitotic centrosomal protein kinase, Aik, revealed two signals in chromosome bands 20q13.2→q13.3 and 1q41→q42. Somatic cell hybrid panel analyses showed the existence of an identical sequence to STK6 cDNA on chromosome 20, and a processed pseudogene on chromosome 1. These results suggest that STK6 is localized at 20q13.2→q13.3 and a pseudogene STK6P at 1sq41→q42.
A novel allelic variant of the human serotonin transporter gene regulatory polymorphism
Authors: Delbrück S.J.W. , Wendel B. , Grunewald I. , Sander T. , Morris-Rosendahl D. , Crocq M.A. , Berrettini W.H. , Hoehe M.R.
Published: May 2008

Allelic variation of the human serotonin transporter gene (SLC6A4) has recently been shown to modulate anxiety-related traits. A tandemly repeated sequence in close proximity to the promoter was found to be represented by a long (L) and short (S) variant, differentially modulating gene expression in vitro. Specifically, allele S, generated by a deletion of 44 bp involving repeats VI to VIII, reduced transcriptional efficiency, gene expression, and 5-hydroxytryptamine uptake and was associated with increased neuroticism scores. We have now identified a novel allelic variant of this promoter-linked polymorphism that is significantly larger than the L allele and which we have designated allele XL (for “extra large”). Sequence analysis revealed that XL arose through duplication of an internal segment composed of repeat elements VI to IX, comprising 85 bp in total, and, most notably, including the segments deleted in the S allele. Additional allelic variants larger than human allele L were observed predominantly in various nonhuman primates. Preliminary data indicated that these variable allelic extensions similarly originate from this specific repeat region. These allelic variants may serve as a valuable model system to further elucidate the relationship between repeat structure, regulatory properties, and behavioral correlates. Finally, allelic variants were found to vary significantly among different human populations, with allele XL being uniquely present in individuals of African origin, allele L most frequent in Africans and Caucasians of Western European descent, and allele S most abundant in East Asians.
Genomic cloning and localization of CTAG, a gene encoding an autoimmunogenic cancer-testis antigen NY-ESO-1 to human chromosome Xq28
Authors: Chen Y.-T. , Boyer A.D. , Viars C.S. , Tsang S. , Old L.J. , Arden K.C.
Published: May 2008

CTAG was initially cloned from an esophageal squamous cell carcinoma cDNA expression library by immunoscreening with autologous patient’s serum. CTAG mRNA is expressed in a proportion of human cancers in a lineage-nonspecific fashion, whereas its expression in normal tissues is restricted to testis and ovary only. This expression pattern suggests that the CTAG product (NY-ESO-1) is an aberrantly activated tumor antigen and can potentially be an antigenic target for tumor vaccination. In the present study, we isolated human genomic clones of CTAG and established its genomic organization. By somatic cell hybrid studies and fluorescence in-situ hybridization, we localized this gene to chromosome Xq28, a region that also contains members of MAGE, a gene family that encodes several immunogenic tumor antigens with the characteristic cancer/testis expression pattern.
Definition of the critical interval for Smith-Magenis syndrome
Authors: Elsea S.H. , Purandare S.M. , Adell R.A. , Juyal R.C. , Davis J.G. , Finucane B. , Magenis R.E. , Patel P.I.
Published: May 2008

Smith-Magenis syndrome (SMS) comprises a complex physical and behavioral phenotype that is associated with an interstitial deletion of chromosome 17p11.2. The deletions observed in patients can range from <2 to >9 megabases of DNA and may include more than 100 genes. In order to determine the critical deletion interval responsible for the syndrome phenotype, we have examined several patients with varying deletions involving 17p11.2 by somatic cell hybrid analyses. We have binned 112 markers along 17p11.2, including 27 markers within the critical interval for SMS, which is bound proximally by D17S29 and distally by cC117–638. In addition, we present two patients who carry deletions involving 17p11.2 but do not exhibit the typical features of SMS. Patients such as these will allow genotype:phenotype correlations to be made and the gene(s) responsible for the SMS phenotype to be determined.
Chromosomal localization of mouse Cenpa gene
Authors: Fowler K.J. , Newson A.J. , MacDonald A.C. , Kalitsis P. , Lyu M.S. , Kozak C.A. , Choo K.H.A.
Published: May 2008

Using a previously isolated mouse centromere protein A (Cenpa) probe, we have localized the gene to the proximal region of mouse Chromosome 5, between the known Il6 and Yes1 loci near [Adra2C-D5H4S43-Hdh]. Comparison of this localization with that of human CENPA, which maps to chromosome 2, is consistent with the presence of a new region of conserved synteny between the two species.
Lentiviral Transduction of ApoAI Into Hematopoietic Progenitor Cells and Macrophages
Authors: Su Yan Ru , Blakemore John L. , Zhang Youmin , Linton MacRae F. , Fazio Sergio
Published: May 2008

Objective—

We used genetically engineered mouse hematopoietic progenitor cells (HPCs) to investigate the therapeutic effects of human apoAI on atherosclerosis in apoE
−/−
mice.

Methods and Results—

Lentiviral constructs expressing either human apoAI (LV-apoAI) or green fluorescent protein (LV-GFP) cDNA under a macrophage specific promoter (CD68) were generated and used for ex vivo transduction of mouse HPCs and macrophages. The transduction efficiency was >25% for HPCs and >70% for macrophages. ApoAI was found in the macrophage culture media, mostly associated with the HDL fraction. Interestingly, a significant increase in mRNA and protein levels for ATP binding cassette A1 (ABCA1) and ABCG1 were found in apoAI-expressing macrophages after acLDL loading. Expression of apoAI significantly increased cholesterol efflux in wild-type and apoE
−/−
macrophages. HPCs transduced with LV-apoAI ex vivo and then transplanted into apoE
−/−
mice caused a 50% reduction in atherosclerotic lesion area compared to GFP controls, without influencing plasma HDL-C levels.

Conclusions—

Lentiviral transduction of apoAI into HPCs reduces atherosclerosis in apoE
−/−
mice. Expression of apoAI in macrophages improves cholesterol trafficking in wild-type apoE-producing macrophages and causes upregulation of ABCA1 and ABCG1. These novel observations set the stage for a cell therapy approach to atherosclerosis regression, exploiting the cooperation between apoE and apoAI to maximize cholesterol exit from the plaque.
RNA Interference for Discoidin Domain Receptor 2 Attenuates Neointimal Formation in Balloon Injured Rat Carotid Artery
Authors: Shyu Kou-Gi , Wang Bao-Wei , Kuan Peiliang , Chang Hang
Published: May 2008

Objectives—

Discoidin domain receptor 2 (DDR2) plays potential roles in the regulation of collagen turnover mediated by smooth muscle cells (SMCs) in atherosclerosis. Little is known about the function of DDR2 in vascular system. We investigated whether inhibition of DDR2 by small interfering RNA (siRNA) can reduce neointimal formation after arterial injury.

Methods and Results—

SMCs from thoracic aorta of adult Wistar rats were cultured. The carotid artery from adult Wistar rats was injured by balloon catheter. DDR2 significantly increased migration and proliferation of SMCs. DDR2 siRNA inhibited 86% of DDR2 protein expression in cultured SMCs. DDR2 protein and mRNA expression significantly increased at 14 days after carotid injury. DDR2 siRNA significantly reduced DDR2 protein and mRNA expression induced by balloon injury. The immunohistochemical stain demonstrated that DDR2 siRNA decreased MMP2 protein labeling induced by balloon injury, a pattern similar to that of DDR2 protein labeling. Neointimal area was significantly increased after carotid injury and was significantly reduced by DDR2 siRNA.

Conclusions—

DDR2 increases migration and proliferation of SMCs, and expression of DDR2 in carotid artery significantly increases after injury. DDR2 siRNA attenuates neointimal formation after carotid injury. DDR2 may play a pivotal role in the pathogenesis of neointimal thickening after mechanical injury.
Counter Regulatory Effects of PKC
βII
and PKC
δ
on Coronary Endothelial Permeability
Authors: Gaudreault Nathalie , Perrin Rachel M. , Guo Mingzang , Clanton Chase P. , Wu Mack H. , Yuan Sarah Y.
Published: May 2008

Objective—

The aim of this study was to examine the endothelial distribution and activity of selected PKC isoforms in coronary vessels with respect to their functional impact on endothelial permeability under the experimental conditions relevant to diabetes.

Methods and Results—

En face immunohistochemistry demonstrated a significant increase of PKC
βΙΙ
and decrease of PKCδ expression in coronary arterial endothelium of Zucker diabetic rats. To test whether changes in PKC expression alter endothelial barrier properties, we measured the transcellular electric resistance in human coronary microvascular endothelial monolayers and found that either PKC
βΙΙ
overexpression or PKCδ inhibition disrupted the cell–cell adhesive barrier. Three-dimensional fluorescence microscopy revealed that hyperpermeability was caused by altered PKC activity in association with distinct translocation of PKC
βΙΙ
to the cell–cell junction and PKCδ localization to the cytosol. Further analyses in fractionated endothelial lysates confirmed the differential redistribution of these isozymes. Additionally, FRET analysis of PKC subcellular dynamics demonstrated a high PKC
βΙΙ
activity at the cell surface and junction, whereas PKCδ activity is concentrated in intracellular membrane organelles.

Conclusion—

Taken together, these data suggest that PKC
βΙΙ
and PKCδ counter-regulate coronary endothelial barrier properties by targeting distinctive subcellular sites. Imbalanced PKC
βΙΙ
/PKCδ expression and activity may contribute to endothelial hyperpermeability and coronary dysfunction in diabetes.
Monocyte Functional Responsiveness After PSGL-1–Mediated Platelet Adhesion Is Dependent on Platelet Activation Status
Authors: Bournazos Stylianos , Rennie Jillian , Hart Simon P. , Fox Keith A.A. , Dransfield Ian
Published: May 2008

Objective—

Acute coronary diseases are characterized by elevated levels of circulating platelet-leukocyte complexes, raising the possibility that proinflammatory processes might be initiated in leukocytes after platelet adhesion. Here we examined the mechanism of platelet binding to polymorphonuclear leukocytes, monocytes, and monocyte subsets and investigated the potential functional consequences of monocyte binding to minimally activated or thrombin-activated platelets.

Methods and Results—

In this article, we describe key differences in terms of stability of PSGL-1–mediated interaction of platelets with monocytes and polymorphonuclear leukocytes and a small but significant difference in platelet binding to monocyte subsets (CD14
high
and CD14
low
/HLA-DR
high
). We also report differential effects of platelet binding on monocyte functional responses between minimally and thrombin-activated platelets. In particular, monocyte CD11b expression and release of proinflammatory cytokines, like interleukin 1β and tumor necrosis factor α, were significantly upregulated on adhesion of stimulated platelets, whereas unstimulated platelets had no effect. Moreover, binding of unstimulated, but not of thrombin-activated, platelets to monocytes had no impact on NF-κB activity, monocyte migration, and induction of apoptosis in the absence of survival factors.

Conclusions—

Our data suggest that in the absence of overt activation, PSGL-1–P-selectin–dependent platelet binding to monocytes represents a normal physiological process with little impact on the potential of monocytes to cause vascular injury.
Arteriolar Genesis and Angiogenesis Induced by Endothelial Nitric Oxide Synthase Overexpression Results in a Mature Vasculature
Authors: Benest Andrew V. , Stone Oliver A. , Miller William H. , Glover Colin P. , Uney James B. , Baker Andrew H. , Harper Steven J. , Bates David O.
Published: May 2008

Background—

Generation of physiologically active vascular beds by delivery of combinations of growth factors offers promise for vascular gene therapy.

Methods and Results—

In a mesenteric model of physiological angiogenesis, combining endothelial nitric oxide synthase (eNOS) (and hence NO production) with VEGF and angiopoietin-1 overexpression resulted in a more functional vascular phenotype than growth factor administration alone. eNOS gene delivery upregulated eNOS, VEGF, and Ang-1 to similar levels as gene transfer with VEGF or Ang-1. eNOS overexpression resulted in neovascularization to a similar extent as VEGF and Ang-1 combined, but not by sprouting angiogenesis. Whereas combining Ang-1 and VEGF increased both exchange vessels and conduit vessels, neither growth factor nor eNOS alone resulted in vessels with smooth muscle cell (SMC) coverage. In contrast, combining all three generated microvessels with SMCs (arteriolar genesis) and further increased functional vessels. Use of a vasodilator, prazosin, in combination with Ang1 and VEGF, but not alone, also generated SMC-positive vessels.

Conclusion—

Coexpression of eNOS, VEGF, and Ang-1 results in a more mature vascularization of connective tissue, and generates new arterioles as well as new capillaries, and provides a more physiological therapeutic approach than single growth factor administration, by combining hemodynamic forces with growth factors.
The LIM Protein Leupaxin Is Enriched in Smooth Muscle and Functions As an Serum Response Factor Cofactor to Induce Smooth Muscle Cell Gene Transcription
Authors: Sundberg-Smith Liisa J. , DiMichele Laura A. , Sayers Rebecca L. , Mack Christopher P. , Taylor Joan M.
Published: May 2008

Leupaxin is a LIM domain–containing adapter protein belonging to the paxillin family that has been previously reported to be preferentially expressed in hematopoietic cells. Herein, we identified leupaxin in a screen for focal adhesion kinase binding partners in aortic smooth muscle, and we show that leupaxin is enriched in human and mouse vascular smooth muscle and that leupaxin expression is dynamically regulated during development. In addition, our studies reveal that leupaxin can undergo cytoplasmic/nuclear shuttling and functions as an serum response factor cofactor in the nucleus. We found that leupaxin forms a complex with serum response factor and associates with CArG-containing regions of smooth muscle promoters and that ectopic expression of leupaxin induces smooth muscle marker gene expression in both 10T1/2 cells and rat aortic smooth muscle cells. Subsequent studies indicated that enhanced focal adhesion kinase activity (induced by fibronectin or expression of constitutively active focal adhesion kinase) attenuates the nuclear accumulation of leupaxin and limits the ability of leupaxin to enhance serum response factor–dependent gene transcription. Thus, these studies indicate that modulation of the subcellular localization of serum response factor cofactors is 1 mechanism by which extracellular matrix–dependent signals may regulate phenotypic switching of smooth muscle cells.
The Trafficking Protein GABARAP Binds to and Enhances Plasma Membrane Expression and Function of the Angiotensin II Type 1 Receptor
Authors: Cook Julia L. , Re Richard N. , deHaro Dawn L. , Abadie Jennifer M. , Peters Michelle , Alam Jawed
Published: May 2008

Proteins that bind to the intracellular expanses, particularly cytoplasmic tail regions, of heptahelical integral membrane receptors are of particular interest in that they can mediate or modulate trafficking or intracellular signaling. In an effort to distinguish new proteins that might promote angiotensin II type 1 (AT
1
) receptor intracellular events, we screened a yeast 2-hybrid mouse brain library with the rat AT
1A
receptor (AT
1
R) carboxyl terminus and identified GABARAP, a protein involved in intracellular trafficking of the GABA
A
receptor, as a binding partner for the AT
1
R. Interaction of GABARAP with the AT
1
R carboxyl terminus was further substantiated using GST pull-down assays, and binding of the full-length tagged AT
1
R to GABARAP was verified using coimmunoprecipitation. Bioluminescence resonance energy transfer assays further confirmed specific interaction of GABARAP with AT
1
R. Moreover, GABARAP clearly increased the steady-state level of plasma membrane-associated AT
1
R in PC-12 cells. Cotransfection of GABARAP with an AT
1
R fluorescent fusion protein increased PC-12 cell surface expression of the AT
1
R more than 6-fold when standardized to the level of intracellular expression. Furthermore, GABARAP overexpression in CHO-K1 cells engineered to express AT
1
R increased angiotensin II binding sites 3.7-fold and angiotensin II–induced phospho–extracellular signal-regulated kinase 1/2 and cellular proliferation significantly over levels obtained with AT
1
R overexpression alone. In addition, small interfering RNA–mediated knockdown of GABARAP reduced the steady-state levels of the AT
1
R fluorescent fusion protein by 43% and its cell surface expression by 84%. Immunoblot analyses confirmed the quantitative image data. We conclude that GABARAP binds to and promotes trafficking of the AT
1
R to the plasma membrane.
Fisheries Enforcement in European Community Waters Since 2002—Developments in Non-Flag Enforcement
Authors: Johnson Constance
Published: May 2008

In 2002 the European Union commenced significant reforms to the Common Fisheries Policy (CFP) which included introduction of a new control and enforcement system. In 2007, the new Community Fisheries Control Agency (CFCA) undertook its first work programme. The establishment of the CFCA was considered to be the final part of the CFP reforms commenced in 2002. This paper examines the course of the 2002 reforms in relation to fisheries control and enforcement in European Community waters, particularly issues concerning non-flag enforcement, and considers whether there has been a shift towards greater reliance on non-flag enforcement measures. The paper examines the two major innovations of the 2002 control and enforcement system, namely: regional cooperation and coordination and the establishment of the CFCA. The paper also examines developments in the European Commission's and Member States' respective roles in relation to control and enforcement. Finally, the paper considers the focus of the 2002 control and enforcement system on technical management tools for the achievement of more effective control and management of fisheries.
Contemporary Exclusive Fishery Zones or Why Some States Still Claim an EFZ
Authors: Kvinikhidze Shalva
Published: May 2008

This article deals with the concept of the Exclusive Fishery Zone (EFZ) which has long existed in the state practice in the law of the sea. It describes the genesis and development of the concept, attempts by coastal states to gain extensive and exclusive control over fishery resources beyond the territorial sea, and the influence of international conferences and the United Nations Convention on the Law of the Sea on EFZs. The main aim of this article is to examine current claims to EFZs, describe the legal nature of contemporary EFZs and analyse the motivation of coastal states for still claiming an EFZ and not an Exclusive Economic Zone (EEZ), which is a multi-functional zone that includes not only exclusive fishing rights of the coastal state but also other rights, jurisdiction and economic activities.
Ocean Fertilisation and Climate Change: The Need to Regulate Emerging High Seas Uses
Authors: Rayfuse Rosemary , Lawrence Mark G. , Gjerde Kristina M.
Published: May 2008

Geo-engineering and environmental modification techniques are increasingly
being proposed as climate change mitigation strategies. Ocean fertilisation
has been promoted as a simple solution to the problem of increasing
atmospheric CO2 levels. However, neither its environmental safety nor its
efficacy has been adequately assessed. This article examines the legality of
ocean fertilisation under the law of the sea and concludes that it is
subject to regulation under the London Convention and London Protocol as its
potential for harm is contrary to the aims of these agreements. Hence, the
sale of carbon offsets to fund ocean fertilisation activities should be
prohibited unless and until an adequate risk assessment based on independent
peer-reviewed science has established that the benefits outweigh the
potential for harm, and appropriate regulation is in place to ensure that
real, measurable, long-term CO2 sequestration can be independently verified.
The initial uncertainties surrounding the appropriate regulatory regime for
ocean fertilisation highlight the need for a comprehensive global regime for
the prior assessment and on-going monitoring of existing, new and emerging
high seas activities and uses to ensure they do not have adverse impacts on
marine biodiversity and the marine environment in areas beyond national
jurisdiction.
Comparison of the Diurnal Cycle of Outgoing Longwave Radiation from a Climate Model with Results from ERBE
Authors: Smith G. Louis , Mlynczak Pamela E. , Rutan David A. , Wong Takmeng
Published: May 2008

The diurnal cycle of outgoing longwave radiation (OLR) computed by a climate model provides a powerful test of the numerical description of various physical processes. Diurnal cycles of OLR computed by version 3 of the Hadley Centre Atmospheric Model (HadAM3) are compared with those observed by the Earth Radiation Budget Satellite (ERBS) for the boreal summer season (June–August). The ERBS observations cover the domain from 55°S to 55°N. To compare the observed and modeled diurnal cycles, the principal component (PC) analysis method is used over this domain. The analysis is performed separately for the land and ocean regions. For land over this domain, the diurnal cycle computed by the model has a root-mean-square (RMS) of 11.4 W m−2, as compared with 13.3 W m−2 for ERBS. PC-1 for ERBS observations and for the model are similar, but the ERBS result has a peak near 1230 LST and decreases very slightly during night, whereas the peak of the model result is an hour later and at night the OLR decreases by 7 W m−2 between 2000 and 0600 LST. Some of the difference between the ERBS and model results is due to the computation of convection too early in the afternoon by the model. PC-2 describes effects of morning/afternoon cloudiness on OLR, depending on the sign. Over ocean in the ERBS domain, the model RMS of the OLR diurnal cycle is 2.8 W m−2, as compared with 5.9 W m−2 for ERBS. Also, for the model, PC-1 accounts for 66% of the variance, while for ERBS, PC-1 accounts for only 16% of the variance. Thus, over ocean, the ERBS results show a greater variety of OLR diurnal cycles than the model does.
Where Inquiry Ends
Authors: Abu-Saad Ismael
Published: May 2008

One of the ongoing struggles of indigenous people against colonization is to be able to exercise the fundamental right to represent themselves and to speak to the dominant society with their own voices and words, rather than to be spoken of or about. This essay discusses the mainstream academic peer review process and the suppression of indigenous standpoints by the dominant culture. The essay goes on to analyze the ways in which one mainstream international academic journal accepted and contained the expression of an indigenous standpoint by then inviting a response from a mainstream scholar who largely delegitimized the indigenous voice, at which point the inquiry ended.
Transdermal Patch Medication Delivery Systems and Pediatric Poisonings, 2002-2006
Authors: Parekh Dina , Miller Michael A. , Borys Doug , Patel Paresh R. , Levsky Marc E.
Published: May 2008

Transdermal drug delivery systems are an increasingly popular method of medication delivery containing large quantities of medication and presenting new opportunities for toxicity. To provide a description of exposures to transdermal medications in a pediatric population, we studied exposures in individuals less than 12 years of age. This is a retrospective database study in which the Texas Poison Center Network database from 2002 to 2006 was reviewed. In all, 336 poison control center records of patch exposures over the 5-year period were identified. Of those, 110 cases involved children less than 12 years old. A majority of cases resulted in no significant clinical effects. One death resulted from opioid toxicity. Although a majority of patch exposures in children less than 12 years of age resulted in no significant clinical toxicity, practitioners and the public must be made aware of the available patch-based medications and their potential for toxicity in children.
Strategy-as-practice and dynamic capabilities: Steps towards a dynamic view of strategy
Authors: Regnér Patrick
Published: May 2008

This article identifies an essential research area where a strategy-as-practice approach can make important contributions to the strategic management field: the dynamic process through which organizational assets are developed. It compares strategy-as-practice and dynamic capabilities perspectives and demonstrates how the former may complement the latter in analyzing activities that underpin and may create and modify organizational assets. Several distinct features of the practice approach are identified as fertile ground for examining the micro-foundations of strategy dynamics. It is demonstrated how activity configurations, socio-cultural embeddedness, co-evolution, social interactions, the inclusion of multiple strategists and an awareness of the importance of imagination can complement the dynamic capabilities perspective and may provide suggestions for a dynamic view more generally.
When They Honor the Voice
Authors: Cummings Melbourne S. , Latta Judi Moore
Published: May 2008

This study assumes a personal and critical perspective and explores the point of view of a few African American ordained women ministers whose stories about their calls to the preaching ministry give clues as to the motivation and inspiration of other women committed to answering a Divine call. It combines thinking of womanist theology, the African American oral tradition, and African Diaspora life storytelling. By using ethnography, theology, and Afrocentrism, it assumes Black women’s “centeredness”—a concept that speaks to one finding one’s own voice in the midst of confusion and uncertainty—in examining the language, voice, and terminology that these women use to place call stories at the center of consideration about their relationship with the Divine.
Analysis of Thermal Stresses in Unsymmetric Cross-ply Composite Strips
Authors: Carbajal N. , Vargas G. , Arrese A. , Mujika F.
Published: May 2008

An approach based on the hypotheses of the classical beam theory for determining thermal stresses in unsymmetric cross-ply strips has been developed. The material behavior is considered linear elastic, and viscoelastic effects are not considered. By supposing linear strain distribution in the cross-sections, the position of the neutral axis, the radius of curvature and the distribution of thermal stresses have been determined. The analysis is valid in the case of large displacements, since the curvature is constant and the deformed shape is an arc of circumference. Five different lay-up configurations of strip geometry specimens have been used for experimental verification. Mid-point deflections have been measured and compared with theoretical values, applying both the proposed approach and the classical laminated plates theory.
The Relationships Between Emerging Adults' Expressed Desire to Marry and Frequency of Participation in Risk-Taking Behaviors
Authors: Willoughby Brian J. , Dworkin Jodi
Published: May 2008

The impact that desire to marry has on risk-taking behaviors during emerging adulthood is examined in the current investigation using nationally representative data from the National Longitudinal Study of Adolescent Health (Add Health). Looking both at the simple relationships between desire to marry and risk-taking behaviors, as well as the possible predictive power desire to marry could have on binge drinking, marijuana use, and sexual behaviors, data revealed that attitudes toward marriage may be an important component of emerging adulthood and risk-taking research. Results indicate that desire to marry was predictive of several risk-taking behaviors for both men and women even after controlling for other known predictors of risk-taking. Implications for future research are discussed.
Novel Trial Design: A Report From the 19th Frontiers Symposium of ACCP
Authors: Ameer Barbara
Published: May 2008

Novel approaches to drug development along with incentives to reward risk undertaken during early drug development have been proposed to improve the medical product development process. The American College of Clinical Pharmacology held its 19th Frontiers Symposium in mid‐2007 to provide a forum to share new knowledge about disease progression models and adaptive and other novel trial designs. These topics reflect the changing paradigm of drug development and regulatory innovation as outlined in the Food and Drug Administration's Critical Path Initiative. This meeting report summarizes some of the presentations and panel discussions among academic, industrial, and regulatory participants regarding clinical trial design, including adaptive Bayesian approaches.